Molecular Technology Research Articles

Molecular characterisation of Histoplasma capsulatum sensu lato from Ethiopian horses reveals two distinct phylogenetic clades.

Epizootic lymphangitis (EL) is a highly prevalent and contagious infectious disease affecting horses in many parts of Ethiopia caused by Histoplasma capsulatum sensu lato ('var. farciminosum'). In this study, 12 suspected isolates of H. capsulatum sensu lato or yeasts unidentified by conventional biochemical tests isolated from Ethiopian horses with EL were characterised by internal transcribed spacer sequencing. Six of the 12 isolates were identified to be members of H. capsulatum sensu lato and the other six were Pichia kudriavzevii (synonym: Candida krusei) (n=3), Trichosporon asahii (n=1), Geotrichum silvicola (n=1) and Moesziomyces aphidis (n=1), respectively. The six H. capsulatum sensu lato isolates were further characterised by multilocus sequence analysis. Four distinct gene loci (arf [462 bases], H-anti [410 bases], ole1 [338 bases] and tub1 [272 bases]) of these six isolates as well as those of two H. capsulatum sensu lato ('var. farciminosum') reference strains (ATCC 58332 and ATCC 28798) were polymerase chain reaction (PCR)-amplified and sequenced. Phylogenetic analyses of their concatenated nucleotide sequences showed that three of the isolates and the reference strain ATCC 58332 were identical and belonged to the Eurasia clade within Latin American (LAm) A (H. suramericanum), and those of the other three isolates and the reference strain ATCC 28798 were identical and belonged to the Africa clade. At least two distinct phylogenetic clades of H. capsulatum sensu lato were circulating in Ethiopian horses with EL. Advanced molecular technologies and bioinformatics tools are crucial for the accurate identification and typing of pathogens as well as the discovery of novel microorganisms in veterinary microbiology.

Review of Pseudomonas aeruginosa and Klebsiella pneumoniae as venereal pathogens in horses.

Three bacteria extensively acknowledged as venereal pathogens with the potential to induce endometritis include Taylorella equigenitalis, the causative agent of contagious equine metritis (CEM), specific strains of Pseudomonas aeruginosa, and certain capsule types of Klebsiella pneumoniae. The United Kingdom's Horserace Betting Levy Board recommends pre-breeding screening for these bacteria in their International Codes of Practice and >20 000 samples are tested per annum in the United Kingdom alone. While the pathogenesis and regulatory importance of CEM are well established, an evaluation of the literature pertaining to venereal transmission of P. aeruginosa and K. pneumoniae was lacking. The aim of this review was to evaluate published literature and determine the significance of P. aeruginosa and K. pneumoniae as venereal pathogens in horses. Literature definitively demonstrating venereal transmission was not available. Instead, application of molecular typing methods suggested that common environmental sources of contamination, such as water, or fomites be considered as modes of transmission. The presence of organisms with pathogenic potential on a horse's external genitalia did not predict venereal transmission with resultant endometritis and reduced fertility. These findings may prompt further investigation using molecular technologies to confirm or exclude venereal spread and investigation of alternative mechanisms of transmission are indicated.

Optical molecular imaging in cancer research: current impact and future prospect

Abstract Cancer has long been a major threat to human health. Recent advancements in molecular imaging have revolutionized cancer research by enabling early and precise disease localization, essential for effective management. In particular, optical molecular imaging is an invaluable cancer detection tool in preoperative planning, intraoperative guidance, and postoperative monitoring owing to its noninvasive nature, rapid turnover, safety, and ease of use. The tumor microenvironment and cells within it express distinct biomarkers. Optical imaging technology leverages these markers to differentiate tumor tissues from surrounding tissues and capture real-time images with high resolution. Nevertheless, a robust understanding of these cancer-related molecules and their dynamic changes is crucial for effectively managing cancer. Recent advancements in optical molecular imaging technologies offer novel approaches for cancer investigation in research and practice. This review investigates the modern optical molecular imaging techniques employed in both preclinical and clinical research, including bioluminescence, fluorescence, chemiluminescence, photoacoustic imaging, and Raman spectroscopy. We explore the current paradigm of optical molecular imaging modalities, their current status in preclinical cancer research and clinical applications, and future perspectives in the fields of cancer research and treatment.

Advances in the selection of functional monomers for molecularly imprinted polymers: A review.

Molecularly imprinted polymers, a type of special polymer materials, are widely used in biosensing and other fields due to their ability to specifically recognize target molecules, often called "artificial receptors.". Nowadays, researchers are constantly exploring new design and synthesis methods for molecularly imprinted materials to improve the selectivity and sensitivity of molecularly imprinted materials. Among them, the selection of functional monomers has attracted great attention. This review comprehensively analyzes and discusses the selection methods of functional monomers. The most commonly used functional monomers among different types of templates are screened based on the structural properties of the template molecules, including the selection of functional monomers among ion-imprinted polymers, protein-imprinted polymers, and bacterial imprinted polymers. The rich binding sites and functional group types of multifunctional monomers are also highlighted to advance the development of molecular imprinting technology. The article further explores the current challenges and prospects in the selection of functional monomers and emphasizes multiplex experiments and computer simulations as important directions for future research. This review provides comprehensive information and constructive guidelines for researchers in selecting functional monomers in areas such as analytical chemistry and biosensors.

Screening and isolation of quorum sensing inhibitors of Pseudomonas aeruginosa from Phellodendron amurense extracts using bio-affinity chromatography.

Drug-resistant bacterial infections pose a significant challenge in the field of bacterial disease treatment. Finding new antibacterial pathways and targets to combat drug-resistant bacteria is crucial. The bacterial quorum sensing (QS) system regulates the expression of bacterial virulence factors. Inhibiting bacterial QS and reducing bacterial virulence can achieve antibacterial therapeutic effects, making QS inhibition an effective strategy to control bacterial pathogenicity. This article mainly focused on the PqsA protein in the QS system of Pseudomonas aeruginosa. An affinity chromatography medium was developed using the SpyTag/SpyCatcher heteropeptide bond system. Berberine, which can interact with the PqsA target, was screened from Phellodendron amurense by affinity chromatography. We characterized its structure, verified its inhibitory activity on P. aeruginosa, and preliminarily analyzed its mechanism using molecular docking technology. This method can also be widely applied to the immobilization of various protein targets and the effective screening of active substances.

Cinnamaldehyde Regulates the Migration and Apoptosis of Gastric Cancer Cells by Inhibiting the Jak2/Stat3 Pathway.

Gastric cancer is a malignant tumor with high morbidity and mortality all around the world. Because of its poor prognosis and low survival rate, the treatment of gastric cancer has received extensive attention. Cinnamaldehyde (CA) is the main single active component of the Chinese herbal medicine cinnamon, which has a variety of pharmacological effects. The inhibitory effect of CA on the growth of some tumor cells has been proven, but its therapeutic effect on gastric cancer has rarely been reported. Through network pharmacology, bioinformatics methods, and molecular docking technology, we predicted the interaction targets of CA and gastric cancer. Moreover, we found that apoptosis is an important mode of action of CA on gastric cancer cells. Subsequently, we validated it in gastric cancer cell lines cultured in vitro. The results showed that in the presence of CA, the Jak2/Stat3 pathway was inhibited, the ratio of Bcl-2/Bax decreased, and the apoptosis of gastric cancer cells was promoted in a concentration-dependent. In conclusion, CA can promote the apoptosis of gastric cancer cells by inhibiting the activity of the Jak2/Stat3 pathway, which may achieve the effect of treating gastric cancer.

Successful development of molecular diagnostic technology combining mini-barcoding and high-resolution melting for traditional Chinese medicine agarwood species based on single-nucleotide polymorphism in the chloroplast genome.

Agarwood is a valuable traditional medicine and fragrance. The production process is a typical injury-induced defense response. Currently, there are approximately 22 known species in the genus Aquilaria Lam., all of which can produce agarwood, whereas there are only two legal species of traditional Chinese medicinal agarwood, Aquilaria sinensis (Lour.) Spreng. and Aquilaria agallocha (Lour.) Roxb. The Taiwan herbal Pharmacopoeia of China stipulates that the medicinal agarwood species are A. sinensis and its relatives in the same genus. Moreover, there are five species of agarwood available for clinical medicinal use in Japan, including A. agallocha and A. sinensis, which are often confused with each other or used in a mixed way in the trade process. Therefore, accurate identification of traditional Chinese medicinal agarwood species is important to ensure the authenticity of traditional medicines and to guide the safety of clinical medication. In this study, 59 specific single-nucleotide polymorphism loci were screened and obtained from the chloroplast genomes of 12 species of the genus Aquilaria Lam. We established an identification method for traditional Chinese medicinal agarwood using mini-barcoding combined with high-resolution melting (HRM) and designed and validated 10 pairs of primers from the psbM-trnD, psbA, rps16, petN, ndhE-psaC, rps4, atpE, ycf1, rps15-trnN, and matK regions. The amplification products were all less than 200 bp, with a high success rate of amplification. The method was applied to successfully identify traditional Chinese medicinal agarwood species from commercial agarwood samples. Overall, the sensitivity of this method was sufficient to detect 1% of adulterants in medicinal agarwood products, proving that mini-barcoding HRM is a powerful and flexible tool. This method can be used as a fast and effective high-throughput method for authenticity testing of traditional Chinese medicinal agarwood and its raw materials containing agarwood-containing proprietary Chinese medicines and is recommended for industrial applications.

The Mechanism of Action of the Active Ingredients of Coptidis rhizoma against Porcine Epidemic Diarrhea Was Investigated Using Network Pharmacology and Molecular Docking Technology.

The objective of this study was to elucidate the mechanism of action of the active components of Coptidis rhizoma against porcine epidemic diarrhea and to provide a theoretical foundation for further development of novel anti-PED therapeutic agents based on Coptidis rhizoma. The potential targets of Coptidis rhizoma against PEDV were identified through a comprehensive literature review and analysis using the TCMSP pharmacological database, SwissDrugDesign database, GeneCards database, and UniProt database. Subsequently, the STRING database and Cytoscape 3.7.1 software were employed to construct a protein-protein interaction (PPI) network and screen key targets. Gene Ontology (GO) function and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were conducted on the identified targets. Molecular docking studies were performed using AutoDock 1.5.7 software to analyze the binding energy and modes of interaction between the active components of Coptidis rhizoma and the target proteins. The PyMOL 2.5.0a0 software was employed to visualize the docking results. Through comprehensive analysis, 74 specific targets of active components of Coptidis rhizoma against PEDV were identified. The core gene targets were screened, and an interaction network diagram was subsequently generated. Ultimately, 14 core targets were identified, with STAT3, ESR1, CASP3, and SRC exhibiting the most significant interactions. GO enrichment analysis revealed a total of 215 molecular items, including 48 biological function items, 139 biological process items, and 28 cellular component items. KEGG enrichment analysis identified 140 signaling pathways. Molecular docking analysis demonstrated that epiberberine and palmatine exhibited high binding affinity with STAT3 protein, worenine showed high binding affinity with ESR1 protein, obacunone exhibited high binding affinity with CASP3 protein, and epiberberine, obacunone, berberine, and berberruine exhibited high binding affinity with SRC protein. A network pharmacology and molecular docking technology approach was employed to screen six important active components of Coptidis rhizoma and four important potential targets against PEDV infection. The findings indicated that the active components of Coptidis rhizoma could serve as promising pharmaceutical agents for the prevention and control of PEDV, with significant potential for clinical application.

Computer simulation‐guided template selection for a molecularly imprinted polymer for selective trifluralin adsorption

AbstractTo meet selective adsorption toward trifluralin, a novel molecularly imprinted polymer (MIP) was fabricated by the dummy template molecular imprinting technology. First, computational simulation was performed to select a suitable dummy template, 3,5‐dinitro‐4‐methylbenzoic acid (T1), based on the maximum basis set superposition error (BSSE)‐corrected binding interaction energy (ΔE) of the monomer N‐vinylpyrrolidone (NVP)‐T1 complex and its structural overlap with trifluralin. Then, the MIP was prepared via the bulk polymerization. The adsorption experiments showed the MIP exhibited a trifluralin adsorption capacity of 5.1 mg g−1, an imprinting factor (IF) of 2.2, and short adsorption equilibrium time of 5 min. The adsorption of trifluralin conformed to the Freundlich adsorption (R2 = 0.985) and pseudo‐second‐order model (R2 = 0.999). In addition, the MIP exhibited selectivity to trifluralin over other adsorbents, including structural analogs (pendimethalin and oryzalin), pesticide (carbendazim), and nitrocompounds (nitrofurantoin, furazolidone, and furaltadone), with the selectivity factor (β) in the range of 1.2–3.0, respectively. In trifluralin/oryzalin mixture, the IF toward oryzalin was still as high as 1.9. The removal rate of the MIP to trifluralin in environmental water samples ranged from 90.08% to 99.04%. This study provides theoretical and experimental insights for the preparation of MIP using dummy templates.

Dual Template Molecularly Imprinted Polymers Targeting Blockade of CD47 for Enhanced Macrophage Phagocytosis and Synergistic Antimetabolic Therapy.

Glycinamide ribonucleotide formyltransferase (GARFT) is an important enzyme in the folate metabolism pathway, and chemical drugs targeting GARFT have been used in tumor treatments over the past few decades. The development of novel antimetabolism drugs that target GARFT with improved performance and superior activity remains an attractive strategy. Herein, we proposed a targeted double-template molecularly imprinted polymer (MIP) for enhancing macrophage phagocytosis and synergistic antimetabolic therapy. The double-template MIP was prepared by imprinting the exposed peptide segment of the extracellular domain of CD47 and the active center of GARFT. Owing to the imprinted cavities on the surface of MIP, it can actively target cancer cells and mask the "do not eat me" signal upon binding to CD47 thereby blocking the CD47-SIRPα pathway and ultimately enhancing phagocytosis by macrophages. In addition, MIP can specifically bind to the active center of GARFT upon entry into the cells, thereby inhibiting its catalytic activity and ultimately interfering with the normal expression of DNA. A series of cell experiments demonstrated that MIP can effectively target CD47 overexpressed 4T1 cancer cells and inhibit the growth of 4T1 cells. The enhanced phagocytosis ability of macrophages-RAW264.7 cells was also clearly observed by confocal imaging experiments. In vivo experiments also showed that the MIP exhibited a satisfactory tumor inhibition effect. Therefore, this study provides a new idea for the application of molecular imprinting technology to antimetabolic therapy in conjunction with macrophage-mediated immunotherapy.

Study on the Effect of Rhubarb and Its Active Components on Pyroptosis in DKD by Regulating STAT3/Caspase-11 Axis

Rhubarb has been found to have a certain protective effect on improving the kidney function. However, the specific mechanism is still unclear. In this study, network pharmacology, molecular docking spontaneous binding technology and molecular biology experiments were used to verify the mechanism of rhubarb and its active ingredients in the treatment of DKD. A total of 10 active compounds and 121 (larger than average) target proteins were collected. The target proteins with higher degree value were screened by PPI according to degree value as follows: AKT1, STAT3, EGFR, NFKB1, SRC, etc. GO and KEGG enrichment analysis suggest that rhubarb therapy for DKD mainly involves Pathways in cancer, Prostate cancer, Proteoglycans in cancer, Chemokine signaling pathway, PI3K-Akt signaling pathway, PD-L1 expression and PD-1 checkpoint pathway in cancer, EGFR tyrosine kinase inhibitor resistance signaling pathway and so on. Furthermore, molecular docking results suggest that hydrogen bonding, salt bridge and hydrophobic interactions contribute to spontaneous binding of the compound to the target protein. Experimental verification shows that rhubarb and aloe emodin affect the mechanism of pyroptosis in diabetic kidney disease by regulating STAT3/Caspase11 axis. In conclusion, this study comprehensively elaborated the active compounds, potential targets and molecular experimental mechanisms of rhubarb to provide the basic experimental theory for clinical treatment of DKD.

Fundamentals, Synthetic Strategies and Applications of Non-Covalently Imprinted Polymers.

Molecular imprinting has emerged as an important and practical technology to create economical and stable synthetic mimics of antibodies and enzymes. It has already found a variety of important applications, such as affinity separation, chemical/biological sensing, disease diagnostics, proteomics, bioimaging, controlled drug release, and catalysis. In the past decade, significant breakthroughs have been made in non-covalently imprinted polymers, from their synthesis through to their applications. In terms of synthesis, quite a few versatile and facile imprinting approaches for preparing MIPs have been invented, which have effectively solved some key issues in molecular imprinting. Additionally, important applications in several areas, such as sensors, proteomics and bioimaging, have been well demonstrated. In this review, we critically and comprehensively survey key recent advances made in the preparation of non-covalently imprinted polymers and their important applications. We focus on the state-of-art of this technology from three different perspectives: fundamentals, synthetic strategies, and applications. We first provide a fundamental basis for molecular imprinting technologies that have been developed, which is extremely helpful for establishing a sound understanding of the challenges in molecular imprinting. Then, we discuss in particular the major breakthroughs within the last ten years (2014-2024), with emphasis on new imprinting approaches, what strengths the breakthroughs can provide, and which new applications the properties of the prepared non-covalently imprinted polymers are fit for.

Protective effects and regulatory mechanisms of Platycodin D against LPS-Induced inflammatory injury in BEAS-2B cells

Platycodin D (PLD), a major bioactive component of triterpene saponins found in Platycodon grandiflora, is renowned for its anti-inflammatory and antioxidant properties. This study aims to explore the protective effects and regulatory mechanisms of PLD in an LPS-induced inflammation injury model of BEAS-2B cells. Initially, PLD was identified from Platycodon grandiflora extracts utilizing UPLC-Q-TOF-MS/MS technology. The effects of PLD on the viability, morphology, ROS levels, and inflammatory factors of LPS-induced BEAS-2B cells were then investigated. The results showed that PLD significantly alleviated LPS-induced oxidative stress and inflammatory injury. Further analysis revealed that PLD positively influenced apoptosis levels, mitochondrial morphology, and related gene expression, indicating its potential to mitigate LPS-induced apoptosis and alleviate mitochondrial dysfunction. Using molecular docking technology, we predicted the binding sites of PLD with mitochondrial autophagy protein. Gene expression levels of autophagy-related proteins were measured to determine the impact of PLD on mitochondrial autophagy. Additionally, the study examined whether the mitochondrial autophagy agonists rapamycin (RAPA) could modulate the upregulation of inflammasome-related factors NLRP3 and Caspase-1 in LPS-induced BEAS-2B cells. This was done to evaluate the regulator mechanisms of PLD in pulmonary inflammatory injury. Our findings suggest that PLD’s mechanism of action involves the regulation of mitochondrial autophagy, which in turn modulates inflammatory responses.

Computational simulation-assisted template selection of magnetic MOFs molecularly imprinted materials applying the adsorption and detection of multiple fluoroquinolones

This study utilized computational simulation and surface molecular imprinting technology to develop a magnetic metal-organic framework molecularly imprinted polymer (Fe3O4@ZIF-8@SMIP) capable of selectively recognizing and detecting multiple fluoroquinolones (FQs). The Fe3O4@ZIF-8@SMIP material was synthesized using the “common” template-ofloxacin, identified by computational simulation, demonstrating notable adsorption capacity (88.61–212.93 mg g−1) and rapid mass-transfer features (equilibration time: 2–3 min) for all tested FQs, consistent with Langmuir adsorption model. Subsequently, this material was employed as a magnetic solid-phase-extraction adsorbent for adsorption and detection of multiple FQs by combining with high performance liquid chromatography. The developed method exhibited good linearity for various FQs within the concentration range of 0.1–500 μg L−1, with low limit of detection (0.0605–0.1529 μg L−1) and limit of quantitation (0.2017–0.5097 μg L−1). Satisfactory recoveries (88.38–103.44%) were obtained when applied to spiked food samples, demonstrating the substantial potential of this Fe3O4@ZIF-8@SMIP material for rapid enrichment and identification for multiple FQs residues.

Integrative Transcriptomic and Single-Cell Protein Characterization of Colorectal Carcinoma Delineates Distinct Tumor Immune Microenvironments Associated with Overall Survival.

Colorectal carcinoma (CRC) is a heterogeneous group of tumors with varying therapeutic response and prognosis, and evidence suggests the tumor immune microenvironment (TIME) plays a pivotal role. Using advanced molecular and spatial biology technologies, we aimed to evaluate the TIME in patients with CRC to determine whether specific alterations in the immune composition correlated with prognosis. We identified primary and metastatic tumor samples from 31 consented patients, which were profiled with whole-exome sequencing and bulk RNA-seq. Immune cell deconvolution followed by gene set enrichment analysis and unsupervised clustering was performed. A subset of tumors underwent in situ analysis of the TIME spatial composition at single-cell resolution through Imaging Mass Mass Cytometry. Gene set enrichment analysis revealed two distinct groups of advanced CRC, one with an immune activated phenotype and the other with a suppressed immune microenvironment. The activated TIME phenotype contained increased Th1 cells, activated dendritic cells, tertiary lymphoid structures, and higher counts of CD8+ T cells whereas the inactive or suppressed TIME contained increased macrophages and a higher M2/M1 ratio. Our findings were further supported by RNA-seq data analysis from the TCGA CRC database, in which unsupervised clustering also identified two separate groups. The immunosuppressed CRC TIME had a lower overall survival probability (HR 1.66, p=0.007). This study supports the pertinent role of the CRC immune microenvironment in tumor progression and patient prognosis. We characterized the immune cell composition to better understand the complexity and vital role that immune activity states of the TIME play in determining patient outcome.

GhWRKY40 Interacts with an Asparaginase GhAPD6 Involved in Fiber Development in Upland Cotton (Gossypium hirsutum L.).

Fiber quality improvement is a primary goal in cotton breeding. Identification of fiber quality-related genes and understanding the underlying molecular mechanisms are essential prerequisites. Previously, studies determined that silencing the gene GhWRKY40 resulted in longer cotton fibers; however, both the underlying mechanisms and whether this transcription factor is additionally involved in the regulation of cotton fiber strength/fineness are unknown. In the current study, we verified that GhWRKY40 influences the fiber strength, fiber fineness, and fiber surface structure by using virus-induced gene silencing (VIGS). Potential proteins that may interact with the nucleus-localized GhWRKY40 were screened in a yeast two-hybrid (Y2H) nuclear-system cDNA library constructed from fibers at 0, 10, and 25 days post-anthesis (DPA) in two near-isogenic lines differing in fiber length and strength. An aspartyl protease/asparaginase-related protein, GhAPD6, was identified and confirmed by Y2H and split-luciferase complementation assays. The expression of GhAPD6 was approximately 30-fold higher in the GhWRKY40-VIGS lines at 10 DPA and aspartyl protease activity was significantly upregulated in the GhWRKY40-VIGS lines at 10-20 DPA. This study suggested that GhWRKY40 may interact with GhAPD6 to regulate fiber development in cotton. The results provide a theoretical reference for the selection and breeding of high-quality cotton fibers assisted by molecular technology.

Six-channel colorimetric sensing of metal ions and advanced molecular information security based on fish scale-derived carbon–gold–silver nanocomposites

Due to its high flexibility and expansibility, nanomaterials-based systems have been constructed and applied from sensing, and medicine to molecular information technology. However, the current challenges include tedious and harsh nanomaterials preparation, the need for a wider range of response channels, new information paradigms, higher information density and length. Herein, fish-scale derived carbon nanomaterials (FSCN) were used as reducing and stabilizing agents for direct, green, and facile synthesis of carbon-bimetallic nanocomposites and applications in six-channel colorimetric sensing of Hg2+, and multi-combination and multi-round information encoding and protection for long text. First, the reaction sequence of three precursors (FSCN, Au3+, Ag+) was optimized to prepare FSCN–Au–Ag nanocomposites with double plasmonic absorption peaks of Au/Ag nanoparticles (NPs). The addition of Hg2+ causes the aggregation of nanoparticles and the formation of Ag/Hg amalgam or Au/Hg amalgam, FSCN–Au–Ag NPs were capable of multi-channel sensing (color, absorbance, and peak shift) of Hg2+. Especially, the detection limit of Hg2+ was 8.9 nM based on absorption response in wide wavelength range (456–546 nm), which was at least 6 times more sensitive than that at single wavelength. Moreover, different long text information can be encrypted and hidden in six-channel selective responses of FSCN–Au–Ag NPs-based colorimetric sensing system. By setting the threshold, these selective responses can be abstracted as a series of binary strings and then divided into shorter strings. Utilizing multi-combination and multi-round information coding, these short binary strings can be decrypted and merged into various long text content. This research promotes preparation and multi-purpose application of carbon-bimetallic nanomaterials, which provides new way to improve the sensing sensitivity of nanosystems and also offers new direction for molecular digitization.

A review on prospect of wheat in Nigeria: Technological Advancement in Focus

The main issue affecting domestic wheat production in Nigeria is environmental incompatibility— the fact that wheat, being a temperate crop, requires cold climates during the growing season to thrive especially during the critical developmental stage between tillering and grain filling. This has limited the wheat growing season in Nigeria, to the cool harmattan period starting from November to March, and the wheat growing areas in Nigeria, along latitude 10°N to 14°N. Unfortunately, the harmattan period itself is also challenged by the absence of a rainy season, which typically starts after the period from April to September with unfavourable high temperature and humidity. Nigerian government policies over the years have centred on accelerating primary production (farming) instead of focusing on addressing the main issues, through research and development of cultivars that will be adaptable in Nigeria’s heat-stressed growing environment, build more irrigation areas along the harmattan latitude, strengthen extension services to ensure farmers adherence to Good Agronomic Practices (GAPs) and ease farmers access to market for fair prices. As a result, the national average ton per hectare output is abysmally 1.3 MT, prepared irrigation area is less than 10% of planned area and a near-complete disconnect between farmers and the off-taking flour milling industry, who are currently fully reliant on imported wheat. The peculiar situation is like in other tropical countries like Egypt, Ethiopia and the warm and humid Bangladesh. The paper recommended the adoption of molecular breeding technologies to strengthen the breeding processes and shorten the seeds generation time.

Discovery of Potent Selective HDAC6 Inhibitors with 5-Phenyl-1H-indole Fragment: Virtual Screening, Rational Design, and Biological Evaluation.

Among the HDACs family, histone deacetylase 6 (HDAC6) has attracted extensive attention due to its unique structure and biological functions. Numerous studies have shown that compared with broad-spectrum HDACs inhibitors, selective HDAC6 inhibitors exert ideal efficacy in tumor treatment with insignificant toxic and side effects, demonstrating promising clinical application prospect. Herein, we carried out rational drug design by integrating a deep learning model, molecular docking, and molecular dynamics simulation technology to construct a virtual screening process. The designed derivatives with 5-phenyl-1H-indole fragment as Cap showed desirable cytotoxicity to the various tumor cell lines, all of which were within 15 μM (ranging from 0.35 to 14.87 μM), among which compound 5i had the best antiproliferative activities against HL-60 (IC50 = 0.35 ± 0.07 μM) and arrested HL-60 cells in the G0/G1 phase. In addition, 5i exhibited better isotype selective inhibitory activities due to the potent potency against HDAC6 (IC50 = 5.16 ± 0.25 nM) and the reduced inhibitory activities against HDAC1 (selective index ≈ 124), which was further verified by immunoblotting results. Moreover, the representative binding conformation of 5i on HDAC6 was revealed and the key residues contributing 5i's binding were also identified via decomposition free-energy analysis. The discovery of lead compound 5i also indicates that virtual screening is still a beneficial tool in drug discovery and can provide more molecular skeletons with research potential for drug design, which is worthy of widespread application.

Study On the Mechanism of Action of Qili Qiangxin Capsule in the Treatment of Heart Failure Based on Network Pharmacology and Molecular Docking Method

Objective: To investigate the pharmacodynamic substances and mechanism of action of Qili Qiangxin Capsule in the treatment of heart failure based on network pharmacology and molecular docking technology. Methods: The active ingredients and drug targets of Qili Qiangxin Capsule were obtained from databases such as TCMSP, GeneCards, OMIM, PharmGkb, TTD, and DrugBank databases were searched for heart failure disease targets. The drug targets and disease targets were corrected by the UniProt database, then the Venn diagram was drawn, and the intersecting genes were screened. Cytoscape 3.8.0 software was used to draw the active ingredient-disease target network for topological analysis. PPI network was constructed in the STRING database platform to predict the core targets. Bioconductor package was used to perform KEGG pathway analysis. autoDock Vina software was used to molecularly dock the core targets with the main active ingredients, and Pymol software visualised the results. Results: Screening of 209 active ingredients of Qili Qiangxin Capsule, including quercetin, luteolin, cryptotanshinone, etc. 11,432 heart failure disease targets, 249 genes intersecting with drugs, including the core targets containing TP53, STAT3, JUN, MAPK1, etc. These genes are mainly involved in the AGE-RAGE, fluid shear stress, and atherosclerosis, PI3K-Akt signaling pathways. Conclusion: This study initially revealed the multi-component, multi-target, multi-pathway mechanism of action of Qili Qiangxin Capsule in the treatment of heart failure, which provides a theoretical basis for further research.