Enantioselective copper-catalyzed azidation/click cascade reaction for access to chiral 1,2,3-triazoles.

Chiral 1,2,3-triazoles are highly attractive motifs in various fields. However, achieving catalytic asymmetric click reactions of azides and alkynes for chiral triazole synthesis remains a significant challenge, mainly due to the limited catalytic systems and substrate scope. Herein, we report an enantioselective azidation/click cascade reaction of N-propargyl-ß-ketoamides with a readily available and potent azido transfer reagent via copper catalysis, which affords a variety of chiral 1,2,3-triazoles with up to 99% yield and 95% ee under mild conditions. Notably, chiral 1,5-disubstituted triazoles that have not been accessed by previous asymmetric click reactions are also prepared with good functional group tolerance.

A phase-separated protein hub modulates resistance to Fusarium head blight in wheat.

Fusarium head blight (FHB) is a devastating wheat disease. Fhb1, the most widely applied genetic locus for FHB resistance, is conferred by TaHRC of an unknown mode of action. Here, we show that TaHRC alleles distinctly drive liquid-liquid phase separation (LLPS) within a proteinaceous complex, determining FHB susceptibility or resistance. TaHRC-S (susceptible) exhibits stronger LLPS ability than TaHRC-R (resistant), and this distinction is further intensified by fungal mycotoxin deoxynivalenol, leading to opposing FHB symptoms. TaHRC recruits a protein class with intrinsic LLPS potentials, referred to as an "HRC-containing hub." TaHRC-S drives condensation of hub components, while TaHRC-R comparatively suppresses hub condensate formation. The function of TaSR45a splicing factor, a hub member, depends on TaHRC-driven condensate state, which in turn differentially directs alternative splicing, switching between susceptibility and resistance to wheat FHB. These findings reveal a mechanism for FHB spread within a spike and shed light on the roles of complex condensates in controlling plant disease.

Targeting PI3K/Akt in Cerebral Ischemia Reperfusion Injury Alleviation: From Signaling Networks to Targeted Therapy.

Ischemia/reperfusion (I/R) injury is a pathological event that results in reperfusion due to low blood flow to an organ. Cerebral ischemia is a common cerebrovascular disease with high mortality, and reperfusion is the current standard intervention. However, reperfusion may further induce cellular damage and dysfunction known as cerebral ischemia/reperfusion injury (CIRI). Currently, strategies for the clinical management of CIRI are limited, necessitating the exploration of novel and efficacious treatment modalities for the benefit of patients. PI3K/Akt signaling pathway is an important cellular process associated with the disease. Stimulation of the PI3K/Akt pathway enhances I/R injury in multiple organs such as heart, brain, lung, and liver. It stands as a pivotal signaling pathway crucial for diminishing cerebral infarction size and safeguarding the functionality of brain tissue after CIRI. During CIRI, activation of the PI3K/Akt pathway exhibits a protective effect on CIRI. Furthermore, activation of the PI3K/Akt pathway has the potential to augment the activity of antioxidant enzymes, resulting in a decrease in reactive oxygen species (ROS) and the associated oxidative stress. Meanwhile, PI3K/Akt plays a neuroprotective role by inhibiting inflammatory responses and apoptosis. For example, PI3K/Akt interacts with NF-κB, Nrf2, and MAPK signaling pathways to mitigate CIRI. This article is aimed to explore the pivotal role and underlying mechanism of PI3K/Akt in ameliorating CIRI and investigate the influence of ischemic preconditioning and post-processing, as well as the impact of pertinent drugs or activators targeting the PI3K/Akt pathway on CIRI. The primary objective is to furnish compelling evidence supporting the activation of PI3K/Akt in the context of CIRI, elucidating its mechanistic intricacies. By doing so, the paper aims to underscore the critical contribution of PI3K/Akt in mitigating CIRI, providing a theoretical foundation for considering the PI3K/Akt pathway as a viable target for CIRI treatment.

Role of traditional Chinese medicine monomers in cerebral ischemia/reperfusion injury:a review of the mechanism.

Ischemia/reperfusion (I/R) injury is a pathological process wherein reperfusion of an ischemic organ or tissue exacerbates the injury, posing a significant health threat and economic burden to patients and their families. I/R triggers a multitude of physiological and pathological events, such as inflammatory responses, oxidative stress, neuronal cell death, and disruption of the blood-brain barrier (BBB). Hence, the development of effective therapeutic strategies targeting the pathological processes resulting from I/R is crucial for the rehabilitation and long-term enhancement of the quality of life in patients with cerebral ischemia/reperfusion injury (CIRI). Traditional Chinese medicine (TCM) monomers refer to bioactive compounds extracted from Chinese herbal medicine, possessing anti-inflammatory and antioxidative effects, and the ability to modulate programmed cell death (PCD). TCM monomers have emerged as promising candidates for the treatment of CIRI and its subsequent complications. Preclinical studies have demonstrated that TCM monomers can enhance the recovery of neurological function following CIRI by mitigating oxidative stress, suppressing inflammatory responses, reducing neuronal cell death and functional impairment, as well as minimizing cerebral infarction volume. The neuroprotective effects of TCM monomers on CIRI have been extensively investigated, and a comprehensive understanding of their mechanisms can pave the way for novel approaches to I/R treatment. This review aims to update and summarize evidence of the protective effects of TCMs in CIRI, with a focus on their role in modulating oxidative stress, inflammation, PCD, glutamate excitotoxicity, Ca2+ overload, as well as promoting blood-brain barrier repairment and angiogenesis. The main objective is to underscore the significant contribution of TCM monomers in alleviating CIRI.

Editorial for the Special Issue "Genetics Studies on Wheat".

Wheat (Triticum aestivum L [...].

Molecular Characterization and Marker Development of the HMW-GS Gene from Thinopyrum elongatum for Improving Wheat Quality.

The quality of wheat primarily depends on its storage protein quality, especially in regards to gluten content and high-molecular-weight glutenin subunits (HMW-GS). The number of HMW-GS alleles is limited in bread wheat (Triticum aestivum L.), whereas it is abundant in wheat relatives. Therefore, HMW-GS alleles from wheat relatives could provide a potential for improving quality in wheat breeding. Thinopyrum elongatum (EE) is one of the relatives of wheat. The E genome is closely related to the ABD genome in wheat; therefore, Th. elongatum is often used as an excellent exogenous gene donor for wheat genetic improvement. In this study, the high-molecular glutenin subunit gene was cloned and sequenced from Th. elongatum. A specific molecular marker for identifying the Glu-1Ey subunit gene was developed and applied to detected wheat-Th. elongatum alien introgression lines. Quality analysis indicated that the substitution and addition lines containing Th. elongatum alleles significantly (p < 0.05) increased grain protein content by 3.76% to 5.11%, wet-gluten content by 6.55% to 8.73%, flour 8-MW by 0.25% to 6.35%, and bread volume value by 33.77 mL to 246.50 mL, in comparing it with Chinese Spring. The GMP content and lactic acid SRC showed significant positive correlations with flour processing quality and might be used as indicators for wheat quality. The results were expected to provide a novel route for improving processing quality in wheat quality breeding.

The putative vacuolar processing enzyme gene TaVPE3cB is a candidate gene for wheat stem pith-thickness.

KEY MESSAGE: The vacuolar processing enzyme gene TaVPE3cB is identified as a candidate gene for a QTL of wheat pith-thickness on chromosome 3B by BSR-seq and differential expression analyses. The high pith-thickness (PT) of the wheat stem could greatly enhance stem mechanical strength, especially the basal internodes which support the heavier upper part, such as upper stems, leaves and spikes. A QTL for PT in wheat was previously discovered on 3BL in a double haploid population of 'Westonia' × 'Kauz'. Here, a bulked segregant RNA-seq analysis was applied to identify candidate genes and develop associated SNP markers for PT. In this study, we aimed at screening differentially expressed genes (DEGs) and SNPs in the 3BL QTL interval. Sixteen DEGs were obtained based on BSR-seq and differential expression analyses. Twenty-four high-probability SNPs in eight genes were identified by comparing the allelic polymorphism in mRNA sequences between the high PT and low PT samples. Among them, six genes were confirmed to be associated with PT by qRT-PCR and sequencing. A putative vacuolar processing enzyme gene TaVPE3cB was screened out as a potential PT candidate gene in Australian wheat 'Westonia'. A robust SNP marker associated with TaVPE3cB was developed, which can assist in the introgression of TaVPE3cB.b in wheat breeding programs. In addition, we also discussed the function of other DEGs which may be related to pith development and programmed cell death (PCD). A five-level hierarchical regulation mechanism of stem pith PCD in wheat was proposed.

Azidobenziodazolones as Azido Sources for the Enantioselective Copper-Catalyzed Azidation of N-Unprotected 3-Trifluoromethylated Oxindoles.

Both azido (N3) and trifluoromethyl (CF3) groups are key moieties of numerous valuable molecules that are extensively applied in drug discovery, chemical biology, and synthetic chemistry. However, the asymmetric construction of chiral quaternary stereocenters bearing both N3 and CF3 groups is still unexplored. Herein, we report a kind of bench-stable and easily adjustable benziodazolone-based azidating reagents. These reagents were used to achieve an enantioselective copper-catalyzed azidation of N-unprotected 3-trifluoromethylated oxindoles to provide diverse enantioenriched 3-N3-3-CF3 oxindoles.

Transfer of the Resistance to Multiple Diseases from a Triticum-Secale-Thinopyrum Trigeneric Hybrid to Ningmai 13 and Yangmai 23 Wheat Using Specific Molecular Markers and GISH.

The middle to lower reaches of the Yangtze River are China's second largest area for wheat production; wheat disease is more serious there than in other areas because of the high humidity and warm weather. However, most cultivated varieties are susceptible to Fusarium head blight (FHB), powdery mildew, and stripe rust, and the lack of disease-resistant germplasm is an obstacle in wheat breeding. Rye and Thinopyrum elongatum, related species of wheat, carry many genes involved in disease resistance. In this study, a trigeneric hybrid, YZU21, with resistance to FHB, powdery mildew, and stripe rust was used to improve two major wheat cultivars, Ningmai 13 (NM13) and Yangmai 23 (YM23). Specific molecular markers and GISH were used to identify hybrid progenies. Five addition or substitution lines and one translocation line of the Triticum-Secale-Thinopyrum trigeneric hybrid were obtained and evaluated for agronomic traits and the resistance to multiple diseases. The results showed that the six trigeneric hybrid lines had desirable agronomic traits and improved resistance to FHB, powdery mildew, and stripe rust; they might be used as parents in wheat breeding for the resistance to multiple disease.

Exploring and applying genes to enhance the resistance to Fusarium head blight in wheat.

Fusarium head blight (FHB) is a destructive disease in wheat worldwide. Fusarium graminearum species complex (FGSC) is the main causal pathogen causing severe damage to wheat with reduction in both grain yield and quality. Additionally, mycotoxins produced by the FHB pathogens are hazardous to the health of human and livestock. Large numbers of genes conferring FHB resistance to date have been characterized from wheat and its relatives, and some of them have been widely used in breeding and significantly improved the resistance to FHB in wheat. However, the disease spreads rapidly and has been severe due to the climate and cropping system changes in the last decade. It is an urgent necessity to explore and apply more genes related to FHB resistant for wheat breeding. In this review, we summarized the genes with FHB resistance and mycotoxin detoxication identified from common wheat and its relatives by using forward- and reverse-genetic approaches, and introduced the effects of such genes and the genes with FHB resistant from other plant species, and host-induced gene silencing (HIGS) in enhancing the resistance to FHB in wheat. We also outlined the molecular rationale of the resistance and the application of the cloned genes for FHB control. Finally, we discussed the future challenges and opportunities in this field.

Premature Termination Codon of 1Dy12 Gene Improves Cookie Quality in Ningmai9 Wheat.

The area between middle and lower reaches of the Yangtze River is the largest region for soft wheat production in China. In soft wheat breeding, the lack of germplasm with desirable quality for end-use products is a barrier. Ningmai9 is the main variety of soft wheat planted in this area. To create germplasm with better quality and yield potential than Ningmai9, mutants of HMW-GSs in Ningmai9 induced by ethylmethanesulfonate (EMS) were obtained. SDS-PAGE showed that two mutants, md10 and md11, were HMW-GS 1Dy deletions. DNA sequencing confirmed that one mutation was caused by a C/T substitution, resulting in the change of CAA encoding glutamine into the termination codon TAA, and another mutation was due to a G/A substitution in the central repetitive domain of the coding region, causing TGG encoding tryptophan to become the termination codon TGA. The premature termination codon of the 1Dy12 gene affected the expression of 1Dy12 and kept the mRNA at a lower transcription level during the kernel development stage in comparison with the wild type. HMW-GS 1Dy12 deletion mutants decreased the content of HMW-GSs and glutenin macropolymers, mixograph envelope peak time and TIMEX width, water solvent retention capacity (WSRC), and lactic acid solvent retention capacity (LASRC). In the HMW-GS 1Dy12 deletion lines, the sugar-snap cookie diameter was 8.70-8.74 cm, which was significantly larger than that in the wild type of 8.0 cm. There were no significant differences in spike number, kernel number, thousand kernel weight, and yield between the deletion lines and wild type. Overall, the study indicated that the knockout of the HMW-GS gene induced by EMS is an effective way to improve wheat quality, and deletion mutants of HMW-GS 1Dy12 decrease gluten strength and increase sugar snap cookie diameter without yield penalty in Ningmai9 wheat.

Effect of Hand Intensive Training on Upper Limb Function of Stroke Patients with Hemiplegia.

Objective: To analyze the effect of hand intensive training on upper limb function of stroke patients with hemiplegia. Methods: 110 stroke patients were randomly divided into two groups: the reference group and the observation group. 55 patients in the reference group were treated with routine rehabilitation treatment, including routine joint activity training, bed training, exercise therapy, and ADL ability training; 55 cases in the observation group received intensive hand training on the basis of routine rehabilitation treatment, including inducing the patient's five finger extension, forcibly pulling the fingers and wrist joints, and suddenly opening his fist after clenching his fist. Results: The treatment period of the two groups was 5 weeks. In the comparison results of Fugl-Meyer (FMA), the exercise effect of the observation group with increased hand intensive training was significantly better than that of the control group with stroke hemiplegia treated with conventional methods. The difference was statistically significant, t < 10.000, P < 0.05; In the comparative analysis of upper limb function test (UEFT), the effect of the observation group was significantly higher than that of the reference group treated with routine rehabilitation nursing (all P < 0.05); In the comprehensive comparison of exercise ability results, the observation group was higher than the reference group in the flexibility, fineness, and fineness of activity behavior after treatment. Conclusion: Strengthening hand intensive training can further improve the upper limb motor function of stroke patients with hemiplegia, reduce the severity of hemiplegia, and improve the recovery effect of stroke patients. It is worthy of clinical promotion and application.

Regulatory Mechanisms of Mitogen-Activated Protein Kinase Cascades in Plants: More than Sequential Phosphorylation.

Mitogen-activated protein kinase (MAPK) cascades play crucial roles in almost all biological processes in plants. They transduce extracellular cues into cells, typically through linear and sequential phosphorylation and activation of members of the signaling cascades. However, accumulating data suggest various regulatory mechanisms of plant MAPK cascades in addition to the traditional phosphorylation pathway, in concert with their large numbers and coordinated roles in plant responses to complex ectocytic signals. Here, we highlight recent studies that describe the uncanonical mechanism of regulation of MAPK cascades, regarding the activation of each tier of the signaling cascades. More particularly, we discuss the unusual role for MAPK kinase kinases (MAPKKKs) in the regulation of MAPK cascades, as accumulating data suggest the non-MAPKKK function of many MAPKKKs. In addition, future work on the biochemical activation of MAPK members that needs attention will be discussed.

Linkage and association mapping and Kompetitive allele-specific PCR marker development for improving grain protein content in wheat.

KEY MESSAGE: Linkage and association mapping identified nine candidate intervals for wheat GPC, and large-scale association mapping based on 9 corresponding KASP markers and 1163 F4 breeding lines revealed 3 significant markers. Wheat grain protein content (GPC) is an important quality indicator. The GPC of wheat grown in the middle and lower reaches of the Yangtze River is often low. Marker-assisted selection (MAS) is an effective tool for improving quantitative traits; however, most markers have not been effectively applied in GPC improvement except Gpc-B1, although many loci associated with GPC were identified. In this study, linkage analysis using a recombinant inbred line population from the cross of core parents of Ningmai 9 and Yangmai 158 and association mapping using the local cultivated varieties were performed and nine candidate intervals were identified. The appropriate kompetitive allele-specific PCR (KASP) markers associated with GPC were successfully developed and screened in 1163 F4 breeding lines. Three markers, Kgpc-2B, Kgpc-2D, and Kgpc-4A, were validated to be significantly related to GPC by large-scale association mapping, and they were combined to achieve the highest efficiency to enhance GPC. We applied these markers in 164 F6 breeding lines and obtained 15 lines with high GPC, indicating their high selective efficiency. Further, strategies for gene exploration in the three significant intervals were proposed. These results were expected to provide a novel route for improving GPC in wheat quality breeding.

Alamandine protects against renal ischaemia-reperfusion injury in rats via inhibiting oxidative stress.

OBJECTIVE: This study was to determine whether alamandine (Ala) could reduce ischaemia and reperfusion (I/R) injury of kidney in rats. METHODS: Renal I/R was induced by an occlusion of bilateral renal arteries for 70 min and a 24-h reperfusion in vivo, and rat kidney proximal tubular epithelial cells NRK52E were exposed to 24 h of hypoxia and followed by 3-h reoxygenation (H/R) in vitro. RESULTS: The elevated serum creatinine (Cr), blood cystatin C (CysC) and blood urea nitrogen (BUN) levels in I/R rats were inhibited by Ala treatment. Tumour necrosis factor alpha (TNF)-α, IL-1ß, IL-6, cleaved caspase-3, cleaved caspase-8 and Bax were increased, and Bcl2 was reduced in the kidney of I/R rats, which were reversed by Ala administration. Ala reversed the increase of TNF-α, IL-1ß, IL-6, cleaved caspase-3, cleaved caspase-8 and Bax and the decrease of Bcl2 in the H/R NRK52E cells. Ala could also inhibit the increase of oxidative stress levels in the kidney of I/R rats. NADPH oxidase 1 (Nox1) overexpression reversed the improving effects of Ala on renal function, inflammation and apoptosis of I/R rats. CONCLUSION: These results indicated that Ala could improve renal function, attenuate inflammation and apoptosis in the kidney of I/R rats via inhibiting oxidative stress.

Characterization of the baking-induced changes in starch molecular and crystalline structures in sugar-snap cookies.

This study aims to understand the starch molecular structural changes from baking sugar-snap cookies. Changes in the whole-molecule size distribution and chain-length distribution of the parent wheat flour and from final cookie products were measured by size-exclusion chromatography with and without enzymatic debranching, and the results fitted by two biosynthesis-based models. Fraction crystallinity was also analyzed. After cooking, there was a significant decrease in average molecular sizes of amylopectin and in the average lengths of amylose chains, and some starch granules lost birefringence. However, the chain-length distributions of amylopectin showed no noticeable difference, resulting in little change in relative crystallinity and gelatinization temperatures. Both the short-range ordered structure and the periodic lamellar structure were disrupted. This study provides new insight into starch structural changes in sugar-snap cookies after baking, which play an important role in determining final cookie quality. For example, a decrease in size of amylose chains influences cookie sensory properties, and thus can be used as an additional tool for choice of grains.

TaUGT6, a Novel UDP-Glycosyltransferase Gene Enhances the Resistance to FHB and DON Accumulation in Wheat.

Fusarium head blight (FHB), a devastating wheat disease, results in loss of yield and production of mycotoxins including deoxynivalenol (DON) in infected grains. DON is harmful to human and animal health and facilitates the spread of FHB symptoms. Its conversion into DON-3-glucoside (D3G) by UDP-glycosyltransferases (UGTs) is correlated with FHB resistance, and only few gene members in wheat have been investigated. Here, Fusarium graminearum and DON-induced TaUGT6 expression in the resistant cultivar Sumai 3 was cloned and characterized. TaUGT6::GFP was subcellularly located throughout cells. Purified TaUGT6 protein could convert DON into D3G to some extent in vitro. Transformation of TaUGT6 into Arabidopsis increased root tolerance when grown on agar plates containing DON. Furthermore, TaUGT6 overexpression in wheat showed improved resistance to Fusarium spread after F. graminearum inoculation. Overall, this study provides useful insight into a novel UGT gene for FHB resistance in wheat.

Phylogeny and gene expression of the complete NITRATE TRANSPORTER 1/PEPTIDE TRANSPORTER FAMILY in Triticum aestivum.

NPF genes encode membrane transporters involved in the transport of a large variety of substrates including nitrate and peptides. The NPF gene family has been described for many plants, but the whole NPF gene family for wheat has not been completely identified. The release of the wheat reference genome has enabled the identification of the entire wheat NPF gene family. A systematic analysis of the whole wheat NPF gene family was performed, including responses of specific gene expression to development and nitrogen supply. A total of 331 NPF genes (113 homoeologous groups) have been identified in wheat. The chromosomal location of the NPF genes is unevenly distributed, with predominant occurrence in the long arms of the chromosomes. The phylogenetic analysis indicated that wheat NPF genes are closely clustered with Arabidopsis, Brachypodium, and rice orthologues, and subdivided into eight subfamilies. The expression profiles of wheat NPF genes were examined using RNA-seq data, and a subset of 44 NPF genes (homoeologous groups) with contrasting expression responses to nitrogen and/or development in different tissues were identified. The systematic identification of gene composition, chromosomal locations, evolutionary relationships, and expression profiles contributes to a better understanding of the roles of the wheat NPF genes and lays the foundation for further functional analysis in wheat.

Sulfate application decreases translocation of arsenic and cadmium within wheat (Triticum aestivum L.) plant.

Arsenic (As) and cadmium (Cd) typically exhibit divergent fates in soil, which complicates efforts to decrease As and Cd accumulation in the edible parts of crops. Here, we performed pot experiments to examine the effect of sulfate application on As and Cd accumulation in the grain of wheat grown in contaminated soil. Compared to the control (no sodium sulfate addition), application of 120 mg kg-1 sodium sulfate decreased the rhizosphere soil pH from 7.27 to 7.10 and increased the soil extractable Cd concentration; however, it did not significantly influence the soil extractable As concentration. However, sodium sulfate addition decreased As and Cd concentrations in wheat grain, in association with decreased As and Cd translocation from root and straw to grain, rather than from soil to root. Furthermore, sodium sulfate addition significantly decreased membrane lipid peroxidation and enhanced photosynthesis, while increasing the uptake of nitrogen, phosphorus, and sulfur. These effects increased the growth and grain weight of plants grown in As and Cd co-contaminated soil. Our findings provide insight into the mechanisms by which sulfate modulates As and Cd uptake and translocation in wheat; moreover, our findings will enable formulation of strategies to decrease As and Cd concentrations in the grain of wheat grown in As and Cd co-contaminated soil.

Yeast two-hybrid screening for proteins that interact with PFT in wheat.

Fusarium head blight (FHB) is a devastating disease of wheat worldwide. Fhb1 is the most consistently reported quantitative trait locus (QTL) for FHB resistance breeding. A pore-forming toxin-like (PFT) gene at Fhb1 was first cloned by map-based cloning and found to confer FHB resistance in wheat. Proteins often interact with each other to execute their functions. Characterization of the proteins interacting with PFT might therefore provide information on the molecular mechanisms of PFT functions. In this study, a high-quality yeast two-hybrid (Y2H) library using RNA extracted from Fusarium graminearum (Fg)-infected wheat spikes of Sumai 3 was constructed. The agglutinin domains of PFT exhibited no self-activation and toxicity to yeast cells and were used as bait to screen the Y2H library. Twenty-three proteins that interact with PFT were obtained, which were mainly involved in the ubiquitination process, clathrin coat assembly, the oxidation-reduction process, and protein phosphorylation. The expression pattern of these interacting genes was analyzed by quantitative real-time PCR. This study clarifies the protein interactions of PFT and raises a regulatory network for PFT regarding FHB resistance in wheat.