Structural optimization and biological evaluation of 1,5-disubstituted pyrazole-3-carboxamines as potent inhibitors of human 5-lipoxygenase

AbstractHuman 5-lipoxygenase (5-LOX) is a well-validated drug target and its inhibitors are potential drugs for treating leukotriene-related disorders. Our previous work on structural optimization of the hit compound 2 from our in-house collection identified two lead compounds, 3a and 3b, exhibiting a potent inhibitory profile against 5-LOX with IC50 values less than 1µmol/L in cell-based assays. Here, we further optimized these compounds to prepare a class of novel pyrazole derivatives by opening the fused-ring system. Several new compounds exhibited more potent inhibitory activity than the lead compounds against 5-LOX. In particular, compound 4e not only suppressed lipopolysaccharide-induced inflammation in brain inflammatory cells and protected neurons from oxidative toxicity, but also significantly decreased infarct damage in a mouse model of cerebral ischemia. Molecular docking analysis further confirmed the consistency of our theoretical results and experimental data. In conclusion, the excellent in vitro and in vivo inhibitory activities of these compounds against 5-LOX suggested that these novel chemical structures have a promising therapeutic potential to treat leukotriene-related disorders

Mechanical overloading induces GPX4-regulated chondrocyte ferroptosis in osteoarthritis via Piezo1 channel facilitated calcium influx

Introductions: Excessive mechanical stress is closely associated with cell death in various conditions. Exposure of chondrocytes to excessive mechanical loading leads to a catabolic response as well as exaggerated cell death. Ferroptosis is a recently identified form of cell death during cell aging and degeneration. However, it's potential association with mechanical stress remains to be illustrated. Objectives: To identify whether excessive mechanical stress can cause ferroptosis. To explore the role of mechanical overloading in chondrocyte ferroptosis. Methods: Chondrocytes were collected from loading and unloading zones of cartilage in patients with osteoarthritis (OA), and the ferroptosis phenotype was analyzed through transmission electron microscope and microarray. Moreover, the relationship between ferroptosis and OA was analyzed by GPX4-conditional knockout (Col2a1-CreERT: GPX4flox/flox) mice OA model and chondrocytes cultured with high strain mechanical stress. Furthermore, the role of Piezo1 ion channel in chondrocyte ferroptosis and OA development was explored by using its inhibitor (GsMTx4) and agonist (Yoda1). Additionally, chondrocyte was cultured in calcium-free medium with mechanical stress, and ferroptosis phenotype was tested. Results: Human cartilage and mouse chondrocyte experiments revealed that mechanical overloading can induce GPX4-associated ferroptosis. Conditional knockout of GPX4 in cartilage aggravated experimental OA process, while additional treatment with ferroptosis suppressor protein (FSP-1) and coenzyme Q10 (CoQ10) abated OA development in GPX4-CKO mice. In mouse OA model and chondrocyte experiments, inhibition of Piezo1 channel activity increased GPX4 expression, attenuated ferroptosis phenotype and reduced the severity of osteoarthritis. Additionally, high strain mechanical stress induced ferroptosis damage in chondrocyte was largely abolished by blocking calcium influx through calcium-free medium. Conclusions: Our findings show that mechanical overloading induces ferroptosis through Piezo1 activation and subsequent calcium influx in chondrocytes, which might provide a potential target for OA treatment

Generation of ESTs for Flowering Gene Discovery and SSR Marker Development in Upland Cotton

BACKGROUND: Upland cotton, Gossypium hirsutum L., is one of the world's most important economic crops. In the absence of the entire genomic sequence, a large number of expressed sequence tag (EST) resources of upland cotton have been generated and used in several studies. However, information about the flower development of this species is rare. METHODOLOGY/PRINCIPAL FINDINGS: To clarify the molecular mechanism of flower development in upland cotton, 22,915 high-quality ESTs were generated and assembled into 14,373 unique sequences consisting of 4,563 contigs and 9,810 singletons from a normalized and full-length cDNA library constructed from pooled RNA isolated from shoot apexes, squares, and flowers. Comparative analysis indicated that 5,352 unique sequences had no high-degree matches to the cotton public database. Functional annotation showed that several upland cotton homologs with flowering-related genes were identified in our library. The majority of these genes were specifically expressed in flowering-related tissues. Three GhSEP (G. hirsutum L. SEPALLATA) genes determining floral organ development were cloned, and quantitative real-time PCR (qRT-PCR) revealed that these genes were expressed preferentially in squares or flowers. Furthermore, 670 new putative microsatellites with flanking sequences sufficient for primer design were identified from the 645 unigenes. Twenty-five EST-simple sequence repeats were randomly selected for validation and transferability testing in 17 Gossypium species. Of these, 23 were identified as true-to-type simple sequence repeat loci and were highly transferable among Gossypium species. CONCLUSIONS/SIGNIFICANCE: A high-quality, normalized, full-length cDNA library with a total of 14,373 unique ESTs was generated to provide sequence information for gene discovery and marker development related to upland cotton flower development. These EST resources form a valuable foundation for gene expression profiling analysis, functional analysis of newly discovered genes, genetic linkage, and quantitative trait loci analysis

FAC-Net: Feedback Attention Network Based on Context Encoder Network for Skin Lesion Segmentation

Considerable research and surveys indicate that skin lesions are an early symptom of skin cancer. Segmentation of skin lesions is still a hot research topic. Dermatological datasets in skin lesion segmentation tasks generated a large number of parameters when data augmented, limiting the application of smart assisted medicine in real life. Hence, this paper proposes an effective feedback attention network (FAC-Net). The network is equipped with the feedback fusion block (FFB) and the attention mechanism block (AMB), through the combination of these two modules, we can obtain richer and more specific feature mapping without data enhancement. Numerous experimental tests were given by us on public datasets (ISIC2018, ISBI2017, ISBI2016), and a good deal of metrics like the Jaccard index (JA) and Dice coefficient (DC) were used to evaluate the results of segmentation. On the ISIC2018 dataset, we obtained results for DC equal to 91.19% and JA equal to 83.99%, compared with the based network. The results of these two main metrics were improved by more than 1%. In addition, the metrics were also improved in the other two datasets. It can be demonstrated through experiments that without any enhancements of the datasets, our lightweight model can achieve better segmentation performance than most deep learning architectures

Magnetic-Field-Induced Orientational Phase Structure Transition

Magnetic field effect on the phase transition at high temperature (from 50 °C) inside the magnetic field has been found in C₁₄G₂ (<i>N</i>-tetradecyllactobionamide)/C₁₂EO₄ (tetraethylene glycol monododecyl ether)/D₂O system. The phase was transited quickly from lamellar phase to isotropic phases [bottom, micellar phase (L₁ phase) and top, sponge phase (L₃ phase)] induced by a magnetic field, which was demonstrated by ²H NMR and FF-TEM measurements. The isotropic phases induced by magnetic field were not stable, and the upper L₃ phase can recover to lamellar phase after being restored in a 55 °C thermostat outside the magnetic field for about one month. During the mechanism study, the C₁₂EO₄ molecule was proved to be the dominant component for the phase transition induced by the magnetic field, while the C₁₄G₂ molecule was the auxiliary and just affected the transition speed. The breaking and rebuilding of hydrogen bonds could play an important role in the phase transition and recovering. Moreover, the surfactant concentration had an effect on the speed of phase transiting and phase recovering. These observations could provide an understanding of the phase transition and also the applications for the controlled drug delivery system of bilayer membranes driving, induced by the magnetic field

Colloidal Wormlike Micelles with Highly Ferromagnetic Properties

For the first time, a new fabrication method for manipulating the ferromagnetic property of molecular magnets by forming wormlike micelles in magnetic-ionic-liquid (mag-IL) complexes is reported. The ferromagnetism of the mag-IL complexes was enhanced 4-fold because of the formation of wormlike micelles, presenting new evidence for the essence of magnetism generation at a molecular level. Characteristics such as morphology and magnetic properties of the wormlike micelle gel were investigated in detail by cryogenic transmission electron microscopy (Cryo-TEM), rheological measurements, circular dichroism (CD), FT-IR spectra, and the superconducting quantum interference device method (SQUID). An explanation of ferromagnetism elevation from the view of the molecular (ionic) distribution is also given. For the changes of magnetic properties (ferromagnetism elevation) in the wormlike micelle systems, the ability of CTAFe in magnetizing AzoNa₄ (or AzoH₄) can be ascribed to an interplay of the magnetic [FeCl₃Br]⁻ ions both in the Stern layer and in the cores of the wormlike micelles. Formation of colloidal aggregates, i.e., wormlike micelles, provides a new strategy to tune the magnetic properties of novel molecular magnets

Controlling the Capture and Release of DNA with a Dual-Responsive Cationic Surfactant

A dual-responsive cationic surfactant, 4-ethoxy-4′-(trimethyl- aminoethoxy) azobenzene trichloromonobromoferrate (azoTAFe), which contains both a light-responsive moiety azobenzene and a paramagnetic counterion, [FeCl₃Br]⁻, was designed and synthesized. Not only does this cationic surfactant abundantly utilize inexhaustible and clean sources, i.e., light and magnetic field, but it also serves as a powerful dual-switch molecule for effectively controlling the capture and release of DNA. Our results could provide potential applications in gene therapy for creating smart and versatile machines to control the transport and delivery of DNA more intelligently and robustly. It was proved that the light switch can independently realize a reversible DNA compaction. The introduction of a magnetic switch can significantly enhance the compaction efficiency, help compact DNA with a lower dosage and achieve a magnetic field-based targeted transport of DNA. In addition, the light switch can make up the irreversibility of magnetic switch. This kind of self-complementation makes the cationic azoTAFe be useful as a potential tool that can be applied to the field of gene therapy and nanomedicine

First Fluorinated Zwitterionic Micelle with Unusually Slow Exchange in an Ionic Liquid

The micellization of a fluorinated zwitterionic surfactant in ethylammonium nitrate (EAN) was investigated. The freeze-fracture transmission electron microscope (FF-TEM) observations confirm the formation of spherical micelles with the average diameter 25.45 ± 3.74 nm. The micellization is an entropy-driven process at low temperature but an enthalpy-driven process at high temperature. Two sets of ¹⁹F NMR signals above the critical micelle concentration (cmc) indicate that the unusually slow exchange between micelles and monomers exists in ionic liquid; meanwhile, surfactant molecules are more inclined to stay in micelle states instead of monomer states at higher concentration. Through the analysis of the half line width (Δν_1/2), we can obtain the kinetic information of fluorinated zwitterionic micellization in an ionic liquid

Magnetic Fullerene-DNA/Hyaluronic Acid Nanovehicles with Magnetism/Reduction Dual-Responsive Triggered Release

We created the dual-responsive nanovehicle that can effectively combine and abundantly utilize magnetic and glutathione (GSH)-reductive triggers to control the drug delivery and achieve more intelligent and powerful targeting. In the nanovehicles, paramagnetic fullerene (C₆₀@CTAF) was prepared via one-step modification of fullerene with magnetic surfactant CTAF by hydrophobic interaction for the first time. The perfect conjugation of C₆₀ and CTAF increased the solubility or dispersity of fullerenes and qualified CTAF with more powerful assembly capability with DNA. DNA molecule in the nanovehicles acted as an electrostatic scaffold to load anticancer drug Dox as well as the important building block for assembly with C₆₀@CTAF into C₆₀@CTAF/DNA. The further combination of deshielding and targeting functions in reduction-responsive disulfide modified HA-SS-COOH coating on C₆₀@CTAF/DNA complexes could reduce the agglomeration and regulate the morphology of C₆₀@CTAF/DNA complexes from irregular microstructures to more uniform ones. More importantly, the introduction of HA-SS-COOH provided a response to a simulating reductive extra-tumoral environment by efficient cleavage of disulfide linkages by GSH and site-specific drug delivery to HepG2 cells. Amazingly, the final nanovehicles presented an increased magnetic susceptibility compared with paramagnetic CTAF, and they “walked” under an applied magnetic field. Because of their facile fabrication, rapid responsiveness to extra tumoral environment, and external automatic controllability by external magnet, the drug delivery nanovehicles constructed by magnetic fullerene-DNA/hyaluronic acid might be of great interest for making new functional nucleic-acid-based drug carriers