Tritylamine as an Ammonia Surrogate in the Ugi Tetrazole Synthesis

The role of tritylamine is introduced as a convenient ammonia substitute in the Ugi tetrazole synthesis. Fifteen examples and their mild cleavage products are described in satisfactory to good yields. <i>N</i>-Unsubstituted α-aminotetrazoles are important compounds with annotated biological activities, and the described two-step synthesis provided an alternative route to otherwise difficult to access derivatives

sj-docx-1-msj-10.1177_13524585231213230 – Supplemental material for The disease-modifying therapy utilisation and cost trend for multiple sclerosis in Australia between 2013 and 2022

Supplemental material, sj-docx-1-msj-10.1177_13524585231213230 for The disease-modifying therapy utilisation and cost trend for multiple sclerosis in Australia between 2013 and 2022 by Ting Zhao, Bruce V Taylor, Julie A Campbell and Andrew J Palmer in Multiple Sclerosis Journal</p

Analyte-Activable Probe for Protease Based on Cytochrome C‑Capped Mn: ZnS Quantum Dots

A new sensor format was proposed here by integrating conjugation of analyte-recognition sites and quenching the luminescence of quantum dots (QDs) in one pot during the synthesis of QDs, with protease as the model analyte. Inherently phosphorescence-attenuated Mn-doped ZnS QDs were prepared with electron transfer protein cytochrome C (Cyt C) as the ligand, which was capable of protease sensing in both label-free and activable format. This detection strategy eliminates the postsynthetic protein conjugation and responses to analyte in the turn-on mode, lowering the signal background. In the presence of protease, the initially “locked” phosphorescence of Mn-doped ZnS QDs could be activated, due to the enzymatic digestion of surface Cyt C ligand and removal of the electron-transfer quenching unit away from the close-proximity of QDs. The proposed probe exhibited good selectivity toward proteases over other proteins and enzymes. Besides, it was also capable of differentiating active and inactive serine proteases. Analytical performance of this probe was evaluated using trypsin as the model serine protease. Limits of detection (LOD) of 2 nM was obtained, which is well below the average urine trypsin level of patients. The analytical application of this probe was demonstrated in determination of trypsin in human pancreatic carcinoma (PANC-1 and 818.4) cells lysates, demonstrating the potential usefulness of this probe in future clinical diagnosis

GFP knock-in does not affect the function of endogenous CHDP-1.

(A-B) Quantification of the number of (A) 2o, 3o and 4o branches in a 100 μm area anterior to the PVD cell body, and (B) the ratio of the intensity in the 2o branches to that of the primary dendritesin wild-type and gfp::chdp-1 knock-in animals. Error bars: SEM. ns: non-significant by Student’s t-test. n = 20–30 for each genotype. (C) Confocal images showing the expression patterns of GFP(7x)::CHDP-1 (left), mCherry (middle) and overlay (right) in the PVD neurons. Scale bar: 20 μm. (D-E) Quantification of (D) the number of 2o, 3o and 4o branches in a 100 μm area anterior to the PVD cell body, and (E) the ratio of the intensity in the primary dendrites to that of the 2o branches in wild-type and gfp(7x)::chdp-1 knock-in animals. Error bars: SEM. ns: non-significant by Student’s t-test. n = 20–30 for each genotype. (TIF)</p

Knockout of <i>sax-1</i> suppresses dendrite development defects in <i>chdp-1</i> mutants.

(A) Confocal images of PVD dendrites in wild-type, chdp-1(tm4947), sax-1(ky491), chdp-1(tm4947); sax-1(ky491), sax-2(ky216) and chdp-1(tm4947); sax-2(ky216). Scale bar: 20 μm. (B-C) Quantification of (B) the number of 2o, 3o and 4o branches, and (C) the ratio of the 2o branches to that of the intensity of the primary dendrites in the 100 μm area anterior to PVD cell body for the genotypes indicated. Error bars, SEM. ***p o branches with F-actin), (middle) TBA-1 (arrows: 2o branches labeled by GFP::TBA-1), and (bottom) NLP-12::Venus and myr-mCherry (arrows: dense-core vesicles in high-ordered branches) in wild-type, chdp-1(tm4947), sax-1(ky491) and chdp-1(tm4947); sax-1(ky491). Scale bar: 10 μm. (E) Quantification of (left) the ratio of the Lifeact::GFP intensity of the 2o dendrite to that of the primary branches, (middle) the ratio of the TBA-1::GFP intensity of the 2o dendrite to that of the primary dendrites, (right) NLP-12::Venus fluorescence intensity in high-ordered dendrites in the 100 μm area anterior to PVD cell body for the genotypes indicated (normalized to WT). Error bars, SEM. ***p < 0.001 by one-way ANOVA with the Tukey correction. ns: non-significant. n = 20–30 for each genotype.</p

CHDP-1 is required for actin assembly during dendrite formation.

(A) Left: Confocal images of Lifeact::GFP in wild-type (upper) and chdp-1(tm4947) mutant (lower) at early L3 stage. Scale bar: 10 μm. Right: Confocal images from time-lapse movies showing actin assembly in wild-type (upper) and chdp-1(tm4947) mutant (lower) during the early L3 stage. Arrows: growth cones of 2o branches labeled by Lifeact::GFP in wild-type, arrowheads: growth cones in chdp-1(tm4947). Scale bar: 5 μm. (B) Quantification of the average growth cones area, and the ratio of the intensity in the growth cones of the 2o branches to that of the primary dendrite in wild-type and chdp-1(tm4947) during the early L3 stage. n = 100 growth cones for each group. (C) Left: Confocal images of Lifeact::GFP in wild-type (upper) and chdp-1(tm4947) mutant (lower) at early L4 stage. Scale bar: 10 μm. Right: Confocal images from time-lapse movies showing actin assembly in wild-type (upper) and chdp-1(tm4947) mutant (lower) during the early L4 stage. Arrows: growth cones of 4o branches labeled by Lifeact::GFP in wild-type, arrowheads: growth cones in chdp-1(tm4947). Scale bar: 5 μm. (D) Quantification of the average growth cones area, and the ratio of the intensity in the growth cones of the 4o branches to that of the primary dendrite in wild-type and chdp-1(tm4947) during the early L4 stage. n = 100 growth cones for each group. (TIFF)</p

Distribution of the NLP-12-positive dense-core vesicles in the primary dendrites.

(A) Confocal images of PVD dendrites (left), NLP-12 (NLP-12::Venus) (middle), overlay (right) in wild-type (upper), chdp-1(tm4947) mutant (lower). Arrows: NLP-12::Venus-positive dense-core vesicles in primary dendrites. Scale bar: 10 μm. (B) Quantification of NLP-12::Venus fluorescence intensity in primary dendrites in a 100 μm area anterior to the PVD cell body (normalized to WT). Error bars: SEM. ***p (TIFF)</p

Identification of cellular proteins from HCV E2 complex.

<p>Details can be found in Experimental Procedures. (A), Genomic organization of the Flag-E2-JFH1 virus. (B), Schematic of the purification strategy. The infection efficiency was nearly 100% in all three replicates. (C), Venn diagram of 89 proteins (including 4 viral proteins) that were identified in all three trials.</p

Endogenous CHDP-1 localizes to the cell cortex.

(A) Left: Confocal images showing the localization of endogenously expressed GFP::CHDP-1 (left), myr-mCherry (middle), and overlay (right) in the embryonic stages. Right: Normalized intensity of GFP::CHDP-1 and myr-mCherry around the cell membrane. Scale bar: 20 μm. (B) Left: Confocal images showing the localization of endogenously expressed GFP::CHDP-1 (left), myr-mCherry (middle), and overlay (right) in the PVD cell body. Right: Normalized intensity of GFP::CHDP-1 and myr-mCherry around the PVD cell body membrane. Scale bar: 5 μm. (TIFF)</p

Microtubule assembly in the growth cones of the anterior primary dendrite in WT and <i>chdp-1</i> mutants.

(A) A cartoon showing the area imaged to analyze the microtubule dynamics in the growth cone of the anterior primary dendrite. Scale bar: 2 μm. (B) Kymographs of EBP-2::GFP in a growth cone of the anterior primary dendrite in wild-type (left), chdp-1(tm4947) (right) during mid to late L2 stage. Scale bar: 2 μm. (C) Quantification of the number of EBP-2::GFP comets either moving away from the cell body (anterograde) or towards the cell body (retrograde) in wild-type and chdp-1(tm4947) mutant animals. Error bars, SEM. ns: non-significant by one-way ANOVA with the Tukey correction. n = 10 worms for each genotype. (D) Quantification of the length of tracks of the EBP-2 comets in wild-type and chdp-1(tm4947) mutant animals. Error bars, SEM. Ns: non-significant by one-way ANOVA with the Tukey correction. n = 10 worms for each genotype. (E) Quantification of the growth duration of tracks of the EBP-2 comets in wild-type and chdp-1(tm4947) mutant animals. Error bars, SEM. ns: non-significant by one-way ANOVA with the Tukey correction. n = 10 worms for each genotype. (F) Quantification of MT pause frequency in wild-type and chdp-1(tm4947) mutant animals. Error bars, SEM. ns: non-significant by one-way ANOVA with the Tukey correction. n = 10 worms for each genotype. (G) Left: Confocal images of PVD (top) dendrites and (down) axon (left), RAB-3 (middle), overlay (right) in wild-type (upper), chdp-1(tm4947) mutant (lower). Right: A cartoon showing the localization of RAB-3 in PVD dendrites and axon in wild-type and chdp-1(tm4947) mutants. Scale bar: 10 μm. (TIFF)</p