RESUMEN
We partially characterized the transferrin-independent iron uptake (Tf-IU) of neuronal and glial cells in the previous report. In the present study, we further examined a mechanism of which glial cells protect neuronal cells against iron stress using neuron-microglia (N-MG) and neuron-astrocyte (N-AS) co-cultures. When each solely purified cell was treated with iron citrate, cell death occurred in N and MG. However, AS proliferated under the same condition. Both N-MG and N-AS co-cultures were effective in resistance to excessive iron. The total and specific Tf-IU activities of N-MG co-cultures similar to those of N did not increase in a density-dependent manner. Contrarily, the total activity of AS was extremely high and the specific activity was extremely low as a result of proliferation. Regarding of effect of co-cultures on H(2)O(2)-induced cell death, N-MG co-cultures were less effective, but N-AS co-cultures were more effective in protecting N from the oxidative stress. These results suggest that N-MG co-cultures suppress the Tf-IU and N-AS co-cultures stimulate AS proliferation to protect neuronal cells. Brain cells from aceruloplasminemia with mutations in the ceruloplasmin gene take up iron by Tf-IU. Therefore, the different mechanisms of neuronal cell protection by MG and AS may explain the pathophysiological observations in the brains of patient with aceruloplasminemia.
Asunto(s)
Astrocitos/fisiología , Ceruloplasmina/deficiencia , Microglía/fisiología , Degeneración Nerviosa/prevención & control , Enfermedades Neurodegenerativas/etiología , Neuronas/fisiología , Estrés Oxidativo/fisiología , Animales , Animales Recién Nacidos , Muerte Celular , Células Cultivadas , Ceruloplasmina/genética , Técnicas de Cocultivo , Citoprotección/fisiología , Embrión de Mamíferos , Hierro/efectos adversos , Hierro/metabolismo , Enfermedades Neurodegenerativas/genética , Enfermedades Neurodegenerativas/patología , Neuronas/metabolismo , Neuronas/patología , RatasRESUMEN
We have developed a passive long-wavelength infrared (LWIR) scattering-type scanning near-field optical microscope (s-SNOM) installed in a helium-free mechanically cooled cryostat, which facilitates cooling of an LWIR detector and optical elements to 4.5 K. To reduce mechanical vibration propagation from a compressor unit, we have introduced a metal bellows damper and a helium gas damper. These dampers ensure the performance of the s-SNOM to be free from mechanical vibration. Furthermore, we have introduced a solid immersion lens to improve the confocal microscope performance. To demonstrate the passive s-SNOM capability, we measured thermally excited surface evanescent waves on Au/SiO2 gratings. A near-field signal-to-noise ratio is 4.5 times the improvement with an acquisition time of 1 s/pixel. These improvements have made the passive s-SNOM a more convenient and higher-performance experimental tool with a higher signal-to-noise ratio for a shorter acquisition time of 0.1 s.
RESUMEN
Fine-pitched microgratings are encoded on fused silica surfaces by a two-beam laser interference technique employing UV femtosecond pulses from the third harmonics of a Ti:sapphire laser. A pump and prove method utilizing a laser-induced optical Kerr effect or transient optical absorption change has been developed to achieve the time coincidence of the two pulses. Use of the UV pulses makes it possible to narrow the grating pitches to an opening as small as 290 nm, and the groove width of the gratings is of nanoscale size. The present technique provides a novel opportunity for the fabrication of periodic nanoscale structures in various materials.
Asunto(s)
Calor , Interferometría/métodos , Rayos Láser , Nanotecnología/métodos , Dióxido de Silicio/efectos de la radiación , Interferometría/instrumentación , Nanotecnología/instrumentación , Dióxido de Silicio/química , Propiedades de Superficie , Rayos UltravioletaRESUMEN
In an attempt to achieve chronopharmacotherapy for asthma, press-coated tablets (250 mg), which contained aminophylline in the core tablet in the form of low-substituted hydroxypropylcellulose (L-HPC) and coated with crystalline cellulose (PH-102) and polyethylene glycol (PEG) at various molecular weights and mixing ratios in the amounts of PH-102 and PEG as the outer shell (press-coating material), were prepared (chronopharmaceutics). Their applicability as timed-release (delayed-release) tablets with a lag time of disintegration and a subsequent rapid drug release phase was investigated. Various types of press-coated tablets were prepared using a tableting machine, and their aminophylline dissolution profiles were evaluated by the JP paddle method. Tablets with the timed-release characteristics could be prepared, and the lag time of disintegration was prolonged as the molecular weight and the amount of PEG, for example PEG 500,000, in the outer shell were increased. The lag time of disintegration could be controlled by the above-mentioned method, however, the pH of the medium had no effect on disintegration of the tablet and dissolution behavior of theophylline. The press-coated tablet (core tablet:aminophylline 50 mg, L-HPC and PEG 6000; outer shell:PH-102:PEG = 8:2 200 mg) with the timed-release characteristics was administered orally to rabbits for an in vivo test. Theophylline was first detected in plasma more than 2 h after administration; thus, this tablet showed a timed-release characteristics in the gastrointestinal tract. The time (tmax) required to reach the maximum plasma theophylline concentration (Cmax) observed after administration of the press-coated tablet was significantly (p < 0.05) delayed compared with that observed after administration of aminophylline solution in the control experiment. However, there was no difference in Cmax and area under the plasma theophylline concentration-time curve (AUC0-->24) between the press-coated tablet and aminophylline solution. These results suggest that the press-coated aminophylline tablet (with the timed-release characteristic) offers a promising forms of theophylline chronotherapy for asthma.
Asunto(s)
Aminofilina , Celulosa , Polietilenglicoles , Aminofilina/farmacocinética , Animales , Asma/tratamiento farmacológico , Cronoterapia , Preparaciones de Acción Retardada , Concentración de Iones de Hidrógeno , Conejos , Solubilidad , Comprimidos RecubiertosRESUMEN
Iron regulatory protein-1 (IRP-1) is known as a cytosolic aconitase and a central regulator of iron (Fe) homeostasis. IRP-1 regulates the expression of Fe metabolism-related proteins by interacting with the Fe-responsive element (IRE) in the untranslated regions of mRNAs of these proteins. However, it is less known whether IRP-1 modulates various non-Fe metals. In the present study, we showed that treatment of homogenously purified IRP-1 with non-Fe metals decreased the affinity to IRE in RNA band shift assays and increased aconitase activity. Non-Fe metals also inhibited (55)Fe incorporation into the fourth labile position of the Fe-S cluster of IRP-1. In PLC hepatoma cells, metal loading inactivated binding activity and activated enzyme activity. It also suppressed transferrin receptor mRNA expression in the cells. These results suggest that various non-Fe metals modulate IRP-1 by conversion of the 3Fe-4S apo-form to a [1 non-Fe metal + 3Fe]-4Fe holo-form.
Asunto(s)
Proteínas Hierro-Azufre/metabolismo , Hierro/metabolismo , Metales/farmacología , Proteínas de Unión al ARN/metabolismo , Aconitato Hidratasa/metabolismo , Animales , Unión Competitiva , Northern Blotting , Cadmio/farmacología , Carcinoma Hepatocelular/metabolismo , Bovinos , Quelantes/farmacología , Cobre/farmacología , Relación Dosis-Respuesta a Droga , Ácido Edético/farmacología , Humanos , Proteína 1 Reguladora de Hierro , Proteínas Reguladoras del Hierro , Cinética , Hígado/metabolismo , Manganeso/farmacología , Mercurio/farmacología , Níquel/farmacología , Unión Proteica , ARN Mensajero/metabolismo , Receptores de Transferrina/metabolismo , Células Tumorales CultivadasRESUMEN
Taxol is an effective anti-tumour drug against a variety of tumour cells. Taxol directly induces apoptosis in addition to a G2/M cell cycle arrest. However, it remains poorly understood how Taxol induces apoptosis in tumour cells. Taxol induces the secretion of inflammatory cytokines in murine macrophages in a toll-like receptor-4 (TLR-4)-dependent manner in addition to its anti-tumour effects, but the effect of Taxol on human macrophages is controversial. In this study, we demonstrated that low doses (less than 1000 nmol/l) of Taxol induced the expression of tumour necrosis factor (TNF)-alpha in human myelomonocytic cells and that the induction of TNF-alpha mRNA was inhibited by dominant-negative myeloid differentiation protein (dnMyD88). Furthermore, we demonstrated that the same doses of Taxol induced apoptosis of the same myelomonocytic cells and that the Taxol-induced apoptosis was also inhibited by dnMyD88. In accordance with the previous reports, Taxol induced the expression of TNF-alpha and apoptosis in a TLR4-independent manner. These results suggest that TNF-alpha expression and apoptosis, both induced by Taxol in human myelomonocytic cells, share the signal transduction molecule MyD88.