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SUMMARY: The accessory proteins Hyponastic‐like 1 (HYL1) and Serrated (SE) enhance the precise and efficient processing of miRNAs by Dicer‐like 1 (DCL1), which is important for proper miRNA function. However, other factors determining the precision and efficiency of miRNA biogenesis are not well‐known. Here, we found that an asymmetric bulge (AB) at the 3′ end of miR‐5p (produced from the 5′ arm of the pre‐miRNA) reduced the precision of the second cleavage, whereas an AB at other sites of miR‐5p mainly affected the accumulation level of miR‐5p in transient expression in Nicotiana benthamiana. In contrast, many ABs in miR‐3p (produced from the 3′ arm of the pre‐miRNA) impose strong negative impact on the processing precision and the accumulation level of miR‐5p in N. benthamiana. Arabidopsis DCL1/SE/HYL1 complex‐mediated miRNA processing was reconstituted in Saccharomyces cerevisiae to further investigate AB‐mediated interference with DCL1 processing. With this system, the positional effect of AB on miRNA processing was tested. The results showed that ABs on the middle of miR‐5p have less of an impact on DCL1 cleavage efficiency and precision, whereas those on miR‐3p or near the ends of miR‐5p strongly reduce DCL1 cleavage activity, precision or both. Studies using the yeast miRNA processing system and transgenic Arabidopsis also revealed the importance of the interaction between the 2‐nt 3′ overhang of pre‐miRNA and the 3′ overhang binding pocket (3′BP) on the precision of the second cleavage reaction for many endogenous miRNAs. These findings provide new insights into the mechanism of miRNA biogenesis. Significance Statement: Factors determining the precision and efficiency of miRNA biogenesis are not well‐known. This study found that an asymmetric bulge at different positions of miR‐5p and miR‐3p had different impacts on miRNA levels and processing precision and revealed the importance of the interaction between the 2‐nt 3′ overhang of pre‐miRNA and the 3′ overhang binding pocket (3'BP) on the precision of the second cleavage reaction for many endogenous miRNAs. [ABSTRACT FROM AUTHOR]

BACKGROUND: Dicer1 plays a crucial role in regulating the development and reproduction of insects. Knockout of Dicer1 causes pupal deformity, low eclosion and low fecundity in Plutella xylostella, but the mechanism behind this phenomenon is not clear. This study aims to identify differentially‐expressed genes and miRNAs in the Dicer1‐knockout strain (ΔPxDcr‐1) and assess their impact on the reproduction and development of P. xylostella. RESULTS: The knockout of Dicer1 affected the expression of genes including the adipokinetic hormone/corazonin‐related peptide receptor (PxACPR). The expression of PxACPR was upregulated, and the expression of miR‐8514‐5p was downregulated in ΔPxDcr‐1 of P. xylostella. The dual luciferase reporter assay and pull‐down assay showed that miR‐8514‐5p bound to PxACPR in vitro and in vivo. The expression profiles demonstrated a negative correlation between PxACPR mRNA and miR‐8514‐5p in different developmental stages of the wild‐type strain. Both the miR‐8514‐5p agomir and double‐stranded RNA of ACPR (dsPxACPR) injected into the pre‐pupae inhibited the mRNA level of PxACPR, causing high mortality and deformity of pupae, and low fecundity and hatching rate, which were consistent with the phenotype of ΔPxDcr‐1. The injection of miR‐8514‐5p antagomir caused a similar phenotype to the injection of miR‐8514‐5p agomir. Additionally, the injection of miR‐8514‐5p antagomir significantly rescued the phenotype caused by dsPxACPR. CONCLUSION: These results indicate that miR‐8514‐5p affects the development and reproduction of P. xylostella by regulating PxACPR, and the homeostasis of PxACPR expression is essential for the development and reproduction of P. xylostella. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

In addition to preventing precocious larval metamorphosis, juvenile hormone (JH), synthesized in corpora allata (CA), is known to stimulate female reproduction of insects. JH titer is extremely low or absent during metamorphosis, but thereafter rapidly increases in the previtellogenic stage and rises to a peak in the vitellogenic phase. However, the mechanisms underlying the biosynthesis of high levels of JH in adults remain unclear. We found in this study that 12 genes involved in JH synthesis pathway were highly expressed in the CA of adult locusts. By transcriptome analysis and quantitative real‐time – polymerase chain reaction validation, a total of 106 evolutionary conserved micro RNAs (miRNAs) and 163 species‐specific miRNAs were identified in locust CA. Dual‐luciferase assay revealed that 17 miRNAs bound to 10 JH synthesis genes (JHSGs) and downregulated their expression. These miRNAs were expressed in low levels during vitellogenic stage, which was oppositive from that of targeting JHSGs. Six miRNAs including miR‐971‐3p, miR‐31a, miR‐9‐5p, miR‐1‐3p, miR‐315, and miR‐282 were selected for function study. Co‐application of agomiRs resulted in significantly decreased levels of targeting JHSGs, accompanied by significantly reduced vitellogenin expression as well as arrested ovarian development. The data suggest that multiple miRNAs expressed synchronously at low levels in the vitellogenic phase, thereby ensuring the high levels of JHSG expression to facilitate JH biosynthesis required for JH‐dependent female reproduction. The findings provide important information for deciphering miRNA—messenger RNA modules for JH biosynthesis as well as JH regulation of insect metamorphosis and reproduction. [ABSTRACT FROM AUTHOR]

As a member of the family of A Disintegrin And Metalloproteinases (ADAM) ADAM8 is preferentially expressed in lymphatic organs, immune cells, and tumor cells. The substrate spectrum for ADAM8 proteolytic activity is not exclusive but is related to effectors of inflammation and signaling in the tumor microenvironment. In addition, complexes of ADAM8 with extracellular binding partners such as integrin β‐1 cause an extensive intracellular signaling in tumor cells, thereby activating kinase pathways with STAT3, ERK1/2, and Akt signaling, which causes increased cell survival and enhanced motility. The cytoplasmic domain of ADAM8 harbors five SRC homology‐3 (SH3) domains that can potentially interact with several proteins involved in actin dynamics and cell motility, including Myosin 1F (MYO1F), which is essential for neutrophil motility. The concept of ADAM8 thus involves immune cell recruitment, in most cases leading to an enhancement of inflammatory (asthma, COPD) and tumor (including pancreatic and breast cancers) pathologies. In this review, we report on available studies that qualify ADAM8 as a therapeutic target in different pathologies. As a signaling hub, ADAM8 controls extracellular, intracellular, and intercellular communication, the latter one mainly mediated by the release of extracellular vesicles with ADAM8 as cargo. Here, we will dissect the contribution of different domains to these distinct ways of communication in several pathologies. We conclude that therapeutic targeting attempts for ADAM8 should consider blocking more than a single domain and that this requires a thorough evaluation of potent molecules targeting ADAM8 in an in vivo setting. [ABSTRACT FROM AUTHOR]

Summary CDKs are the master regulator of cell division and their activity is controlled by the regulatory subunit cyclins and phosphorylation by the CAKs. However, the role of MAP kinases in regulating plant cell cycle or CDKs have not been explored. Here, we report that the MAP kinases OsMPK3, OsMPK4, and OsMPK6 physically interact and phosphorylate OsCDKD and its regulatory subunit OsCYCH in rice. MAP kinases phosphorylate CDKD at Ser‐168 and Thr‐235 residues in OsCDKD. The MAP kinase‐mediated phosphorylation of OsCDKD is required for its activation to control the small RNA biogenesis. The phosphodead version of OsCDKD fails to activate the C‐terminal domain of RNA Polymerase II, thereby negatively impacting small RNA transcription. Further, the overexpression lines of wild‐type (WT) OsCDKD and phosphomimic OsCDKD show increased root growth, plant height, tiller number, panicle number, and seed number in comparison to WT, phosphodead OsCDKD‐OE, and kinase‐dead OsCDKD‐OE plants. In a nutshell, our study establishes a novel regulation of OsCDKD by MAPK‐mediated phosphorylation in rice. The phosphorylation of OsCDKD by MAPKs imparts a positive effect on rice growth and development by regulating miRNAs transcription. [ABSTRACT FROM AUTHOR]

Eukaryotic cells respond to stress by altering coding and non‐coding gene expression programs. Alongside many approaches and regulatory mechanisms, long non‐coding RNAs (lncRNA) are finding a significant place in gene regulation, suggesting an involvement in various cellular processes and pathophysiology. LncRNAs are regulated by many transcription factors, including SMAR1 and p53, which are tumor suppressor genes. SMAR1 inhibits cancer cell metastasis and invasion and is also known to inhibit apoptosis during low‐dose stress in coordination with p53. Data mining analysis suggested that these tumor suppressor genes might coregulate the lncRNA RP11‐431M3.1 in colon cancer cells. Importantly, RP11‐431M3.1 expression was found to be negatively correlated with patient survival rates in a number of cancers. Oxidative stress occurs when an imbalance in the body is caused by reactive oxygen species (ROS). This imbalance is known to be important in the development/pathogenesis of colon cancer. We are researching the role and control of this lncRNA in HCT116 cells under conditions of oxidative stress. We observed a dose‐dependent differential expression of lncRNA upon H2O2 treatment and found that p53 and SMAR1 bind differentially to the promoter in response to the dose of stress inducer used. RP11‐431M3.1 was observed to sponge miR‐138 which has an important target gene, hypoxia‐inducible factor (HIF1A). miR‐138 was observed to bind differentially to RP11‐431M3.1 and HIF1A RNA depending on the dose of oxidative stress. Furthermore, the knockdown of RP11‐431M3.1 decreased the migration and proliferation of colon cancer cells. Our results suggest a previously undescribed regulatory mechanism through which RP11‐431M3.1 is transcriptionally regulated by SMAR1 and p53, target HIF1A through miR‐138, and highlight its potential as a therapeutic and diagnostic marker for cancer. [ABSTRACT FROM AUTHOR]

In melanoma, carcinoma‐associated fibroblasts (CAFs) are important cellular components in the tumour microenvironment due to their potential to promote tumour growth and metastatic spread of malignant cells. Melanoma cells have the ability to affect non‐tumour cells in the microenvironment by releasing extracellular vesicles (EVs). The mechanisms responsible for reprogramming normal dermal fibroblasts (NHDFs) into CAFs remain incompletely understood. However, it is likely thought to be mediated by melanoma‐specific miRNAs, which are transported by EVs derived from melanoma cells. Therefore, we wondered if one of the most enriched miRNAs in EVs secreted by melanoma cells, miR‐92b‐3p, is involved in the conversion of normal fibroblasts into CAFs. We observed that melanoma cell‐derived EVs indeed delivered miR‐92b‐3p into NHDFs and that its accumulation correlated with CAF formation, as demonstrated by enhanced expression of CAF marker genes and increased proliferation and migration. Overexpression of miR‐92b‐3p in NHDFs revealed similar results, while EVs deficient of miR‐92b‐3p did not induce a CAF phenotype. As a target we identified PTEN, whose repression led to increased expression of CAF markers. We thus provide a novel pathway of intercellular communication by which melanoma cells control the transformation of CAFs by virtue of EV‐transported miRNAs. [ABSTRACT FROM AUTHOR]

5‐Fluorouracil (5‐FU) has been used for chemotherapy for colorectal and other cancers for over 50 years. The prevailing view of its mechanism of action is inhibition of thymidine synthase leading to defects in DNA replication and repair. However, 5‐FU is also incorporated into RNA causing defects in RNA metabolism, inhibition of pseudouridine modification, and altered ribosome function. We examined the impact of 5‐FU on post‐transcriptional small RNA modifications (PTxMs) and the expression and export of RNA into small extracellular vesicles (sEVs). EVs are secreted by all cells and contain a variety of proteins and RNAs that can function in cell‐cell communication. We found that treatment of colorectal cancer (CRC) cells with 5‐FU represses sEV export of miRNA and snRNA‐derived RNAs, but promotes export of snoRNA‐derived RNAs. Strikingly, 5‐FU treatment significantly decreased the levels of pseudouridine on both cellular and sEV small RNA profiles. In contrast, 5‐FU exposure led to increased levels of cellular small RNAs containing a variety of methyl‐modified bases. These unexpected findings show that 5‐FU exposure leads to altered RNA expression, base modification, and aberrant trafficking and localization of small RNAs. [ABSTRACT FROM AUTHOR]

Summary: Securing agricultural supplies for the increasing population without negative impacts on environment demands new crop varieties with higher yields, better quality, and stronger stress resilience. But breeding such super crop varieties is restrained by growth–defense (G–D) trade‐off. MicroRNAs (miRNAs) are versatile regulators of plant growth and immune responses, with several being demonstrated to simultaneously regulate crop growth and defense against biotic stresses and to balance G–D trade‐off. Increasing evidence also links miRNAs to the metabolism and signaling of phytohormones, another type of master regulator of plant growth and defense. Here, we synthesize the reported functions of miRNAs in crop growth, development, and responses to bio‐stressors, summarize the regulatory scenarios of miRNAs based on their relationship with target(s), and discuss how miRNAs, particularly those involved in crosstalk with phytohormones, can be applied in balancing G–D trade‐off in crops. We also propose several open questions to be addressed for adopting miRNAs in balancing crop G–D trade‐off. [ABSTRACT FROM AUTHOR]

BACKGROUND: Bombyx mori nuclear polyhedrosis virus (BmNPV), as a typical baculovirus, is the primary pathogen that infects the silkworm B. mori, a lepidopteran species. Owing to the high biological safety of BmNPV in infecting insects, it is commonly utilized as a biological insecticide for pest control. Apoptosis is important in the interaction between the host and pathogenic microorganisms. MicroRNAs (miRNAs) influence immune responses and promote stability of the immune system via apoptosis. Therefore, the study of apoptosis‐related miRNA in silkworms during virus infection can not only provide support for standardizing the prevention and control of diseases and insect pests, but also reduce the economic losses to sericulture caused by the misuse of biological pesticides. RESULTS: Through transcriptome sequencing, we identified a miRNA, miR‐31‐5p, and demonstrated that it can inhibit apoptosis in silkworm cells and promote the proliferation of BmNPV in BmE‐SWU1 cells. We identified a target gene of miR‐31‐5p, B. mori cytochrome P450 9e2 (BmCYP9e2), and demonstrated that it can promote apoptosis in silkworm cells and inhibit the proliferation of BmNPV. Moreover, we constructed transgenic silkworm strains with miR‐31‐5p knockout and confirmed that they can inhibit the proliferation of BmNPV. CONCLUSION: These data indicate that miR‐31‐5p may exert functions of inhibiting apoptosis and promoting virus proliferation by regulating BmCYP9e2. The findings demonstrate how miRNAs influence host cell apoptosis and how they are involved in the host immune system response to viruses, providing important insights into the applications of biological insecticides for pest control. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

The functional integration of biological switches with synthetic building blocks enables the design of modular, stimulus‐responsive biohybrid materials. By connecting the individual modules via diffusible signals, information‐processing circuits can be designed. Such systems are, however, mostly limited to respond to either small molecules, proteins, or optical input thus limiting the sensing and application scope of the material circuits. Here, a highly modular biohybrid material is design based on CRISPR/Cas13a to translate arbitrary single‐stranded RNAs into a biomolecular material response. This system exemplified by the development of a cascade of communicating materials that can detect the tumor biomarker microRNA miR19b in patient samples or sequences specific for SARS‐CoV. Specificity of the system is further demonstrated by discriminating between input miRNA sequences with single‐nucleotide differences. To quantitatively understand information processing in the materials cascade, a mathematical model is developed. The model is used to guide systems design for enhancing signal amplification functionality of the overall materials system. The newly designed modular materials can be used to interface desired RNA input with stimulus‐responsive and information‐processing materials for building point‐of‐care suitable sensors as well as multi‐input diagnostic systems with integrated data processing and interpretation. [ABSTRACT FROM AUTHOR]

Aim Methodology Results Conclusion The aim of this study is to investigate the expression of inflammatory biomarkers (TNF‐α, IL‐10, IL‐1β) and the pulpitis‐associated miRNA (miR‐30a‐5p and miR‐128‐3p) in pulp tissue samples from unrestored teeth with a vital normal pulp (NP), teeth with symptomatic irreversible pulpitis (IP) and in unrestored teeth with periodontal disease, unresponsive to periodontal therapy, and a vital pulp (EP).Thirty patients were included in this observational study (10 teeth with NP, 10 teeth with IP, 10 teeth with EP). Dental pulp tissues samples were collected from patients during root canal treatment (RCT). RNA was extracted and qRT‐PCR of target genes (tumour necrosis factor [TNF]‐α, interleukin [IL]‐1β, IL‐10) and miRNAs (has‐miR‐30a‐5p, has‐miR‐128‐3p) performed to assess the expression profile. Fold‐change in expression was calculated using the formula 2−(ΔCt(Exp)−ΔCt(Ctrl)). One‐way anova with post‐hoc Tukey's was used to determine significant differences between groups. The significance level was set at 5% (p < .05). All teeth were also followed up clinically for 1 year and evaluated for a range of endodontic and periodontal‐related outcomes.All investigated genes significantly increased in expression and miRNAs significantly decreased in expression in the IP and EP groups compared with the NP group (p < .05). With regards to TNF‐α and IL‐1β there were no significant differences in expression between the IP and EP groups (p > .05), whereas IL‐10 expression levels were significantly reduced in the EP compared with the IP group (p < .05). Both miR‐30a‐5p and miR‐128‐3p showed significantly reduced expression in both IP and EP lesions, compared with NP (p < .05); however, no significant differences in miRNA expression were observed between IP and EP groups (p > .05). One year after root canal treatment and periodontal maintenance, tooth mobility and probing depth were significantly reduced in the EP group (p < .05).Pulp tissues from teeth with IP and EP presented similar levels of altered inflammatory markers compared with NP. TNF‐α, IL‐10, IL‐1β cytokines and miRNAs (miR‐30a‐5p and miR‐128‐3p) are potential objective biomarkers to indicate pulpal inflammatory status, aiding diagnosis and directing clinical decision‐making. RCT may be beneficial to improve stage III periodontitis unresponsive to non‐surgical periodontal treatment, but further research is required. [ABSTRACT FROM AUTHOR]

As "non‐cellular organisms", viruses need to infect living cells to survive themselves. The virus infection must alter host's metabolisms. However, the influence of the metabolites from the altered metabolisms of virus‐infected host cells on virus‐host interactions remains largely unclear. To address this issue, shrimp, a representative species of crustaceans, is challenged with white spot syndrome virus (WSSV) in this study. The in vivo results presented that the WSSV infection enhanced shrimp glycolysis, leading to the accumulation of lactate. The lactate accumulation in turn promoted the site‐specific histone lactylation (H3K18la and H4K12la) in a p300/HDAC1/HDAC3‐dependent manner. H3K18la and H4K12la are enriched in the promoters of 75 target genes, of which the H3K18la and H4K12la modification upregulated the expression of ribosomal protein S6 kinases 2 (S6K2) in the virus‐infected hosts to promote the virus infection. Further data revealed that the virus‐encoded miR‐N20 targeted hypoxia inducible factor‐1α (HIF‐1α) to inhibit the host glycolysis, leading to the suppression of H3K18la and H4K12la. Therefore, the findings contributed novel insights into the effects and the underlying mechanism of the virus‐induced histone lactylation on the virus‐host interactions, providing new targets for the control of virus infection. [ABSTRACT FROM AUTHOR]

Background: Prostate cancer (PCa) patients with elevated level of androgen receptor (AR) correlate with higher metastatic incidence. Protein expression of AR and its target gene prostate‐specific antigen (PSA) are elevated in metastatic prostate tumors as compared to organ‐confined tumors. Androgen treatment or elevation of AR promotes metastasis of PCa in cell culture and murine model. However, under androgen depleted condition, AR suppressed cell mobility and invasiveness of PCa cells. Androgen deprivation therapy in PCa patients is associated with higher risk of cancer metastasis. We therefore investigated the dual roles of AR and miRNAs on PCa metastasis. Methods: The PC‐3AR (PC‐3 cells re‐expressing AR) and LNCaP cells were used as PCa cell model. Transwell migration and invasion assay, wound‐healing assay, zebrafish xenotransplantation assay, and zebrafish vascular exit assay were used to investigate the role of AR and androgen on PCa metastasis. Micro‐Western Array, co‐immunoprecipitation and Immunofluorescence were applied to dissect the molecular mechanism lying underneath. The miRNA array, miRNA inhibitors or plasmid, and chromatin immunoprecipitation assay were used to study the role of miRNAs on PCa metastasis. Results: In the absence of androgen, AR repressed the migration and invasion of PCa cells. When androgen was present, AR stimulated the migration and invasion of PCa cells both in vitro and in zebrafish xenotransplantation model. Androgen increased phospho‐AR Ser81 and yes‐associated protein 1 (YAP), decreased phospho‐YAP Ser217, and altered epithelial‐mesenchymal transition (EMT) proteins in PCa cells. Co‐IP assay demonstrated that androgen augmented the interaction between YAP and AR in nucleus. Knockdown of YAP or treatment with YAP inhibitor abolished the androgen‐induced migration and invasion of PCa cells, while overexpression of YAP showed opposite effects. The miRNA array revealed that androgen decreased hsa‐miR‐5001‐5p but increased hsa‐miR‐203a and hsa‐miR‐210‐3p in PC‐3AR cells but not PC‐3 cells. Treatment with inhibitors targeting hsa‐miR‐203a/hsa‐miR‐210‐3p, or overexpression of hsa‐miR‐5001‐5p decreased YAP expression as well as suppressed the androgen‐induced migration and invasion of PCa cells. Chromatin immunoprecipitation (ChIP) assay demonstrated that AR binds with promoter region of has‐miR‐210‐3p in the presence of androgen. Conclusions: Our observations indicated that miRNAs 203a/210‐3p/5001‐5p regulate the androgen/AR/YAP‐induced PCa metastasis. [ABSTRACT FROM AUTHOR]

Summary: Branch number is one of the most important agronomic traits of fruit trees such as peach. Little is known about how LncRNA and/or miRNA modules regulate branching through transcription factors.Here, we used molecular and genetic tools to clarify the molecular mechanisms underlying brassinosteroid (BR) altering plant branching.We found that the number of sylleptic branch and BR content in pillar peach ('Zhaoshouhong') was lower than those of standard type ('Okubo'), and exogenous BR application could significantly promote branching. PpTCP4 expressed great differentially comparing 'Zhaoshouhong' with 'Okubo'. PpTCP4 could directly bind to DWARF2 (PpD2) and inhibited its expression. PpD2 was the only one differentially expressed key gene in the path of BR biosynthesis. At the same time, PpTCP4 was identified as a target of miR6288b‐3p. LncRNA1 could act as the endogenous target mimic of miR6288b‐3p and repress expression of miR6288b‐3p. Three deletions and five SNP sites of lncRNA1 promoter were found in 'Zhaoshouhong', which was an important cause of different mRNA level of PpTCP4 and BR content. Moreover, overexpressed PpTCP4 significantly inhibited branching.A novel mechanism in which the lncRNA1‐miR6288b‐3p‐PpTCP4‐PpD2 module regulates peach branching number was proposed. [ABSTRACT FROM AUTHOR]

Aim: MicroRNAs (miRNAs) regulate β‐cell function, and β‐cell mitochondria and insulin secretion are perturbed in diabetes. We aimed to identify key miRNAs regulating β‐cell mitochondrial metabolism and novel β‐cell miRNA‐mitochondrial pathways. Methods: TargetScan (http://www.targetscan.org/) was used to predict if 16 miRNAs implicated in β‐cell function target 27 cis‐eGenes implicated in mitochondrial activity. The expression of candidate miRNAs and insulin secretion after 24 and 1 h pre‐incubation in 2.8, 11.1‐ and 16.7‐mM glucose was measured in clonal INS‐1 832/13 β‐cells. MiR‐29 silenced INS‐1 832/13 cells were assessed for insulin secretion (glucose, pyruvate, and K+), target cis‐eGene expression (Ndufv3 and Ndufa10 components of mitochondrial complex I (CI)), OXPHOS (CI‐V) protein expression, and mitochondrial OXPHOS respiration/activity. The expression of differentially expressed miR‐29 miRNAs was evaluated in Goto‐Kakizaki (GK) rat, db/db mouse and type 2 diabetic (T2D) human islets, as well as NMRI mouse islets cultured under glucolipotoxic conditions. Results: MiR‐29, miR‐15 and miR‐124 were predicted to regulate ~20 cis‐eGenes, while miR‐29 alone was predicted to regulate ≥12 of these in rat and human species. MiR‐29 expression and insulin secretion were reduced in INS‐1 832/13 cells after 24 h in elevated glucose. MiR‐29 knockdown increased all tested insulin secretory responses, Nudfv3, Ndufa10, complex I and II expression, and cellular mitochondrial OXPHOS. MiR‐29 expression was reduced in db/db islets but increased in GK rat and T2D human islets. Conclusion: We conclude miR‐29 is a key miRNA in regulating β‐cell mitochondrial metabolism and insulin secretion via underlying miR‐29‐OXPHOS complex pathways. Furthermore, we infer reduced miR‐29 expression compensatorily enhances insulin secretion under glucotoxicity. [ABSTRACT FROM AUTHOR]

Positron Emission Tomography (PET) is an important part of the medical imaging field which is continually exploring novel biological targets as exemplified by PET imaging of neuroinflammation. Due to limitations stemming from either sub‐optimal biological targets or a lack of available selective radiotracers, alternative biomarkers and PET imaging agent candidates are considered. One such possible target is microRNA (miRNA) and herein, we discuss the potential of miRNA for PET imaging. With the aim of addressing key strategies for imaging miRNA with PET, we identify three distinct approaches as follows: small molecules directly targeting miRNA, small molecules indirectly targeting Argonaute 2 (AGO2)‐protein complexes, and direct chemical modification of antisense oligonucleotides. The radiosynthetic approaches are based on the methods of direct radiolabelling of respective antisense oligonucleotides and several examples are described herein, showcasing the potential of miRNA in PET imaging. Whilst these approaches offer different radiolabelling strategies, application of these radiolabelled molecules towards PET imaging of miRNA are scarce with only one, limited example applied to bone remodeling reported in the literature. [ABSTRACT FROM AUTHOR]

Background and Objectives: Aged red blood cell (RBC) transfusions in lung cancer patients are often related to cancer recurrence and shorter lifespans. Extracellular vesicles (EVs) accumulated in stored RBC suspensions may be one of the important influential factors. This study aims to investigate how EVs derived from RBC suspensions affect the progress of lung cancer through the most enriched microRNAs (miRNAs) previously reported in our research. Study Design and Methods: EVs derived from stored RBC suspensions in Weeks 1, 3 and 5 were harvested via ultracentrifugation. Lung adenocarcinoma H1975 cells were co‐cultured with EVs and transfected with miR1246 and miR150‐3p mimics to evaluate alterations in their proliferation, invasion and migration abilities in vitro. Proteomics and bioinformatics were performed to predict the signalling pathway related to invasion and migration of H1975, which were verified by western blotting (WB) and flow cytometry. Results: EVs derived from stored RBC suspensions in Weeks 3 and 5 could significantly enhance the invasion and migration ability of H1975 cells and also increase the expression of miR1246 and miR150‐3p. After transfection with miR1246 and miR150‐3p mimics, invasion, migration and proliferation of H1975 cells were obviously enhanced. Proteomics analysis demonstrated that EVs co‐cultivation and miRNA transfection groups were both enriched in cell adhesion molecules. WB and cytometry indicated that integrin beta‐1 (ITGB1) and Rap1b were increased. Conclusions: EVs derived from stored RBC suspensions can enhance invasion and migration ability of lung cancer cells via the most accumulated miR1246 and miR150‐3p, which may increase the expression of ITGB1 through Rap1 signalling pathway. [ABSTRACT FROM AUTHOR]

Exosomes from cancer cells function as carriers to spread or transport specific microRNAs (miRNAs) to distant sites to exert their effects, but the mechanism of exosomal miRNA action in esophageal squamous cell carcinoma (ESCC) has not been fully explained. Therefore, in this study, we were interested in the impact of exosomal miR‐196a‐5p in ESCC progression. We found that miR‐196a‐5p was expressed enriched in clinical tissues, ESCC cells, and exosomes. Functionally, depletion of miR‐196a‐5p impeded ESCC cell growth, migration, and invasion, whereas overexpression of miR‐196a‐5p produced the opposite results. Moreover, enhancement of exosomal miR‐196a‐5p in recipient ESCC cells triggered more intense proliferation and migration. Mechanistically, we identified integral membrane protein 2B (ITM2B) as a direct target of miR‐196a‐5p. Silencing of ITM2B partially counteracted the inhibitory effect of miR‐196a‐5p inhibitors on the malignant phenotype of ESCC. Furthermore, in vivo, lower miR‐196a‐5p levels triggered by the introduction of antagomiR‐196a‐5p resulted in the generation of smaller volume and weight xenograft tumors. Thus, our results demonstrated novel mechanisms of exosomal and intracellular miR‐196a‐5p‐mediated ESCC growth and migration and identify the interaction of miR‐196a‐5p with ITM2B. These works might provide new targets and basis for the development of clinical treatment options for ESCC. [ABSTRACT FROM AUTHOR]

Currently, human papillomavirus tests and cytology are used to screen for cervical cancer. However, more accurate ancillary screening tests are needed. MicroRNAs (miRNAs) and cytokines are promising biomarkers that are aberrantly expressed in cervical cancer. Therefore, the potential of developing new screening markers based on the levels of miRNAs and cytokines in serum and local mucus samples from the same patients with cervical neoplasia was investigated. miRNA screening was performed by microarray and measurement using real‐time reverse‐transcriptase PCR. Cytokine were measured using multiplex bead assay, and changes in expressions were analyzed based on disease severity. As lesions progressed, miR‐20b‐5p, −155‐5p, −144‐3p, −451a, and −126‐3p expression levels were increased in mucus, and miR‐16‐5p, −223‐3p, and ‐451a expression levels were decreased in serum. Regarding cytokines, IL‐6, IL‐8, monocyte chemoattractant protein‐1, Eotaxin, interferon‐γ, and RANTES were increased, whereas granulocyte–colony‐stimulating factor (G‐CSF) was significantly decreased in mucus. miRNAs and cytokines in serum did not have high diagnostic accuracy. However, a combination of miR‐20b‐5p, ‐451a, ‐126‐3p, Eotaxin, as well as G‐CSF in mucus samples, had high diagnostic accuracy with an area under the receiver operating characteristic curve of 0.989 (0.979–0.999). Our results suggest that using mucus for this ancillary test is more beneficial than serum. [ABSTRACT FROM AUTHOR]

Objective: To analyze the correlation between the miRNA expression profile in vestibular schwannoma (VS) tumor tissue and preoperative patient's hearing status, using the RNA‐seq technique. Methods: Nineteen tumor samples were collected from patients operated for VS in a Tertiary Academic Center. Samples were classified into "good hearing" and "poor hearing" study group based on the results of audiometric studies. Tumor miRNA expression was analyzed using high‐throughput RNA sequencing (RNA‐seq) technique, using NovaSeq 6000 Illumina system. Functional analysis was performed with the use of DIANA miRpath v. 4.0 online tool. Results: The most overexpressed miRNAs in VS samples derived from poor hearing patients belonged to miR 449a/b, miR 15/16–1, and hypoxamiR families. Functional analysis showed that the differentially expressed miRNAs regulate cellular pathways associated with hypoxia, adherence junction functions, and signaling pathways such as Hippo, FOXO, MAPK, and Wnt signaling pathway. Conclusion: Our study identified a specific miRNA expression profile in VS tumor tissues that correlates with hearing impairment. These results suggest potential new molecular mechanisms related to hearing loss in the course of VS. Level of Evidence: 3 (cohort study) Laryngoscope, 134:3778–3785, 2024 [ABSTRACT FROM AUTHOR]

Mechanisms regulating the gene expression program at different hypoxia severity levels in patient tumors are not understood. We aimed to determine microRNA (miRNA) regulation of this program at defined hypoxia levels from moderate to severe in prostate cancer. Biopsies from 95 patients were used, where 83 patients received the hypoxia marker pimonidazole before prostatectomy. Forty hypoxia levels were extracted from pimonidazole‐stained histological sections and correlated with miRNA and gene expression profiles determined by RNA sequencing and Illumina bead arrays. This identified miRNAs associated with moderate (n = 7) and severe (n = 28) hypoxia and predicted their target genes. The scores of miRNAs or target genes showed prognostic significance, as validated in an external cohort of 417 patients. The target genes showed enrichment of gene sets for cell proliferation and MYC activation at all hypoxia levels and PTEN inactivation at severe hypoxia. This was confirmed by RT‐qPCR for MYC and PTEN, by Ki67 immunohistochemistry, and by gene set analysis in an external cohort. To assess whether miRNA regulation occurred within the predicted hypoxic regions, a method to quantify co‐localization of multiple histopathology parameters at defined hypoxia levels was applied. A high Ki67 proliferation index co‐localized significantly with hypoxia at all levels. The co‐localization index was strongly associated with poor prognosis. Absence of PTEN staining co‐localized significantly with severe hypoxia. The scores for miRNAs correlated with the co‐localization index for Ki67 staining and hypoxia, consistent with miRNA regulation within the overlapping regions. This was confirmed by showing miR‐210‐3p expression within severe hypoxia by in situ hybridization. Cell line experiments (22Rv1, PC3) were conducted to determine whether miRNAs and target genes were regulated directly by hypoxia. Most of them were hypoxia‐unresponsive, and probably regulated by other mechanisms such as MYC activation. In conclusion, in aggressive, hypoxic prostate tumors, cancer cells exhibit different proliferative gene expression programs that is regulated by miRNAs and depend on whether the cells reside in moderate or severe hypoxic regions. © 2024 The Author(s). The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland. [ABSTRACT FROM AUTHOR]

Dicer, a multi‐domain ribonuclease III (RNase III) protein, is crucial for gene regulation via RNA interference. It processes hairpin‐like precursors into microRNAs (miRNAs) and long double‐stranded RNAs (dsRNAs) into small interfering RNAs (siRNAs). During the "dicing" process, the miRNA or siRNA substrate is stably anchored and cleaved by Dicer's RNase III domain. Although numerous studies have investigated long dsRNA cleavage by Dicer, the specific mechanism by which human Dicer (hDICER) processes pre‐miRNA remains unelucidated. This review introduces the recently revealed hDICER structure bound to pre‐miRNA uncovered through cryo‐electron microscopy and compares it with previous reports describing Dicer. The domain‐wise movements of the helicase and dsRNA‐binding domain (dsRBD) and specific residues involved in substrate sequence recognition have been identified. During RNA substrate binding, the hDICER apical domains and dsRBD recognize the pre‐miRNA termini and cleavage site, respectively. Residue rearrangements in positively charged pockets within the apical domain influence substrate recognition and cleavage site determination. The specific interactions between dsRBD positively charged residues and nucleotide bases near the cleavage site emphasize the significance of cis‐acting elements in the hDICER processing mechanism. These findings provide valuable insights for understanding hDICER‐related diseases. [ABSTRACT FROM AUTHOR]

Increased malignancy and poor treatability associated with solid tumour cancers have commonly been attributed to mitochondrial calcium (Ca2+) dysregulation. The mitochondrial Ca2+ uniporter complex (mtCU) is the predominant mode of Ca2+ uptake into the mitochondrial matrix. The main components of mtCU are the pore‐forming mitochondrial Ca2+ uniporter (MCU) subunit, MCU dominant‐negative beta (MCUb) subunit, essential MCU regulator (EMRE) and the gatekeeping mitochondrial Ca2+ uptake 1 and 2 (MICU1 and MICU2) proteins. In this review, we describe mtCU‐mediated mitochondrial Ca2+ dysregulation in solid tumour cancer types, finding enhanced mtCU activity observed in colorectal cancer, breast cancer, oral squamous cell carcinoma, pancreatic cancer, hepatocellular carcinoma and embryonal rhabdomyosarcoma. By contrast, decreased mtCU activity is associated with melanoma, whereas the nature of mtCU dysregulation remains unclear in glioblastoma. Furthermore, we show that numerous polymorphisms associated with cancer may alter phosphorylation sites on the pore forming MCU and MCUb subunits, which cluster at interfaces with EMRE. We highlight downstream/upstream biomolecular modulators of MCU and MCUb that alter mtCU‐mediated mitochondrial Ca2+ uptake and may be used as biomarkers or to aid in the development of novel cancer therapeutics. Additionally, we provide an overview of the current small molecule inhibitors of mtCU that interact with the Asp residue of the critical Asp‐Ile‐Met‐Glu motif or through other allosteric regulatory mechanisms to block Ca2+ permeation. Finally, we describe the relationship between MCU‐ and MCUb‐mediating microRNAs and mitochondrial Ca2+ uptake that should be considered in the discovery of new treatment approaches for cancer. [ABSTRACT FROM AUTHOR]

The circular RNA (circRNA) Cdr1as is conserved across mammals and highly expressed in neurons, where it directly interacts with microRNA miR-7. However, the biological function of this interaction is unknown. Here, using primary cortical murine neurons, we demonstrate that stimulating neurons by sustained depolarization rapidly induces two-fold transcriptional upregulation of Cdr1as and strong post-transcriptional stabilization of miR-7. Cdr1as loss causes doubling of glutamate release from stimulated synapses and increased frequency and duration of local neuronal bursts. Moreover, the periodicity of neuronal networks increases, and synchronicity is impaired. Strikingly, these effects are reverted by sustained expression of miR-7, which also clears Cdr1as molecules from neuronal projections. Consistently, without Cdr1as, transcriptomic changes caused by miR-7 overexpression are stronger (including miR-7-targets downregulation) and enriched in secretion/synaptic plasticity pathways. Altogether, our results suggest that in cortical neurons Cdr1as buffers miR-7 activity to control glutamatergic excitatory transmission and neuronal connectivity important for long-lasting synaptic adaptations. Synopsis: In mouse cortical neurons, miR-7 regulates glutamate release, controlled by the buffering action of Cdr1as. These molecules together govern glutamatergic transmission and neuronal connectivity, influencing synaptic adaptations at transcriptional and neuronal activity levels. Neuronal depolarization transcriptionally induces Cdr1as and post-transcriptionally stabilizes miR-7 Cdr1as-KO neurons show chronically elevated neuronal activity, higher asynchrony and oscillation, increased pre-synaptic glutamate release, and inhibition of synaptic adaptation. Cdr1as:miR-7 interaction exerts a synergetic gene regulation of the neuronal transcriptome, compared to their individual mutations. Additionally, miR-7 up-regulation is sufficient to restore local and network neural activity and glutamate secretion, but it depends on Cdr1as expression. In mouse cortical neurons, miR-7 regulates glutamate release, controlled by the buffering action of Cdr1as. These molecules together govern glutamatergic transmission and neuronal connectivity, influencing synaptic adaptations at transcriptional and neuronal activity levels. [ABSTRACT FROM AUTHOR]

Chronic thromboembolic pulmonary hypertension (CTEPH) is marked by persistent blood clots in pulmonary arteries, leading to significant morbidity and mortality. Emerging evidence highlights the role of microRNAs (miRNAs) in pulmonary hypertension, though findings on miRNA expression in CTEPH remain limited and inconsistent. This systematic review evaluates miRNA expression changes in CTEPH and their direction. Following Preferred Reporting Items for Systematic Reviews and Meta‐Analyses guidelines, we registered our protocol in International Prospective Register of Systematic Reviews (CRD42024524469). We included studies on miRNA expression in CTEPH with comparative or analytical designs, excluding nonhuman studies, interventions, non‐English texts, conference abstracts, and editorials. Databases searched included PubMed, EMBASE, Scopus, CENTRAL, and ProQuest. The Quality Assessment of Diagnostic Accuracy Studies‐2 tool assessed bias risk, and results were synthesized narratively. Of 313 unique studies, 39 full texts were reviewed, and 9 met inclusion criteria, totaling 235 participants. Blood samples were analysed using quantitative real time polymerase chain reaction. Seven miRNAs (miR‐665, miR‐3202, miR‐382, miR‐127, miR‐664, miR‐376c, miR‐30) were uniformly upregulated, while nine (miR‐20a‐5p13, miR‐17‐5p, miR‐93‐5p, miR‐22, let‐7b, miR‐106b‐5p, miR‐3148, miR‐320‐a, miR‐320b) were downregulated in CTEPH patients. Two upregulated miRNAs (miR‐127 and miR‐30a) were consistently associated with previous evidence in the mechanism inducing the development of CTEPH, and five downregulated miRNAs (miR‐20‐a, miR‐17‐5p, miR‐93‐5p, let‐7b, miR‐106b‐5p) were associated with a protective effect against CTEPH. We also identified gaps in the literature where the evidence for five upregulated miRNAs (miR‐665, miR‐3202, miR‐382, miR‐664 and miR‐376c) and four downregulated miRNAs (miR‐22, miR‐3148, miR‐320‐a, and miR‐320b) in CTEPH is conflicting. Our findings offer insights into the role of miRNAs in CTEPH and underscore the need for further research to validate these miRNAs as biomarkers or therapeutic targets. [ABSTRACT FROM AUTHOR]

Background: Personalized medicine represents a novel and integrative approach that focuses on an individual's genetics and epigenetics, precision medicine, lifestyle and exposures as key players of health status and disease phenotypes. Methods: In this narrative review, we aim to carefully discuss the current knowledge on gender disparities in cardiometabolic diseases, and we consider the sex‐ specific expression of miRNAs and their role as promising tool in precision medicine. Results: Personalised medicine overcomes the restricted care of patient based on a binomial sex approach, by enriching itself with a holistic and dynamic gender integration. Recognized as a major worldwide health emergency, cardiometabolic disorders continue to rise, impacting on health systems and requiring more effective and targeted strategies. Several sex and gender drivers might affect the onset and progression of cardiometabolic disorders in males and females at multiple levels. In this respect, distinct contribution of genetic and epigenetic mechanisms, molecular and physiological pathways, sex hormones, visceral fat and subcutaneous fat and lifestyle lead to differences in disease burden and outcomes in males and females. Conclusions: Sex and gender play a pivotal role in precision medicine because the influence the physiology of each individual and the way they interact with environment from intrauterine life. [ABSTRACT FROM AUTHOR]

The hypothalamic–pituitary axis is central to the functioning of the neuroendocrine system and essential for regulating physiological and behavioral homeostasis and coordinating fundamental body functions. The expanding line of evidence shows the indispensable role of the microRNA pathway in regulating the gene expression profile in the developing and adult hypothalamus and pituitary gland. Experiments provoking a depletion of miRNA maturation in the context of the hypothalamic–pituitary axis brought into focus a prominent involvement of miRNAs in neuroendocrine functions. There are also a few individual miRNAs and miRNA families that have been studied in depth revealing their crucial role in mediating the regulation of fundamental processes such as temporal precision of puberty timing, hormone production, fertility and reproduction capacity, and energy balance. Among these miRNAs, miR‐7 was shown to be hypothalamus‐enriched and the top one highly expressed in the pituitary gland, where it has a profound impact on gene expression regulation. Here, we review miRNA profiles, knockout phenotypes, and miRNA interaction (targets) in the hypothalamic–pituitary axis that advance our understanding of the roles of miRNAs in mammalian neurosecretion and related physiology. [ABSTRACT FROM AUTHOR]

Achieving seedlessness in citrus varieties is one of the important objectives of citrus breeding. Male sterility associated with abnormal pollen development is an important factor in seedlessness. However, our understanding of the regulatory mechanism underlying the seedlessness phenotype in citrus is still limited. Here, we determined that the miR159a‐DUO1 module played an important role in regulating pollen development in citrus, which further indirectly modulated seed development and fruit size. Both the overexpression of csi‐miR159a and the knocking out of DUO1 in Hong Kong kumquat (Fortunella hindsii) resulted in small and seedless fruit phenotypes. Moreover, pollen was severely aborted in both transgenic lines, with arrested pollen mitotic I and abnormal pollen starch metabolism. Through additional cross‐pollination experiments, DUO1 was proven to be the key target gene for miR159a to regulate male sterility in citrus. Based on DNA affinity purification sequencing (DAP‐seq), RNA‐seq, and verified interaction assays, YUC2/YUC6, SS4 and STP8 were identified as downstream target genes of DUO1, those were all positively regulated by DUO1. In transgenic F. hindsii lines, the miR159a‐DUO1 module down‐regulated the expression of YUC2/YUC6, which decreased indoleacetic acid (IAA) levels and modulated auxin signaling to repress pollen mitotic I. The miR159a‐DUO1 module reduced the expression of the starch synthesis gene SS4 and sugar transport gene STP8 to disrupt starch metabolism in pollen. Overall, this work reveals a new mechanism by which the miR159a‐DUO1 module regulates pollen development and elucidates the molecular regulatory network underlying male sterility in citrus. [ABSTRACT FROM AUTHOR]

The growth of liquid biopsy, i. e., the possibility of obtaining health information by analysing circulating species (nucleic acids, cells, proteins, and vesicles) in peripheric biofluids, is pushing the field of sensors and biosensors beyond the limit to provide decentralised solutions for nonspecialists. In particular, among all the circulating species that can be adopted in managing cancer evolution, both for diagnostic and prognostic applications, microRNAs have been highly studied and detected. The development of electrochemical devices is particularly relevant for liquid biopsy purposes, and the screen‐printed electrodes (SPEs) represent one of the building blocks for producing novel portable devices. In this work, we have taken miR‐2115‐3p as model target (it is related to lung cancer), and we have developed a biosensor by exploiting the use of a complementary DNA probe modified with methylene blue as redox mediator. In particular, the chosen sensing architecture was applied to serum measurements of the selected miRNA, obtaining a detection limit within the low nanomolar range; in addition, various platforms were interrogated, namely commercial and hand‐made SPEs, with the aim of providing the reader with some insights about the optimal platform to be used by considering both the cost and the analytical performance. [ABSTRACT FROM AUTHOR]

Back pain is a global epidemiological and socioeconomic problem affecting up to 80% of people at some stage during their life and is often due to degeneration of the intervertebral disc (IVD). Therapies aimed at restoring the intradiscal space have predominantly focused on delivery of biomaterials, cells, or growth factors, among others, with variable degrees of success. While viral gene delivery strategies have emerged as promising therapeutic options in recent years, these approaches often have off‐target effects and are associated with immunogenicity risks and other comorbidities. Consequently, nonviral methods have gained traction as potential avenues for gene delivery. Herein, enhanced cell‐penetrating peptide (CPP) systems are used to deliver microRNAs in an in vitro and ex vivo model of disc degeneration. The data suggest that nanoparticle complexation of CPPs with (miR‐221‐inhibitor + miR‐149‐mimic) promotes protective effects in nucleus pulposus cells challenged with inflammatory cytokines TNF‐α and IL‐1β. Specifically, increases in matrix deposition, significant decreases in the secretion of an array of inflammatory cytokines, and decreased expression of matrix degradation enzymes MMP13 and ADAMTS5 are observed. These miR‐CPP nanocomplexes can be further employed for targeting of the pericellular matrix space through homing, thus providing a promising approach for therapies of the intradiscal space. [ABSTRACT FROM AUTHOR]

Pardosa pseudoannulata is key predator in rice ecosystems; it plays a crucial role in rice pest biological control, effectively managing major rice pests such as rice planthopper. At present, there are few related studies on spiders' microRNA (miRNA). Consequently, we initiate a study on miR‐6826‐3p in P. pseudoannulata in response to high temperatures. First, we identified a total of 686 target genes of miR‐6826‐3p, 439 target genes had annotated results in the database, primarily involved in transcription and metabolism functions. Subsequently, quantitative real‐time PCR was employed to assess the relative expression levels of miR‐6826‐3p in P. pseudoannulata under different temperatures (31, 34, 37, 40°C) and various durations (1, 2, 4, 8, 12 h). The results revealed that compared to the control group (25°C), the relative expression level of miR‐6826‐3p in female spiders significantly increased under the same temperature and different duration stresses. The expression level of miR‐6826‐3p in male spiders showed an initial increase followed by a decrease trend, with relatively lower expression levels after 1 and 12 h at the same temperature conditions. Furthermore, under the same duration of stress, compared to the control group (25°C), the relative expression level of miR‐6826‐3p in female spiders significantly increased at different temperatures. These results demonstrate that P. pseudoannulata miR‐6826‐3p plays a significant role in response to high‐temperature stress. The research findings can provide a basis for future in‐depth studies on the miRNA functional of P. pseudoannulata, aiding in the elucidation of the regulatory mechanisms of P. pseudoannulata miRNAs in response to high‐temperature environments. [ABSTRACT FROM AUTHOR]

Cardiac fibrosis is the hallmark of cardiovascular disease (CVD), which is leading cause of death worldwide. Previously, we have shown that interleukin‐10 (IL10) reduces pressure overload (PO)‐induced cardiac fibrosis by inhibiting the recruitment of bone marrow fibroblast progenitor cells (FPCs) to the heart. However, the precise mechanism of FPC involvement in cardiac fibrosis remains unclear. Recently, exosomes and small extracellular vesicles (sEVs) have been linked to CVD progression. Thus, we hypothesized that pro‐fibrotic miRNAs enriched in sEV‐derived from IL10 KO FPCs promote cardiac fibrosis in pressure‐overloaded myocardium. Small EVs were isolated from FPCs cultured media and characterized as per MISEV‐2018 guidelines. Small EV's miRNA profiling was performed using Qiagen fibrosis‐associated miRNA profiler kit. For functional analysis, sEVs were injected in the heart following TAC surgery. Interestingly, TGFβ‐treated IL10‐KO‐FPCs sEV increased profibrotic genes expression in cardiac fibroblasts. The exosomal miRNA profiling identified miR‐21a‐5p as the key player, and its inhibition with antagomir prevented profibrotic signalling and fibrosis. At mechanistic level, miR‐21a‐5p binds and stabilizes ITGAV (integrin av) mRNA. Finally, miR‐21a‐5p‐silenced in sEV reduced PO‐induced cardiac fibrosis and improved cardiac function. Our study elucidates the mechanism by which inflammatory FPC‐derived sEV exacerbate cardiac fibrosis through the miR‐21a‐5p/ITGAV/Col1α signalling pathway, suggesting miR‐21a‐5p as a potential therapeutic target for treating hypertrophic cardiac remodelling and heart failure. [ABSTRACT FROM AUTHOR]

Background: Altered T‐cell repertoire with an aberrant T‐cell activation and imbalance of the Th17/Treg cells has been reported in acquired aplastic anemia (aAA). miRNAs are well known to orchestrate T‐cell activation and differentiation, however, their role in aAA is poorly characterized. The study aimed at identifying the profile of miRNAs likely to be involved in T‐cell activation and the Th17/Treg‐cell imbalance in aAA, to explore newer therapeutic targets. Methods: Five milliliters peripheral blood samples from 30 patients of aAA and 15 healthy controls were subjected to flow cytometry for evaluating Th17‐ and Treg‐cell subsets. The differential expression of 7 selected miRNAs viz; hsa‐miR‐126‐3p, miR‐146b‐5p, miR‐155‐5p, miR‐16, miR‐17, miR‐326, and miR‐181c was evaluated in the PB‐MNCs. Expression analysis of the miRNAs was performed using qRT‐PCR and fold change was calculated by 2−ΔΔCt method. The alterations in the target genes of deregulated miRNAs were assessed by qRT‐PCR. The targets studied included various transcription factors, cytokines, and downstream proteins. Results: The absolute CD3+ lymphocytes were significantly elevated in the PB of aAA patients when compared with healthy controls (p < 0.0035), however, the CD4:CD8 ratio was unperturbed. Th17: Treg‐cell ratio was altered in aAA patients (9.1 vs. 3.7%, p value <0.05), which correlated positively with disease severity and the PNH positive aAA. Across all severities of aAA, altered expression of the 07 miRNAs was noted in comparison to controls; upregulation of miR‐155 (FC—2.174, p‐value‐0.0001), miR‐146 (FC—2.006, p‐value‐0.0001), and miR‐17 (FC—3.1, p‐value‐0.0001), and downregulation of miR‐126 (FC—0.329, p‐value‐0.0001), miR‐181c (FC—0.317, p‐value‐0.0001), miR‐16 (FC—0.348, p‐value‐0.0001), and miR‐326 (FC—0.334, p‐value‐0.0001). Target study for these miRNAs revealed an increased expression of transcription factors responsible for Th1 and Th17 differentiation (T‐bet, RORϒt, IL‐17, IL‐6, and IFN‐ϒ), T‐cell activation (NFκB, MYC, and PIK3R2), downregulation of FOX‐P3, and other regulatory downstream molecules like SHIP‐1, ETS‐1, IRAK‐1, TRAF‐6, and PTEN. Conclusion: The study for the first time highlights the plausible role of different miRNAs in deregulating the Th17/Treg‐cell imbalance in aAA, and comprehensively suggest the role of altered NF‐kB and mTOR pathways in aAA. The axis may be actively explored for development of newer therapeutic targets in aAA. [ABSTRACT FROM AUTHOR]

Neutrophils are considered 'first‐line defence' cells as they can be rapidly recruited to the site of the immune response. As key components of non‐specific immune mechanisms, neutrophils use phagocytosis, degranulation, and formation of neutrophil extracellular traps (NETs) to fight pathogens. Recently, immunoregulatory abilities of neutrophils associated with the secretion of several mediators, including cytokines and extracellular vesicles (EVs) containing, among other components, microRNAs (miRNAs), have also been reported. EVs are small structures released by cells into the extracellular space and are present in all body fluids. Microvesicles show the composition and status of the releasing cell, its physiological state, and pathological changes. Currently, EVs have gained immense scientific interest as they act as transporters of epigenetic information in intercellular communication. This review summarises findings from recent scientific reports that have evaluated the utility of miRNA molecules as biomarkers for effective diagnostics or even as start‐points for new therapeutic strategies in neutrophil‐mediated immune reactions. In addition, this review describes the current state of knowledge on miRNA molecules, which are endogenous regulators of gene expression besides being involved in the regulation of the immune response. [ABSTRACT FROM AUTHOR]

Introduction: Identification of molecular biomarkers in the saliva and serum of oral cavity cancer patients represents a first step in the development of essential and efficient clinical tools for early detection and post‐treatment monitoring. We hypothesized that molecular analyses of paired saliva and serum samples from an individual would likely yield better results than analyses of either serum or saliva alone. Materials and Methods: We performed whole‐transcriptome and small non‐coding RNA sequencing analyses on 32 samples of saliva and serum collected from the same patients with oral squamous cell carcinoma (OSCC) and healthy controls (HC). Results: We identified 12 novel saliva and serum miRNAs and a panel of unique miRNA and mRNA signatures, significantly differentially expressed in OSCC patients relative to HC (log2 fold change: 2.6–26.8; DE: 0.02–0.000001). We utilized a combined panel of the 10 top‐deregulated miRNAs and mRNAs and evaluated their putative diagnostic potential (>87% sensitivity; 100% specificity), recommending seven of them for further validation. We also identified unique saliva and serum miRNAs associated with OSCC and smoking history (OSCC smokers vs. never‐smokers or HC: log2 fold change: 22–23; DE: 0.00003–0.000000001). Functional and pathway analyses indicated interactions between the discovered OSCC‐related non‐invasive miRNAs and mRNAs and their targets, through PI3K/AKT/mTOR signaling. Conclusion: Our data support our hypothesis that using paired saliva and serum from the same individuals and deep sequencing analyses can provide unique combined mRNA and miRNA signatures associated with canonical pathways that may have a diagnostic advantage relative to saliva or serum alone and may be useful for clinical testing. We believe this data will contribute to effective preventive care by post‐treatment monitoring of patients, as well as suggesting potential targets for therapeutic approaches. [ABSTRACT FROM AUTHOR]

Protein phosphatase 2A (PP2A), a heterotrimeric holoenzyme (scaffolding, catalytic, and regulatory subunits), regulates dephosphorylation for more than half of serine/threonine phosphosites and exhibits diverse cellular functions. Although several studies on natural products and miRNAs have emphasized their impacts on PP2A regulation, their connections lack systemic organization. Moreover, only part of the PP2A family has been investigated. This review focuses on the PP2A‐modulating effects of natural products and miRNAs' interactions with potential PP2A targets in cancer and non‐cancer cells. PP2A‐modulating natural products and miRNAs were retrieved through a literature search. Utilizing the miRDB database, potential PP2A targets of these PP2A‐modulating miRNAs for the whole set (17 members) of the PP2A family were retrieved. Finally, PP2A‐modulating natural products and miRNAs were linked via a literature search. This review provides systemic directions for assessing natural products and miRNAs relating to the PP2A‐modulating functions in cancer and disease treatments. [ABSTRACT FROM AUTHOR]

Podocyte injury plays a critical role in the progression of diabetic kidney disease (DKD), but the underlying cellular and molecular mechanisms remain poorly understanding. MicroRNAs (miRNAs) can disrupt gene expression by inducing translation inhibition and mRNA degradation, and recent evidence has shown that miRNAs may play a key role in many kidney diseases. In this study, we identified miR‐4645‐3p by global transcriptome expression profiling as one of the major downregulated miRNAs in high glucose‐cultured podocytes. Moreover, whether DKD patients or STZ‐induced diabetic mice, expression of miR‐4645‐3p was also significantly decreased in kidney. In the podocytes cultured by normal glucose, inhibition of miR‐4645‐3p expression promoted mitochondrial damage and podocyte apoptosis. In the podocytes cultured by high glucose (30 mM glucose), overexpression of miR‐4645‐3p significantly attenuated mitochondrial dysfunction and podocyte apoptosis induced by high glucose. Furthermore, we found that miR‐4645‐3p exerted protective roles by targeting Cdk5 inhibition. In vitro, miR‐4645‐3p obviously antagonized podocyte injury by inhibiting overexpression of Cdk5. In vivo of diabetic mice, podocyte injury, proteinuria, and impaired renal function were all effectively ameliorated by treatment with exogenous miR‐4645‐3p. Collectively, these findings demonstrate that miR‐4645‐3p can attenuate podocyte injury and mitochondrial dysfunction in DKD by targeting Cdk5. Sustaining the expression of miR‐4645‐3p in podocytes may be a novel strategy to treat DKD. [ABSTRACT FROM AUTHOR]

Ago2 differentially regulates oncogenic and tumor-suppressive miRNAs in cancer cells. This discrepancy suggests a secondary event regulating Ago2/miRNA action in a context-dependent manner. We show here that a positive charge of Ago2 K212, that is preserved by SIR2-mediated Ago2 deacetylation in cancer cells, is responsible for the direct interaction between Ago2 and Caveolin-1 (CAV1). Through this interaction, CAV1 sequesters Ago2 on the plasma membranes and regulates miRNA-mediated translational repression in a compartment-dependent manner. Ago2/CAV1 interaction plays a role in miRNA-mediated mRNA suppression and in miRNA release via extracellular vesicles (EVs) from tumors into the circulation, which can be used as a biomarker of tumor progression. Increased Ago2/CAV1 interaction with tumor progression promotes aggressive cancer behaviors, including metastasis. Ago2/CAV1 interaction acts as a secondary event in miRNA-mediated suppression and increases the complexity of miRNA actions in cancer. Synopsis: A direct interaction between Ago2 and CAV1, mediated by the positive charge of Ago2 K212, increases the complexity of miRNA actions and modulates exosomal cargo sorting of metastatic tumors. Ago2 directly binds to the CSD of CAV1 through W199FGFHQSVRPSLWK212, the CBM of Ago2 in cancer cells. The positive charge of Ago2 lysine 212, which is preserved by SIRT-2-mediated deacetylation in cancer cells, in the CBM is required for Ago2/CAV1 interaction. Elevated Ago2/CAV1 interaction in metastatic tumors increases the plasma membrane-association of Ago2, complexity of miRNA-mediated mRNA expression, and Ago2/miRNA release via exosomes. A direct interaction between Ago2 and CAV1, mediated by the positive charge of Ago2 K212, increases the complexity of miRNA actions and modulates exosomal cargo sorting of metastatic tumors. [ABSTRACT FROM AUTHOR]

Background: Radical prostatectomy, radiotherapy, chemotherapy, and androgen‐deprivation therapy are among the most common treatment options for different forms of prostate cancer (PCa). However, making therapeutic decisions is difficult due to the lack of reliable prediction markers indicating therapy outcomes in clinical practice. The involvement of miRNAs in all mechanisms of the PCa development and their easy detection characterize them as attractive PCa biomarkers. Although there are extensive data on the role of miRNAs in PCa therapy resistance and sensitivity development, the issues of whether they could be used as a guide for therapy choice and, if so, how we can progress toward this goal, remain unclear. Thus, generalizable reviews and studies which summarize, compare, and analyze data on miRNA involvement in responses to different types of PCa therapies are required. Objectives: Data on the involvement of miRNAs in therapy responses, on the role of cross‐miRNA expression in different therapies, and on miRNA targets were analyzed in order to determine the miRNA‐related factors which can lend perspective to the future development of personalized predictors of PCa sensitivity/resistance to therapies. Materials and methods: The data available on the miRNAs associated with different PCa therapies (resistance and sensitivity therapies) are summarized and analyzed in this study, including analyses using bioinformatics resources. Special attention was dedicated to the mechanisms of the development of therapy resistance. Results and discussion: A comprehensive combined analysis of the current data revealed a panel of miRNAs that were shown to be most closely associated with the PCa therapy response and were found to regulate the genes involved in PCa development via cell proliferation regulation, epithelial‐mesenchymal transition (EMT), apoptosis, cell‐cycle progression, angiogenesis, metastasis and invasion regulation, androgen‐independent development, and colony formation. Conclusion: The selected miRNA‐based panel has the potential to be a guide for therapeutic decision making in the effective treatment of PCa. [ABSTRACT FROM AUTHOR]

MicroRNAs (miRNAs) are small noncoding RNAs that play a vital role in plant responses to abiotic and biotic stresses. Recently, it has been discovered that some primary miRNAs (pri‐miRNAs) encode regulatory short peptides called miPEPs. However, the presence of miPEPs in rice, and their functions in response to abiotic stresses, particularly stress induced by heavy metals, remain poorly understood. Here, we identified a functional small peptide (miPEP156e) encoded by pri‐miR156e that regulates the expression of miR156 and its target SPL genes, thereby affecting miR156‐mediated cadmium (Cd) tolerance in rice. Overexpression of miPEP156e led to decreased uptake and accumulation of Cd and reactive oxygen species (ROS) levels in plants under Cd stress, resulting in improved rice Cd tolerance, as observed in miR156‐overexpressing lines. Conversely, miPEP156e mutants displayed sensitivity to Cd stress due to the elevated accumulation of Cd and ROS. Transcriptome analysis further revealed that miPEP156e improved rice Cd tolerance by modulating Cd transporter genes and ROS scavenging genes. Our study provides insights into the regulatory mechanism of miPEP156e in rice response to Cd stress and demonstrates the potential of miPEPs as an effective tool for improving crop abiotic stress tolerance. Summary statement: The 5' end of the pri‐miR156e contains a short open reading frame that encodes a small peptide called miPEP156e. This peptide plays a crucial role in regulating the activity of miR156 and has an impact on cadmium tolerance and accumulation by modulating cadmium uptake and maintaining redox homoeostasis in Oryza sativa. [ABSTRACT FROM AUTHOR]

Maternal stress and glucocorticoid exposure during pregnancy have multigenerational effects on neuroendocrine function and behaviours in offspring. Importantly, effects are transmitted through the paternal lineage. Altered phenotypes are associated with profound differences in transcription and DNA methylation in the brain. In the present study, we hypothesized that maternal prenatal synthetic glucocorticoid (sGC) exposure in the F0 pregnancy will result in differences in miRNA levels in testes germ cells and sperm across multiple generations, and that these changes will associate with modified microRNA (miRNA) profiles and gene expression in the prefrontal cortex (PFC) of subsequent generations. Pregnant guinea‐pigs (F0) were treated with multiple courses of the sGC betamethasone (Beta) (1 mg kg–1; gestational days 40, 41, 50, 51, 60 and 61) in late gestation. miRNA levels were assessed in testes germ cells and in F2 PFC using the GeneChip miRNA 4.0 Array and candidate miRNA measured in epididymal sperm by quantitative real‐time PCR. Maternal Beta exposure did not alter miRNA levels in germ cells derived from the testes of adult male offspring. However, there were significant differences in the levels of four candidate miRNAs in the sperm of F1 and F2 adult males. There were no changes in miRNA levels in the PFC of juvenile F2 female offspring. The present study has identified that maternal Beta exposure leads to altered miRNA levels in sperm that are apparent for at least two generations. The fact that differences were confined to epididymal sperm suggests that the intergenerational effects of Beta may target the epididymis. Key points: Paternal glucocorticoid exposure prior to conception leads to profound epigenetic changes in the brain and somatic tissues in offspring, and microRNAs (miRNAs) in sperm may mediate these changes.We show that there were significant differences in the miRNA profile of epididymal sperm in two generations following prenatal glucocorticoid exposure that were not observed in germ cells derived from the testes.The epididymis is a probable target for intergenerational programming.The effects of prenatal glucocorticoid treatment may span multiple generations. [ABSTRACT FROM AUTHOR]

In this study, Holstein dairy cows raised in Ningxia were selected as the research object. Mammary epithelial cells (BMECs) were extracted from the milk of eight Holstein cows with significantly different milk fat expression rates and transcribed for sequencing. Bioinformatics analysis was used to analyse the correlation of fat milk percentage, and the critical miR‐2285f regulating milk fat was screened out. The target gene binding sites were predicted, and 293T cells and mammary epithelial cells were used as miRNA and target gene models for functional verification in vitro. The tissue difference of miR‐2285f Holstein cows was quantitatively analysed by transfecting miR‐2285f mimic and inhibitor. Assay (dual luciferase reporter gene assay) and quantitative real‐time PCR (quantitative real‐time PCR, qRT‐PCR), triglyceride (TAG) detection, oil red O detection of lipid droplets, Western Blot assay, Edu and Flow cytometry, The molecular regulatory effects of miR‐2285f and target gene MAP2K2 on milk fat metabolism of Holstein dairy cows were studied. The wild‐type vector and mutant vector of map2k2‐3′utr were constructed, and double luciferase reporting experiments were conducted to verify that MAP2K2 was one of the target genes of miR‐2285f. According to qRT‐PCR and Western Blot analysis, miR‐2285f mainly regulates the expression of MAP2K2 protein in BMECs at the translation level. Bta‐miR‐2285f can promote cell proliferation and slow cell apoptosis by regulating MAP2K2. Bta‐miR‐2285f can promote triglyceride (TAG) and lipid droplet accumulation in mammary epithelial cells by targeting MAP2K2. Bta‐miR‐2285f can regulate protein levels of fat milk marker gene PPARG by targeting MAP2K2. In conclusion, miR‐2285f can target the expression of the MAP2K2 gene, promote the proliferation of dairy mammary epithelial cells, inhibit cell apoptosis and regulate the milk fat metabolism in dairy mammary epithelial cells. The results of this study revealed the function of miR‐2285f in regulating the differential expression of fat milk in Holstein dairy cows at the cellular level. They provided a theoretical and experimental basis for analysing the regulation network of milk fat synthesis of Holstein dairy cows and the molecular breeding of dairy cows. [ABSTRACT FROM AUTHOR]

Hepatitis E virus (HEV) infection in pregnant women is associated with a wide spectrum of adverse consequences for both mother and fetus. The high mortality in this population appears to be associated with hormonal changes and consequent immunological changes. This study conducted an analysis of immune responses in pregnant women infected with HEV manifesting varying severity. Data mining analysis of the GSE79197 was utilized to examine differentially biological functions in pregnant women with HEV infection (P‐HEV) versus without HEV infection (P‐nHEV), P‐HEV progressing to ALF (P‐ALF) versus P‐HEV, and P‐HEV versus non‐pregnant women with HEV infection (nP‐HEV). We found cellular response to interleukin and immune response‐regulating signalings were activated in P‐HEV compared with P‐nHEV. However, there was a significant decrease of immune responses, such as T cell activation, leukocyte cell‐cell adhesion, regulation of lymphocyte activation, and immune response‐regulating signaling pathway in P‐ALF patient than P‐HEV patient. Compared with nP‐HEV, MHC protein complex binding function was inhibited in P‐HEV. Further microRNA enrichment analysis showed that MAPK and T cell receptor signaling pathways were inhibited in P‐HEV compared with nP‐HEV. In summary, immune responses were activated during HEV infection while being suppressed when developing ALF during pregnancy, heightening the importance of immune mediation in the pathogenesis of severe outcome in HEV infected pregnant women. [ABSTRACT FROM AUTHOR]

CircRNAs represent a new class of non‐coding RNAs which show aberrant expression in diverse cancers, such as gastric cancer (GC). circSTRBP, for instance, is suggested to be overexpressed in GC cells and tissues. However, the biological role of circSTRBP in the progression of GC and the potential mechanisms have not been investigated. circSTRBP levels within GC cells and tissues were measured by RT‐qPCR. The stability of circSTRBP was assessed by actinomycin D and Ribonuclease R treatment. Cell proliferation, migration, invasion and in vitro angiogenic abilities after circSTRBP knockdown were analysed through CCK‐8 assay, transwell culture system and the tube formation assay. The interaction of circSTRBP with the predicted target microRNA (miRNA) was examined by RNA immunoprecipitation and luciferase reporter assays. Xenograft tumour model was established to evaluate the role of exosomal circSTRBP in the tumour formation of GC cells. circSTRBP was upregulated in GC cells and tissues, and there was an increased level of circSTRBP in GC‐derived exosomes. circSTRBP in the exosomes enhanced GC cell growth and migration in vitro, which modulates E2F Transcription Factor 2 (E2F2) expression through targeting miR‐1294 and miR‐593‐3p. Additionally, exosomal circSTRBP promoted the tumour growth of GC cells in the xenograft model. Exosomal circSTRBP is implicated in the progression of GC by modulating the activity of miR‐1294/miR‐593‐3p/E2F2 axis. [ABSTRACT FROM AUTHOR]

Breast cancer (BC) poses a persistent global health challenge, particularly in countries with elevated human development indices linked to factors such as increased life expectancy, education, and wealth. Despite therapeutic progress, challenges persist, and the role of epitranscriptomic RNA modifications in BC remains inadequately understood. The epitranscriptome, comprising diverse posttranscriptional modifications on RNA molecules, holds the potential to intricately modulate RNA function and regulation, implicating dysregulation in various diseases, including BC. Noncoding RNAs (ncRNAs), acting as posttranscriptional regulators, influence physiological and pathological processes, including cancer. RNA modifications in long noncoding RNAs (lncRNAs) and microRNAs (miRNAs) add an extra layer to gene expression control. This review delves into recent insights into epitranscriptomic RNA modifications, such as N‐6‐methyladenosine (m6A), adenine‐to‐inosine (A‐to‐I) editing, and 5‐methylcytosine (m5C), specifically in the context of lncRNA and miRNAs in BC, highlighting their potential implications in BC development and progression. Understanding this intricate regulatory landscape is vital for deciphering the molecular mechanisms underlying BC and identifying potential therapeutic targets. Significance Statement: The role of epitranscriptomic RNA modifications in breast cancer (BC) remains inadequately understood. This review delves into recent insights into epitranscriptomic RNA modifications, such as N‐6‐methyladenosine (m6A), adenine‐to‐inosine (A‐to‐I) editing, and 5‐methylcytosine (m5C), specifically in the context of lncRNA and miRNAs in BC, highlighting their potential implications in BC development and progression. Understanding this intricate regulatory landscape is vital for deciphering the molecular mechanisms underlying BC and identifying potential therapeutic targets. [ABSTRACT FROM AUTHOR]

We found that GroEL in Porphyromonas gingivalis accelerated tumor growth and increased mortality in tumor‐bearing mice; GroEL promoted proangiogenic function, which may be the reason for promoting tumor growth. To understand the regulatory mechanisms by which GroEL increases the proangiogenic function of endothelial progenitor cells (EPCs), we explored in this study. In EPCs, MTT assay, wound‐healing assay, and tube formation assay were performed to analyze its activity. Western blot and immunoprecipitation were used to study the protein expression along with next‐generation sequencing for miRNA expression. Finally, a murine tumorigenesis animal model was used to confirm the results of in vitro. The results indicated that thrombomodulin (TM) direct interacts with PI3K/Akt to inhibit the activation of signaling pathways. When the expression of TM is decreased by GroEL stimulation, molecules in the PI3K/Akt signaling axis are released and activated, resulting in increased migration and tube formation of EPCs. In addition, GroEL inhibits TM mRNA expression by activating miR‐1248, miR‐1291, and miR‐5701. Losing the functions of miR‐1248, miR‐1291, and miR‐5701 can effectively alleviate the GroEL‐induced decrease in TM protein levels and inhibit the proangiogenic abilities of EPCs. These results were also confirmed in animal experiments. In conclusion, the intracellular domain of the TM of EPCs plays a negative regulatory role in the proangiogenic capabilities of EPCs, mainly through direct interaction between TM and PI3K/Akt to inhibit the activation of signaling pathways. The effects of GroEL on tumor growth can be reduced by inhibiting the proangiogenic properties of EPCs through the inhibition of the expression of specific miRNAs. [ABSTRACT FROM AUTHOR]

Aim: Force expression is characterized by an interplay of biological and molecular determinants that are expected to differentiate males and females in terms of maximal performance. These include muscle characteristics (muscle size, fiber type, contractility), neuromuscular regulation (central and peripheral factors of force expression), and individual genetic factors (miRNAs and gene/protein expression). This research aims to comprehensively assess these physiological variables and their role as determinants of maximal force difference between sexes. Methods: Experimental evaluations include neuromuscular components of isometric contraction, intrinsic muscle characteristics (proteins and fiber type), and some biomarkers associated with muscle function (circulating miRNAs and gut microbiome) in 12 young and healthy males and 12 females. Results: Male strength superiority appears to stem primarily from muscle size while muscle fiber‐type distribution plays a crucial role in contractile properties. Moderate‐to‐strong pooled correlations between these muscle parameters were established with specific circulating miRNAs, as well as muscle and plasma proteins. Conclusion: Muscle size is crucial in explaining the differences in maximal voluntary isometric force generation between males and females with similar fiber type distribution. Potential physiological mechanisms are seen from associations between maximal force, skeletal muscle contractile properties, and biological markers. [ABSTRACT FROM AUTHOR]

Objective: Studies have shown that the levels of pleiotrophin (PTN) are greatly elevated in the synovial fluid and cartilage in osteoarthritis. Therefore, the purpose of this study was to investigate the effect and mechanism of PTN on the chondrogenic differentiation of DPSCs in inflammatory and normal microenvironments. Materials and Methods: A lentiviral vector was used to deplete or overexpress PTN in DPSCs. The inflammatory microenvironment was simulated in vitro by the addition of IL‐1β to the culture medium. The chondrogenic differentiation potential was assessed using Alcian Blue staining and the main chondrogenic markers. A dual‐luciferase reporter assay was used to explore the relationship between miR‐137 and PTN. Results: The results showed that 0.1 ng/mL IL‐1β treatment during chondrogenic induction greatly impaired the chondrogenic differentiation of DPSCs. Supplementation with PTN and PTN overexpression inhibited chondrogenic differentiation of DPSCs, while PTN depletion promoted chondrogenic differentiation. MiR‐137 negatively regulated the expression of PTN by binding to the 3′UTR of its mRNA. Moreover, miR‐137 promoted chondrogenic differentiation of DPSCs in normal and inflammatory microenvironments. Conclusion: Our results suggest that PTN may play an inhibitory role in the chondrogenic differentiation of DPSCs in normal and inflammatory microenvironments, which is regulated by miR‐137. [ABSTRACT FROM AUTHOR]

SUMMARY: Small RNAs play important roles in regulation of plant development and response to various stresses. Northern blot is an important technique in small RNA research. Isotope‐ and biotin‐ (or digoxigenin) labeled probes are frequently used in small RNA northern blot. However, isotope‐based probe is limited by strict environmental regulation and availability in many places in the world while biotin‐based probe is usually suffered from low sensitivity. In this study, we developed a T4 DNA polymerase‐based method for incorporation of a cluster of 33 biotin‐labeled C in small RNA probe (T4BC33 probe). T4BC33 probe reaches similar sensitivity as 32P‐labeled probe in dot blot and small RNA northern blot experiments. Addition of locked nucleic acids in T4BC33 probe further enhanced its sensitivity in detecting low‐abundance miRNAs. With newly developed northern blot method, expression of miR6027 and miR6149 family members was validated. Northern blot analysis also confirmed the successful application of virus‐based miRNA silencing in pepper, knocking down accumulation of Can‐miR6027a and Can‐miR6149L. Importantly, further analysis showed that knocking‐down Can‐miR6027a led to upregulation of a nucleotide binding‐leucine rich repeat domain protein coding gene (CaRLb1) and increased immunity against Phytophthora capsici in pepper leaves. Our study provided a highly sensitive and convenient method for sRNA research and identified new targets for genetic improvement of pepper immunity against P. capsici. Significance Statement: A highly sensitive and convenient method is developed for synthesis of biotin‐based probe and validated in small RNA northern blot. Can‐miR6027a is identified as a negative regulator of immunity against Phytophthora capsici in pepper using virus‐based miRNA silencing technology in conjunction with newly developed northern blot method. [ABSTRACT FROM AUTHOR]