Secondary Pathways Research Articles

Comparative analysis of the molecular response characteristics in Platycodon grandiflorus irradiated with heavy ion beams and X-rays

The response of plants to radiation is an essential topic in both space plant cultivation and mutation breeding by radiation. In this study, heavy ion beams (HIB) generated by the ground accelerator and X-rays (XR) were used as models of high linear energy transfer (LET) and low LET radiation to study the molecular response mechanism of Platycodon grandiflorus (P. grandiflorus) seedlings after irradiation. The gene and protein expression profiles of P. grandiflorus after 15 Gy HIB and 20 Gy XR radiation were analyzed by transcriptome and proteome. The results showed that the number of differentially expressed genes (DEGs) induced by HIB radiation was less than that of XR group, but HIB radiation induced more differentially expressed proteins (DEPs). Both HIB and XR radiation activated genes of RNA silencing, double-strand break repair and cell catabolic process. DNA replication and cell cycle related genes were down-regulated. The genes of cell wall and external encapsulating structure were up-regulated after HIB radiation. The gene expression of protein folding and glucan biosynthesis increased after XR radiation. Protein enrichment analysis indicated that HIB radiation resulted in differential protein enriched in photosynthesis and secondary metabolite biosynthesis pathways, while XR radiation induced differential protein of glyoxylate and dicarboxylate metabolism and carbon metabolism. After HIB and XR radiation, the genes of antioxidant system and terpenoid and polyketide metabolic pathways presented different expression patterns. HIB radiation led to the enrichment of non-homologous end-joining pathway. The results will contribute to understanding the biological effects of plants under space radiation.

Transcriptome analysis of flower colour reveals the correlation between SNP and differential expression genes in Phalaenopsis.

Phalaenopsis is an important ornamental plant that has great economic value in the world flower market as one of the most popular flower resources. To investigate the flower colour formation of Phalaenopsis at the transcription level, the genes involved in flower color formation were identified from RNA-seq in this study. In this study, white and purple petals of Phalaenopsis were collected and analyzed to obtained (1) differential expression genes (DEGs) between white and purple flower color and (2) the association between single nucleotide polymorphisms (SNP) mutations and DEGs at the transcriptome level. The results indicated that a total of 1,175 DEGs were identified, and 718 and 457 of them were up- and down-regulated genes, respectively. Gene Ontology and pathway enrichment showed that the biosynthesis of the secondary metabolites pathway was key to color formation, and the expression of 12 crucial genes (C4H, CCoAOMT, F3'H, UA3'5'GT, PAL, 4CL, CCR, CAD, CALDH, bglx, SGTase, and E1.11.17) that are involved in the regulation of flower color in Phalaenopsis. This study reported the association between the SNP mutations and DEGs for color formation at RNA level, and provides a new insight to further investigate the gene expression and its relationship with genetic variants from RNA-seq data in other species.

Amorphophallus muelleri activates ferulic acid and phenylpropane biosynthesis pathways to defend against Fusarium solani infection.

Amorphophallus sp. is an economically important crop for rural revitalization in southwest China. However, Fusarium solani often infects Amorphophallus sp. corms during storage, damaging the corm quality and affecting leaf elongation and flowering in the subsequent crop. In this study, the mechanism of resistance to F. solani was investigated in the leaf bud and flower bud corms of Amorphophallus muelleri through transcriptome and metabolome analyses. A total of 42.52 Gb clean reads and 1,525 metabolites were detected in a total of 12 samples including 3 samples each of disease-free leaf bud corms (LC), leaf bud corms inoculated with F. solani for three days (LD), disease-free flower bud corms (FC), and flower bud corms inoculated with F. solani for three days (FD). Transcriptome, metabolome, and conjoint analyses showed that 'MAPK signal transduction', 'plant-pathogen interaction', 'plant hormone signal transduction', and other secondary metabolite biosynthesis pathways, including 'phenylpropane biosynthesis', 'arachidonic acid metabolism', 'stilbene, diarylheptane and gingerolin biosynthesis', and 'isoquinoline alkaloids biosynthesis', among others, were involved in the defense response of A. muelleri to F. solani. Ultimately, the expression of six genes of interest (AmCDPK20, AmRBOH, AmWRKY33, Am4CL, Am POD and AmCYP73A1) was validated by real-time fluorescence quantitative polymerase chain reaction, and the results indicated that these genes were involved in the response of A. muelleri to F. solani. Ferulic acid inhibited the growth of F. solani, reducing the harm caused by F. solani to A. muelleri corms to a certain extent. Overall, this study lays a strong foundation for further investigation of the interaction between A. muelleri and F. solani, and provides a list of genes for the future breeding of F. solani-resistant A. muelleri cultivars.

Metabolomics analysis of mycelial exudates provides insights into fungal antagonists of Armillaria

ABSTRACT The genus Armillaria has high edible and medical values, with zones of antagonism often occurring when different species are paired in culture on agar media, while the antagonism-induced metabolic alteration remains unclear. Here, the metabolome of mycelial exudates of two Chinese Armillaria biological species, C and G, co-cultured or cultured separately was analysed to discover the candidate biomarkers and the key metabolic pathways involved in Armillaria antagonists. A total of 2,377 metabolites were identified, mainly organic acids and derivatives, lipids and lipid-like molecules, and organoheterocyclic compounds. There were 248 and 142 differentially expressed metabolites between group C-G and C, C-G, and G, respectively, and fourteen common differentially expressed metabolites including malate, uracil, Leu-Gln-Arg, etc. Metabolic pathways like TCA cycle and pyrimidine metabolism were significantly affected by C-G co-culture. Additionally, 156 new metabolites (largely organic acids and derivatives) including 32 potential antifungal compounds, primarily enriched into biosynthesis of secondary metabolites pathways were identified in C-G co-culture mode. We concluded that malate and uracil could be used as the candidate biomarkers, and TCA cycle and pyrimidine metabolism were the key metabolic pathways involved in Armillaria antagonists. The metabolic changes revealed in this study provide insights into the mechanisms underlying fungal antagonists.

Metabolic profiling and expression analysis of key genetic factors in the biosynthetic pathways of antioxidant metabolites in mungbean sprouts.

Mungbeans (Vigna radiata L.), a major legume crop in Asia, contain higher amounts of functional substances than other legumes, such as catechin, chlorogenic acid, and vitexin. Germination can improve the nutritional value of legume seeds. Here, 20 functional substances were profiled in germinated mungbeans and the expression levels of the transcripts of key enzymes in targeted secondary metabolite biosynthetic pathways were identified. VC1973A, a reference mungbean elite cultivar, had the highest amount of gallic acid (99.93 ± 0.13 mg/100 g DW) but showed lower contents of most metabolites than the other genotypes. Wild mungbeans contained a large amount of isoflavones compared with cultivated genotypes, especially for daidzin, genistin and glycitin. The expression of key genes involved in biosynthetic pathways had significant positive or negative correlations with the target secondary metabolite contents. The results indicate that functional substance contents are regulated at the transcriptional level, which can be applied to improve the nutritional value of mungbean sprouts in molecular breeding or genetic engineering, and wild mungbeans are a useful resource to improve the quality of mungbean sprouts.

Exploring changing attitudes to non-invasive liver fibrosis tests in secondary care pathways: comparison of two national surveys

IntroductionThe increasing availability of non-invasive tests (NITs) has created the opportunity to explore their use in improving risk stratification of advanced liver disease. The study aimed to determine the attitudes...

Pandemic Job Separation and Psychological Distress: Modeling Chains of Adversity

Although recent studies have linked pandemic unemployment with poorer mental health, the mechanisms underlying this association remain understudied. In this paper, we develop a mediation model to explain why pandemic job separation might undermine mental health. Using national data from the 2021 Crime, Health, and Politics Survey (n = 1,258), we test the indirect effects of pandemic job separation on psychological distress through several mechanisms. Mediation analyses reveal compound indirect effects of pandemic job separation on psychological distress through the primary pathway of financial strain and the secondary pathways of social support, self-esteem, mastery, religious struggles, and sleep disturbance. Absent the indirect effect of pandemic job separation through financial strain, we would have failed to observe any simple indirect effects through the other proposed mechanisms. Formal moderated mediation analyses also indicate that our observed indirect effects are invariant to subgroup differences in current employment status, education, and household income. In short, our indirect effects are observed for those respondents who were able to regain employment, those with college degrees, and those with the most financial resources. Our results suggest that the temporary expansion of public assistance has been insufficient to offset widespread unemployment and financial hardship during a global pandemic.

SAT-130 - Exploring attitudes towards non-invasive liver fibrosis tests in secondary care pathways: comparing national surveys between 2014 and 2021

SAT-130 - Exploring attitudes towards non-invasive liver fibrosis tests in secondary care pathways: comparing national surveys between 2014 and 2021

Browning mechanism of tobacco leaves during flue-curing process: Proteomics and metabolomics analysis reveals the changes in materials

Tobacco leaves, as raw materials, undergo complex processing before the final product is obtained. During the flue-curing process, inadequate control of environmental factors such as temperature, humidity or water loss rate, may lead to enzymatic browning occur caused by starvation metabolism of tobacco leaves, and therefore large amount of internal chemical substances consumed and changed, leading to excessive darker color and affecting the quality of products. This study aimed to investigate the mechanisms and material basis of browning, as well as the relationship between browning and internal substances changes. The microstructural changes of tobacco leaves under transmission electron microscopy were observed, quantitative proteomic and untargeted metabonomic methods were conducted in browning and normal tobacco leaves. In the results, 106 differential proteins and 576 differential metabolites were identified, with differential proteins mainly involved in biosynthesis and metabolism of polysaccharides, lipid, as well as the regulation of chlorophyll degradation and polyphenol oxidation related proteins. Differential metabolites were mainly enriched in purine metabolism, histidine metabolism and isoflavone biosynthesis pathway, with more secondary metabolites biosynthesis pathway enrichment. Moreover, tyrosine metabolism and phenylpropane biosynthesis metabolism were identified as the mainly differential proteins and metabolites pathways. Additionally, two metabolites, resveratrol and iridin, that exhibited strong free radicals scavenging activity and good antioxidant properties were uncovered. Taken together, these findings demonstrated changes of chemical substances and differences of proteins and metabolites between browning and normal tobacco leaves under specific conditions, and provided a theoretical reference for exploring optimized conditions and controlling material changes during the processing of tobacco to inhibit browning.

Transcriptomic and metabolic studies on the role of inorganic and organic iodine compounds in lettuce plants

Iodine (I) is considered a beneficial element or even micronutrient for plants. The aim of this study was to determine the molecular and physiological processes of uptake, transport, and metabolism of I applied to lettuce plants. KIO3, KIO3 + salicylic acid, 5-iodosalicylic acid and 3,5-diiodosalicylic acid were applied. RNA-sequencing was executed using 18 cDNA libraries constructed separately for leaves and roots from KIO3, SA and control plants. De novo transcriptome assembly generated 1937.76 million sequence reads resulting in 27,163 transcripts with N50 of 1638 bp. 329 differentially expressed genes (DEGs) in roots were detected after application of KIO3, out of which 252 genes were up-regulated, and 77 were down-regulated. In leaves, 9 genes revealed differential expression pattern. DEGs analysis indicated its involvement in such metabolic pathways and processes as: chloride transmembrane transport, phenylpropanoid metabolism, positive regulation of defense response and leaf abscission, and also ubiquinone and other terpenoid-quinone biosynthesis, protein processing in endoplasmic reticulum, circadian rhythm including flowering induction as well as a putative PDTHA (i.e. Plant Derived Thyroid Hormone Analogs) metabolic pathway. qRT-PCR of selected genes suggested their participation in the transport and metabolism of iodine compounds, biosynthesis of primary and secondary metabolites, PDTHA pathway and flowering induction.

Inhibition of nitrogen assimilation promotes carbon-based secondary metabolism in callus of Cyclocarya paliurus

The biosynthesis and accumulation of secondary metabolites are critical important to quality formation of medicinal plants, which are usually give way to primary processes and growth. Here, methionine sulfoximine (MSO) was used to inhibit the nitrogen assimilation in callus of Cyclocarya paliurus. The newly assimilated nitrogen characterized by 15N atom percentage excess, and the levels of amino acid and protein were reduced. The other primary processes such as carbohydrate metabolism and lipid metabolism were also repressed. In addition, the expression of the growth-related target of rapamycin (TOR) signaling was repressed, indicating nitrogen assimilation inhibition led to a systematic down-regulated primary metabolisms and resulted in a disruption of growth. In contrast, the biosynthesis of flavonoids and triterpenoids, antioxidase system, and the SnRK2-mediated abscisic acid (ABA) and jasmonic acid (JA) signaling were induced, which can improve plant stress resistance and defense. Nitrogen assimilation inhibition led to the carbon metabolic flux redirection from primary processes to secondary pathways, and facilitated the biosynthesis of flavonoids and triterpenoids in calluses of C. paliurus. Our results provide a comprehensive understanding of metabolic flux redirection between primary and secondary metabolic pathways and a potential means to improve the quality of medicinal plants.

Biosynthetic Potential of Hypogymnia Holobionts: Insights into Secondary Metabolite Pathways.

Lichens are symbiotic associations consisting of a photobiont (algae or cyanobacteria) and a mycobiont (fungus). They are known to produce a variety of unique secondary metabolites. To access this biosynthetic potential for biotechnological applications, deeper insights into the biosynthetic pathways and corresponding gene clusters are necessary. Here we provide a comprehensive view of the biosynthetic gene clusters of all organisms comprising a lichen thallus: fungi, green algae, and bacteria. We present two high-quality PacBio metagenomes, in which we identified a total of 460 biosynthetic gene clusters. Lichen mycobionts yielded 73-114 clusters, other lichen associated ascomycetes 8-40, green algae of the genus Trebouxia 14-19, and lichen-associated bacteria 101-105 clusters. The mycobionts contained mainly T1PKSs, followed by NRPSs, and terpenes; Trebouxia reads harbored mainly clusters linked to terpenes, followed by NRPSs and T3PKSs. Other lichen-associated ascomycetes and bacteria contained a mix of diverse biosynthetic gene clusters. In this study, we identified for the first time the biosynthetic gene clusters of entire lichen holobionts. The yet untapped biosynthetic potential of two species of the genus Hypogymnia is made accessible for further research.

Comparative Transcriptome and Widely Targeted Metabolome Analysis Reveals the Molecular Mechanism of Powdery Mildew Resistance in Tomato.

Powdery mildew is a serious problem in tomato production; therefore, the PM-resistant tomato inbred line, '63187', and the susceptible tomato variety, 'Moneymaker (MM)', were used as experimental materials for the combined analysis of transcriptome and widely targeted metabolome on tomato leaves at 0 h post inoculation (hpi), 12 hpi, and 48 hpi. The results indicated that 276 genes were expressed in all treatments, and the K-means cluster analysis showed that these genes were divided into eight classes in '63187' and ten classes in 'MM'. KEGG enrichment showed that amino acid metabolism, signal transduction, energy metabolism, and other secondary metabolites biosynthesis pathways were significantly enriched. Interestingly, the analysis of WRKY family transcription factors (TFs) showed that the expression of four TFs in '63187' increased with no obvious change in 'MM'; and the expression of one TF in 'MM' increased with no obvious change in '63187'. The combined analysis revealed that both phenylpropanoid biosynthesis and flavonoid biosynthesis pathways were enriched in '63187' and 'MM'. In '63187', six metabolites involved in this pathway were downregulated, and four genes were highly expressed, while in 'MM', three metabolites were upregulated, four metabolites were downregulated, and ten genes were highly expressed. These metabolites and genes might be candidates for PM resistance or susceptibility in subsequent studies. These results provide favorable molecular information for the study of the different resistances of tomatoes to PM, and they provide a basis for the breeding of tomato varieties resistant to PM.

Advanced practice physiotherapists in Scottish primary care: Axial Spondyloarthropathy epidemiology, time to diagnosis, and referrals to rheumatology.

(1) Generate empirical knowledge of a Musculoskeletal (MSK) Advanced Practice Physiotherapist (APP) Service in Scottish Primary Care; (2) Identify the incidence and baseline time to diagnosis of Axial Spondyloarthropathy (AxSpA); (3) Identify APP Rheumatology referral fulfilment of the NICE 2017 Guidelines and Spondylarthritis Diagnosis Evaluation (SPADE) Tool; (4) Calculate APP Rheumatology referral conversion rates for AxSpA diagnosis and further investigation; (5) Contribute towards the current body of literature for informing analysis of MSK APP services within Scottish Primary Care. An audit and evaluation approach was undertaken over a 3-year period (May 2019-April 2022). Relevant clinical cases from the whole-service data-set were identified and analysed, using retrospective electronic healthcare record review and descriptive statistical techniques. A total of 37,656 primary care MSK APP consultations took place, with N=19 suspected AxSpA referrals made to Rheumatology. N=6 cases of AxSpA were diagnosed by a Rheumatologist (31.6%). The mean age of individuals diagnosed with AxSpA was 39.6±8.8, and 66.7% (4/6) were female. Mean time to diagnosis was 3.4years, and incidence per-10,000 person-years was 1.6. Compliance of referrals with the NICE 2017 Guidelines and SPADE Tool Criteria was 78.9%. Of those diagnosed with AxSpA, 66.7% met both referral criterion sets. Those referred by an MSK APP from primary care had a 5.1year shorter time to diagnosis than the previous reported UK average of 8.5years. APPs identified relevant AxSpA features in referring to Rheumatology, and supported effective implementation of the local secondary care pathway.

Transcriptome analysis reveals upregulated secondary metabolite pathways in micropropagated Lawsonia inermis L.

Transcriptome analysis reveals upregulated secondary metabolite pathways in micropropagated Lawsonia inermis L.

Transcriptome profiling, physiological, and biochemical analyses provide new insights towards drought stress response in sugar maple (Acer saccharum Marshall) saplings.

Sugar maple (Acer saccharum Marshall) is a temperate tree species in the northeastern parts of the United States and is economically important for its hardwood and syrup production. Sugar maple trees are highly vulnerable to changing climatic conditions, especially drought, so understanding the physiological, biochemical, and molecular responses is critical. The sugar maple saplings were subjected to drought stress for 7, 14, and 21 days and physiological data collected at 7, 14, and 21 days after stress (DAS) showed significantly reduced chlorophyll and Normalized Difference Vegetation Index with increasing drought stress time. The drought stress-induced biochemical changes revealed a higher accumulation of malondialdehyde, proline, and peroxidase activity in response to drought stress. Transcriptome analysis identified a total of 14,099 differentially expressed genes (DEGs); 328 were common among all stress periods. Among the DEGs, transcription factors (including NAC, HSF, ZFPs, GRFs, and ERF), chloroplast-related and stress-responsive genes such as peroxidases, membrane transporters, kinases, and protein detoxifiers were predominant. GO enrichment and KEGG pathway analysis revealed significantly enriched processes related to protein phosphorylation, transmembrane transport, nucleic acids, and metabolic, secondary metabolite biosynthesis pathways, circadian rhythm-plant, and carotenoid biosynthesis in response to drought stress. Time-series transcriptomic analysis revealed changes in gene regulation patterns in eight different clusters, and pathway analysis by individual clusters revealed a hub of stress-responsive pathways. In addition, qRT-PCR validation of selected DEGs revealed that the expression patterns were consistent with transcriptome analysis. The results from this study provide insights into the dynamics of physiological, biochemical, and gene responses to progressive drought stress and reveal the important stress-adaptive mechanisms of sugar maple saplings in response to drought stress.

Molecular cloning and functional characterization of BcTSA in the biosynthesis of indole alkaloids in Baphicacanthus cusia.

Baphicacanthus cusia (Nees) Bremek (B. cusia) is an essential traditional Chinese herb that is commonly used to treat colds, fever, and influenza. Indole alkaloids, such as indigo and indirubin, are the primary active constituents of B. cusia. The indole-producing reaction is crucial for regulating the flow of indole alkaloids metabolites along the pathways and coordinating primary and secondary product biosynthesis in plants. The tryptophan synthase alpha-subunit (TSA) can catalyse a process that produces indole, which is free to enter secondary metabolite pathways; however, the underlying potential mechanism of regulating indigo alkaloids synthesis remains unknown. Here, a BcTSA was cloned from the transcriptome of B. cusia. The BcTSA has a significant degree of similarity with other plant TSAs according to bioinformatics and phylogenetic analyses. Quantitative real-time PCR (RT-qPCR) research showed that BcTSA was dramatically enhanced in response to treatment with methyl jasmonate (MeJA), salicylic acid (SA), and abscisic acid (ABA), and was predominantly expressed in the stems as opposed to the leaves and rhizomes. Subcellular localization revealed that BcTSA is localized in chloroplasts, which is compatible with the fact that the conversion of indole-3-glycerol phosphate (IGP) to indole occurs in chloroplasts. The complementation assay results showed that BcTSA was functional, demonstrating that it was capable of catalyzing the conversion of IGP to indole. BcTSA was shown to stimulate the manufacture of indigo alkaloids including isatin, indigo, and indirubin when the gene was overexpressed in the hairy roots of Isatis indigotica. In conclusion, our research provides novel perspectives that might be applied to manipulating the indole alkaloid composition of B. cusia.

Effect of Six Eye Exercises Protocol on Oculomotor Control, Anxiety and Emotional Intelligence in Hypertensive Subjects - A Randomized Clinical Trial

Hypertension has a larger impact on the various systems of the body along with poor quality of life. After a span of almost one year, alteration in the oculomotor movements can be seen which is a lesser known fact . Research suggests that there is correlation of between oculomotor movements which is going to cause anxiety prevailing emotional distress. This association is linked with elevation of BP , hampering daily activities as well as work calibre. To address all these issues, currently there are various interventions for hypertensive patients at surgical, medical and advanced level but there is a lack of study on the community level. This highlights the need for a patient friendly approach which will focus on these specific concerns .the need of this study is to create a community friendly intervention for hypertensive patients whose oculomotor movements are affected due to which anxiety and emotional intelligence may get impaired. This study will give a secondary pathway to manage all these symptoms in a very convenient way . Six Eye Exercise protocol being a completely patient friendly protocol will not only allow the patient to perform it with ease but will also create a community friendly approach of treating the above issues.

Abstract 4872: DCC-3116, a first-in-class selective ULK1/2 inhibitor of autophagy, in combination with the KIT inhibitor ripretinib induces complete regressions in GIST preclinical models

Abstract Background: Cancer cells activate autophagy as an adaptive stress response (ASR) mechanism to therapies targeting the RTK/RAS/MAPK/PI3K pathways, limiting antitumor response. Autophagy is initiated through ULK1/2 kinases and is triggered by inhibitors of the MAPK and PI3K pathways. Most gastrointestinal stromal tumors (GIST) are driven by mutations in KIT kinase. KIT signals through MAPK/PI3K pathways, suppressing ULK1/2 kinases and autophagy1,2,3. Inhibition of mutant KIT reverses this suppression, activating autophagy and cancer cell survival. Approved therapies for GIST include imatinib, sunitinib, regorafenib, ripretinib, and avapritinib. Treatment with these inhibitors is initially successful, but drug resistance can develop either through KIT secondary mutations or ASR pathways including autophagy. DCC-3116 is a selective, potent, first-in-class investigational inhibitor of the ULK1/2 in clinical development in combination with targeted therapies that activate the autophagic ASR pathway. Herein we demonstrate that ULK1/2 and autophagy are activated upon treatment with ripretinib in KIT mutant GIST models A combination of ripretinib with DCC-3116 inhibits autophagy in vitro and leads to complete tumor regressions in preclinical models of GIST. Methods: Inhibition of ULK1/2 in cell assays was measured using an ELISA for the ULK substrate phospho-ATG13 (pATG13). Autophagic flux was measured by monitoring mCherry/GFP tagged LC3 protein in GIST cells. Xenograft studies were performed at CROs. Results: Ripretinib treatment led to the activation of ULK1/2 by 2-3-fold in mutant KIT GIST cell lines. DCC-3116 inhibited both ripretinib-induced and basal pATG13 with IC50 values of 12-32 nM. Treatment of GIST-T1 cells with ripretinib increased autophagic flux 3-fold. DCC-3116 potently inhibited flux with an IC50 value of 38 nM. Ripretinib also induced pATG13 and autophagic flux (1.5-2.5-fold) in multiple imatinib-resistant cell lines, which was inhibited by DCC-3116 with IC50 values between 8-189 nM. In a GIST T1 PK/PD model, DCC-3116 inhibited ULK1/2-mediated pATG13. The combination of DCC-3116 with ripretinib resulted in complete tumor regressions in comparison to single agent treatment in GIST preclinical models. Conclusions: These data demonstrate preclinically that, like other receptor tyrosine kinase inhibitors1, ripretinib activates ULK1/2-mediated autophagy as an ASR resistance mechanism which is inhibited by DCC-3116, providing the rationale to study the combination of DCC-3116 with ripretinib in GIST patients. DCC-3116 is currently in a Phase 1 clinical trial in patients with advanced solid tumors (NCT04892017).

Mechanistic study of Coriandrum sativum on neuritogenesis and synaptogenesis based on computationally guided in vitro analyses.

Acceleration of neurite outgrowth and halting neurodegeneration are the most critical factors that are negatively regulated in various neurodegenerative diseases or injuries in the central nervous system (CNS). Functional foods or nutrients are considered alternative sources of bioactive components to alleviate various CNS injuries by promoting neuritogenesis and synaptogenesis, while their exact molecular mechanism remains unexplored. Coriandrum sativum L. (CS) is one of the popular herbs in the Apiaceae family, of which CNS modulating action is a well-documented traditionally but detailed study on memory boosting function yet remains unexplored. Consequently, this study aims to analyze the neurogenic and synaptogenic modulation of CS aqueous ethanol (CSAE) extract in the primary hippocampal neurons. Primary hippocampal neurons were cultured and allowed to incubate with CSAE or vehicle. To observe the early neuronal differentiation, axonal and dendritic arborization, and synapse formation, neurons were immune-stained against indicated antibody or stained directly with a lipophilic dye (1, 1'-dioctadecyl-3, 3, 3', 3'-tetramethyl indocarbocyanine perchlorate, DiL). Meanwhile, Western blot was used to validate the synaptogenesis effect of CSAE compared to vehicle. Additionally, molecular docking and system pharmacology approach were applied to confirm the possible secondary metabolites and pathways in which CSAE play a role in neuritogenesis. Results show that CSAE can induce neuritogenesis and synaptogenesis at 30 μg/mL concentrations. The treatment impacts early neuronal polarization, axonal and dendritic arborization, synaptogenesis, and synaptic plasticity via NMDARs expressions in primary neurons. In silico network pharmacology of CS metabolites show that the CSAE-mediated neurogenic effect is likely dependent on the NTRK2 (TrkB) mediated neurotrophin signaling pathway. Indeed, the observed neurogenic activity of CSAE is markedly reduced upon the co-treatment with a TrkB-specific inhibitor. Furthermore, molecular docking following binding energy calculation shows that one of the CS metabolites, scoparone, has a high affinity to bind in the BDNF mimetic site of TrkB, suggesting its role in TrkB activation. Scoparone was found to enhance neuritogenesis, but not to the same extent as CSAE. Moreover, the expression of TrkB signaling-related proteins (BCL2, CASP3, GSK3, and BDNF), which was found to be modulated by scoparone, was significantly affected by the co-treatment of TrkB inhibitor (ANA-12). These results further suggest that the modulation of neuritogenesis by scoparone is TrkB-dependent. This study provides deeper insights into the molecular mechanism of CS in boosting neuronal growth and memory function, which might implicate the prevention of many neurological disorders.