Reduced on- and off-transients in <i>dmn</i> and <i>rab6</i> mutants.

<p>ERG response of (A) control, (B) <i>dmn</i>^{<i>K16109</i>}<i>/hid</i> (<i>ey-flp Rh1</i>::<i>GFP; FRT42D dmn</i>^{<i>K16109</i>}<i>/ FRT42D GMR-hid CL</i>), (C) precise p-element excised <i>dmn</i>^<i>ex</i><i>/hid</i> (<i>ey-flp Rh1</i>::<i>GFP; FRT42D dmn</i>^<i>ex</i><i>/ FRT42D GMR-hid CL</i>), (D) <i>dmn</i>^{<i>K16109</i>}<i>/hid;GMR>dmn</i> (<i>ey-flp Rh1</i>::<i>GFP; FRT42D dmn</i>^{<i>K16109</i>}<i>/ FRT42D GMR-hid CL;GMR-gal4/UAS-dmn</i>), (E) <i>rab6</i>^{<i>EP2397</i>}<i>/hid</i> (<i>ey-flp Rh1</i>::<i>GFP; rab6</i>^{<i>EP2397</i>}<i>FRT40A/GMR-hid CL FRT40A</i>), (F) precise p-element excised <i>rab6</i>^<i>ex</i><i>/hid</i> (<i>ey-flp Rh1</i>::<i>GFP; rab6</i>^<i>ex</i><i>FRT40A/GMR-hid CL FRT40A</i>) flies in response to a 2-s orange light stimuli.</p

Rhodopsin levels and the integrity of photoreceptor cells using Rh1-GFP.

<p>Representative images of the GFP fluorescence in intact eyes are shown. (A-E) The green fluorescing deep pseudopupil of flies with different genotypes expressing Rh1-GFP (upper panel). (A’-E’) GFP-fluorescence was detected in intact eyes after cornea optical neutralization by water immersion. (A, A’) wild type (<i>Rh1</i>::<i>GFP</i>), (B, B’) <i>Rh1</i>::<i>GFP</i> flies raised in vitamin A-free food, (C, C’) <i>ninaA</i>^<i>1</i> (<i>Rh1</i>::<i>GFP;ninaA</i>^<i>1</i>), (D, D’) <i>rdgA</i>^<i>BS12</i> (<i>rdgA</i>^<i>BS12</i><i>Rh1</i>::<i>GFP</i>), (E, E’) <i>rdgA</i>^<i>BS12</i> 5 day-old. With the exception of the <i>rdgA</i>^<i>BS12</i> flies in E and E’, flies depicted in this figure were 1 day old. Scale bar on upper panels, 50 μm; on lower panel, 2 μm.</p

<i>roh</i> mutation results in Rh1 decrease.

<p>(A) GFP fluorescence of one-day old wild-type (left panel) and <i>roh</i>^{<i>EY04039</i>} (right panel) flies showing a reduction in Rh1-GFP. Scale bar, 50 μm. (B) Western blotting showing decreased Rh1 levels in the <i>roh</i>^{<i>EY04039</i>} mutant (<i>ey-flp Rh1</i>::<i>GFP; FRT42D roh</i>^{<i>EY04039</i>}<i>/ FRT42D GMR-hid CL</i>); the INAD and TRP levels are not affected compared to the wild-type. The reduction of Rh1 levels could be rescued in <i>roh</i>^<i>ex</i> (<i>ey-flp Rh1</i>::<i>GFP; FRT42D roh</i>^<i>ex</i><i>/FRT42D GMR-hid CL</i>) and <i>roh</i>^{<i>EY04039</i>};<i>GMR>roh</i> (<i>ey-flp Rh1</i>::<i>GFP; FRT42D roh</i>^{<i>EY04039</i>}<i>/ FRT42D GMR-hid CL;GMR-gal4/UAS-roh</i>) flies. Flies less than 1 day old were used. (C) Quantification of relative Rh1 level in various genotypes: wt, <i>ninaA</i>^<i>1</i>, <i>roh</i>^{<i>EY04039</i>}, <i>roh</i>^<i>ex</i> and <i>roh</i>^{<i>EY04039</i>};<i>GMR>roh</i>. The Rh1 levels were normalized to tubulin. (D) Quantitative real-time PCR of wild-type (<i>ey-flp Rh1</i>::<i>GFP; FRT42D/ FRT42D GMR-hid CL</i>) and <i>roh</i>^{<i>EY04039</i>} fly head with <i>rh1</i>-specific primers. The results are normalized to the expression level of <i>gpdh</i>. The graphs represent the means ± SD of three independent experiments. Error bars represent the SD. (E) Average rhabdomere numbers per ommatidia of the <i>roh</i>^{<i>EY04039</i>} mutant under the indicated conditions. Each data point was based on examination of >60 ommatidia from >3 flies. Error bars represent the SDs. Asterisks indicate statistically-significant differences (one-way ANOVA and post-hoc Dunnett’s test; ns: not significant, **p < 0.01).</p

The <i>scox</i> and <i>porin</i> mutations lead to light-dependent photoreceptor cell degeneration.

<p>(A-B) Average rhabdomere numbers per ommatidia of (A) the <i>scox</i> mutant flies and (B) the <i>porin</i> mutant flies under the indicated conditions. Each data point was based on examination of >60 ommatidia from >3 flies. Error bars represent the SD. Asterisks indicate statistically-significant differences (one-way ANOVA and post-hoc Dunnett’s test, **p < 0.01). (C-K) Transmission electron microscopy sections of single ommatidia of fly compound eyes with the indicated genotype and conditions. (C) 10 day-old wild-type, (D) 1 day-old <i>scox</i>^{<i>EY05333</i>}<i>/hid</i> (<i>ey-flp Rh1</i>::<i>GFP; scox</i>^{<i>EY05333</i>}<i>FRT40A/GMR-hid CL FRT40A</i>), (E) 10 day-old <i>scox</i>^{<i>EY05333</i>}<i>/hid</i>, (F) 10 day-old <i>scox</i>^{<i>EY05333</i>}<i>/hid</i> under dark condition, (G) 10 day-old P-element excised <i>scox</i>^<i>ex</i><i>/hid</i> (<i>ey-flp Rh1</i>::<i>GFP; scox</i>^<i>ex</i><i>FRT40A/GMR-hid CL FRT40A</i>), (H) 1 day-old <i>porin</i>^{<i>k05123</i>}<i>/hid</i> (<i>ey-flp Rh1</i>::<i>GFP; porin</i>^{<i>k05123</i>}<i>FRT40A/GMR-hid CL FRT40A</i>), (I) 10 day-old <i>porin</i>^{<i>k05123</i>}<i>/hid</i>, (J) 10 day-old <i>porin</i>^{<i>k05123</i>}<i>/hid</i> under dark condition, (K) 10 day-old p-element excised <i>porin</i>^<i>ex</i><i>/hid</i> (<i>ey-flp Rh1</i>::<i>GFP; porin</i>^<i>ex</i><i>FRT40A/GMR-hid CL FRT40A</i>). Scale bar, 2 μm. With the exception of the dark-reared (F) <i>scox</i>^{<i>EY05333</i>}<i>/hid</i> and (J) <i>porin</i>^{<i>k05123</i>}<i>/hid</i> flies, flies were maintained under a 12 hr light/12 hr dark cycle. (L) ERG responses of wild-type, <i>scox</i>^{<i>EY05333</i>}<i>/hid</i>, <i>scox</i>^<i>ex</i><i>/hid</i>, <i>porin</i>^{<i>k05123</i>}<i>/hid</i>, and p-element excised <i>porin</i>^<i>ex</i><i>/hid</i> flies in response to a 10-s orange light stimulus as indicated. Flies used were less than 2 days old.</p

Analysis of mutants of rhodopsin homeostasis, retinal degeneration, and phototransduction with the <i>Rh1</i>::<i>GFP ey-flp/hid</i> method.

<p>(A-E) Detection of fluorescence in the deep pseudopupil (left panels) and by cornea optical neutralization (right panel). (A) <i>FRT40A/hid</i>, (B) <i>ninaA</i>^<i>1</i><i>/hid</i> (<i>ey-flp Rh1</i>::<i>GFP;ninaA</i>^<i>1</i><i>FRT40A/GMR-hid CL FRT40A</i>), (C) <i>trp</i>^<i>P343</i><i>/hid</i> (<i>ey-flp Rh1</i>::<i>GFP;FRT82B trp</i>^<i>P343</i><i>/ FRT82B GMR-hid CL</i>), (D) <i>trp</i>^<i>P343</i><i>/hid</i> 10 day-old. 1 day-old flies were used, with the exception of the <i>trp</i>^<i>P343</i><i>/hid</i> flies, which were 10 day-old (D). Scale bar in right panels, 50 μm; in the left panels, 2μm. (E-H) ERG recordings of (E) wild type and (F) <i>ninaA</i>^<i>1</i><i>/hid</i> flies. Flies were exposed to 5 s pulses of orange light (O) or blue light (B), interspersed by 7 s, as indicated. A PDA was induced in the wild-type by blue light and terminated by orange light (arrows). (G-J) ERG response of (G) wild-type, (H) <i>Hdc</i>^<i>P217</i><i>/hid</i>, (I) <i>trp</i>^<i>P343</i>, and (J) <i>trp</i>^<i>P343</i><i>/hid</i> flies in response to a 5-s orange light stimulus.</p

Brain response to luminance-based and motion-based stimulation using inter-modulation frequencies

<div><p>Steady state visual evoked potential (SSVEP)-based brain computer interface (BCI) has advantages of high information transfer rate (ITR), less electrodes and little training. So it has been widely investigated. However, the available stimulus frequencies are limited by brain responses. Simultaneous modulation of stimulus luminance is a novel method to resolve this problem. In this study, three experiments were devised to gain a deeper understanding of the brain response to the stimulation using inter-modulation frequencies. First, luminance-based stimulation using one to five inter-modulation frequencies was analyzed for the first time. The characteristics of the brain responses to the proposed stimulation were reported. Second, the motion-based stimulation with equal luminance using inter-modulation frequencies was also proposed for the first time. The response of the brain under these conditions were similar to that of luminance-based stimulation which can induce combination frequencies. And an elementary analysis was conducted to explain the reason of the occurrence of combination frequencies. Finally, the online test demonstrated the efficacy of our proposed two stimulation methods for BCI. The average ITRs reached 34.7836 bits/min and 39.2856 bits/min for luminance-based and motion-based stimulation respectively. This study demonstrated that the simultaneous modulation of stimulus luminance could extend to at least five frequencies to induce SSVEP and the brain response to the stimulus still maintained a certain positive correlation with luminance. And not only luminance-based stimulation, but also motion-based stimulation with equal luminance can elicit inter-modulation frequencies to effectively increase the number of targets for multi-class SSVEP.</p></div

<i>De novo</i> Transcriptome Assembly and the Putative Biosynthetic Pathway of Steroidal Sapogenins of <i>Dioscorea composita</i>

<div><p>The plant <i>Dioscorea composita</i> has important applications in the medical and energy industries, and can be used for the extraction of steroidal sapogenins (important raw materials for the synthesis of steroidal drugs) and bioethanol production. However, little is known at the genetic level about how sapogenins are biosynthesized in this plant. Using Illumina deep sequencing, 62,341 unigenes were obtained by assembling its transcriptome, and 27,720 unigenes were annotated. Of these, 8,022 unigenes were mapped to 243 specific pathways, and 531 unigenes were identified to be involved in 24 secondary metabolic pathways. 35 enzymes, which were encoded by 79 unigenes, were related to the biosynthesis of steroidal sapogenins in this transcriptome database, covering almost all the nodes in the steroidal pathway. The results of real-time PCR experiments on ten related transcripts (<i>HMGR</i>, <i>MK</i>, <i>SQLE</i>, <i>FPPS</i>, <i>DXS</i>, <i>CAS</i>, <i>HMED</i>, <i>CYP51</i>, <i>DHCR7</i>, and <i>DHCR24</i>) indicated that sapogenins were mainly biosynthesized by the mevalonate pathway. The expression of these ten transcripts in the tuber and leaves was found to be much higher than in the stem. Also, expression in the shoots was low. The nucleotide and protein sequences and conserved domains of four related genes (<i>HMGR</i>, <i>CAS</i>, <i>SQS</i>, and <i>SMT1</i>) were highly conserved between <i>D</i>. <i>composita</i> and <i>D</i>. <i>zingiberensis</i>; but expression of these four genes is greater in <i>D</i>. <i>composita</i>. However, there is no expression of these key enzymes in potato and no steroidal sapogenins are synthesized.</p></div

Online classification accuracy of luminance-based stimulation according to stimulation duration.

<p>Online classification accuracy of luminance-based stimulation according to stimulation duration.</p

Shape Evolution of “Multibranched” Mn–Zn Ferrite Nanostructures with High Performance: A Transformation of Nanocrystals into Nanoclusters

Monodisperse magnetic Mn–Zn ferrite nanostructures with various morphologies have been successfully synthesized via high-temperature decomposition of metal acetylacetonate (acac) in the presence of oleic acid (OA) and oleyamine (OAm). In a classical crystal nucleation/growth process, differential stabilization of OA on specific crystal facets may alter relative crystal growth rates, resulting in the formation of zero-dimensional (0-D) spherical, cubical, and starlike nanocrystals (ca. 9, 11, 16 nm), respectively. Furthermore, shortening nucleation duration might bring a deficient nucleation and a rapid increase in monomer concentration, which accelerates the subsequent growth process of nanocrystals, leading to the formation of the starlike nanocrystals with larger size (ca. 19–23 nm). They are further oriented to assemble reciprocally, gradually forming initial three-dimensional (3-D) “branched” nanoclusters (ca. 30–40 nm) to minimize the magnetostatic energy, owing to their size-dependent magnetic dipolar interaction. In addition, the surface-defect-induced secondary growth of the “branched” nanoclusters may considerably improve their uniformity, accompanied by the size increase in the presence of the monomers, resulting in the final “multibranched” nanoclusters with formation of sharp or obtuse edges (ca. 45–50 nm). Our study reveals the transformation of 0-D nanocrystals to 3-D nanoclusters as well as the shape evolution mechanism, which provide a versatile synthetic strategy for shape-controlled nanostructure. The multibranched nanoclusters have the higher magnetization and magnetically induced heating efficiency in an alternating current magnetic field, which can be used as promising heating agents for biomedical application

Expression of four transcripts related to steroidal sapogenin in the tuber of <i>D</i>. <i>composita</i>, <i>D</i>. <i>zingiberensis</i>, and potato.

<p>The <i>D</i>. <i>composita</i> and <i>D</i>. <i>zingiberensis</i> samples came from 18-month-old plants, and the samples of potato from 6-month-old plants.</p