RESUMEN
Nanoscopic synchrotron X-ray imaging was performed on scalp hair samples of patients with breast cancer and healthy individuals to investigate any structural differences as diagnostic tool. Hair strands were divided into 2-3 segments along the strands from root to tip, followed by imaging either in projection or in CT scanning with a monochromatic 6.78-keV X-ray using zone-plate optics with a resolving power of 60 nm. All the examined cancer hairs exhibited medulla loss with cancer stage-dependent pattern; complete loss, discontinuous or trace along the strands. In contrast, medullas were well retained without complete loss in the healthy hair. In the CT-scanned axial images, the cortical spindle compartments had no contrast in the healthy hair, but appeared hypointense in contrast to the surrounding hyperintense cortical membrane complex in the cancer hair. In conclusion, observation of medulla loss and cortical membrane enhancements in the hair strands of breast cancer patients demonstrated structural variations in the cancer hair, providing a new platform for further synchrotron X-ray imaging study of screening breast cancer patients.
Asunto(s)
Neoplasias de la Mama/diagnóstico por imagen , Cabello/diagnóstico por imagen , Cuero Cabelludo/diagnóstico por imagen , Sincrotrones , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Persona de Mediana Edad , Tomografía Computarizada por Rayos XRESUMEN
We investigated the relationship between oct4 gene expression patterns and CpG sites methylation profiles during ES cell differentiation into neurons, and identified relevant binding factor. The oct4 gene expression level gradually declined as ES cell differentiation progressed, and the CpG sites in the oct4 proximal enhancer (PE) and promoter regions were methylated in concert with ES cell differentiation. An electro-mobility shift assay (EMSA) showed that putative proteins bind to CpG sites in the oct4 PE/promoter. We purified CpG binding proteins with DNAbinding purification method, and NonO was identified by liquid chromatography-mass spectrometry. EMSA with specific competitors revealed that NonO specifically binds to the conserved CCGGTGAC sequence in the oct4 promoter. Methylation at a specific cytosine residue (CC* GGTGAC) reduced the binding affinity of NonO for the recognition sequence. Chromatin immunoprecipitation analysis confirmed that NonO binds to the unmethylated oct4 promoter. There were no changes in the NonO mRNA and protein levels between ES cells and differentiated cells. The transcriptional role of NonO in oct4 gene expression was evaluated by luciferase assays and knockdown experiments. The luciferase activity significantly increased threefold when the NonO expression vector was cotransfected with the NonO recognition sequence, indicating that NonO has a transcription activator effect on oct4 gene expression. In accordance with this effect, when NonO expression was inhibited by siRNA treatment, oct4 expression was also significantly reduced. In summary, we purified NonO, a novel protein that binds to the CpG island of oct4 promoter, and positively regulates oct4 gene expression in ES cells.
Asunto(s)
Islas de CpG/genética , Metilación de ADN , Proteínas de Unión al ADN/metabolismo , Células Madre Embrionarias/metabolismo , Regulación de la Expresión Génica , Factor 3 de Transcripción de Unión a Octámeros/genética , Regiones Promotoras Genéticas/genética , Animales , Western Blotting , Diferenciación Celular , Células Cultivadas , Inmunoprecipitación de Cromatina , Proteínas de Unión al ADN/antagonistas & inhibidores , Proteínas de Unión al ADN/genética , Ensayo de Cambio de Movilidad Electroforética , Células Madre Embrionarias/citología , Técnicas para Inmunoenzimas , Luciferasas , Ratones , Factor 3 de Transcripción de Unión a Octámeros/metabolismo , ARN Mensajero/genética , ARN Interferente Pequeño/genética , Proteínas de Unión al ARN , Reacción en Cadena en Tiempo Real de la Polimerasa , Secuencias Reguladoras de Ácidos Nucleicos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Activación TranscripcionalRESUMEN
We demonstrate an enhanced photo-sensitivity (PS) through an increased light-trapping using surface nano-structuring technique by inductively coupled plasma (ICP) etching on multi-walled carbon nanotube (MWCNT) etch masked Si with hexamethyl-disilazane (HMDS) dispersion. In order for a systematic comparison, four samples are prepared, respectively, by conventional photolithography and ICP etching using MWCNT as a etch mask. MWCNT-etched Si with HMDS dispersion shows the highest RMS roughness and the lowest reflectance of the four. Two test device structures are fabricated with active regions of bare-Si as a reference and MWCNT etch masked Si with HMDS dispersion. The increased light-trapping was most significant at mid-UV, somewhat less at visible and less noticeable at infrared. With an ICP-etched Si using CNT HMDS dispersion, PS is very sharply increased. This result can lead to applications in optoelectronics where the enhancement in light-trapping is important.
