RESUMEN
BACKGROUND: This study aimed to systematically evaluate the effect of intense pulsed light (IPL) therapy in patients harboring dry eye disease caused by meibomian gland dysfunction (MGD) based on qualified studies. METHODS: The electronic databases, including PubMed, Cochrane, and Embase, were searched using keywords to identify available publications updated to November 2021. Relative risk or weighted mean difference combined with 95% confidence interval was used to synthesize the outcomes of included studies. The meta-analysis included 15 randomized controlled trials with 1,142 patients (2,284 eyes). RESULTS: The results revealed that IPL could significantly decrease the ocular surface disease index (OSDI), standard patient evaluation of eye dryness (SPEED), artificial tear usage, tear film lipid layer, meibomian gland quality (MGQ), meibomian gland expression (MGX), and corneal fluorescein staining (CFS) while increase tear break-up time (TBUT) and noninvasive tear break-up time (NIBUT) compared with sham. Compared with MGX, IPL+MGX markedly decreased the SPEED, CFS, and tear meniscus height (TMH), but with increased TBUT. Compared with MGX, IPL showed significant effect in increasing the OSDI and TBUT, but decreasing the TMH and NIBUT. However, no significant differences were seen between IP+MGX and MGX in OSDI, MGQ, and MGX, nor between IPL and MGX in OSDI, SPEED, and TBUT. CONCLUSION: We identified that the application of IPL alone or IPL combined with MGX elicited superior clinical effect for improving the eye function and symptoms in the treatment of MGD-related dry eye disease, which is considered available for wide clinical application.
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Síndromes de Ojo Seco , Tratamiento de Luz Pulsada Intensa , Disfunción de la Glándula de Meibomio , Síndromes de Ojo Seco/etiología , Síndromes de Ojo Seco/metabolismo , Síndromes de Ojo Seco/terapia , Fluoresceína/metabolismo , Humanos , Tratamiento de Luz Pulsada Intensa/métodos , Lípidos , Gotas Lubricantes para Ojos , Disfunción de la Glándula de Meibomio/complicaciones , Disfunción de la Glándula de Meibomio/terapia , Glándulas Tarsales/metabolismo , Lágrimas/metabolismoRESUMEN
OBJECTIVE: Interstitial fluid in extracellular matrices may not be totally fixed but partially flow through long-distance oriented fibrous connective tissues via physical mechanisms. We hypothesized there is a long-distance interstitial fluid transport network beyond vascular circulations. MATERIALS AND METHODS: We first used 20 volunteers to determine hypodermic entrant points to visualize long-distance extravascular pathway by MRI. We then investigated the extravascular pathways initiating from the point of thumb in cadavers by chest compressor. The distributions and structures of long-distance pathways from extremity ending to associated visceral structures were identified. RESULTS: Using fluorescent tracer, the pathways from right thumb to right atrium wall near chest were visualized in seven of 10 subjects. The cutaneous pathways were found in dermic, hypodermic and fascial tissues of hand and forearm. The perivascular pathways were along the veins of arm, axillary sheath, superior vena cava and into the superficial tissues on right atrium. Histological and micro-CT data showed these pathways were neither blood nor lymphatic vessels but long-distance oriented fibrous matrices, which contained the longitudinally assembled micro-scale fibres consistently from thumb to superficial tissues on right atrium. CONCLUSIONS: These data revealed the structural framework of the fibrous extracellular matrices in oriented fibrous connective tissues was of the long-distance assembled fibres throughout human body. Along fibres, interstitial fluid can systemically transport by certain driving-transfer mechanisms beyond vascular circulations.
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Tejido Conectivo/metabolismo , Matriz Extracelular/metabolismo , Puntos de Acupuntura , Adulto , Cadáver , Tejido Conectivo/química , Tejido Conectivo/patología , Medios de Contraste/química , Medios de Contraste/metabolismo , Femenino , Fluoresceína/química , Fluoresceína/metabolismo , Pie/diagnóstico por imagen , Mano/diagnóstico por imagen , Humanos , Vasos Linfáticos/anatomía & histología , Vasos Linfáticos/diagnóstico por imagen , Imagen por Resonancia Magnética , Masculino , Microscopía Confocal , Puntos Cuánticos/química , Puntos Cuánticos/metabolismo , Adulto JovenRESUMEN
PURPOSE: To assess whether omega-3 fatty acid (FA) supplementation is more efficacious than placebo in amelioration of signs and symptoms of dry eye disease. METHODS: We performed a systematic literature search in PubMed, Scopus, Web of Science, and Cochrane Central Register of Controlled Trials databases. We included randomized clinical trials comparing omega-3 FA supplementation with placebo in patients with dry eye disease. The outcome measures were dry eye symptoms, breakup time (BUT), Schirmer test, and corneal fluorescein staining. The pooled effect sizes were estimated using a random-effects model. Heterogeneity was evaluated using the Q and I tests. Sensitivity analysis and assessment of publication bias were performed. Meta-regression was performed to evaluate the source of heterogeneity. RESULTS: Seventeen randomized clinical trials involving 3363 patients were included. Compared with placebo, omega-3 FA supplementation decreased dry eye symptoms [standardized difference in mean values (SDM) = 0.968; 95% confidence interval (CI) 0.554-1.383; P < 0.001] and corneal fluorescein staining (SDM = 0.517; 95% CI, 0.043-0.991; P = 0.032), whereas it increased the BUT (SDM = 0.905; 95% CI, 0.564-1.246; P < 0.001) and Schirmer test values (SDM = 0.905; 95% CI, 0.564-1.246; P < 0.001). No evidence of publication bias was observed, and sensitivity analyses indicated the robustness of results obtained. Meta-regression analysis showed a higher improvement of dry eye symptoms and BUT in studies conducted in India. CONCLUSIONS: This meta-analysis provides evidence that omega-3 FA supplementation significantly improves dry eye symptoms and signs in patients with dry eye disease. Therefore, our findings indicate that omega-3 FA supplementation may be an effective treatment for dry eye disease.
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Suplementos Dietéticos , Síndromes de Ojo Seco/terapia , Ácidos Grasos Omega-3/administración & dosificación , Córnea/metabolismo , Síndromes de Ojo Seco/metabolismo , Fluoresceína/metabolismo , Humanos , Ensayos Clínicos Controlados Aleatorios como Asunto , Lágrimas/fisiologíaRESUMEN
Our investigations on extracellular biochemical events to find readily and sensitively detectable/measurable molecular targets for developing easier, simpler, and quicker diagnostic methods and tools for bacterial pathogens led to the observation that bacteria grown in the presence of glucose produced a compound capable of quenching fluorescein. Under the experimental conditions, among various sugars, glucose was found to induce maximum amount of the quencher when Escherichia coli was grown in presence of 50 mM glucose in rarified LB. The release of quencher closely following bacterial growth significantly from fourth hour after moderate inoculation. This fluorescein-quencher was purified using TLC and HPLC and identified using GC-MS as 13-docosenamide or erucamide, originally known as plant lipid, is a neuroactive compound in human and animals. Fluorescence and UV-absorption spectral analysis showed that the compound formed stable adduct with fluorescein in the ground state. Commercial 13-docosonamide enabled quantitation of the compound produced in micromolar quantities during glucose utilization from the medium. Twenty-seven different commonly encountered bacteria, pathogens or otherwise, could produce the quencher. A simple microplate-based growth monitoring method was developed exploiting quenching as an easily and readily measurable signal, either using a reader or an imager. While 13-docosenamide release by bacteria may be relevant in host-bacteria interactions, especially when growing under conditions that provide glucose, the new approach with inexpensive reagents can provide a new antibiogram technique.
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Ácidos Erucicos/metabolismo , Escherichia coli/efectos de los fármacos , Fluoresceína/metabolismo , Glucosa/farmacología , Pruebas de Sensibilidad Microbiana/métodos , Escherichia coli/crecimiento & desarrolloRESUMEN
A polymerase ribozyme can be used to label the 3' end of RNA or DNA molecules by incorporating a variety of functionalized nucleotide analogs. Guided by a complementary template, the ribozyme adds a single nucleotide that may contain a fluorophore, biotin, azide or alkyne moiety, thus enabling the detection and/or capture of selectively labeled materials. Employing a variety of commercially available nucleotide analogs, efficient labeling was demonstrated for model RNAs and DNAs, human microRNAs and natural tRNA.
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Región de Flanqueo 3' , ARN Polimerasas Dirigidas por ADN/metabolismo , Ácidos Nucleicos/metabolismo , ARN Catalítico/metabolismo , Coloración y Etiquetado/métodos , Biotina/química , Biotina/metabolismo , ADN/química , ADN/metabolismo , Fluoresceína/química , Fluoresceína/metabolismo , Colorantes Fluorescentes/química , Colorantes Fluorescentes/metabolismo , MicroARNs/química , MicroARNs/metabolismo , Conformación de Ácido Nucleico , Ácidos Nucleicos/química , Nucleotidiltransferasas/metabolismo , ARN/química , ARN/metabolismo , ARN de Transferencia/química , ARN de Transferencia/metabolismo , Rodaminas/química , Rodaminas/metabolismoRESUMEN
The transdermal delivery of 2 fluorescent probes with similar molecular weight but different lipophilicity, into and through the skin from 2 commercially available transdermal bases, pluronic lecithin organogel, and Lipoderm® has been evaluated. First, in vitro penetration of fluorescein sodium and fluorescein (free acid) through porcine skin was evaluated. Retention and depth distribution profiles in skin were obtained by tape stripping and then followed by optical sectioning using multiphoton microscopy. The results showed that Lipoderm® led to an enhanced penetration of the hydrophilic compound, fluorescein sodium. For the lipophilic compound fluorescein (free acid), Lipoderm® performed similar to pluronic lecithin organogel base, where minimal drug was detected in either receptor phase. The skin retention and depth distribution results also showed that the hydrophilic fluorescein sodium had high skin retention with Lipoderm®, whereas fluorescein (free acid) had very low penetration and retention with increasing skin depth. Moreover, optical sectioning by multiphoton microscopy revealed an uneven distribution of probes across the skin in the x-y plane for both transdermal bases. This work showed that a hydrophilic compound has significantly increased skin penetration and retention when formulated with Lipoderm®, and the skin retention of the probe was the main determinant of its skin flux.
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Lecitinas/administración & dosificación , Lecitinas/metabolismo , Poloxámero/administración & dosificación , Poloxámero/metabolismo , Piel/metabolismo , Administración Cutánea , Animales , Química Farmacéutica/métodos , Sistemas de Liberación de Medicamentos/métodos , Fluoresceína/metabolismo , Lecitinas/química , Permeabilidad , Poloxámero/química , Absorción Cutánea/fisiología , PorcinosRESUMEN
In a previous study, we demonstrated that topical D-beta-hydroxybutyrate ameliorates corneal epithelial erosion and superficial punctate keratopathy in a rat model of dry eye disease. In the current investigation, we performed a prospective, randomized, multicentre, double-blind, placebo-controlled study to assess the safety and efficacy of 1% D-3-hydroxybutyrate eye drops in patients with dry eye disease. A total of 65 patients were randomly assigned to either the placebo group or the 1% D-3-hydroxybutyrate group, and the treatments were administered 6 times a day for 4 weeks. We then evaluated corneal fluorescein staining, corneal and conjunctival rose Bengal staining, tear film break-up time (BUT), Schirmer score, and subjective symptoms. At both 2 and 4 weeks, the corneal rose Bengal score was significantly better in the 1% D-3-hydroxybutyrate group than in the placebo group. Among patients with an initial Schirmer score of ≤5 mm, the corneal fluorescein staining score was significantly better in the 1% D-3-hydroxybutyrate group than in the placebo group at two weeks. Mild ocular symptoms occurred in both groups, and these spontaneously resolved. The present study suggested that 1% D-3-hydroxybutyrate eye drops are safe and effective in treating ocular surface disorders in patients with tear-deficient dry eye disease.
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Ácido 3-Hidroxibutírico/uso terapéutico , Conjuntiva/efectos de los fármacos , Córnea/efectos de los fármacos , Síndromes de Ojo Seco/tratamiento farmacológico , Soluciones Oftálmicas/uso terapéutico , Lágrimas/efectos de los fármacos , Adulto , Anciano , Animales , Conjuntiva/fisiopatología , Córnea/fisiopatología , Método Doble Ciego , Síndromes de Ojo Seco/fisiopatología , Femenino , Fluoresceína/química , Fluoresceína/metabolismo , Colorantes Fluorescentes/química , Colorantes Fluorescentes/metabolismo , Humanos , Masculino , Persona de Mediana Edad , Seguridad del Paciente , Estudios Prospectivos , Ratas , Rosa Bengala/química , Rosa Bengala/metabolismo , Coloración y EtiquetadoRESUMEN
A 24-amino acid leader peptide of a new human recombinant manganese superoxide dismutase can enter cells and carry molecules. Here, we demonstrated that six of the 24 amino acids penetrate cells through a particular gate represented by a specific amino acid sequence of the oestrogen receptor (ER). We analysed the internalization of the synthetic hexapeptide and the cytotoxic activity of the hexapeptide conjugated to cisplatin on a cell line panel. In most cell lines, the hexapeptide delivered an amount of cisplatin that was 2 to 8 times greater than that released by cisplatin when the drug was used alone. This increased delivery increases the therapeutic index of cisplatin and reduces side effects caused by a high dosage or long-term treatment times. We may consider this hexapeptide a new molecular carrier to deliver molecules with therapeutic activity into ER(+) cells for diagnostic purposes and clinical or immune therapy.
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Portadores de Fármacos/química , Fragmentos de Péptidos/química , Secuencia de Aminoácidos , Antineoplásicos/química , Antineoplásicos/metabolismo , Antineoplásicos/farmacología , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Cisplatino/química , Cisplatino/metabolismo , Cisplatino/farmacología , Portadores de Fármacos/metabolismo , Portadores de Fármacos/farmacología , Ensayos de Selección de Medicamentos Antitumorales , Retículo Endoplásmico/metabolismo , Fluoresceína/química , Fluoresceína/metabolismo , Colorantes Fluorescentes/química , Colorantes Fluorescentes/metabolismo , Humanos , Concentración 50 Inhibidora , Fragmentos de Péptidos/metabolismo , Fragmentos de Péptidos/farmacología , Señales de Clasificación de Proteína , Receptores de Estrógenos/metabolismo , Proteínas Recombinantes/química , Superóxido Dismutasa/químicaRESUMEN
PURPOSE: To test our hypothesis about whether there is water migration in the horizontal corneal plane and investigate its developmental mechanism. METHODS: A fluorescein solution was intrastromally injected into normal and edematous corneas of rabbits, and the movement of the fluorescein solution was observed and recorded over time. RESULTS: In normal corneas, the water flow was characterized by a swirling movement from the center to the periphery in the stroma. The fluorescein solution ultimately spread and occupied the entire cornea, indicating horizontal intracorneal swirling of water. In contrast, when the corneal endothelia were injured by intracameral injection of a preservative to create corneal edema, no water migration occurred, suggesting that the integrity of the corneal endothelial function is essential for water migration. The water migration stopped with injection of a sodium-potassium pump inhibitor, indicating that the enzyme is necessary for physiologic water migration in the cornea. With recovery of corneal endothelial function, the water migration began, and focal edema remained in the periphery with no water migration in this edematous area. CONCLUSIONS: We report for the first time the presence of horizontal water migration in the cornea in a swirling pattern (i.e., intracorneal swirling migration of water, generated by the pump function in the corneal endothelial cells), which may supplement the conventional concept of development of corneal edema in the vertical plane. This dynamic water circulatory system may be involved in increasing the efficiency of the water transfer in the entire cornea.
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Córnea/metabolismo , Edema Corneal/metabolismo , Lesiones de la Cornea/metabolismo , Endotelio Corneal/fisiología , Agua/metabolismo , Animales , Compuestos de Benzalconio/farmacología , Córnea/efectos de los fármacos , Sustancia Propia/metabolismo , Detergentes/farmacología , Modelos Animales de Enfermedad , Inhibidores Enzimáticos/farmacología , Femenino , Fluoresceína/metabolismo , Inyecciones Intraoculares , ConejosRESUMEN
PURPOSE: The aim of this work was to gather preliminary data in different conditions of healthy eyes, aqueous tear deficient dry eyes, obstructive meibomian gland disease (MGD) and non-obvious obstructive MGD (NOMGD) individuals, using a new, contrast-enhanced optical coherence tomography (OCT) imaging method to evaluate the clearance of lipids in human tears. METHODS: Eighty-two adult patients presenting with complaints of ocular irritation were studied for abnormalities of the ocular surface and classified as healthy (n = 21), aqueous tear deficient dry eyes (n = 20), obstructive MGD (n = 15) and NOMGD (n = 26) individuals. A lipid-based tracer, containing an oil-in-water emulsion, was used to obtain an enhanced OCT imaging of the lower tear meniscus. After instillation, a dramatic initial increase of reflectivity of the lower tear meniscus was detected by OCT, followed by a decay back to baseline values over time. Based on this finding, the clearance of lipids was measured in real-time by Fourier-domain anterior segment OCT. RESULTS: The differences in the clearance of lipids among the four groups as well as the correlations between symptom questionnaire score, standardized visual scale test, fluorescein break-up time, ocular surface fluorescein staining score, Schirmer I test scores were found to be statistically significant. The individual areas under the curve of the clearance of lipids calculated by the receiver operating characteristic curve technique ranged from 0.66 to 0.98, suggesting reliable sensitivity and specificity of lipid-enhanced OCT imaging. CONCLUSIONS: This new technique of contrast-enhanced OCT imaging of the tear film following lipid-based tracer instillation provides a measure of the clearance of lipids. The quantitative values found are in agreement with other methods of evaluation of the lacrimal system. An improvement of the clinician's ability in the diagnosis and understanding of abnormalities of the ocular surface may be achieved by this simple approach.
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Medios de Contraste , Enfermedades de los Párpados/diagnóstico , Lípidos/química , Lágrimas/química , Adulto , Anciano , Área Bajo la Curva , Aceite de Ricino/química , Medios de Contraste/química , Enfermedades de los Párpados/diagnóstico por imagen , Femenino , Fluoresceína/metabolismo , Humanos , Masculino , Glándulas Tarsales/metabolismo , Persona de Mediana Edad , Curva ROC , Radiografía , Lágrimas/metabolismo , Tomografía de Coherencia Óptica , Agua/químicaRESUMEN
Active substances in traditional Chinese Medicine (TCM) contain not only a variety of small molecules, but also many other macromolecules (TCMMs), such as proteins, peptides and polysaccharides. Active TCMM can achieve good therapeutic effects by regulating the body's overall function with lower side effects. This review summarized the literatures published in recent years on the application of fluorescently labeled tracer technique for detection of natural active macromolecules in TCM. Classified by fluorescent markers, applications of fluorescein, rhodamine, and quantum dots (QDs) in TCMM active tracer are reviewed, and the methods and principles of TCMM fluorescent marker are illustrated. Studies on active TCMMs and their action mechanism are quite difficult due to a multitarget, multicomponent, and multipath system of TCM. However, the development of fluorescently labeled active tracer technique (FLATT) provides this research with new tools. Traditional fluorescent markers have many deficiencies, such as easily quenched, short luminous cycle, and intrinsic toxicity. Relatively, FLATT has many obvious advantages, and its application in TCMM is still at the early stage. In order to improve the overall level of fluorescence labeling in TCMM active tracer, the improvement on FLATT's detection sensitivity and biological affinity is urgent and critical to allow study of these interesting molecules.
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Técnicas Bacteriológicas/métodos , Fluorescencia , Medicina Tradicional China , Coloración y Etiquetado/métodos , Animales , Fluoresceína/metabolismo , Humanos , Puntos Cuánticos/metabolismo , Rodaminas/metabolismoRESUMEN
Handling and mixing ultrasmall volumes of reactants in parallel can increase the throughput and complexity of screening assays while simultaneously reducing reagent consumption. Microfabricated silicon and plastic can provide reliable fluidic devices, but cannot typically handle total volumes smaller than â¼1 × 10(-12) l. Self-assembled soft matter nanocontainers can in principle significantly improve miniaturization and biocompatibility, but exploiting their full potential is a challenge due to their small dimensions. Here, we show that small unilamellar lipid vesicles can be used to mix volumes as small as 1 × 10(-19) l in a reproducible and highly parallelized fashion. The self-enclosed nanoreactors are functionalized with lipids of opposite charge to achieve reliable fusion. Single vesicles encapsulating one set of reactants are immobilized on a glass surface and then fused with diffusing vesicles of opposite charge that carry a complementary set of reactants. We find that â¼85% of the â¼1 × 10(6) cm(-2) surface-tethered nanoreactors undergo non-deterministic fusion, which is leakage-free in all cases, and the system allows up to three to four consecutive mixing events per nanoreactor.
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Reactores Biológicos , Técnicas Analíticas Microfluídicas/instrumentación , Técnicas Analíticas Microfluídicas/métodos , Nanoestructuras/química , Nanotecnología/instrumentación , Nanotecnología/métodos , Fosfatasa Alcalina/química , Fosfatasa Alcalina/metabolismo , Fluoresceína/química , Fluoresceína/metabolismo , Reproducibilidad de los Resultados , Espectrometría de Fluorescencia , Liposomas Unilamelares/químicaRESUMEN
AIM: To investigate colorectal uptake of solid lipid nanoparticles (SLNs) in mice receiving different doses of 1,2-dimethylhydrazine (DMH) using magnetic resonance (MR) and laser-scanning confocal fluorescence microscope (LSCFM) imaging. METHODS: Eight mice were sacrificed in a pilot study to establish the experimental protocol and to visualize colorectal uptake of SLNs in normal mice. Gadopentetate dimeglumine and fluorescein isothiocyanate (FITC)-loaded SLN (Gd-FITC-SLN) enemas were performed on mice receiving DMH for 10 wk (group 1, n = 9) or 16 wk (group 2, n = 7) and FITC-SLN enema was performed on 4 DMH-treated mice (group 3). Pre- and post-enema MR examinations were made to visualize the air-inflated distal colorectum. Histological and LSCFM examinations were performed to verify colorectal malignancy and to track the distribution of SLNs. RESULTS: Homogeneous enhancement and dense fluorescence (FITC) deposition in colorectal wall were observed in normal mice and 1 DMH-treated mouse (group 1) on fluid attenuated inversion recovery (FLAIR) and LSCFM images, respectively. Heterogeneous mural enhancement was found in 6 mice (4 in group 1; 2 in group 2). No visible mural enhancement was observed in the other mice. LSCFM imaging revealed linear fluorescence deposition along the colorectal mucosa in all groups. Nine intraluminal masses and one prolapsed mass were detected by MR imaging with different enhancement modes and pathologies. Interstitial FITC deposition was identified where obvious enhancement was observed in FLAIR images. Bladder imaging agent accumulations were observed in 11 of 16 DMH-treated mice of groups 1 and 2. CONCLUSION: There are significant differences in colorectal uptake and distribution of SLNs between normal and DMH-treated mice, which may provide a new mechanism of contrast for MR colonography.
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1,2-Dimetilhidrazina/farmacología , Carcinógenos/farmacología , Colon/efectos de los fármacos , Colon/metabolismo , Nanopartículas , Recto/efectos de los fármacos , Recto/metabolismo , 1,2-Dimetilhidrazina/administración & dosificación , Adenocarcinoma/inducido químicamente , Adenocarcinoma/metabolismo , Adenocarcinoma/patología , Animales , Colon/patología , Neoplasias Colorrectales/inducido químicamente , Neoplasias Colorrectales/metabolismo , Neoplasias Colorrectales/patología , Enema , Fluoresceína/metabolismo , Colorantes Fluorescentes/metabolismo , Imagen por Resonancia Magnética/métodos , Masculino , Ratones , Microscopía Confocal/métodos , Proyectos Piloto , Recto/patologíaRESUMEN
In this paper, we describe a new continuous fluorescence turn-on method for trypsin assay and inhibitor screening in situ. This assay is designed based on the following assumptions: (1) It is expected that the fluorescein-labeled peptide composed of six arginine residues (Arg(6)-FAM) with positive charges will interact with the negatively charged edge of water-soluble graphene oxide (GO) because of electrostatic interactions to form a GO/Arg(6)-FAM complex. As a result, the fluorescence of fluorescein will be quenched because of the energy transfer from fluorescein to GO. (2) Arg(6)-FAM can be hydrolyzed into small fragments in the presence of trypsin, and accordingly, the GO/Arg(6)-FAM complex will be dissociated, gradually leading to fluorescence recovery for the solution. In this way, the trypsin activity can be easily assayed with the ensemble of Arg(6)-FAM and GO. Additionally, the ensemble can be employed for screening of the inhibitors of trypsin.
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Evaluación Preclínica de Medicamentos/métodos , Fluorescencia , Grafito/metabolismo , Óxidos/metabolismo , Inhibidores de Tripsina/aislamiento & purificación , Inhibidores de Tripsina/farmacología , Tripsina/metabolismo , Fluoresceína/metabolismo , Coloración y Etiquetado/métodosRESUMEN
This work evaluates the protective effects of Chinese herbs against ONOO(-)-induced biomolecule damage. Thirty-two Chinese traditional herbs were preliminarily screened for their ONOO(-)-scavenging activity by the fluorometric method. The potency of scavenging activity was in the following order: Apis cerana Fabricius (Propolis) > Rosmarinus officinalis L (Rosemary) > Pseudolarix amabilis (Nelson) Rehd. (Pine Bark PE) > Echinacea Moenck. (Echinacea); the remaining twenty-eight herbs performed unsatisfactorily for their scavenging activity. The four extracts with high ONOO(-) scavenging activity were selected for further characterization of their inhibiting effects on DNA single breaks, tyrosine nitration and LDL oxidation. A further study of the origin of this scavenging activity on the major active components showed cichoric acid and rosemary acid to both have strong ability to scavenge ONOO(-). These scavengers might be developed as therapeutic drugs for preventing ONOO(-)-involved diseases.
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Medicamentos Herbarios Chinos/farmacología , Depuradores de Radicales Libres/farmacología , Ácido Peroxinitroso/toxicidad , Roturas del ADN/efectos de los fármacos , Evaluación Preclínica de Medicamentos , Fluoresceína/metabolismo , Fluorometría , Humanos , Lipoproteínas LDL/metabolismo , Oxidación-Reducción/efectos de los fármacos , Tirosina/metabolismoRESUMEN
Numerous studies using Caenorhabditis elegans have used a protocol in which chemicals are orally delivered by incorporating them into the nematode growth media or mixing them with the food bacteria. However, actual exposure levels are difficult to estimate as they are influenced by both the rates of ingestion into the intestine as well as absorption from the intestinal lumen. We used liposomes loaded with the hydrophilic fluorescent reagent uranin to test oral administration of water-soluble substances to C. elegans. Ingestion of liposomes loaded with fluorescent dye resulted in successful oral delivery of chemicals into the intestines of C. elegans. Using liposomes, oral administration of hydrophilic antioxidants (ascorbic acid, N-acetyl-cysteine, reduced glutathione, and thioproline) prolonged the lifespan of the nematodes, whereas the conventional method of delivery showed neither fluorescence nor longevity effects. Our method efficiently and quantitatively delivers solutes to nematodes.
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Antioxidantes/farmacología , Caenorhabditis elegans/efectos de los fármacos , Longevidad/efectos de los fármacos , Nematodos/metabolismo , Administración Oral , Animales , Caenorhabditis elegans/genética , Relación Dosis-Respuesta a Droga , Fluoresceína/metabolismo , Colorantes Fluorescentes/metabolismo , Liposomas/metabolismo , Solubilidad , Factores de Tiempo , Agua/químicaRESUMEN
Cytochrome P450 enzyme (CYP450s) assays are critical enzymes in early-stage lead discovery and optimization in drug development. Currently available fluorescence-based reaction assays provide a rapid and reliable method for monitoring CYP450 enzyme activity but are confined to medium-throughput well-plate systems. The authors present a high-throughput, integrated screening platform for CYP450 assays combining enzyme encapsulation techniques, microarraying methods, and wide-field imaging. Alginate-containing microarrays consisting of up to 1134 CYP450 reaction elements were fabricated on functionalized glass slides (reaction volumes 20 to 80 nL, total enzyme content in pg) and imaged to yield endpoint activity, stability, and kinetic data. A charge-coupled device imager acquired quantitative, high-resolution images of a 20x20 mm area/snapshot using custom-built wide-field optics with telecentric lenses and easily interchangeable filter sets. The imaging system offered a broad dynamic intensity range (linear over 3 orders of magnitude) and sensitivity down to fluorochrome quantities of <5 fmols, with read accuracy similar to a laser scanner or a fluorescence plate reader but with higher throughput. Rapid image acquisition enabled analysis of CYP450 kinetics. Fluorogenic assays with CYP3A4, CYP2C9, and CYP2D6 on the alginate microarrays exhibited Z' factors ranging from 0.75 to 0.85, sensitive detection of inhibitory compounds, and reactivity comparable to that in solution, thereby demonstrating the reliability and accuracy of the microarray platform. This system enables for the first time a significant miniaturization of CYP enzyme assays with significant conservation of assay reagents, greatly increased throughput, and no apparent loss of enzyme activity or assay sensitivity.
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Sistema Enzimático del Citocromo P-450/metabolismo , Evaluación Preclínica de Medicamentos/métodos , Análisis por Micromatrices/métodos , Alginatos/metabolismo , Calibración , Inhibidores Enzimáticos del Citocromo P-450 , Inhibidores Enzimáticos/farmacología , Estabilidad de Enzimas/efectos de los fármacos , Fluoresceína/metabolismo , Fluorescencia , Ácido Glucurónico/metabolismo , Ácidos Hexurónicos/metabolismo , Humanos , Concentración 50 Inhibidora , Cinética , Especificidad por Sustrato/efectos de los fármacosRESUMEN
PURPOSE: To examine the effects of berberine, an alkaloid isolated from some medicinal herbs, on the disruption of the barrier function in a human retinal pigment epithelial cell line (ARPE-19) stimulated with interleukin-1beta (IL-1beta). METHODS: ARPE-19 cells were cultured to confluence. Berberine and IL-1beta were added to the medium. Barrier functions were evaluated by measuring transepithelial electrical resistance (TER) and the permeability to horseradish peroxidase (HRP) and sodium fluorescein (SF). RESULTS: Berberine dose-dependently inhibited decreased TER and increased the permeability to HRP and SF in the cells stimulated with IL-1beta. CONCLUSIONS: Berberine dose-dependently inhibited the disruption of the barrier function in the ARPE-19 cell line induced by IL-1beta.
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Berberina/farmacología , Barrera Hematorretinal/fisiología , Epitelio Pigmentado Ocular/efectos de los fármacos , Transporte Biológico/efectos de los fármacos , Línea Celular , Permeabilidad de la Membrana Celular , Relación Dosis-Respuesta a Droga , Impedancia Eléctrica , Electrofisiología , Fluoresceína/metabolismo , Peroxidasa de Rábano Silvestre/metabolismo , Humanos , Interleucina-1beta/farmacología , Epitelio Pigmentado Ocular/metabolismoRESUMEN
PURPOSE: To evaluate the efficacy of a thermo sensitive punctum plug, the Smart Plug, in the treatment of dry eyes. DESIGN: Observational case-series study. METHODS: Eighteen eyes of 10 dry eye patients who previously failed conventional plug insertion were enrolled. Schirmer 1 test, vital staining scores, and tear clearance tests were performed before plug insertion, at 2 weeks, 1 month, and 3 months after insertion. RESULTS: Although Schirmer test values were not significantly different before and after plug insertion, the tear clearance rate considerably decreased (Before insertion: 14.8+/-11.4; After insertion: 8.2+/-6.6) (P<.05). Rose-Bengal and fluorescein scores improved significantly after plug insertion (Rose-Bengal: Before insertion: 6.4+/-2.0; After insertion: 3.3+/-1.6 points, Fluorescein: Before insertion: 4.8+/-2.3; After insertion: 2.1+/-1.3) (P<.05). No complications were observed. CONCLUSIONS: Smart Plug proved to be a safe and efficient option in the treatment of dry eye.
Asunto(s)
Síndromes de Ojo Seco/cirugía , Aparato Lagrimal/cirugía , Prótesis e Implantes , Implantación de Prótesis , Síndromes de Ojo Seco/metabolismo , Fluoresceína/metabolismo , Colorantes Fluorescentes/metabolismo , Humanos , Aparato Lagrimal/metabolismo , Rosa Bengala/metabolismo , Lágrimas/metabolismoRESUMEN
PURPOSE: Trehalose eyedrops were found by a previous study to be safe and effective compared with saline in the treatment of moderate-to-severe dry eye syndrome. The present study was designed to compare the efficacy of trehalose eyedrops with that of the commercially available eyedrops containing hyaluronan or cellulose now used in the treatment of moderate-to-severe dry eye syndrome. METHODS: In a randomized, double-masked, 4-week crossover, controlled clinical trial, 36 patients with moderate-to-severe dry eye syndrome were divided into two groups: the hyaluronan (Hyalein)-comparison group (18 patients) and the hydroxyethylcellulose (Mytear)-comparison group (18 patients). Each group used either trehalose or one of the commercially available medications contained in a masked eyedrop container for the first 4 weeks, and then for the second 4 weeks, switched to either trehalose or the commercial eyedrop not used for the first 4 weeks. Symptoms and signs in both eyes were recorded at the baseline, at 4 weeks, and at 8 weeks. RESULTS: At 4 weeks after the treatment, fluorescein and rose bengal staining scores of the ocular surface as well as the tear film breakup time had improved significantly with trehalose eyedrops compared with the commercially available eyedrops containing either hyaluronan or hydroxyethylcellulose (P < 0.001, Wilcoxon signed ranks test). In addition, all the objective signs were significantly better in patients who finished with trehalose at the end of the 8-week trial compared with those who finished with either of the two commercially available drugs. A larger number of patients evaluated trehalose as a better treatment than the commercially available eyedrops. CONCLUSIONS: Trehalose solution was a better treatment for moderate-to-severe dry eye syndrome in comparison with two commercially available eyedrops containing hyaluronan or hydroxyethylcellulose.