Expression Of Homolog Research Articles

Nucleoporin PNET1 coordinates mitotic nuclear pore complex dynamics for rapid cell division.

The nuclear pore complex (NPC) is a cornerstone of eukaryotic cell functionality, orchestrating the nucleocytoplasmic shuttling of macromolecules. Here we report that Plant Nuclear Envelope Transmembrane 1 (PNET1), a transmembrane nucleoporin, is an adaptable NPC component that is mainly expressed in actively dividing cells. PNET1's selective incorporation into the NPC is required for rapid cell growth in highly proliferative meristem and callus tissues in Arabidopsis. We demonstrate that the cell cycle-dependent phosphorylation of PNET1 coordinates mitotic disassembly and post-mitotic reassembly of NPCs during the cell cycle. PNET1 hyperphosphorylation disrupts its interaction with the NPC scaffold, facilitating NPC dismantling and nuclear membrane breakdown to trigger mitosis. In contrast, nascent, unphosphorylated PNET1 is incorporated into the nuclear pore membrane in the daughter cells, where it restores interactions with scaffolding nucleoporins for NPC reassembly. The expression of the human PNET1 homologue is required for and markedly upregulated during cancer cell growth, suggesting that PNET1 plays a conserved role in facilitating rapid cell division during open mitosis in highly proliferative tissues.

Feedback loop centered on MAF1 reduces blood–brain barrier damage in sepsis-associated encephalopathy

BackgroundA previous study found that MAF1 homolog, a negative regulator of RNA polymerase III (MAF1), protects the blood–brain barrier (BBB) in sepsis-associated encephalopathy (SAE); however, the related molecular mechanisms remain unclear.Subjects and methodsIn this study, a rat sepsis model was constructed using the cecum ligation and puncture (CLP) method. In vitro, rat brain microvascular endothelial cells and astrocytes were stimulated with serum from the sepsis model rats. The loss of MAF1 protein levels and the molecular mechanisms leading to cell damage were investigated.ResultsIt was shown in the SAE models that MAF1 was expressed at low levels. Knockdown of Cullin 2 (CUL2) stimulated the accumulation of MAF1 protein, attenuated the RNA sensor RIG-I/interferon regulatory factor 3 (IRF3) signaling pathway, and reduced cell apoptosis. Furthermore, it increased phosphatase and tensin homolog (PTEN) expression and inactivated the serine/threonine kinase (AKT)/mechanistic target of the rapamycin kinase (mTOR) signaling pathway. Interference with forkhead box O1 (FOXO1) inhibited MAF1 expression and activated the RIG-I/IRF3 signaling pathway, while MAF1 overexpression promoted PTEN expression, decreased cell apoptosis, and normalized autophagy.ConclusionsThese findings demonstrate that CUL2 promoted MAF1 ubiquitination and caused BBB injury in SAE. Through the regulatory loop of PTEN/AKT/FOXO1/MAF1, CUL2 initiated the gradual downregulation of MAF1, which subsequently regulated polymerase III (Pol III)-dependent transcription and played essential roles in cell apoptosis in SAE.Clinical trial number: not applicable.Graphical

Calcium Chloride Enhances Defense Response Against Diaporthe nobilis by Eliciting Reactive Oxygen Species in Kiwifruit

Kiwifruit (Actinidia spp.) rot, caused by pathogenic fungi, represents the most severe disease during postharvest storage. The utilization of induced resistance against fungal pathogens in harvested fruit has been demonstrated as a promising strategy for controlling postharvest losses. In this study, the effect of calcium chloride (CaCl2) on induction of resistance to Diaporthe nobilis, one of the most epidemic pathogens of soft rot in kiwifruit, and the underlying mechanisms were investigated. Our data showed that the application of 3% CaCl2 resulted in smaller rotten lesion diameters after inoculation with the pathogen, induced expression of the respiratory burst oxidase homolog (RBOH) gene family, and increased accumulation of reactive oxygen species (ROS). Moreover, we found that the activity of three pathogenesis‐related antioxidant enzymes—peroxidase (POD), catalase (CAT), and superoxide dismutase (SOD)—as well as the expression of important defense‐related genes, was enhanced by CaCl2 treatment. The results of our study suggest that the application of CaCl2 treatment holds promise as a green and viable method for effectively preventing and safeguarding kiwifruit against postharvest decay.

Comprehensive evaluation of genomic and functional assays for homologous recombination deficiency with high-grade epithelial ovarian cancer: Platinum sensitivityand prognosis.

Homologous recombination deficiency assays, guiding treatment of poly (adenosine diphosphate ribose) polymerase inhibitors, are increasingly applied in clinics. This study aimed to evaluate the predictive performance of homologous recombination deficiency status at genomic and functional perspective on the efficacy of platinum-based chemotherapy in ovarian cancer. Between 2016 and 2019, 134 patients with high-grade ovarian cancer were retrospectively analyzed. Formalin-fixed paraffin-embedded tissues were subjected to DNA sequencing using the AmoyDx HRD Complete Panel. The genomic scar score and the genomic instability score were calculated based on copy number variation events. Furthermore, the RAD51 and SLFN11 protein levels in tumors were assessed by immunohistochemistry. Of all patients, 106 of 134 (79.1%) were homologous recombination deficiency (genomic scar score)-positive, with a higher platinum sensitivity rate than those who were homologous recombination deficiency (genomic scar score)-negative (78.3% vs 57.1%, p = .023). Similarly, 104 of 134 (77.6%) were homologous recombination deficiency (genomic instability score)-positive, with increased platinum sensitivity compared with homologous recombination deficiency (genomic instability score)-negative (77.9% vs 60.0%, p = .049). The overall concordance rate of homologous recombination deficiency status defined by the 2 scores was 98.5%. Genomic scar score and genomic instability score determined homologous recombination deficiency-positive statuses correlated with better progression-free survival (p = .0019, p = .0041) and overall survival (p = .018, p = .031). Patients with nuclear RAD51-loss or SLFN11-positive expression were likely to be homologous recombination deficiency-positive by genomic scar score/genomic instability score (94.1% and 97.6%; 94.1% and 95.2%, respectively). Patients with nuclear RAD51-loss and SLFN11-positive expression had better overall survival than those with RAD51-positive and SLFN11-negative expression. Among homologous recombination deficiency statuses, RAD51 and SLFN11 expressions, homologous recombination deficiency (genomic scar score)-positive was most associated with progression-free survival and platinum sensitivity. Multivariate regression analysis showed that homologous recombination deficiency (genomic scar score)-positive status was a good prognostic factor, implying a higher possibility of platinum sensitivity. Genomic scar score, given by AmoyDx HRD Complete Panel, was most associated with the efficacy of platinum treatment in patients with high-grade ovarian cancer. Validation is warranted via prospective studies.

Exosomes derived from cancer-associated fibrolasts mediated ciplatin resistance

Objective: Nasopharyngeal carcinoma (NPC), a highly invasive form of head and neck cancer, carries a significant risk of distant metastasis. NPC is particularly prevalent in Asia and has a high incidence in southern China. Cisplatin-diamminedichloroplatinum (DDP), a chemotherapy agent, is commonly employed in NPC treatment. Despite DDP’s efficacy, many patients eventually develop resistance to it over the course of their therapy, which significantly hinders treatment outcomes. Cancer-associated fibroblasts (CAFs) are key components of the tumor micro-environment and contribute to tumor progression and chemotherapy resistance. Exosomes secreted by CAFs serve as crucial mediators of intercellular communication and participate in modulating diverse biological processes. This study aimed to explore how exosomes derived from CAFs contribute to DDP resistance in NPC. Material and Methods: An in vitro coculture system was used to simulate the interaction between CAFs and NPC cells, and exosomes secreted by CAFs were isolated and characterized. The expression of autophagy hallmark proteins was detected by Western blot and quantitative real-time polymerase chain reaction. Autophagy intensity was quantified using monodansylcadaverine staining, and cell proliferation was assessed by colony formation assays and methylthiazolyldiphenyl–tetrazolium assays. NPC cells were treated with autophagy inducers (rapamycin), and the expression of Ras homologue enriched in brain (Rheb), mammalian target of rapamycin complex (mTORC1), and UNC51-like kinase was detected. Immunofluorescence was used to determine the cellular localization and expression intensity of mTORC1, and the effect on DDP sensitivity was evaluated through cell proliferation rates. In addition, the exosome-mediated resistance mechanism was further validated using an in vivo xenograft tumor model. Results: Coculture of CAFs with NPC cells significantly promoted the proliferation of NPC cells (P < 0.01), significantly elevated the IC50 value of DDP (P < 0.01), and elevated the resistance of NPC cells to DDP. CAF-derived exosomes elevated autophagy hallmark proteins light chain 3B-II, Beclin, and increased the autophagy intensity (P < 0.01). CAF-derived exosomes promoted autophagy by inhibiting mTORC1 (P < 0.01). In the in vitro model, exosomes promoted the growth of tumor tissues (P < 0.01), and the inhibition of exosome secretion reversed the promotion effect of autophagy (P < 0.01) and elevated the sensitivity of NPC cells to DDP. Conclusion: CAF-derived exosomes promote protective autophagy in NPC cells through the Rheb/mTOR axis, and result in DDP resistance in NPC.

Mutations in tumor suppressor genes Vhl and Rassf1a cause DNA damage, chromosomal instability and induce gene expression changes characteristic of clear cell renal cell carcinoma.

RASSF1A is frequently biallelically inactivated in clear cell renal cell carcinoma (ccRCC) due to loss of chromosome 3p and promoter hypermethylation. Here we investigated the cellular and molecular consequences of single and combined deletion of the Rassf1a and Vhl tumor suppressor genes to model the common ccRCC genotype of combined loss of function of RASSF1A and VHL. In mouse embryonic fibroblasts and in primary kidney epithelial cells, double deletion of Rassf1a and Vhl caused chromosomal segregation defects and increased formation of micronuclei, demonstrating that pVHL and RASSF1A function to maintain genomic integrity. Combined Rassf1a and Vhl deletion in kidney epithelial cells in vivo increased proliferation and caused mild tubular disorganization, but did not lead to the development of kidney tumors. Single cell RNA-sequencing unexpectedly revealed that Rassf1a or Vhl deletion both induce the expression of an overlapping set of genes in a sub-population of proximal tubule cells. Many of these genes are also upregulated in the Vhl/Trp53/Rb1 deficient mouse model of ccRCC. In other subsets of proximal tubule cells, combined Vhl/Rassf1a deletion induced the expression of additional genes that were not upregulated in each of the single knockouts. The expression of the human homologues of Rassf1a-regulated genes correlate negatively with RASSF1 expression levels in human ccRCC. Our results suggest that the loss of RASSF1A function establishes a ccRCC-characteristic gene expression pattern.

The palmitoyl-CoA ligase Fum16 is part of a Fusarium verticillioides fumonisin subcluster involved in self-protection.

Fusarium verticillioides produces the mycotoxin fumonisin B1 (FB1), which disrupts sphingolipid biosynthesis by inhibiting ceramide synthase and affects the health of plants, animals, and humans. The means by which F. verticillioides protects itself from its own mycotoxin are not completely understood. Some fumonisin (FUM) cluster genes do not contribute to the biosynthesis of the compound, but their function has remained enigmatic. Recently, we showed that FUM17, FUM18, and FUM19 encode two ceramide synthases and an ATP-binding cassette transporter, respectively, which play a role in antagonizing the toxicity mediated by FB1. In the present work, we uncovered functions of two adjacent genes, FUM15 and FUM16. Using homologous and heterologous expression systems, in F. verticillioides and Saccharomyces cerevisiae, respectively, we provide evidence that both contribute to protection against FB1. Our data indicate a potential role for the P450 monooxygenase Fum15 in the modification and detoxification of FB1 since the deletion and overexpression of the respective gene affected extracellular FB1 levels in both hosts. Furthermore, relative quantification of ceramide intermediates and an in vitro enzyme assay revealed that Fum16 is a functional palmitoyl-CoA ligase. It co-localizes together with the ceramide synthase Fum18 to the endoplasmic reticulum, where they contribute to sphingolipid biosynthesis. Thereby, FUM15-19 constitute a subcluster within the FUM biosynthetic gene cluster dedicated to the fungal self-protection against FB1.IMPORTANCEThe study identifies a five-gene FUM subcluster (FUM15-19) in Fusarium verticillioides involved in self-protection against FB1. FUM16 (palmitoyl-CoA ligase), FUM17, and FUM18 (ceramide synthases) enzymatically supplement ceramide biosynthesis, while FUM19 (ATP-binding cassette transporter) acts as a repressor of the FUM cluster. The evolutionary conservation of FUM15 (P450 monooxygenase) in Fusarium and Aspergillus FUM clusters is discussed, and its effect on extracellular FB1 levels in both native (F. verticillioides) and heterologous (Saccharomyces cerevisiae) hosts is highlighted. These findings enhance our understanding of mycotoxin self-protection mechanisms and could inform strategies for predicting biological activity of unknown secondary metabolites, managing mycotoxin contamination, and developing resistant crop cultivars.

A 2-oxoglutarate-dependent dioxygenase, GLUCORAPHASATIN SYNTHASE 1(GRS1) is a major determinant for different aliphatic glucosinolates between radish and Chinese cabbage.

Glucosinolates (GSLs) are secondary metabolites in Brassicaceae plants and play a defensive role against a variety of abiotic and biotic stresses. Also, it exhibits anti-cancer activity against cancer cell in human. Different profiles of aliphatic GSL compounds between radish and Chinese cabbage were previously reported. However, molecular details underlying the divergent profile between two species were not clearly understood. In this study, we found that major difference of aliphatic GSLs profiles between two species is determined by the dominantly expressed genes in first step of the secondary modification phase, which are responsible for enzymatic catalysis of methylthioalkyl-glucosinolate. For instance, active expression of GLUCORAPHASATIN SYNTHASE 1 (GRS1) gene in radish play an important role in the production of glucoraphasatin (GRH) and glucoraphenin (GRE), a major aliphatic GSLs in radish. Meanwhile, Chinese cabbage was found to merely produce glucoraphasatin (GRH), instead producing glucoraphanin (GRA) and gluconapin (GNP) due to the mere expression of GRS1 homologs and abundant expressions of FLAVIN-CONTAINING MONOOXYGENASES (FMO GS-OX) homologs in Chinese cabbage. In addition, we noticed that wounding treatment on leaf tissues substantially enhanced the production of aliphatic and benzenic GSLs in both Chinese cabbage and radish, indicating that GSLs are wound-induced defensive compounds in both Chinese cabbage and radish plants.

Evaluation of Selected Folk Herbs on the Fertility of Sprague Dawley Male Rats: Biochemical, Histological, and Molecular Investigations.

Scientists have shown great interest in traditional plant extracts, particularly Lepidium sativum (LS), Origanum majorana (OM), Ferula hermonis (FH), and Eruca sativa (ES), which are frequ ently used to improve health. Recently, attention has been directed toward their influence on spermatogenesis and male fertility. Hence, the objective of this study was to explore their impact on male rats' fertility. Antioxidant activity and total phenolic content (TPC) were determined, along with the identification and quantification of phenolic compounds. Oral administration of aqueous extracts was performed individually or as a mixture (MIX) at a dose of 100 mg/kg in 28 male Sprague Dawley rats over a 60-day period. Organ weight, sex hormone concentrations, sperm parameters, oxidative stress markers, histological and morphometric analysis, and protein expression levels were investigated. OM and MIX showed the highest TPC and antioxidant activities, and MIX possessed the highest polyphenolic constituents. Elevated serum testosterone, epididymal sperm concentration, testes glutathione levels, and histomorphometric parameters were manifested in all groups, especially in MIX. MIX group also displayed elevated levels of vimentin, protein kinase B, and mTOR expression in the testes, complemented by declined expression of Phosphatase and Tensin Homolog (PTEN). In conclusion, these findings propose that these extracts, especially MIX followed by OM, enhance fertility by stimulating spermatogenesis.

Characterization of a family IV esterase from extremely halophilic archaeon Haloarcula japonica

The novel esterase gene lipP1, which encodes HjEstP1, was discovered in the genome of the extremely halophilic archaeon Haloarcula japonica. A homology search and sequence alignment revealed that HjEstP1 is a member of family IV esterases with conserved GXSXG and HGGG motifs. lipP1 was expressed in its parental strain, and recombinant HjEstP1 was purified and characterized. Optimal pH and temperature of HjEstP1 were 6.0 and > 60 °C, respectively. HjEstP1 showed higher activity with increasing NaCl concentration, and optimal NaCl concentration was > 4.5 M. Furthermore, HjEstP1 preferentially hydrolyzed pNP and glycerol esters with short chain fatty acids. To our knowledge, this is the first report of an esterase from an extremely halophilic archaeon obtained via homologous expression.

Disruption of lipid raft reverses drug resistance in colorectal cancer cells through the phosphatase and tensin homolog/phosphoinositide 3-kinase/protein kinase B pathway and P-glycoprotein.

Regarding flotillin knockdown, drug resistance is reversed in colorectal cancer (CRC) cell lines; this is associated with the phosphoinositide 3-kinase/protein kinase B (PI3K/AKT) pathway, as our previous experimental results indicated. However, the exact mechanism underlying this pathway remains unclear. PI3K inhibitor and activator were added separately to clarify the role of the PI3K pathway in reversing drug resistance. The results showed decreased resistance after inhibiting PI3K activity. Furthermore, the reduced resistance due to flotillin knockdown was restored after adding the PI3K activator. Additional results showed no changes in PI3K molecules. However, p-AKT expression was downregulated. Further results suggested that the phosphatidylinositol (3,4,5)-trisphosphate/phosphatidylinositol 4,5-bisphosphate (PIP3/PIP2) ratio was downregulated, whereas the phosphatase and tensin homolog (PTEN) expression was upregulated. In addition, we also found that P-gp activity inhibition resulted in increased adriamycin accumulation and reversal of resistance, and flotillin knockdown was accompanied by a downregulation of P-gp expression in CRC cells. In conclusion, our study demonstrated that flotillin knockdown could reverse drug resistance in CRC cells by downregulating the PTEN/PI3K/AKT pathway and P-gp.

Autism-Linked Mutations in α2δ-1 and α2δ-3 Reduce Protein Membrane Expression but Affect Neither Calcium Channels nor Trans-Synaptic Signaling.

α2δ proteins regulate membrane trafficking and biophysical properties of voltage-gated calcium channels. Moreover, they modulate axonal wiring, synapse formation, and trans-synaptic signaling. Several rare missense variants in CACNA2D1 (coding for α2δ-1) and CACNA2D3 (coding for α2δ-3) genes were identified in patients with autism spectrum disorder (ASD). However, the pathogenicity of these variants is not known, and the molecular mechanism by which α2δ proteins may contribute to the pathophysiology of autism is, as of today, not understood. Therefore, in this study we functionally characterized two heterozygous missense variants in α2δ-1 (p.R351T) and α2δ-3 (p.A275T), previously identified in patients with ASD. Electrophysiological recordings in transfected tsA201 cells were used to study specific channel-dependent functions of mutated α2δ proteins. Membrane expression, presynaptic targeting, and trans-synaptic signaling of mutated α2δ proteins were studied upon expression in murine cultured hippocampal neurons. Homologous expression of both mutated α2δ proteins revealed a strongly reduced membrane expression and synaptic localization compared to the corresponding wild type α2δ proteins. Moreover, the A275T mutation in α2δ-3 resulted in an altered glycosylation pattern upon heterologous expression. However, neither of the mutations compromised the biophysical properties of postsynaptic L-type (CaV1.2 and CaV1.3) and presynaptic P/Q-type (CaV2.1) channels when co-expressed in tsA201 cells. Furthermore, presynaptic expression of p.R351T in the α2δ-1 splice variant lacking exon 23 did not affect trans-synaptic signaling to postsynaptic GABAA receptors. Our data provide evidence that the pathophysiological mechanisms of ASD-causing mutations of α2δ proteins may not involve their classical channel-dependent and trans-synaptic functions. Alternatively, these mutations may induce subtle changes in synapse formation or neuronal network function, highlighting the need for future α2δ protein-linked disease models.

YTHDF2 contributes to psoriasis by promoting proliferation and inflammatory response through regulation of the Wnt signaling pathway

YT521-B homology domain family 2 (YTHDF2), a pivotal m6A-binding protein, is now understood to significantly influence a diverse array of biological functions, including cell migration, proliferation, differentiation, and inflammatory responses. Additionally, YTHDF2 participates in mRNA decay and pre-rRNA processing. This study explored the specific role of YTHDF2 in the pathogenesis of psoriasis and its underlying mechanisms. Our preliminary findings revealed upregulation of YTHDF2 expression in psoriasis. Subsequent silencing of YTHDF2 in a psoriatic cell model resulted in a marked decrease in mRNA expression of IL-17A, S100A8, and S100A9, accompanied by a reduction in cell proliferation. Conversely, overexpression of YTHDF2 led to the opposite effects. Treatment with DC-Y27-13, a YTHDF2 inhibitor, demonstrated a therapeutic effect in psoriasis mice. Next, mRNA sequencing analysis identified significant enrichment of differentially expressed genes within the Wnt signaling pathway. Further investigation revealed that deletion of YTHDF2 increased the half-life and expression of Dickkopf homolog 3 (DKK3), a potent inhibitor of the Wnt signaling pathway. Consequently, the inhibition of Wnt signaling attenuated the inflammatory response and inhibited cell proliferation.

PGC-1α drives small cell neuroendocrine cancer progression toward an ASCL1-expressing subtype with increased mitochondrial capacity

Adenocarcinomas from multiple tissues can converge to treatment-resistant small cell neuroendocrine (SCN) cancers composed of ASCL1, POU2F3, NEUROD1, and YAP1 subtypes. We investigated how mitochondrial metabolism influences SCN cancer (SCNC) progression. Extensive bioinformatics analyses encompassing thousands of patient tumors and human cancer cell lines uncovered enhanced expression of proliferator-activatedreceptor gamma coactivator 1-alpha (PGC-1α), a potent regulator of mitochondrial oxidative phosphorylation (OXPHOS), across several SCNCs. PGC-1α correlated tightly with increased expression of the lineage marker Achaete-scute homolog 1, (ASCL1) through a positive feedback mechanism. Analyses using a human prostate tissue-based SCN transformation system showed that the ASCL1 subtype has heightened PGC-1α expression and OXPHOS activity. PGC-1α inhibition diminished OXPHOS, reduced SCNC cell proliferation, and blocked SCN prostate tumor formation. Conversely, PGC-1α overexpression enhanced OXPHOS, validated by small-animal Positron Emission Tomography mitochondrial imaging, tripled the SCN prostate tumor formation rate, and promoted commitment to the ASCL1 lineage. These results establish PGC-1α as a driver of SCNC progression and subtype determination, highlighting metabolic vulnerabilities in SCNCs across different tissues.

Nitric oxide-mediated PpRAP2.2 regulates the transcription of PpRbohA/D to enhance resistance to Monilinia fructicola in peach fruit

Nitric oxide (NO), a critical signaling molecule, plays key roles in plant cell physiological processes, particularly in redox system. To investigate the resistance of exogenous NO to brown rot (Monilinia fructicola) in peach fruit during storage, postharvest peaches were treated with NO and c-PTIO (an NO scavenger), exploring the mechanism of NO enhancing disease resistance in peach fruit through reactive oxygen species (ROS) metabolism. The results indicated that NO significantly inhibited brown rot in postharvest peach fruit, evidenced by reduced disease incidence and smaller lesion diameters. Additionally, exogenous NO treatment decreased the expression of respiratory burst oxidase homologs (PpRbohs), which is a crucial factor in ROS metabolism. The transcription factor PpRAP2.2 was identified through yeast one-hybrid (Y1H) screening with the promoter of PpRbohD. The expression pattern of PpRAP2.2 was closely associated with superoxide anion (O2-) content and peach fruit disease resistance. PpRAP2.2, a nuclear-localized transcription factor, activated the expression of PpRbohA and PpRbohD by directly binding to their promoter. The virus-induced gene silencing (VIGS) assay showed that silencing of PpRAP2.2 facilitated peach disease resistance by reducing the transcription of PpRbohA and PpRbohD. Furthermore, the overexpression of PpRAP2.2 resulted in a significant reduction in disease incidence and conidiophores in transgenic tomatoes relative to control group under M. fructicola, suggesting that PpRAP2.2 effectively enhanced the resistance of tomatoes to M. fructicola. These results indicate that PpRAP2.2 enhances NO-mediated disease resistance in peach by regulating the expression of PpRbohA and PpRbohD. These finding provide a basis for further studies on function and regulatory mechanisms of peach fruit disease resistance.

A mesocorticolimbic insulin receptor gene co-expression network moderates the association between early life adversity and food approach eating behaviour style in childhood

Insulin receptors, located in brain regions associated with reward sensitivity and decision-making, facilitate insulin action in the brain, modulating intracellular signaling cascades, gene expression, and neural activity. Here, we tested if variations in the expression of the insulin receptor gene network in the prefrontal cortex (PFC) and striatum (STR) moderate the association between early life adversity and eating behaviour in childhood and if this moderation is sex-specific. Participants from the Maternal Adversity, Vulnerability and Neurodevelopment (MAVAN) and Basal Influences on the Baby's Development (BIBO) were included as two independent cohorts. A biologically-informed polygenic score reflecting functional variation of the mesocorticolimbic insulin receptor gene network was created by using insulin receptor co-expression data from the PFC and STR in mice, and validated in humans through filtering by homologous expression in PFC using well-known databases. Early life adversity exposure was measured as a composite score. Eating behaviour was characterized using the Child Eating Behaviour Questionnaire administered to mothers of children aged 4 and 6 years in MAVAN, and 6 years in BIBO. We found that only in those with high expression of the mesocorticolimbic insulin receptor gene network a higher early adversity score associated with a higher desire to drink in 4-year boys and 6-year girls, as well as a higher food approach score and food approach/food avoidance ratio in 4-year girls. Also, a higher early life adversity was associated with higher food responsiveness, food approach score and food approach/food avoidance ratio at 6 years in the MAVAN full sample. The moderation observed on desire to drink was partially replicated in BIBO children aged 6 years. Identifying individual differences in response to early adversity may help to prioritize individuals at high risk for long-term disease and design suitable interventions.

The Ubiquitin Ligase HERC2 promotes Ang II-induced cardiac hypertrophy via destabilization of MeCP2 to enhance Lin28a expression.

HECT-type E3 ubiquitin ligases (HECT E3s) participate in the progression of cardiovascular diseases. HERC2 has been reported to play critical roles in many pathological processes, but its role in cardiac hypertrophy remains unclear. In this study, we observed that the expression and activity of HERC2 was significantly upregulated in hypertrophic hearts and angiotensin II (Ang II)-stimulated primary cardiomyocytes. Knockdown of HERC2 in cardiomyocytes significantly alleviated the myocardial hypertrophy induced by Ang II. Conversely, cardiac specific overexpression of HERC2 aggravated Ang II-induced cardiac hypertrophy in vitro and in vivo. Furthermore, we demonstrated that HERC2 promoted cardiac hypertrophy via increasing the expression of lin-28 homologue A (Lin28a), an RNA-binding protein that regulates pathological cardiac hypertrophic. Knocking down Lin28a attenuated Ang II-induced myocardial hypertrophy and abolished the increase in myocardial hypertrophy by overexpression of HERC2. Further investigation indicated that HERC2 promoted the expression level of Lin28a by reducing MeCP2, a transcriptional suppressor of Lin28a. We also showed that the pro-hypertrophic effect of HERC2 was partially dependent on MeCP2 inhibition. Mechanistically, HERC2 directly bound with MeCP2, and promoting its K48-linked polyubiquitination and degradation. Combined, these findings demonstrate HERC2 plays a crucial role in pathological cardiac hypertrophy, thereby indicating that targeting the HERC2/MeCP2/Lin28a axis is a potential strategy for heart failure therapy.

High-throughput screening and identification of lignin peroxidase based on spore surface display of Bacillus subtilis.

Lignin peroxidase is closely related to agriculture and food as it improves the quality of feedstuffs, facilitates the degradation of lignin in agricultural wastes, and degrades azo dyes that have similar complex structures to lignin. However, the current status of homologous or heterologous expression of lignin peroxidase is unsatisfactory and needs to be modified with the help of immobilization and directed evolution to maximize its potential. Directed evolution technology is an effective strategy for designing and improving enzyme characteristics, and Bacillus subtilis spore surface display technology is an efficient method for preparing immobilized enzymes. A colorimetric dye decolorization assay using Congo red as a substrate was developed and optimized for high-throughput screening of spore surface display in a 96-well plate. After two rounds of screening, a superior mutant strain was selected from 2700 mutants. Its highest catalytic activity was 196.36%. The amino acid substitution sites were identified as N120D and I242T. The mechanism for the enhanced catalytic activity was explained using protein modeling and functional analysis software. This study provides insights into the rational design of lignin peroxidase and its application in food and agriculture. © 2024 Society of Chemical Industry.

Benzylurea Protects hPDLFs Against LPS-Induced Mitochondrial Dysfunction Through MTCH2.

The aim of this study is to explore the mechanism of benzylurea in the inflammatory injury of human periodontal ligament fibroblasts (hPDLFs). An inflammation model of hPDLFs was established using LPS. Nuclear transport of nuclear transcription factor-κB (NF-κB), secretion of cytokines, and the morphology and distribution of F-actin were determined. Mitochondrial function was assessed by measuring mitochondrial membrane potential (MMP), mitochondrial permeability transition pore (mPTP), and reactive oxygen species (ROS) levels. The expression of mitochondrial carrier homolog 2 (MTCH2) and Cytochrome b5 type B (CYB5B) was detected. Benzylurea alleviated the effects of lipopolysaccharide (LPS) on the proliferation and apoptosis of hPDLFs. It reduced the release of inflammatory cytokines and inhibited NF-κB nuclear translocation. Benzylurea improved mitochondrial function by regulating MMP and preventing excessive mPTP opening. Furthermore, LPS elevated the expression of MTCH2 and reduced the expression of CYB5B in hPDLFs. However, these effects can be inhibited by benzylurea. The altered expression of MTCH2 directly affected CYB5B expression, the release of inflammatory cytokines, and the activation of nuclear translocation of NF-κB. CYB5B may act as an effector of MTCH2, with benzylurea enhancing mitochondrial function and protecting hPDLFs from LPS-induced injury through MTCH2.

Ginsenoside Rg1 promotes non-rapid eye movement sleep via inhibition of orexin neurons of the lateral hypothalamus and corticotropin-releasing hormone neurons of the paraventricular hypothalamic nucleus

ObjectiveThis study investigates the sleep-modulating effects of ginsenoside Rg1 (Rg1, C42H72O14), a key bioactive component of ginseng, and elucidates its underlying mechanisms. MethodsC57BL/6J mice were intraperitoneally administered doses of Rg1 ranging from 12.5 to 100 mg/kg. Sleep parameters were assessed to determine the average duration of each sleep stage by monitoring the electrical activity of the brain and muscles. Further, orexin neurons in the lateral hypothalamus (LH) and corticotropin-releasing hormone (CRH) neurons in the paraventricular hypothalamic nucleus (PVH) were ablated using viral vector surgery and electrode embedding. The excitability of LHorexin and PVHCRH neurons was evaluated through the measurement of cellular Finkel-Biskis-Jinkins murine osteosarcoma viral oncogene homolog (c-Fos) expression. ResultsRg1 (12.5–100 mg/kg) augmented the duration of non-rapid eye movement (NREM) sleep phases, while reducing the duration of wakefulness, in a dose dependent manner. The reduced latency from wakefulness to NREM sleep indicates an accelerated sleep initiation time. We found that these sleep-promoting effects were weakened in the LHorexin and PVHCRH neuron ablation groups, and disappeared in the orexin and CRH double-ablation group. Decreased c-Fos protein expression in the LH and PVH confirmed that Rg1 promoted NREM sleep by inhibiting orexin and CRH neurons. ConclusionRg1 increases the duration of NREM sleep, underscoring the essential roles of LHorexin and PVHCRH neurons in facilitating the sleep-promoting effects of Rg1.Please cite this article as: Wang YY, Wu Y, Yu KW, Xie HY, Gui Y, Chen CR, Wang NH. Ginsenoside Rg1 promotes non-rapid eye movement sleep via inhibition of orexin neurons of the lateral hypothalamus and corticotropin-releasing hormone neurons of the paraventricular hypothalamic nucleus. J Integr Med . 2024; Epub ahead of print.