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1.
J Pharm (Cairo) ; 2013: 848275, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-26555997

RESUMO

We have developed submicron-sized liposomes modified with a mucoadhesive polymer to enhance peptide drug absorption after oral administration. Liposomal behavior in the gastrointestinal tract is a critical factor for effective peptide drug delivery. The purpose of this study was to prepare quantum dot- (QD-) loaded submicron-sized liposomes and examine liposomal behavior in the body after oral administration using in vivo fluorescence imaging. Two types of CdSe/CdZnS QDs with different surface properties were used: hydrophobic (unmodified) QDs and hydrophilic QDs with glutathione (GSH) surface modifications. QD- and GSH-QD-loaded liposomes were prepared by a thin film hydration method. Transmission electron microscopy revealed that QDs were embedded in the liposomal lipid bilayer. Conversely, GSH-QDs were present in the inner aqueous phase. Some of the GSH-QDs were electrostatically associated with the lipid membrane of stearylamine-bearing cationic liposomes. QD-loaded liposomes were detected in Caco-2 cells after exposure to the liposomes, and these liposomes were not toxic to the Caco-2 cells. Furthermore, we evaluated the in vivo bioadhesion and intestinal penetration of orally administered QD-loaded liposomes by observing the intestinal segment using confocal laser scanning microscopy.

2.
Artigo em Japonês | MEDLINE | ID: mdl-22186198

RESUMO

The half-value layer (HVL) is an important index of the image quality or radiation risk in mammography. Radiation risk of the breast tissue is evaluated with the average glandular dose. The HVL index is indispensable for the average glandular dose computations. We investigated the influence of multiple factors that affect HVL value, such as thickness or purity of the aluminum attenuator, detector material of dosimeter, fluctuation of X-ray output, detector location in X-ray field and so on, for accurate average glandular dose computations. We found some aluminum plates about 20% thicker than nominal thickness. The HVL values between seven filter sets were different in about 5% at the maximum. In addition, we reduced a fluctuation of X-ray output with dose monitoring. Then, the standard deviation of HVL value decreased from 1.114% to 0.105%. HVL value obtained from a solid-state detector was statistically thicker than that measured by ionization chamber. It has been reported that there was a difference in the half-value layer under the influence of a heel effect by location of the measurement. Accompanied with alternation of detector location, HVL value of PCM (Konica Minolta) had a significant difference, while Novation (Siemens) and Senographe 2000D (GE) had no change.


Assuntos
Mamografia/instrumentação , Mamografia/métodos , Alumínio , Filtração , Mamografia/efeitos adversos , Doses de Radiação , Radiometria/instrumentação , Risco , Sensibilidade e Especificidade
3.
Eur J Pharm Biopharm ; 77(2): 216-24, 2011 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-21147220

RESUMO

Surface modification of liposomal nanocarriers with a novel polymer-lectin conjugate was proposed for enhancing the systemic uptake of encapsulated peptide and protein therapeutics after oral administration. Wheat germ agglutinin (WGA) was covalently attached to carbopol (CP) using the carbodiimide method. The prepared WGA-CP conjugate retained the biological cell binding activity of WGA without any evidence of cytotoxicity to Caco-2 monolayers. Cationic liposomes in the size range of 100 nm were prepared by the lipid film hydration method followed by probe sonication and surface modification with negatively charged WGA-CP. The uptake of WGA-CP liposomes by Caco-2 cells was significantly higher than that of non-modified or CP liposomes. The uptake was dependent on the surface concentration of WGA, temperature, and incubation period and was significantly inhibited in the presence of chlorpromazine and 10-fold excess of free WGA. These results suggest the involvement of active transport mechanism for the cellular uptake of the modified liposomes, mediated mainly by binding of WGA to its specific cell membrane receptors. Dual channel confocal microscopy confirmed the simultaneous association and internalization of the polymer conjugate and the liposomal carrier by Caco-2 cells and intestinal membrane of rats. In addition, the pharmacological efficacy of calcitonin, a model peptide drug, was enhanced by more than 20- and 3-fold following peroral administration of calcitonin-loaded WGA-CP liposomes when compared to non-modified and CP liposomes, respectively.


Assuntos
Calcitonina/administração & dosagem , Cálcio/sangue , Lipossomos , Polivinil , Aglutininas do Germe de Trigo , Resinas Acrílicas , Administração Oral , Animais , Células CACO-2 , Calcitonina/farmacocinética , Sobrevivência Celular , Humanos , Intestino Delgado/efeitos dos fármacos , Intestino Delgado/metabolismo , Lectinas/química , Lipossomos/química , Lipossomos/metabolismo , Masculino , Tamanho da Partícula , Polímeros/química , Polivinil/química , Ratos , Ratos Wistar , Aglutininas do Germe de Trigo/química , Aglutininas do Germe de Trigo/metabolismo
4.
Mol Cell Biol ; 23(2): 474-81, 2003 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-12509447

RESUMO

In lower eukaryotes, Rad18 plays a crucial role in postreplication repair. Previously, we isolated a human homologue of RAD18 (hRAD18) and showed that human cells overexpressing hRad18 protein with a mutation in the RING finger motif are defective in postreplication repair. Here, we report the construction of RAD18-knockout mouse embryonic stem cells by gene targeting. These cells had almost the same growth rate as wild-type cells and manifested phenotypes similar to those of human cells expressing mutant Rad18 protein: hypersensitivity to multiple DNA damaging agents and a defect in postreplication repair. Mutation was not induced in the knockout cells with any higher frequencies than in wild-type cells, as shown by ouabain resistance. In the knockout cells, spontaneous sister chromatid exchange (SCE) occurred with twice the frequency observed in normal cells. After mild DNA damage, SCE was threefold higher in the knockout cells, while no increase was observed in normal cells. Stable transformation efficiencies were approximately 20-fold higher in knockout cells, and gene targeting occurred with approximately 40-fold-higher frequency than in wild-type cells at the Oct3/4 locus. These results indicate that dysfunction of Rad18 greatly increases both the frequency of homologous as well as illegitimate recombination, and that RAD18 contributes to maintenance of genomic stability through postreplication repair.


Assuntos
Reparo do DNA , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/fisiologia , Proteínas de Saccharomyces cerevisiae , Motivos de Aminoácidos , Animais , Células COS , DNA/metabolismo , Dano ao DNA , DNA Complementar/metabolismo , Proteínas de Ligação a DNA/metabolismo , Relação Dose-Resposta a Droga , Glutationa Transferase/metabolismo , Humanos , Imuno-Histoquímica , Ligases/metabolismo , Camundongos , Camundongos Knockout , Modelos Genéticos , Mutagênicos/farmacologia , Mutação , Plasmídeos/metabolismo , Testes de Precipitina , Recombinação Genética , Troca de Cromátide Irmã , Células-Tronco/citologia , Fatores de Tempo , Transfecção , Enzimas de Conjugação de Ubiquitina , Ubiquitina-Proteína Ligases , Raios Ultravioleta , Raios X
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