Graphic representation of the thickness (taken at 1000μm from the optic nerve head) of different retinal layers: RPE (A); SL (B); ONL (C); OPL (D); INL (E); IPL (F); RGC (G) and total retina (H) for the control (blue lines) and the light exposed (pink lines) groups in both the superior (solid lines) and the inferior retina (dashed lines).

<p>Measurements were obtained at different experimental postnatal periods (from P30 to P400). Results are given as the mean thickness (μm) ± 1SD. Asterisks: Between P30-P35 and at P400, the values reported are those of the thickness of the subretinal space as the length of the outer and inner segments could not be quantifiable (only debris occupied the subretinal space during this period).</p

Sequencing and Validation of Reference Genes to Analyze Endogenous Gene Expression and Quantify Yellow Dwarf Viruses Using RT-qPCR in Viruliferous <i>Rhopalosiphum padi</i>

<div><p>The bird cherry-oat aphid (<i>Rhopalosiphum padi</i>), an important pest of cereal crops, not only directly sucks sap from plants, but also transmits a number of plant viruses, collectively the yellow dwarf viruses (YDVs). For quantifying changes in gene expression in vector aphids, reverse transcription-quantitative polymerase chain reaction (RT-qPCR) is a touchstone method, but the selection and validation of housekeeping genes (HKGs) as reference genes to normalize the expression level of endogenous genes of the vector and for exogenous genes of the virus in the aphids is critical to obtaining valid results. Such an assessment has not been done, however, for <i>R. padi</i> and YDVs. Here, we tested three algorithms (GeNorm, NormFinder and BestKeeper) to assess the suitability of candidate reference genes (EF-1α, ACT1, GAPDH, 18S rRNA) in 6 combinations of YDV and vector aphid morph. EF-1α and ACT1 together or in combination with GAPDH or with GAPDH and 18S rRNA could confidently be used to normalize virus titre and expression levels of endogenous genes in winged or wingless <i>R. padi</i> infected with Barley yellow dwarf virus isolates (BYDV)-PAV and BYDV-GAV. The use of only one reference gene, whether the most stably expressed (EF-1α) or the least stably expressed (18S rRNA), was not adequate for obtaining valid relative expression data from the RT-qPCR. Because of discrepancies among values for changes in relative expression obtained using 3 regions of the same gene, different regions of an endogenous aphid gene, including each terminus and the middle, should be analyzed at the same time with RT-qPCR. Our results highlight the necessity of choosing the best reference genes to obtain valid experimental data and provide several HKGs for relative quantification of virus titre in YDV-viruliferous aphids.</p></div

High-Performance Microsupercapacitors Based on Two-Dimensional Graphene/Manganese Dioxide/Silver Nanowire Ternary Hybrid Film

Microsupercapacitors (MSCs), as one type of significant power source or energy storage unit in microelectronic devices, have attracted more and more attention. However, how to reasonably design electrode structures and exploit the active materials to endow the MSCs with excellent performances in a limited surface area still remains a challenge. Here, a reduced graphene oxide (RGO)/manganese dioxide (MnO₂)/silver nanowire (AgNW) ternary hybrid film (RGMA ternary hybrid film) is successfully fabricated by a facile vacuum filtration and subsequent thermal reduction, and is used directly as a binder-free electrode for MSCs. Additionally, a flexible, transparent, all-solid state RMGA-MSC is also built, and its electrochemical performance in an ionic liquid gel electrolyte are investigated in depth. Notably, the RGMA-MSCs display superior electrochemical properties, including exceptionally high rate capability (up to 50000 mV·s^–1), high frequency response (very short corresponding time constant τ₀ = 0.14 ms), and excellent cycle stability (90.3% of the initial capacitance after 6000 cycles in ionic liquid gel electrolyte). Importantly, the electrochemical performance of RGMA-MSCs shows a strong dependence on the geometric parameters including the interspace between adjacent fingers and the width of the finger of MSCs. These encouraging results may not only provide important references for the design and fabrication of high-performance MSCs, but also make the RGMA ternary hybrid film promising for the next generation film lithium ion batteries and other energy storage devices

Influence of YDV entry and wing morph on raw Cq value for individual genes.

<p>Two-way ANOVA analysis: ns, nonsignificant, <i>p</i>>0.05; * <i>p</i><0.05, ** <i>p</i><0.01; *** <i>p</i><0.001.</p

Box and whisker plots of Cq values for the 4 candidate reference genes in 6 experimental groups (one of three viruses in either winged or wingless adults of <i>Rhopalosiphum padi</i>).

<p>Each box shows the lower 25^th and upper 75^th percentiles with median Cq values; the whiskers mark the lower 5^th and upper 95^th percentiles of the Cq values in each data set. Experimental groups from left to right: WYDV-GPV in winged adult, BYDV-PAV in winged adult, BYDV-GAV in winged adult, WYDV-GPV and wingless adult, BYDV-PAV and wingless adult, and BYDV-GAV and wingless adult.</p

Paired comparisons for reference genes between the WYDV-GPV (GPV), BYDV-PAV (PAV) and BYDV-GAV (GAV)-viruliferous aphids (<i>Rhopalosiphum padi</i>).

<p>Tukey’s HSD post hoc tests: ns, nonsignificant, <i>p</i>>0.05; * <i>p</i><0.05; ** <i>p</i><0.01; *** <i>p</i><0.001.</p

Mean relative titre of YDVs (± SE, <i>n</i> = 3) in viruliferous winged adults of <i>Rhopalosiphum padi</i> after different virus-feeding durations.

<p>Virus titre is illustrated by relative expression values of the CP (A, C and E) and RTD (B, D and F) gene of YDVs. Expression values for the CP or RTD gene at each duration were normalized with the reference gene(s) selected by GeNorm and then compared with a one-way ANOVA (<i>p</i>) among these durations with each normalization condition. A and B: WYDV-GPV; C and D: BYDV-PAV; E and F: BYDV-GAV. 2re = 2 best reference genes; 3re = 3 best reference genes; 4re = all 4 reference genes; bad2re = 2 least stable reference genes; EF-1α = EF-1α as the reference gene; 18S = 18S rRNA as the reference gene.</p

Stability rankings according to three software tools for individual endogenous reference genes in winged adults of <i>Rhopalosiphum padi</i> carrying one of three YDVs.

a<p>According to GeNorm, the smallest <i>M</i> value indicates the most stable gene expression, the largest <i>M</i> value the most unstable; expression of a gene with <i>M</i> >1.5 is considered inconsistent.</p>b<p>Like GeNorm, the most stable gene has the lowest stability value, and vice versa.</p>c<p>Genes with SD <1 can be ranked according to their Cq SD values. The gene most stably expressed has the lowest SD value; the highest SD value indicates the gene with the most unstable expression.</p

Mean relative titre of YDVs (± SE, <i>n = </i>3) in viruliferous wingless adults of <i>Rhopalosiphum padi</i> after different virus-feeding durations.

<p>Virus titre is illustrated by relative expression values of the CP (A, C and E) and RTD (B, D and F) gene of YDVs. Expression values for the CP or RTD gene at each duration were normalized with the reference gene(s) selected by GeNorm and then compared with a one-way ANOVA (<i>p</i>) among these durations with each normalization condition. A and B: WYDV-GPV; C and D: BYDV-PAV; E and F: BYDV-GAV. 2re = 2 best reference genes; 3re = 3 best reference genes; 4re = all 4 reference genes; bad2re = 2 least stable reference genes; EF-1α = EF-1α as the reference gene; 18S = 18S rRNA as the reference gene.</p

Lack of Association between NLGN3, NLGN4, SHANK2 and SHANK3 Gene Variants and Autism Spectrum Disorder in a Chinese Population

<div><p>Autism spectrum disorder (ASD) is a neurodevelopmental disorder characterized by deficits in social communication, absence or delay in language development, and stereotyped or repetitive behaviors. Genetic studies show that neurexin-neuroligin (NRXN-NLGN) pathway genes contribute susceptibility to ASD, which include cell adhesion molecules <i>NLGN3</i>, <i>NLGN4</i> and scaffolding proteins <i>SHANK2</i> and <i>SHANK3</i>. Neuroligin proteins play an important role in synaptic function and trans-synaptic signaling by interacting with presynaptic neurexins. Shank proteins are scaffolding molecules of excitatory synapses, which function as central organizers of the postsynaptic density. Sequence level mutations and structural variations in these genes have been identified in ASD cases, while few studies were performed in Chinese population. In this study, we examined the copy numbers of four genes <i>NLGN4, NLGN3, SHANK2,</i> and <i>SHANK3</i> in 285 ASD cases using multiplex fluorescence competitive polymerase chain reaction (PCR). We also screened the regulatory region including the promoter region and 5′/3′ untranslated regions (UTR) and the entire coding region of <i>NLGN4</i> in a cohort of 285 ASD patients and 384 controls by direct sequencing of genomic DNA using the Sanger method. DNA copy number calculation in four genes showed no deletion or duplication in our cases. No missense mutations in <i>NLGN4</i> were identified in our cohort. Association analysis of 6 common SNPs in <i>NLGN4</i> did not find significant difference between ASD cases and controls. These findings showed that these genes may not be major disease genes in Chinese ASD cases.</p> </div