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Ligand-Controlled C2- or C3-Selectivity Switching in the Palladium-Catalyzed C–H Arylation of Nonsubstituted 1H-Pyrrole
No full textapplication/pdfThe C2- or C3-selective direct C–H arylation of nonsubstituted 1H-pyrrole with aryl chlorides/nonaflates was achieved using catalysts derived from palladium and appropriate phosphine ligands. The site selectivity of the arylation can be switched by changing the ligands, and the C3-selective arylation of nonsubstituted 1H-pyrrole was realized for the first time. tBuOLi played an important role in suppressing N-arylation and accelerating C2- or C3-arylation.journal articl
The importin α proteins IMPA1, IMPA2, and IMPA4 play redundant roles in suppressing autoimmunity in Arabidopsis thaliana
No full textapplication/pdfSUMMARY
Proteins in the importin α (IMPA) family play pivotal roles inintracellular nucleocytoplasmic transport. Arabidopsis thalianapossesses nine IMPA members, with diverse tissue-specific expressionpatterns. Among these nine IMPAS, IMPA1, IMPA2, and IMPA4 clustertogether phylogenetically, suggesting potential functional redundancy. Toexplore this redundancy, we analyzed single and multiple T-DNA mutants forthese genes and discovered severe growth defects in the impa1impa2impa4tripleknockout mutant but not in the single or double mutants. Complementationwith IMPA1, IMPA2, or IMPA4 fused to green fluorescent protein (GFP)rescued the growth defectsobservedintheimpa1impa2impa4mutant,
indicating the functional redundancy of these three IMPAs. The IMPA-GFPfusion proteins were localized in the nucleus andnuclearenvelope,suggestingtheirinvolvementinnucleocytoplasmictransport processes. Comparativetranscriptomics revealed that salicylic acid (SA)- responsive genes were
significantly upregulated in the impa1 impa2 impa4 triple mutant. Consistentwith this observation, impa1 impa2 impa4 mutant plants accumulated SA andreactive oxygen speciestohighlevelscompared with wild-type plants. Wealso found enhanced resistance to the anthracnose pathogen Colletotrichum
higginsianum in the impa1 impa2 impa4 mutants, suggesting thatdefenseresponseswereconstitutively activated in the impa1 impa2 impa4 mutant. Ourfindings shed light on the redundant roles of IMPA1, IMPA2, and IMPA4 insuppressing the autoimmune responses and suggest avenues of research to
clarify their potentially unique roles.journal articl
Simple and fast one-step FRET assay of therapeutic mAb bevacizumab using anti-idiotype DNA aptamer for process analytical technology
No full textapplication/pdfWe developed an aptamer-based fluorescence resonance energy transfer (FRET) assay capable of recognizing therapeutic monoclonal antibody bevacizumab and rapidly quantifying its concentration with just one mixing step. In this assay, two fluorescent dyes (fluorescein and tetramethylrhodamine) labeled aptamers bind to two Fab regions on bevacizumab, and FRET fluorescence is observed when both dyes come into close proximity. We optimized this assay in three different formats, catering to a wide range of analytical needs. When applied to hybridoma culture samples in practical settings, this assay exhibited a signal response that was concentration-dependent, falling within the range of 50–2000 μg/mL. The coefficients of determination (r2) ranged from 0.998 to 0.999, and bias and precision results were within ±24.0 % and 20.3 %, respectively. Additionally, during thermal and UV stress testing, this assay demonstrated the ability to detect denatured samples in a manner comparable to conventional Size Exclusion Chromatography. Notably, it offers the added advantage of detecting decreases in binding activity without changes in molecular weight. In contrast to many existing process analytical technology tools, this assay not only identifies bevacizumab but also directly measures the quality attributes related to mAb efficacy, such as the binding activity. As a result, this assay holds great potential as a valuable platform for providing highly reliable quality attribute information in real-time. We consider this will make a significant contribution to the worldwide distribution of high-quality therapeutic mAbs in various aspects of antibody manufacturing, including production monitoring, quality control, commercial lot release, and stability testing.journal articl
Weaving Stories: The Mutual Growth of People with Intellectual Disabilities and Diverse Others
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A Study on the Establishment of "Standards" for Elementary Schools in the first half of the Meiji Era
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