Selective Boryl Silyl Ether Formation in the Photoreaction of Bisboryloxide/Boroxine with Hydrosilane Catalyzed by a Transition-Metal Carbonyl Complex

Selective B–O–Si bond formation was achieved in the reaction of bisboryloxide O­(Bpin)₂ (pin = (OCMe₂)₂)/boroxine (MeBO)₃ system with tertiary silane R₃SiH in the presence of stoichiometric water and a catalytic amount of [M]­(CO)₅ ([M] = Mo­(CO), W­(CO), Fe) to give boryl silyl ethers. Moreover, this reaction can be applied to various hydrosilanes (disilyl compounds and secondary silanes) and hydrogermane. Some of the boryl silyl ethers thus formed were confirmed by X-ray analysis

Selective Double Hydroboration and Dihydroborylsilylation of Organonitriles by an Iron–indium Cooperative Catalytic System

Organonitriles (RCN) were selectively converted into the corresponding diborylamines (RCH₂N­(Bpin)₂) in reactions with pinacolborane (pinBH) in the presence of a catalytic amount of the iron–indium complex [Fe­(CH₃CN)₆]­[<i>cis</i>-Fe­(CO)₄(InCl₃)₂]. The catalytic reaction mechanism was tentatively proposed. In addition, this catalytic system was found to be applicable for the synthesis of borylsilylamine in high yield when organonitrile was treated with hydroborane and hydrosilane simultaneously

<i>Agrobacterium rhizogenes</i> strains.

<p><i>Agrobacterium rhizogenes</i> strains.</p

Detection of transgenes by PCR and RT-PCR.

<p><b>A-B</b> PCR analysis of genomic DNA isolated from <i>P. japonicum</i> fluorescent hairy roots (F) and non-transformed tissues (NT). <b>A</b>. Amplification of <i>GFP</i> and <i>rolB</i> fragments with expected sizes (380 bp and 780 bp, respectively). <b>B</b>. No amplification of <i>virD1</i> fragment (450 bp) in fluorescent tissues (F), as positive control a diluted ATCC15834 bacterial suspension (Ar) was used. <b>C</b>. Southern blot of genomic DNA extracted from fluorescent (F) and non-transformed (NT) <i>P. japonicum</i> roots. DNA was digested with <i>Eco</i>RI. The positive control (Pl) corresponds to linearised pBCR101 plasmid (30 ng). <b>D</b>. RT-PCR analysis of the <i>rolB</i> gene using total RNAs extracted from <i>P. japonicum</i> fluorescent roots (F) and non-transformed tissues (NT).</p

Optimization of factors influencing stable transformation of <i>P. japonicum</i>.

<p>Frequency of stable transformation in <i>P. japonicum s</i>eedlings submitted to different treatments. <b>A</b> Transformation efficiency in 3-day-old seedlings infected with strains ATCC15834, LBA1334 and AR1193 and co-cultivated for 2 or 7 days. The data show representative results from one of two independent experiments using 20 to 60 plants each. <b>B</b> Effect of acetosyringone (AS) on transformation efficiency. <i>S</i>eedlings were co-cultivated with <i>A. rhizogenes</i> strains ATCC15834 and LBA1334 in media with or without 100 µM AS for 2 days for LBA1334, and 7 days for ATCC15834. 4–5 weeks after the inoculation the transformation frequency was scored. The data show representative results from one of at least two independent experiments using 20 to 60 plants each.</p

Stable and transient transformation of <i>P. japonicum</i> mediated by <i>A. rhizogenes</i>.

<p>Transformation induced by <i>A. rhizogenes</i> ATCC15834. <b>A</b>. Five-week-old plants showing accumulation of black substance(s) at the wound site after hypocotyl-cutting infection method. <b>B</b>. Transformed roots emerged from cotyledons 5 weeks after <i>A. rhizogenes</i> inoculation by the SAAT method. The black arrow indicates transformed roots. <b>C</b> and <b>D</b> GFP-fluorescing transformed roots observed under bright field (C) or fluorescent (D) microscopy. The black arrows point to fluorescent roots and the white arrow to non-fluorescent root. <b>E</b> and <b>F</b> Transiently-transformed cotyledons observed under bright field (E) or fluorescent (F) microscopy. <b>G</b> and <b>H</b> Confocal micrograph of cotyledon leaves. Non-transformed (G) and transiently-transformed (H). Red color corresponds to autofluorescence from chlorophyll. White bars correspond to 2 mm and yellow bars to 20 µm.</p

Efficiency of stable transformation after the addition of Silwet L-77 and/or NAA into bacterial suspension.

<p>Efficiency of stable transformation after the addition of Silwet L-77 and/or NAA into bacterial suspension.</p

Transgenic root retains parasitic competence.

<p><b>A</b> and <b>B</b>. Haustorium development in transgenic hairy roots following 2-day exposure to 10 µM DMBQ observed under bright field (A) and fluorescent (B) microscopy. The white arrows point to haustoria developing on a transformed root and the black arrow points to a haustorium in a non-transformed root. <b>C</b> and <b>D</b>. Haustorial connection with host rice observed under bright field (C) and fluorescence (D) microscopy. <b>E</b> and <b>F</b>. Haustorial connection with host maize observed under bright field (E) and fluorescence (F) microscopy. White arrows indicate haustorial connection of transgenic roots to hosts. H: host, P: parasite. Bars correspond to 0.5 mm.</p

A flowchart for hairy root transformation in <i>P. japonicum</i>.

<p>A flowchart for hairy root transformation in <i>P. japonicum</i>.</p

Encryption of agonistic motifs for TLR4 into artificial antigens augmented the maturation of antigen-presenting cells

<div><p>Adjuvants are indispensable for achieving a sufficient immune response from vaccinations. From a functional viewpoint, adjuvants are classified into two categories: “physical adjuvants” increase the efficacy of antigen presentation by antigen-presenting cells (APC) and “signal adjuvants” induce the maturation of APC. Our previous study has demonstrated that a physical adjuvant can be encrypted into proteinous antigens by creating artificial proteins from combinatorial assemblages of epitope peptides and those peptide sequences having propensities to form certain protein structures (motif programming). However, the artificial antigens still require a signal adjuvant to maturate the APC; for example, co-administration of the Toll-like receptor 4 (TLR4) agonist monophosphoryl lipid A (MPLA) was required to induce an <i>in vivo</i> immunoreaction. In this study, we further modified the previous artificial antigens by appending the peptide motifs, which have been reported to have agonistic activity for TLR4, to create “adjuvant-free” antigens. The created antigens with triple TLR4 agonistic motifs in their C-terminus have activated NF-κB signaling pathways through TLR4. These proteins also induced the production of the inflammatory cytokine TNF-α, and the expression of the co-stimulatory molecule CD40 in APC, supporting the maturation of APC <i>in vitro</i>. Unexpectedly, these signal adjuvant-encrypted proteins have lost their ability to be physical adjuvants because they did not induce cytotoxic T lymphocytes (CTL) <i>in vivo</i>, while the parental proteins induced CTL. These results confirmed that the manifestation of a motif’s function is context-dependent and simple addition does not always work for motif-programing. Further optimization of the molecular context of the TLR4 agonistic motifs in antigens should be required to create adjuvant-free antigens.</p></div