RGMa peptide 4 (aa 284–293) inhibits RGMa-induced growth cone collapse.

<p>(A) Effects of RGMa peptide 4 on RGMa-induced growth cone collapse. Cortical neurons were treated with RGMa and/or RGMa peptides for 30 min. The results were quantified from 3 independent experiments, and the percentage of collapsed growth cones is shown (**<i>P</i><0.01). Pep4, peptide 4; PepA, Peptide A; PepB, peptide B. (B) Representative images of growth cones. Red: Phalloidin, Green: Tuj1. Scale bar: 10 µm.</p

The RGMa domain required for binding to neogenin is within aa 259–295.

<p>(A and C) Schematic representation of RGMa and its deletion mutants are shown with their domain structures. Arrowhead shows potential cleavage site. SP: signal peptide, vWD: von Willebrand factor type-D domain, HD: hydrophobic domain, GPI: GPI-anchor. (B, D, and E) Co-immunoprecipitation of full-length neogenin-VSV-G with the deletion mutants of RGMa-Myc. HEK293T cells were transiently transfected with the indicated plasmids. Cell lysates were immunoprecipitated with the anti-VSV-G antibody. The immunoprecipitates (IP) and cell lysates (Lysates) were analyzed by western blotting with anti-Myc and anti-VSV-G antibodies.</p

RGMa aa 284–293 are critical for the interaction with neogenin.

<p>(A) Sequence alignment of the synthesized RGMa peptide. Peptides 1–6 were designed within aa 257–310, which is the region required for the interaction between RGMa and neogenin as shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0032791#pone-0032791-g001" target="_blank">Fig. 1</a>. (B) RGMa peptide 4 (aa 284–293) directly binds to neogenin's ECD. Relative binding of each RGMa peptide to ELISA plates coated with neogenin-ECD-Fc is shown. p75NTR-Fc was used as a control. Results (%) are shown as the mean ± SEM of 3 independent experiments (**<i>P</i><0.01, *<i>P</i><0.05).</p

Alkoxy-Substituted Anthra[1,2‑<i>c</i>:5,6‑<i>c</i>′]bis([1,2,5]thiadiazole) (ATz): A New Electron-Acceptor Unit in the Semiconducting Polymers for Organic Electronics

A new type of thiadiazole-based acceptor unit and its donor–acceptor copolymers were synthesized and characterized to develop the high-performance semiconducting polymers for organic field-effect transistors (OFETs) and organic photovoltaics (OPVs). We successfully synthesized an anthra­[1,2-<i>c</i>:5,6-<i>c</i>′]­bis­([1,2,5]­thiadiazole) (ATz) core and ATz-quaterthiophene copolymers. These copolymers possess a wide energy gap of ca. 1.8 eV and a deeper HOMO energy levels around −5.4 eV than that of typical thiadiazole–oligothiophene copolymers. Such weak electron-accepting nature may be due to the decreased electron affinity of the ATz core by an existence of alkoxy groups with strong electron-donating ability. The ATz copolymers exhibited good semiconducting properties with hole mobility of up to 0.03 cm² V^–1 s^–1 and photovoltaic response with PCE of up to 5.7%, despite the unfavorable molecular orders, thin-film structure, and/or amorphous structure

Induced Production of Halogenated Epidithiodiketopiperazines by a Marine-Derived <i>Trichoderma</i> cf. <i>brevicompactum</i> with Sodium Halides

Marine-derived <i>Trichoderma</i> sp. TPU199 (cf. <i>T. brevicompactum</i>) produced gliovirin (<b>1</b>), pretrichodermamide A (<b>2</b>), and trichodermamide A (<b>3</b>) in a freshwater medium. Compounds <b>1</b> and <b>2</b> are rare epidithiodiketopiperazines possessing an unusual disulfide linkage. In the seawater medium, the strain biosynthesized the 5-chloro-5-deoxy derivatives (<b>4</b> and <b>5</b>) of <b>3</b> and <b>2</b>. The production of <b>5</b> was proportional to the concentration of seawater (NaCl). Therefore, 5-bromo-5-deoxy (<b>6</b>) and 5-deoxy-5-iodo (<b>7</b>) derivatives were biosynthesized in the freshwater media supplemented with NaBr and NaI, respectively. The structure of a new iodo derivative (<b>7</b>) was elucidated on the basis of its spectroscopic data

Additional file 5: Figure S4. of Brainstem tau pathology in Alzheimer’s disease is characterized by increase of three repeat tau and independent of amyloid β

Predominance of RD3+ NFTs over RD4+ NFTs is shared between brainstem and hippocampus. To see whether the same trend of RD3 dominance for the neurofibrillary changes over RD4 is also demonstrated by the conventional DAB immunohistochemistry, we performed quantification of the RD3 and RD4 levels on representative adjacent pontine and hippocampal sections. After thresholding of the DAB labeling by RGB values, the counts and sizes of RD3 and RD4 were calculated on CellSens software (Olympus). RD3/RD4 ratio of total NFT counts and area on the pontine section were 6.00 and 4.49, respectively, indicating more intense deposition of RD3-positive neurofibrillary changes than RD4 (a, c, case 17). While the difference in the methods used make direct comparisons difficult, this observation of RD3 dominance agreed with our double-immunofluorolabeling of the same case using anti-4R tau antibody and RD3. RD3/RD4 ratio of total NFT counts and area on the hippocampal sections were 25.6 and 34.3, respectively. Thus, the dominance of 3R tau was also detected for the neurofibrillary changes in the hippocampal area of the same case (b, d, case 17). (TIFF 3761 kb

Additional file 2: of Inoculation of α-synuclein preformed fibrils into the mouse gastrointestinal tract induces Lewy body-like aggregates in the brainstem via the vagus nerve

Figure S2. No phosphorylated α-synuclein (p-α-Syn) pathology is seen in either mouse brain or stomach 45 days after phosphate-buffered saline inoculation into a mouse gastric wall. a P-α-Syn (EP1536Y) immunohistochemistry of a section around bregma − 7.08 mm. Scale bar 100 μm. 4V, fourth ventricle. b P-α-Syn (EP1536Y) immunohistochemistry of a section around bregma − 7.48 mm. Scale bar 100 μm. AP, area postrema; cc, central canal. c P-α-Syn (EP1536Y) immunohistochemistry in the stomach. Scale bar 100 μm. (TIF 7863 kb

Additional file 1: of Brainstem tau pathology in Alzheimer’s disease is characterized by increase of three repeat tau and independent of amyloid β

This file contains additional description of the image acquisition, and concomitant Lewy pathology of the investigated cases. The file also contains the results of additional statistical studies, including the regional difference in the proportion of 3R tau positive neurofibrillary changes on the same horizontal levels of the brainstem. (DOCX 25 kb

Phenanthrodithiophene (PDT)–Difluorobenzothiadiazole (DFBT) Copolymers: Effect on Molecular Orientation and Solar Cell Performance of Alkyl Substitution onto a PDT Core

Synthesis, characterization, and solar cell application of three 4,7-dialkylated phenanthro­[1,2-<i>b</i>:8,7-<i>b</i>′]­dithiophene (PDT)–difluorobenzothiadiazole (DFBT copolymers (<b>P1</b>–<b>P3</b>) with different linear alkyl side chains to improve solubility, molecular weight, and molecular orientation are described. The utilization of Ir-catalyzed direct borylation and sequential functionalization can selectively afford the target 4,7-dialkylated PDT as the monomers. Migita–Kosugi–Stille coupling in the presence of CuI can accelerate polymerization to afford high-molecular-weight polymers along with their improved solubility. The effect of alkyl substitution at the 4,7-positions on the electronic structure of PDT–DFBT copolymers is negligible. By installation of additional alkyl chains at the 4,7-positions of PDT, the synthesized polymers <b>P1</b>–<b>P3</b> have lower intermolecular interaction than that of nonalkylated <b>P0</b>, but they still maintained aggregation behavior in solution. In addition, they formed a favorable face-on orientation with a short π-stacking distance of 3.6 Å, which can enhance their carrier transport ability, resulting in high <i>J</i>_sc and FF. As a result, their fabricated solar cells reached a PCE exceeding 6%, which are about 1.7-fold higher than that of <b>P0</b>. Comparison of alkyl side chain length at the 4,7-positions of PDT revealed that all polymers formed a predominantly face-on orientation and have a similar face-on ratio in blended films, but their crystallinity was decreased as the carbon chains at the 4,7-positions of PDT became shorter. On the other hand, the polymers with short alkyl side chains tended to have low surface roughness and small domain size of active layers, which is an ideal phase separation structure for high-performance PSCs. From these results, it could be seen that the polymers have a trade-off relationship between their domain size and crystallinity, but the impact of alkyl side chain length on their photovoltaic properties is rather small. Thus, the construction of face-on orientation is highly important to achieve a high PCE. Among three polymers, the <b>P3</b>/PC₆₁BM-based solar cell with an optimal nanoscale phase separation structure with bicontinuous domain showed the highest PCE of up to 6.6%

Additional file 2: Figure S1. of Brainstem tau pathology in Alzheimer’s disease is characterized by increase of three repeat tau and independent of amyloid β

Quenching of autofluorescence after Sudan Black B treatment. Autofluorescence of intraneuronal lipofuscin is known to be prominent in the brainstem when quenching treatment is not performed. To clarify the effect of Sudan Black B treatment, we measured fluorescence emission spectra of intraneuronal lipofuscin on formalin-fixed, paraffin-embedded midbrain sections using Zeiss LSM780 lambda mode with or without the Sudan Black B treatment. The fluorescence spectrum of lipofuscin with excitation at 488 nm was broad and gently sloping (a-e, emission peak at 591 nm). This very intense fluorescence with broad spectrum agreed with the preceding studies on the fluorescence of lipofuscin. Sudan Black B treatment eliminated this autofluorescence of the lipofuscin (f-j, adjacent section). The fluorescence spectrum of lipofuscin did not overlap with the fluorescence spectra of Alexa 488- and Alexa 568-conjugated secondary antibodies labeling anti-4R tau antibody and RD3, respectively, on the pontine section, which underwent the Sudan Black B treatment (k-o). The small peak around 600 nm with excitation at 488 nm corresponded to colocalized Alexa 568 signal, which was blocked by the dichroic mirror on image capturing of the Alexa 488 signal. Therefore, we concluded that the autofluorescence of lipofuscin was effectively quenched by Sudan black treatment and did not affect the result of our study. Em: emission, Ex: excitation. (TIFF 7768 kb