No abstract

We established a protocol of the SuperSAGE technology combined with next-generation sequencing, coined “High-Throughput (HT-) SuperSAGE”. SuperSAGE is a method of digital gene expression profiling that allows isolation of 26-bp tag fragments from expressed transcripts. In the present protocol, index (barcode) sequences are employed to discriminate tags from different samples. Such barcodes allow researchers to analyze digital tags from transcriptomes of many samples in a single sequencing run by simply pooling the libraries. Here, we demonstrated that HT-SuperSAGE provided highly sensitive, reproducible and accurate digital gene expression data. By increasing throughput for analysis in HT-SuperSAGE, various applications are foreseen and several examples are provided in the present study, including analyses of laser-microdissected cells, biological replicates and tag extraction using different anchoring enzymes.

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We report a new approach for fabricating layer-by-layer (LBL) structured ultrathin hybrid films on electrospun nanofibres. Oppositely charged anatase TiO2 nanoparticles and poly(acrylic acid) (PAA) were alternately deposited on the surface of negatively charged cellulose acetate (CA) nanofibres using the electrostatic LBL self-assembly technique. The fibrous mats were characterized by wide-angle x-ray diffraction, Fourier transform infrared spectroscopy, scanning electron microscopy, transmission electron microscopy and Brunauer–Emmett–Teller (BET) surface area techniques. The crystalline phase of anatase TiO2 remained unchanged in the resultant TiO2/PAA films coated on CA fibrous mats. Moreover, the TiO2/PAA film coated fibres showed rough surfaces with grains due to the deposition of aggregated TiO2 particles. The average diameter of the fibres increased from 344 to 584 nm and the BET surface area of the fibrous mats increased from 2.5 to 6.0 m2 g−1 after coating with five bilayers of TiO2/PAA films.

We investigated whether cones are the only photosensitive process mediating the photopic pupillary light reflex. New analyses were performed on previously published recordings, focusing on those evoked by the onset of photopically equated short- and long-wavelength stimuli. Comparisons between responses revealed contraction differences that slowly grew to a peak and gradually declined. The late contraction was associated with short wavelengths and appeared mostly at the higher stimulus intensities. We conclude that cones are not the only photoreception process mediating the photopic ON-reflex and infer that melanopsin is another. Melanopsin contributes to the steady-state pupil size in daylight illumination.