Palladium-Catalyzed Alkenylation via sp² C–H Bond Activation Using Phenolic Hydroxyl as the Directing Group

This note describes the efficient and highly regioselective synthesis of 2-(2′-alkenylphenyl)­phenol derivatives via palladium-catalyzed 2′-alkenylation of 2-arylphenols directed by the phenolic hydroxyl group using benzoquinone as the oxidant in an atmosphere of air. This reaction can tolerate a series of functional groups and provides the alkenylation products regio- and stereoselectively in moderate to good yields

Image1_Effect of Bufalin-PLGA Microspheres in the Alleviation of Neuropathic Pain via the CCI Model.TIF

The treatment of neuropathic pain (NPP) is considered challenging, while the search for alternative medication is striving. NPP pathology is related with the expression of both the purinergic 2X7 (P2X7) receptor and the transient receptor potential vanilloid 1 receptor (TRPV1). Bufalin is a traditional Chinese medication derived from toad venom with pronounced antitumor, analgesic, and anti-inflammatory properties. However, poor solubility, rapid metabolism, and the knowledge gap on its pain alleviation mechanism have limited the clinical application of bufalin. Hence, the purpose of this study is to illustrate the NPP alleviation mechanism of bufalin via chronic constriction injury (CCI). To address the concern on fast metabolism, bufalin-PLGA microspheres (MS) were prepared via membrane emulsification to achieve prolonged pain-relieving effects. Western blot, real-time PCR, immunofluorescence, and molecular docking were employed to demonstrate the therapeutic action of bufalin on NPP. The results showed enhanced thermal withdrawal latency (TWL) and mechanical withdrawal threshold (MWT) after the administration of both bufalin and bufalin-PLGA MS in the CCI rats. Prolonged pain-relieving effects for up to 3 days with reduced dose frequency was achieved via bufalin-PLGA MS. In the CCI rats treated with bufalin-PLGA MS, the expression levels of protein and mRNA in TRPV1 and P2X7, both localized in the dorsal root ganglion (DRG), were reduced. Moreover, bufalin-PLGA MS effectively reduced the levels of IL-1β, IL-18, IL-6, and TNF-α in the CCI group. The results from molecular docking suggested a possible mechanism of NPP alleviation of bufalin through binding to P2X7 receptors directly. The administration of bufalin-PLGA MS prepared by membrane emulsification demonstrated promising applications for sustained effect on the alleviation of NPP.</p

Table2_N6-Methyladenosine Methylation Regulator RBM15 is a Potential Prognostic Biomarker and Promotes Cell Proliferation in Pancreatic Adenocarcinoma.XLSX

RNA binding motif protein 15 (RBM15) is a key regulatory factor involved in N6-methyladenosine (m6A) methylation. It has been reported that RBM15 plays an important role in the progress of laryngeal squamous cell carcinoma (LSCC), promoting LSCC migration and invasion. However, the role of RBM15 in human different cancers remains unknown. This study aims to analyze the prognostic value of RBM15, and to demonstrate the correlation between RBM15 expression and tumor immunity, as well as to provide clues for further mechanism research. The results showed that RBM15 was mutated or copy number varied in 25 types of cancer. RBM15 mRNA was abnormally up-regulated across various cancers. Survival analysis suggested high expression of RBM15 was associated with poor prognosis in many cancer types. Among these, it affected patients’ overall survival (OS) in 10 cancer types, disease-free interval (DFI) in 8 cancer types, progression-free interval (PFI) in 12 cancer types and disease-specific survival (DSS) in 7 cancer types. Importantly, in pancreatic adenocarcinoma (PAAD), overexpression of RBM15 is associated with patients’ OS, DFI, PFI, or DSS. In addition, RBM15 expression was positively correlated with immune infiltrating cells in kidney renal clear cell carcinoma (KIRC), brain lower grade glioma (LGG), and PAAD. Moreover, RBM15 expression showed a strong correlation with immune checkpoint markers in PAAD. Cell counting kit-8 (CCK-8) assay showed that knockdown of RBM15 significantly inhibited the proliferation of pancreatic cancer cells. PPI analysis showed USP10, USP24, SMG1, NRAS were closely connected with RBM15 alterations. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that many biological processes (BP), cellular components (CC), molecular functions (MF), cancer related pathways including “sister chromatid cohesion”, “peptidyl-serine phosphorylation”, “cell division”, “nucleoplasm”, “nucleus”, “protein binding”, “protein serine/threonine kinase activity”, “T cell receptor signaling pathway”, “Cell cycle” were regulated by RBM15 alterations. Taken together, pan-cancer analysis of RBM15 suggested it may be served as a prognostic biomarker and immunotherapeutic target for PAAD.</p

Systematic and Quantitative Investigation of the Mechanism of Carbon Nanotubes’ Toxicity toward Algae

Concurrent with the increasing production and application of carbon nanotubes (CNTs) comes an increasing likelihood of CNTs presenting in the aquatic environment, and thereby potentially threatening aquatic organisms via toxic mechanisms that are, at present, poorly understood. This study systematically investigated the toxicity of three multiwalled CNT (MWCNT) samples toward a green alga (<i>Chlorella</i> sp.), focusing on examining and quantifying the contributions of five possible mechanisms to the algal growth inhibition. The results showed that the MWCNTs significantly inhibited the algal growth. The contribution of metal catalyst residues in the MWCNTs to the algal growth inhibition was negligible, as was the contribution from the MWCNTs’ adsorption of nutrient elements. The algal toxicity of MWCNTs could mainly be explained by the combined effects of oxidative stress, agglomeration and physical interactions, and shading effects, with the quantitative contributions from these mechanisms depending on the MWCNT size and concentration. At MWCNT concentrations around 96 h IC₅₀, the oxidative stress accounted for approximately 50% of the algal growth inhibition, whereas the agglomeration and physical interactions, and the shading effects each took approximately 25% of the responsibility

Table1_N6-Methyladenosine Methylation Regulator RBM15 is a Potential Prognostic Biomarker and Promotes Cell Proliferation in Pancreatic Adenocarcinoma.XLSX

RNA binding motif protein 15 (RBM15) is a key regulatory factor involved in N6-methyladenosine (m6A) methylation. It has been reported that RBM15 plays an important role in the progress of laryngeal squamous cell carcinoma (LSCC), promoting LSCC migration and invasion. However, the role of RBM15 in human different cancers remains unknown. This study aims to analyze the prognostic value of RBM15, and to demonstrate the correlation between RBM15 expression and tumor immunity, as well as to provide clues for further mechanism research. The results showed that RBM15 was mutated or copy number varied in 25 types of cancer. RBM15 mRNA was abnormally up-regulated across various cancers. Survival analysis suggested high expression of RBM15 was associated with poor prognosis in many cancer types. Among these, it affected patients’ overall survival (OS) in 10 cancer types, disease-free interval (DFI) in 8 cancer types, progression-free interval (PFI) in 12 cancer types and disease-specific survival (DSS) in 7 cancer types. Importantly, in pancreatic adenocarcinoma (PAAD), overexpression of RBM15 is associated with patients’ OS, DFI, PFI, or DSS. In addition, RBM15 expression was positively correlated with immune infiltrating cells in kidney renal clear cell carcinoma (KIRC), brain lower grade glioma (LGG), and PAAD. Moreover, RBM15 expression showed a strong correlation with immune checkpoint markers in PAAD. Cell counting kit-8 (CCK-8) assay showed that knockdown of RBM15 significantly inhibited the proliferation of pancreatic cancer cells. PPI analysis showed USP10, USP24, SMG1, NRAS were closely connected with RBM15 alterations. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that many biological processes (BP), cellular components (CC), molecular functions (MF), cancer related pathways including “sister chromatid cohesion”, “peptidyl-serine phosphorylation”, “cell division”, “nucleoplasm”, “nucleus”, “protein binding”, “protein serine/threonine kinase activity”, “T cell receptor signaling pathway”, “Cell cycle” were regulated by RBM15 alterations. Taken together, pan-cancer analysis of RBM15 suggested it may be served as a prognostic biomarker and immunotherapeutic target for PAAD.</p

Synthesis, crystal structures and urease inhibition of copper, nickel and zinc complexes derived from 4-chloro-2-((pyridin-2-ylmethylene)amino)phenol

A series of copper, nickel and zinc complexes, [CuClL] (1), [CuBrL] (2), [NiL2] (3), [ZnL2] (4) and [Zn2I2L2] (5), where L is the deprotonated form of 4-chloro-2-((pyridin-2-ylmethylene)amino)phenol, have been synthesized and characterized by elemental analysis, IR and UV-Vis spectra. The structures of the complexes were further confirmed by single crystal X-ray diffraction. The Cu ions in 1 and 2 are in square planar coordination. The Ni and Zn ions in 3 and 4 are in octahedral coordination. The coordination of the Zn ions in centrosymmetric complex 5 is between square pyramidal and trigonal bipyramidal. The copper complexes have excellent urease inhibitory activity. </p

Table_1_Over-Expression of LcPDS, LcZDS, and LcCRTISO, Genes From Wolfberry for Carotenoid Biosynthesis, Enhanced Carotenoid Accumulation, and Salt Tolerance in Tobacco.docx

It is of great importance to combine stress tolerance and plant quality for breeding research. In this study, the role of phytoene desaturase (PDS), ζ-carotene desaturase (ZDS) and carotene isomerase (CRTISO) in the carotenoid biosynthesis are correlated and compared. The three genes were derived from Lycium chinenses and involved in the desaturation of tetraterpene. Their over-expression significantly increased carotenoid accumulation and enhanced photosynthesis and salt tolerance in transgenic tobacco. Up-regulation of almost all the genes involved in the carotenoid biosynthesis pathway and only significant down-regulation of lycopene ε-cyclase (ε-LCY) gene were detected in those transgenic plants. Under salt stress, proline content, and activities of catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD) were significantly increased, whereas malonaldehyde (MDA) and hydrogen peroxide (H2O2) accumulated less in the transgenic plants. The genes encoding ascorbate peroxidase (APX), CAT, POD, SOD, and pyrroline-5-carboxylate reductase (P5CR) were shown to responsive up-regulated significantly under the salt stress in the transgenic plants. This study indicated that LcPDS, LcZDS, and LcCRTISO have the potential to improve carotenoid content and salt tolerance in higher plant breeding.</p

DataSheet_1_Multiplex CRISPR/Cas9-mediated raffinose synthase gene editing reduces raffinose family oligosaccharides in soybean.docx

Soybean [Glycine max (L.) Merr.] is an important world economic crop. It is rich in oil, protein, and starch, and soluble carbohydrates in soybean seeds are also important for human and livestock consumption. The predominant soluble carbohydrate in soybean seed is composed of sucrose and raffinose family oligosaccharides (RFOs). Among these carbohydrates, only sucrose can be digested by humans and monogastric animals and is beneficial for metabolizable energy, while RFOs are anti-nutritional factors in diets, usually leading to flatulence and indigestion, ultimately reducing energy efficiency. Hence, breeding efforts to remove RFOs from soybean seeds can increase metabolizable energy and improve nutritional quality. The objective of this research is to use the multiplex Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9‐mediated gene editing system to induce the knockout of soybean raffinose synthase (RS) genes RS2 and RS3 simultaneously to reduce RFOs in mature seeds. First, we constructed five types of multiplex gene editing systems and compared their editing efficiency in soybean hairy roots. We confirmed that the two-component transcriptional unit (TCTU) and single transcriptional unit (STU) systems with transfer RNA (tRNA) as the cleavage site performed better than other systems. The average editing efficiency at the four targets with TCTU-tRNA and STU-tRNA was 50.5% and 46.7%, respectively. Then, we designed four single-guide RNA (sgRNA) targets to induce mutations at RS2 and RS3 by using the TCTU-tRNA system. After the soybean transformation, we obtained several RS2 and RS3 mutation plants, and a subset of alleles was successfully transferred to the progeny. We identified null single and double mutants at the T2 generation and analyzed the seed carbohydrate content of their progeny. The RS2 and RS3 double mutants and the RS2 single mutant exhibited dramatically reduced levels of raffinose and stachyose in mature seeds. Further analysis of the growth and development of these mutants showed that there were no penalties on these phenotypes. Our results indicate that knocking out RS genes by multiplex CRISPR/Cas9-mediated gene editing is an efficient way to reduce RFOs in soybean. This research demonstrates the potential of using elite soybean cultivars to improve the soybean meal trait by multiplex CRISPR(Clustered Regularly Interspaced Short Palindromic Repeats)/Cas9-mediated gene editing.</p

Image_2_Over-Expression of LcPDS, LcZDS, and LcCRTISO, Genes From Wolfberry for Carotenoid Biosynthesis, Enhanced Carotenoid Accumulation, and Salt Tolerance in Tobacco.tiff

It is of great importance to combine stress tolerance and plant quality for breeding research. In this study, the role of phytoene desaturase (PDS), ζ-carotene desaturase (ZDS) and carotene isomerase (CRTISO) in the carotenoid biosynthesis are correlated and compared. The three genes were derived from Lycium chinenses and involved in the desaturation of tetraterpene. Their over-expression significantly increased carotenoid accumulation and enhanced photosynthesis and salt tolerance in transgenic tobacco. Up-regulation of almost all the genes involved in the carotenoid biosynthesis pathway and only significant down-regulation of lycopene ε-cyclase (ε-LCY) gene were detected in those transgenic plants. Under salt stress, proline content, and activities of catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD) were significantly increased, whereas malonaldehyde (MDA) and hydrogen peroxide (H2O2) accumulated less in the transgenic plants. The genes encoding ascorbate peroxidase (APX), CAT, POD, SOD, and pyrroline-5-carboxylate reductase (P5CR) were shown to responsive up-regulated significantly under the salt stress in the transgenic plants. This study indicated that LcPDS, LcZDS, and LcCRTISO have the potential to improve carotenoid content and salt tolerance in higher plant breeding.</p

Data_Sheet_1_Dual effects of serum urate on stroke risk and prognosis: insights from Mendelian randomization.xlsx

BackgroundTo investigate the causal associations of serum urate (SUA) with stroke risk and prognosis using Mendelian randomization (MR) and the potential mediating role of stroke risk factors in the causal pathways.MethodsWe used the random-effects inverse variance weighting (IVW) as our primary method. We initially performed two-sample univariable MR (UVMR) to identify the causal associations of SUA (n = 437,354) with any stroke (AS, FinnGen: n = 311,635; MEGASTROKE: n = 446,696), ischemic stroke (IS, FinnGen: n = 212,774; MEGASTROKE: n = 440,328), intracranial hemorrhage (ICH, FinnGen: n = 343,663; ISGC: n = 3,026), functional outcome after ischemic stroke at 90d (n = 4,363), and motor recovery within 24 months after stroke (n = 488), and then multivariable MR (MVMR) to estimate the direct causal effects of SUA on these outcomes, adjusting for potential confounders. Finally, we further conducted a two-step MR to investigate the potential mediating role of body mass index (BMI), systolic blood pressure (SBP), diastolic blood pressure (DBP), and estimated glomerular filtration rate (eGFR) in the identified causal pathways.ResultsGenetically predicted elevated SUA levels were significantly associated with increased risk of AS (meta-analysis: OR = 1.09, 95% CI [1.04–1.13], p = 3.69e-05) and IS (meta-analysis: OR = 1.10, 95% CI [1.01–1.19], p = 0.021) and with improved poor functional outcome after ischemic stroke at 90d (OR = 0.81, 95% CI [0.72–0.90], p = 1.79e-04) and motor recovery within 24 months after stroke (OR = 1.42, 95% CI [1.23–1.64], p = 2.15e-06). In MVMR, SBP and DBP significantly attenuated the causal effects of SUA on AS, IS, and functional outcome after ischemic stroke at 90d and motor recovery within 24 months after stroke. Further mediation analyses showed that SBP mediated 52.4 and 34.5% of the effects of SUA on AS and IS, while DBP mediated 28.5 and 23.4% of the causal effects, respectively.ConclusionThis study supports the dual role of genetically predicted SUA in increasing stroke risk, especially ischemic stroke risk, and in improving functional outcome and motor recovery. SBP and DBP are key mediators lying on the causal pathways of SUA with AS and IS.</p