MiRNA expression profiles may serve as valuable biomarkers for early cancer diagnosis. However, the detection of miRNA remains greatly challenging due to its shorter length and lower abundance in cells. Electrochemical biosensors based on DNA nanostructures have attracted significant attention due to their improved capture efficiency, high sensitivity, and ease of miniaturization, but the complex construction process, non-specific interactions among DNA probes, and their low stability continue to severely restrict their widespread application in clinical diagnostics. Therefore, developing robust and sensitive methods for miRNA analysis is still of great value.

We describe an ultra-sensitive electrochemical biosensor for miRNA by integrating the PNA-DNA2 three-way junction (3WJ) nanostructure with target-recycling catalytic hairpin assembly (CHA) and hybridization chain reaction (HCR) dual cascade isothermal amplification. The target miRNA triggers CHA amplification, resulting in the generation of substantial quantities of double-stranded DNA (dsDNA) products. These dsDNA are subsequently captured by PNA probes immobilized on the electrode surface, leading to the formation of a densely packed layer of PNA-DNA2 3WJ nanostructure. Subsequently, the single-stranded termini of the two branches extending from 3WJ function as promoters to initiate the HCR, resulting in the formation of a layer of intricately intertwined branched long dsDNA molecules, which could adsorb substantial quantities of [Ru(NH3)6]3+, thereby significantly amplifying the electrochemical signal. This electrochemical biosensor exhibits exceptional sensitivity towards miRNA-21, achieving a detection limit as low as 2.9 aM while effectively discriminating single base mutations.

This represents the first electrochemical system for miRNA detection by integrating the PNA-DNA2 3WJ with CHA-HCR dual cascade isothermal amplification. The robust, specific and ultrasensitive feature makes cancer cell miRNA monitoring possible, suggesting its great application prospect as a promising sensing platform for monitoring various miRNA biomarkers in cancer diagnostics.

Vitiligo is a common skin depigmentation condition caused by selective destruction of melanocytes. It is regarded as a polygenic disorder. In addition to protein-coding loci, non-coding regions of the genome contribute to the pathogenesis of vitiligo. A bulk of evidence highlights contribution of different classes of non-coding RNAs in this condition. Expression profile of different non-coding RNAs has been evaluated in the plasma, serum, blood cells and skin samples of patients with vitiligo. Notably, these transcripts not only partake the pathogenesis of vitiligo, but also are regarded as putative targets for prospective treatment strategies for this disorder. The current review focuses on depicting the role of miRNAs, long non-coding RNAs and circular RNAs in the etiology of vitiligo. Moreover, we discuss the shared functions of these transcripts in the pathogenesis of vitiligo and melanoma.

Hepatocellular carcinoma (HCC) is the most fatal cancer that has affected both male and female populations globally. With poor diagnosis and patient survival rates, it has become a global need for scientists to come to the aid. The main objective of the study was to profile the miRNAs in the serum of Control and DEN-treated mice at different time intervals (4 Weeks, 8 Weeks, 12 Weeks, and 16 Weeks) and identify HCC-associated miRNA as putative early biomarkers along with the miRNA regulated candidate gene which may be involved in HCC. Our study group involves 4,8,12, & 16 weeks 16-week-old treated male mice. Each group was sacrificed and analyzed for the stages of HCC. We employed in silico techniques for the small RNA-Seq and bioinformatics pipeline for further analysis. Our analysis revealed over 400 differentially expressed miRNAs in each treated sample and 10 novel miRNAs. The downstream analysis of these differentially expressed miRNAs, and their target genes opened an arena of different biological processes and pathways that these miRNAs affect during the development of HCC. The work has a promising role as the miRNAs predicted through this study can be used as biomarkers for early detection of HCC.

Alzheimer's disease (AD) is a neurological disorder that affects cognition and behavior, accounting for 60-70 % of dementia cases. Its mechanisms involve amyloid aggregates, hyperphosphorylated tau tangles, and loss of neural connections. Current treatments have limited efficacy due to a lack of specific targets. Recently, microRNAs (miRNAs) have emerged as key modulators in AD, regulating gene expression through interactions with mRNA. Dysregulation of specific miRNAs contributes to disease progression by disrupting clearance pathways. Antisense oligonucleotide (ASO)-based therapies show promise for AD treatment, particularly when combined with miRNA mimics or antagonists, targeting complex regulatory networks. However, miRNAs can interact with each other, complicating cellular processes and potentially leading to side effects. Our review emphasizes the role of miRNAs in regulating amyloid-beta (Aβ) clearance and highlights their potential as therapeutic targets and early biomarkers for AD, underscoring the need for further research to enhance their efficacy and safety.

Endometritis is a significant cause of infertility in mare. Some infectious agents disrupt the endometrium's innate immune system, resulting in a prolonged systemic inflammatory response that circulates via the blood or cellular degeneration, which ultimately leads to endometritis from bacterial endotoxins. Numerous biological processes use various small, non-coding RNA molecules called MicroRNAs. MicroRNAs (miRNAs) regulate gene expression after transcription by blocking transcription and translation. This manuscript examines patho-morphological discoveries in equine endometritis, the expression and effects of eca-miR-17, eca-miR-223, eca-miR-200a, eca-miR-155, and eca-miR-205, and the therapeutic function of miRNA in endometritis. MiRNAs play a crucial role in controlling inflammatory disorders by modulating cytokine signaling pathways. This review emphasizes the demand for cutting-edge genetic technologies and the development of novel pharmaceutical preparations to improve our understanding of the genes encoding by these miRNAs. It also focuses on the efficacy of miRNAs for control, early diagnosis, and prevention of endometritis.

Background: Cocaine and illicit amphetamines (disguised as "Adderall") are being laced with fentanyl and producing accidental and intentional fatal overdoses. Vaccines can prevent these overdoses, but 33% of humans generate insufficient anti-drug antibody (AB) levels. Plasma microRNAs (miRs) can be used to predict non-responders. We have plasma stored from 152 cocaine vaccine trial participants following three vaccinations over 9 weeks and examined miRs as potential response biomarkers. Methods: We compared 2517 miRs before anti-cocaine vaccination in participants with the highest (n = 25) to the lowest (n = 23) antibody levels. False Discovery Rates (FDRs) were applied to identify differentially expressed (DE) miRs. We used miR target prediction pipelines to identify the miR-regulated genes. Results: Using a DE-FDR < 0.05 and a >3-fold difference between high- and low-AB responders yielded 12 miRs down and 3 miRs up compared to low-AB patients. Furthermore, 11 among 1673 genes were targeted by 3 or more of the 12 down DE-miRs. Conclusions: A significant DE-miR for identifying optimal antibody responders replicated previous vaccine study predictors (miR-150), and several more miRs appear to be strong candidates for future consideration in replications based upon significance of individual DE-miRs and upon multiple miRs converging on individual genes.

Radon, a radioactive gas, is a significant risk factor for lung cancer, especially in non-smokers. This study examines the expression of exosomal microRNAs (miRNAs) as potential biomarkers for radon-induced effects.

A total of 109 participants from high- and low-radon areas in Kazakhstan were included. Exosomal hsa-miR-125b-5p and hsa-miR-320c levels were quantified using real-time PCR.

Results revealed a 25.4-fold increase in hsa-miR-125b-5p and a 12.5-fold decrease in hsa-miR-320c in participants exposed to high-radon levels compared to controls. Bioinformatic analysis identified key target genes, such as PRDM1 and IRF4, which are implicated in cancer development.

These findings suggest that exosomal miRNAs could serve as non-invasive biomarkers for radon exposure, offering potential for early diagnosis and monitoring of radon-induced lung cancer. The study underscores the need for further research to validate these miRNAs as reliable diagnostic tools.

MicroRNAs are abundant in serum and have emerged as important regulators of gene expression, implicating them in a wide range of diseases. The purpose of this study was to discover and validate serum miRNAs in prediabetes associated with alcohol dependence syndrome (ADS).

Serum samples from ADS patients with or without prediabetes and normoglycemic controls were subjected to microarray. Validation of identified candidate miRNAs was performed by RT-qPCR. Additionally, GO and KEGG pathway analyses were carried out to uncover target genes anticipated to be controlled by the candidate miRNAs.

Notably, 198, and 172 miRNAs were differentially expressed in ADS-patients with or without prediabetes compared to healthy controls, and 7 miRNAs in ADS-patients with prediabetes compared to ADS-normoglycemic patients, respectively. Furthermore, hsa-miR-320b and hsa-miR-3135b were differentially expressed exclusively in ADS-patients with prediabetes, and this was further validated. Interestingly, GO and KEGG pathway analysis revealed that genes predicted to be modulated by the candidates were considerably enriched in numerous diabetes-related biological processes and pathways.

Our findings revealed that ADS-patients with or without prediabetes have different sets of miRNAs compared to normoglycemic healthy subjects. We propose serum hsa-miR-320b and hsa-miR-3135b as potential biomarkers for the diagnosis of prediabetes in ADS-patients.

Non-coding small molecule RNAs are associated with a variety of diseases, including infectious diseases. However, small RNA-related studies in onychomycosis have not been reported. The aim of the present study was to conduct an initial investigation of small RNA in onychomycosis. The present study collected a total of 33 affected nail samples from patients with onychomycosis and 18 normal nail samples from healthy people. Through RNA sequencing, 37 differentially expressed microRNAs (miRNAs or miRs), including 15 upregulated and 22 downregulated miRNAs, were identified in 3 patients with onychomycosis compared with 3 healthy controls. Moreover, three differentially expressed miRNAs were analyzed for further verification by RT-qPCR in other 30 affected nail and 15 healthy nail samples. Among the three verified miRNAs, a significant difference between the downregulated hsa-miR-1-3p and hsa-miR-361-3p was observed (P<0.05). A total of 14,511 target genes of 37 differentially expressed miRNAs were predicted by the miRanda and RNAhybrid databases, while the Kyoto Encyclopedia of Genes and Genomes and Gene Ontology analysis showed that these target genes were enriched in multiple signaling pathways. The present study indicated that hsa-miR-1-3p and hsa-miR-361-3p may be potential biomarkers for onychomycosis. Furthermore, the findings of the present study can be used in future research on RNA in onychomycosis.

Early diagnosis and timely management are critical for determining disease outcomes and prognoses. To date, certain methods for developing disease-specific biomarkers have been reported; however, strategies for musculoskeletal disease-specific biomarker development have rarely been studied. Recent studies have highlighted the potential application of extracellular vesicles (EVs) as disease-specific biomarkers. EVs encapsulate proteins, lipids, messenger RNAs, and microRNAs derived from their cellular origin; these constituents remain stable within the EVs and can traverse the blood-brain barrier. Because of these distinctive characteristics, EVs have been actively investigated as diagnostic tools for various conditions, including cancer, inflammatory diseases, and musculoskeletal disorders. Although EVs have many advantages for biomarker development, they have not yet been fully researched in the context of musculoskeletal pathologies. The current review aimed to highlight the potential of EVs in the development of disease-specific biomarkers, summarize the processes of EV biomarkers, and discuss current limitations and future perspectives of EVs as biomarkers.

MicroRNAs (miRNAs), which are small non-coding RNAs, are recognized as important significant endogenous bio-molecules that regulate the post-transcriptional processes of target genes. However, predictive methods for significantly working miRNAs are poorly understood. The present study aimed to establish a novel method, miRNA protein analysis of integrative relationship (miR-PAIR), for the identification of effectively working miRNAs involved in physiological or pathological events. To establish the miR-PAIR, comprehensive expression data of miRNAs and proteins were obtained using small RNA-sequence and quantitative proteomics approach in the alveolar epithelial cell line, A549 treated with bleomycin (BLM) and methotrexate (MTX) as pulmonary toxic drugs. Differentially expressed miRNAs and proteins were integrated using TargetScan, a freely available web tool for predicting the target gene of miRNAs. Next, the enrichment of the integrated miRNA-protein pairs was analyzed, followed by the determination of significantly working miRNAs in BLM- and MTX-induced protein expression changes. The miR-PAIR method identified 22 downregulated and 9 upregulated miRNAs. Among them, miR-493-5p (p = 1.71E-05), an upregulated miRNA, suppressed approximately 70 % of the target proteins, and miR-598-3p (p = 1.1E-03), a downregulated miRNA, canceled 50 % of the target protein expression changes induced by BLM and MTX. Thus, a miR-PAIR could be an effective method to identify significantly working miRNAs associated with biological events such as drug-induced lung injury.

microRNA-22 (miR-22) plays a pivotal role in the regulation of metabolic processes and has emerged as a therapeutic target in metabolic disorders, including obesity, type 2 diabetes, and metabolic-associated liver diseases. While miR-22 exhibits context-dependent effects, promoting or inhibiting metabolic pathways depending on tissue and condition, current research highlights its therapeutic potential, particularly through inhibition strategies using chemically modified antisense oligonucleotides. This review examines the dual regulatory functions of miR-22 across key metabolic pathways, offering perspectives on its integration into next-generation diagnostic and therapeutic approaches while acknowledging the complexities of its roles in metabolic homeostasis.

Cholesterol is vital for nerve processes. Changes in cholesterol homeostasis lead to neurodegeneration and Alzheimer's disease (AD). In recent years, extensive research has confirmed the influential role of adipose tissue mesenchymal stem cells (MSCs) in managing AD. The present study aims is to investigate a new approach concerning AD by MSCs with particular reference to the cholesterol homeostasis pathway and its regulatory miRNAs in an AD-like rat model. Three groups of 24 male Wistar rats have been divided: healthy rats (control), Alzheimer's rats (AD), and Alzheimer's rats that received MSCs (AD + MSC). Cholesterol level was measured using the GC-mass technique. The mRNA and expression levels of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), apolipoprotein E (APOE), ATP-binding cassette transporter A1 (ABCA1), and CYP46A1 genes, as well as their regulating miRNAs, were assessed using real-time polymerase chain reaction (RT-PCR) and western blotting techniques, respectively. Intraventricular transplantation of MSCs improved behavioral disorders and decreased the count of Aβ plaques in brain tissue. Transplantation of these cells also led to a significant decrease in cholesterol levels and HMGCR, ApoE, and ABCA1 and a remarkable increase in CYP46A1 mRNAs and protein expression. These cells considerably changed the expression of microRNAs regulating these genes. These results indicated that the examined miRNAs could be used as promising biomarkers for AD management. Additionally, the potential therapeutic role of MSCs in improving cholesterol levels the expression levels of the targeted miRNAs and their related genes in the cholesterol homeostasis pathway was established.

Gliomas, particularly glioblastomas, have grim prognoses, necessitating early diagnostic and prognostic indicators. MicroRNAs (miRNAs), influential in cancer research, show potential as glioma biomarkers. This systematic review aimed to examine the efficacy of miRNAs in the diagnosis and prognosis of high-grade glioma.

A comprehensive search was conducted of PubMed, EMBASE, Cochrane Library, and Web of Science for studies published from 2013 to 2023. The eligibility criteria included high-grade glioma, histopathological confirmation, miRNA samples from cerebrospinal fluid or plasma, and relevant outcome data. Studies were excluded if they were experimental or reviews and not in English.

Of the 1120 initial results, 8 studies involving 660 subjects met the inclusion criteria. Several studies have assessed miRNA expression and its association with diagnosis and prognosis of high-grade gliomas. Overexpression of miR-221, miR-222, miR-210, miR-21, miR-125b, and miR-223 and under-expression of miR-15b and miR-124-3p showed significant potential in differentiating high-grade glioma patients from controls. Additionally, miRNAs are associated with distinct tumorigenic pathways.

Elevated or depressed expression levels of specific circulating miRNAs hold significant promise as noninvasive biomarkers for the diagnosis and prognosis of high-grade glioma. These miRNAs offer valuable insights into disease progression and patient outcome. Further validation through extensive clinical trials and in-depth mechanistic studies is essential to realize their full clinical utility.

Alzheimer's disease (AD) lacks a less invasive and early detectable biomarker. Here, we investigated the biomarker potential of miR-501-3p and miR-502-3p using different AD sources.

MiR-501-3p and miR-502-3p expressions were evaluated in AD CSF exosomes, serum exosomes, familial and sporadic AD fibroblasts and B-lymphocytes by qRT-PCR analysis. Further, miR-501-3p and miR-502-3p expressions were analyzed in APP, Tau cells and media exosomes.

MiR-501-3p and miR-502-3p expressions were significantly upregulated in AD CSF exosomes relative to controls. MiRNA levels were high in accordance with amyloid plaque and NFT density in multiple brain regions. Similarly, both miRNAs were elevated in AD and MCI serum exosomes compared to controls. MiR-502-3p expression was high in fAD and sAD B-lymphocytes. Finally, miR-501-3p and miR-502-3p expression were elevated intracellularly and secreted extracellularly in response to APP and Tau pathology.

These results suggest that miR-501-3p and miR-502-3p could be promising biomarkers for AD.

Hirschsprung's disease (HSCR) is characterized by congenital absence of ganglion cells in the gastrointestinal tract, which leads to impaired defecation, constipation and intestinal obstruction. The current diagnosis of HSCR is based on Rectal Suction Biopsies (RSBs), which could be complex in newborns. Occasionally, there is a delay in diagnosis that can increase the risk of clinical complications. Consequently, there is room for new non-invasive diagnostic methods that are objective, more logistically feasible and also deliver a far earlier base for a potential surgical intervention. In recent years, microRNA (miRNA) has come into the focus as a relevant early marker that could provide more insights into the etiology and progression of diseases. Therefore, in the search of a non-invasive HSCR biomarker, we analyzed miRNA expression in urine samples of HSCR patients. Results from 5 HSCR patients using microarrays, revealed hsa-miR-378 h, hsa-miR-210-5p, hsa-miR-6876-3p, hsa-miR-634 and hsa-miR-6883-3p as the most upregulated miRNAs; while hsa-miR-4443, hsa-miR-22-3p, hsa-miR-4732-5p, hsa-miR-3187-5p, and hsa-miR-371b-5p where the most downregulated miRNAs. Further search in miRNAwalk and miRDB databases showed that certainly most of these dysregulated miRNAs identified target HSCR associated genes, such as RET, GDNF, BDNF, EDN3, EDNRB, ERBB, NRG1, SOX10; and other genes implied in neuronal migration and neurogenesis. Finally, we could also validate some of these miRNA changes in HSCR urine by RT-qPCR. Altogether, our analyzed HSCR cohort presents a dysregulated miRNA expression presents that can be detected in urine. Our findings open the possibility of using specific urine miRNA signatures as non-invasive HSCR diagnosis method in the future.

There is no diagnostic test for primary immune thrombocytopenia (ITP). Certain microRNAs have shown to have diagnostic potential in ITP. We validated 12 microRNAs identified from two previous studies to find a diagnostic biomarker. The study included two ITP cohorts (n = 61) and healthy controls (n = 28). The first ITP cohort involved 24 patients from the Prolong study, patients with newly diagnosed/persistent ITP (<1 year) treated with corticosteroids ± IVIG but relapsed/failed to respond. The second cohort comprised 37 patients from ITP biobank, Østfold Hospital, Norway, patients had different disease stages and therapies. Twelve microRNAs were measured: miR-199a-5p, miR-33a-5p, miR-195-5p, miR-130a-3p, miR-144-3p, miR-146a-5p, miR-222-3p, miR-374b-5p, miR-486-5p, miR-1341-5p, miR-766-3p and miR-409-3p. miR-199a-5p, miR-33a-5p, miR-374b-5p, miR-146a-5p and miR-409-3p were expressed differentially in the entire ITP cohort compared to controls; of those only miR-199a-5p showed good discriminative ability between ITP and controls with area under the curve (AUC) of 0.718 (95% CI: 0.599-0.836). In the Prolong cohort (ITP < 1 year), miR-199a-5p and miR-374b-5p showed very good discriminative ability between ITP and controls with AUC of 0.824 (0.708-0.940) and 0.806 (0.688-0.924) respectively. This study confirmed that miR-199a-5p has good discriminative ability between primary ITP and healthy controls, thus may be a diagnostic biomarker of ITP.

Patients with estrogen receptor-positive breast cancer undergoing continuous adjuvant hormone therapy often experience delayed recurrence with tamoxifen use, potentially causing adverse effects. However, the lack of biomarkers hampers patient selection for extended endocrine therapy. This study aimed to elucidate the molecular mechanisms underlying delayed recurrence and identify biomarkers. When miRNA expression was assessed in luminal breast cancer tissues with and without delayed recurrence using NanoString, a significant increase in the expression of miR483-3p was observed in samples from patients with delayed recurrence compared with those without. miR483-3p expression was elevated in tamoxifen resistant (TAMR) EFM19 cells than in non-resistant EFM19 cells. Notably, genes associated with cancer metastasis (AMOTL2, ANKRD1, CTGF, and VEGF) were upregulated in TAMR EFM19 cells, although cell motility and proliferation were reduced. Transfection of miR483-3p mimics into both non-resistant EFM19 and MCF7 cells resulted in increased expression of cancer metastasis-related genes, but decreased proliferation and migration. Given that miR483-3p can bind to the 3'UTR region of O-GlcNAc transferase (OGT) and potentially affect its protein expression, we examined OGT protein levels and found that transfection with miR483-3p mimics selectively reduced OGT expression. Overall, breast cancer cells subjected to long-term hormone therapy displayed elevated miR483-3p expression, reducing motility and dormancy induction via decreased OGT expression. These findings suggest that miR483-3p is a potential biomarker for long-term endocrine therapy.

Dysregulated levels of cytokines may lead to cytokine storm, which has been implicated in the immunopathogenesis of coronavirus disease 2019 (COVID-19). Here in the current study, the role of microRNA (miR)-155-5p, miR-146a, and miR-221-3p in the regulation of the immune responses and inflammatory state in patients with COVID-19 was investigated.

In this case-control study, peripheral blood samples were obtained from 75 COVID-19 subjects and 100 healthy controls. From the plasma samples, RNA was extracted and cDNA was synthesized, and subsequently the transcript level of miRNAs was measured by Real-time PCR. The plasma levels of interleukin (IL)-4 and interferon (IFN)-γ were determined using ELISA.

miR-155-5p (fold change = 1.87, P = 0.020) and miR-221-3p (fold change = 2.26, P = 0.008), but not miR-146a, was upregulated in the plasma sample of COVID-19 cases compared to controls. The level of IFN-γ (but not IL-4) was significantly higher in the plasma samples of COVID-19 patients compared to control group. The expression level of miR-155-5p (r = 0.35, corrected P = 0.066) and miR-221-3p (r = 0.25, corrected P = 0.066) had positive correlation with the plasma levels of IFN-γ.

IFN-γ pathway in involved in the pathogenesis of COVID-19 that is regulated through miR-155-5p and miR-221-3p. These miRNAs showed potential utility as biomarkers for predicting the severity of COVID-19.

Pregnancy complicated by maternal obesity contributes to an increased cardiovascular risk in offspring, which is increasingly concerning as the rates of obesity and cardiovascular disease are higher than ever before and still growing. There has been much research in humans and preclinical animal models to understand the impact of maternal obesity on offspring health. This review summarizes what is known about the offspring cardiovascular phenotype, describing a mechanistic role for oxidative stress, metabolic inflexibility, and mitochondrial dysfunction in mediating these impairments. It also discusses the impact of secondary postnatal insults, which may reveal latent cardiovascular deficits that originated in utero. Finally, current interventional efforts and gaps of knowledge to limit the developmental origins of cardiovascular dysfunction in offspring of obese pregnancy are highlighted.

The coronavirus disease of 2019 (COVID-19), caused by the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), can alter the expression levels of host microRNAs (miRNAs). Increasing evidence suggests that circulating miRNAs can potentially play an important role in the diagnosis and prognosis of respiratory infectious diseases, especially COVID-19, and might serve as sensitive indicators of disease before the emergence of clinical symptoms. Here, we review the potential of circulatory microRNAs as novel biomarkers for different aspects of COVID-19. Recent studies have suggested that they can be useful not only for COVID-19 prognosis but also for prediction of disease severity and mortality among intensive care unit (ICU) and ward patients. Moreover, extracellular vesicle (EV) miRNAs can be associated with antibody titer after COVID-19 vaccination. This review provides an overview of miRNA-based biomarkers.

In the personalized medicine era, affordable and portable devices for quicker cancer monitoring, even in remote areas, are crucial. To address this need, we have developed an enzyme-assisted electrochemical point-of-care (POC) test for application toward liquid biopsy. In particular, miR-200a-5p has been taken into account as the model target due to its correlation for triple negative breast cancer (TNBC) prognosis. The proposed platform has been conceived as signal-ON, and the detection architecture is based on the presence of a duplex-specific nuclease (DSN) that is selective for DNA-RNA heteroduplexes. When the miRNA is recognized by an ad-hoc designed DNA probe, the DSN enzyme enables for the isothermal target recycling and signal enhancement, which is essential for detecting the miRNA trace in biofluids. Introducing a methylene blue (MB) modification on the DNA probe, a single miRNA strand is capable of triggering multiple DSN cleavage circles, increasing the free MB and thus the electrochemical signal recorded. All the optimization studies have been carried out using a screen-printed strip, resulting in a dynamic range comprised between 0.1 pM and 100 nM and a detection limit down to the fM level. A satisfactory selectivity was highlighted by interrogating the system toward random miRNA target mixtures, and the platform was also tested in spiked commercial serum samples.

Vascular aging refers to a series of processes where the elasticity of blood vessels diminishes, leading to stiffening, and deposition of fat components on the vessel walls, causing inflammation. Cardiovascular diseases, such as stroke and hypertension, play significant roles in morbidity and mortality rates among the elderly population. In this study, the Reactive Hyperemia Index (RHI) was measured to assess vascular endothelial function and aging-induced pathogenesis of vascular diseases in Korean subjects. We aimed to identify extracellular vesicle microRNAs (EV-miRNAs) with differential abundance between groups of individuals at the ends of a continuum in vascular aging acceleration, revealing miRNAs regulating genes in endocrine hormone regulation and tumor-related pathways. We also discovered that the principal component characterizing the global miRNA expression profile is significantly associated with clinical traits including cholesterol levels. Together, these data provide a foundation for understanding the role of miRNAs as modulators of longevity and for developing age-specific epigenetic biomarkers.

Placenta previa is the abnormal implantation of the placenta into the lower segment of the uterus, is associated with adverse maternal and fetal outcomes such as placenta accreta spectrum disorders, antepartum and postpartum hemorrhage, fetal growth restriction, prematurity, stillbirth and neonatal death, thrombophlebitis, and septicemia. The aim of the study was to assess retrospectively how the later onset of placenta previa affects the microRNA expression profile in the whole peripheral blood during the first trimester of gestation.

Regarding the occurrence of the association between aberrant microRNA expression profiles at early stages of gestation and later onset of various pregnancy-related complications, we selected for the study pregnancies developing placenta previa as the only pregnancy-related disorder. In total, 24 singleton pregnancies diagnosed with placenta previa that underwent first-trimester prenatal screening and delivered on-site within the period November 2012-May 2018 were included in the study. Overall, 80 normal pregnancies that delivered appropriate-for-gestational age newborns after completing 37 weeks of gestation were selected as the control group based on the equality of the length of biological sample storage.

Downregulation of multiple microRNAs (miR-20b-5p, miR-24-3p, miR-26a-5p, miR-92a-3p, miR-103a-3p, miR-130b-3p, miR-133a-3p, miR-145-5p, miR-146a-5p, miR-155-5p, miR-181a-5p, miR-195-5p, miR-210-3p, miR-342-3p, and miR-574-3p) was observed in pregnancies destined to develop placenta previa. The combination of seven microRNAs (miR-130b-3p, miR-145-5p, miR-155-5p, miR-181a-5p, miR-210-3p, miR-342-3p, and miR-574-3p) showed the highest accuracy (AUC 0.937, p < 0.001, 100.0% sensitivity, 83.75% specificity) to differentiate, at early stages of gestation, between pregnancies with a normal course of gestation and those with placenta previa diagnosed in the second half of pregnancy. Overall, 75% of pregnancies destined to develop placenta previa were correctly identified at 10.0% FPR.

Consecutive large-scale analyses must be performed to verify the reliability of the proposed novel early predictive model for placenta previa occurring as the only pregnancy-related disorder.

Circulating plasma miRNAs have emerged as potential early predictors of glucometabolic disorders. However, their biomarker potential remains unvalidated in populations with diverse genetic backgrounds, races, and ethnicities. This study aims to validate the biomarker potential of plasma miR-9, miR-29a, miR-192, and miR-375 for early detection of prediabetes and type 2 diabetes mellitus (T2DM) in Nepali populations that represent distinct genetic backgrounds, races, and ethnicities. A total of 46 adults, categorized into healthy controls (n = 25), prediabetes (n = 9), and T2DM (n = 12) groups, were enrolled. Baseline sociodemographic, anthropometric, and clinical characteristics were collected. Fold change in plasma expression of all four miRNAs was quantified using RT-qPCR against the RNU6B reference gene. Their biomarker potential was determined by receiver operating characteristic (ROC) curve analysis. Multivariate discriminant function and hierarchical cluster analyses were used to evaluate the effectiveness of the miRNA panel in reclassifying study participants who were initially categorized according to their glucose tolerance status. Plasma expression of all four miRNAs was significantly upregulated in T2DM patients compared to normoglycemic controls. Furthermore, the expression of only miR-29a and miR-375 was upregulated in T2DM patients than in prediabetic individuals. Notably, only miR-192 expression was significantly upregulated in prediabetic individuals than in the normoglycemic controls. The miRNA expression profiles had the potential of reclassifying the participants into three original groups with an accuracy of 69.6 %. ROC curve analysis identified miR-192 as the predictor for both prediabetes and T2DM, while miR-9, miR-29a, miR-192, and miR-375 were predictive only for T2DM. The specific set of miRNA combinations significantly improved their predictive accuracy. This study validates the early predictive biomarker potential of plasma miR-9, miR-29a, miR-192, and miR-375 also in the Nepali population and paves the way for future translational studies to validate their utility in clinical laboratories.

Disorders of glucose and lipid metabolism are an important cause of type 2 diabetes mellitus (T2DM). Identifying the molecular mechanism of metabolic disorders is key to the treatment of T2DM. The study was to investigate the effect of circRNA PIP5K1A (circPIP5K1A) on glucose and lipid metabolism and inflammation in T2DM rats. A T2DM rat model was established, and then the T2DM rats were injected with lentiviral vectors that interfere with circPIP5K1A, miR-552-3p, or ENO1 expression. Fasting blood glucose (FBG) and fasting insulin (FINS) levels of rats were detected by an automatic analyzer and insulin detection kit, and HOMA-IR was calculated. Lipid metabolism was assessed by measuring serum levels of TG, TC, LDL-C, leptin, and resistin. Serum levels of inflammatory factors (TNF-α and IL-6) were detected by ELISA. The pathological conditions of pancreatic tissue were observed by HE staining. circPIP5K1A, miR-552-3p and ENO1 levels were recorded. The experimental results showed that circPIP5K1A and ENO1 were up-regulated, and miR-552-3p was down-regulated in T2DM rats. Down-regulating circPIP5K1A or up-regulating miR-552-3p reduced blood glucose and lipid levels, inhibited inflammation, and improved pancreatic histopathological changes in T2DM rats. In addition, up-regulating ENO1 rescued the ameliorating effects of down-regulated circPIP5K1A on T2DM rats. In general, downregulating circPIP5K1A improves insulin resistance and lipid metabolism disorders and inhibits inflammation by targeting miR-552-3p to mediate ENO1 expression.

To investigate the role of microRNA-378 (miR-378) in the regulation of mucin gene expression and inflammatory response in human middle ear epithelial cells (HMEEC) during bacterial infection by non-typeable Haemophilus influenzae (NTHi).

Human middle ear epithelial cells (HMEEC) were cultured and transfected with miR-378 or control miRNA. Post-transfection, cells were exposed to NTHi lysates. mRNA levels of MUC5B, MUC5AC, and IL-8 were quantified using RT-qPCR, and promoter activity was measured via luciferase assays. The effects of miR-378 on mucin and cytokine gene expression were analyzed.

Transfection with miR-378 significantly increased the expression of MUC5B (3.6 fold, p < 0.01), MUC5AC (19.1 fold, p < 0.01), and IL-8 (2.01 fold, p < 0.05) in HMEEC. NTHi exposure reduced MUC5B (1.385 fold, p < 0.05) and MUC5AC (1.61 fold, p < 0.05) gene expression in miR-378 transfected cells but significantly increased IL-8 levels (1.32 fold, p < 0.05). Luciferase assays showed that miR-378 upregulated the promoter activity of MUC5B (1.4 fold, p < 0.01) and MUC5AC (1.6 fold, p < 0.01) genes, indicating its role in transcriptional regulation.

miR-378 plays a crucial role in promoting mucin overproduction and an inflammatory response in the middle ear epithelium during OM. Targeting miR-378 could offer a novel therapeutic strategy for preventing the progression from AOM to COM.

na Laryngoscope, 2024.

Johne's disease, caused by Mycobacterium avium subspecies paratuberculosis (MAP), is a chronic enteritis that adversely affects welfare and productivity in cattle. Screening and subsequent removal of affected animals is a common approach for disease management, but efforts are hindered by low diagnostic sensitivity. Expression levels of small non-coding RNA molecules involved in gene regulation (microRNAs), which may be altered during mycobacterial infection, may present an alternative diagnostic method.

The expression levels of 24 microRNAs affected by mycobacterial infection were measured in sera from MAP-positive (n = 66) and MAP-negative cattle (n = 65). They were then used within a machine learning approach to build an optimal classifier for MAP diagnosis.

The method provided 72% accuracy, 73% sensitivity and 71% specificity on average, with an area under the curve of 78%.

Although control samples were collected from farms nominally MAP-free, the low sensitivity of current diagnostics means some animals may have been misclassified.

MicroRNA profiling combined with advanced predictive modelling enables rapid and accurate diagnosis of Johne's disease in cattle.

Bronchodilator response (BDR) is a measure of improvement in airway smooth muscle tone, inhibition of liquid accumulation and mucus section into the lumen in response to short-acting beta-2 agonists that varies among asthmatic patients. MicroRNAs (miRNAs) are well-known post-translational regulators. Identifying miRNAs associated with BDR could lead to a better understanding of the underlying complex pathophysiology.

The purpose of this study is to identify circulating miRNAs associated with bronchodilator response in asthma and decipher possible mechanism of bronchodilator response variation.

We used available small RNA sequencing on blood serum from 1,134 asthmatic children aged 6 to 14 years who participated in the Genetics of Asthma in Costa Rica Study (GACRS). We filtered the participants into the highest and lowest bronchodilator response (BDR) quartiles and used DeSeq2 to identify miRNAs with differential expression (DE) in high (N = 277) vs. low (N = 278) BDR group. Replication was carried out in the Leukotriene modifier Or Corticosteroids or Corticosteroid-Salmeterol trial (LOCCS), an adult asthma cohort. The putative target genes of DE miRNAs were identified, and pathway enrichment analysis was performed.

We identified 10 down-regulated miRNAs having odds ratios (OR) between 0.37 and 0.76 for a doubling of miRNA counts and one up-regulated miRNA (OR = 2.26) between high and low BDR group. These were assessed for replication in the LOCCS cohort, where two miRNAs (miR-200b-3p and miR-1246) were associated. Further, functional annotation of 11 DE miRNAs were performed as well as of two replicated miRs. Target genes of these miRs were enriched in regulation of cholesterol biosynthesis by SREBPs, ESR-mediated signaling, G1/S transition, RHO GTPase cycle, and signaling by TGFB family pathways.

MiRNAs miR-1246 and miR-200b-3p are associated with both childhood and adult asthma BDR. Our findings add to the growing body of evidence that miRNAs play a significant role in the difference of asthma treatment response among patients as it points to genomic regulatory machinery underlying difference in bronchodilator response among patients.

LOCCS cohort [ClinicalTrials.gov number NCT00156819, Registration date 20050912], GACRS cohort [ClinicalTrials.gov number NCT00021840].

Background and Objectives: Type 1 Diabetes Mellitus (T1DM) is an autoimmune disease with T cell-mediated pathogenesis of pancreatic β-cell destruction, leading to insulin deficiency. MicroRNAs such as miR-223 and miR-106b, along with PTEN, have been reported to participate in the pathophysiology of diabetes and its complications. The current study has explored the expression of miR-223, miR-106b, and PTEN and their association with various clinical and biochemical parameters in subjects diagnosed with T1DM. Materials and Methods: Sixty T1DM patients (two groups as uncomplicated/ with microalbuminuria) and fifty healthy volunteers, age- and sex-matched, were enrolled in this study. The fasting venous blood samples were collected, and PTEN and miRNAs (miR-223 and miR-106b) levels were measured by ELISA and real-time PCR, respectively. Results: The PTEN levels of patients with microalbuminuria were significantly lower than those of patients without microalbuminuria, while those of miR-223 and miR-106b were significantly increased in the T1DM group compared with the healthy control group (p < 0.001). ROC analysis indicated that PTEN, miR-223, and miR-106b could be potential biomarkers for diagnosing T1DM with high specificity but with variable sensitivities. Also, PTEN and miR-223 were negatively correlated with r =-0.398 and p < 0.0001, indicating that they were interrelated in their role within the T1DM pathophysiology. Conclusions: In the current study, it has been shown that the circulating levels of PTEN, miR-223, and miR-106b are significantly changed in T1DM patients and may back their potential to be used as non-invasive biomarkers for the diagnosis and monitoring of T1DM. Low PTEN protein expression was related to high miR-223 expression, indicating involvement of these miRNA in the regulation of PTEN. Further studies should be performed to clarify the exact mechanisms and possible clinical applications of these molecules.

Erythropoiesis is regulated by the differential expression of many genes. Besides being transcriptionally regulated, these genes are also with the oath of epigenetic regulation by the microRNAs (miRNAs), in particular. Various miRNAs appear to be very important for the normal process of erythropoiesis and various hematological abnormalities in humans. Therefore, the review aims to summarize the significance of miRNAs in erythropoiesis and different hematological diseases with clinical importance. Our analysis indicates that specific miRNAs regulate erythropoiesis in a stage-specific manner from hematopoietic stem cells to differentiated erythrocytes. Further, many miRNAs have been reported to be linked with various hematological diseases. The importance of miRNAs as biomarkers or therapeutic drug targets for various hematological disorders like anemia, β-thalassemia, and leukemia has been revealed through various clinical studies and clinical trials. The miR-34a mimic and miR-155 inhibitor demonstrate promising therapeutic effects in various hematological malignancies. Additionally, miR-34a, miR-538e, miR-193e, and miR-198 exhibit diagnostic potential in acute myeloid leukemia, while miR-451, miR-151-5p, and miR-1290 show diagnostic potential in B-cell acute lymphoblastic leukemia. Thus, this review encompasses the latest observations and implications of specific miRNAs in erythropoiesis and various hematological disorders. However, challenges persist in developing safe and efficient delivery strategies to target miRNAs specifically, minimizing off-target effects and enhancing therapeutic outcomes. Future mechanistic pre-clinical and clinical research would contribute to overcoming these challenges.

Per- and polyfluoroalkyl substances (PFAS) are synthetic chemicals that persist in the environment and can accumulate in humans, leading to adverse health effects. MicroRNAs (miRNAs) are emerging biomarkers that can advance the understanding of the mechanisms of PFAS effects on human health. However, little is known about the associations between PFAS exposures and miRNA alterations in humans.

To investigate associations between PFAS concentrations and miRNA levels in children.

Data from two distinct cohorts were utilized: 176 participants (average age 17.1 years; 75.6% female) from the Teen-Longitudinal Assessment of Bariatric Surgery (Teen-LABS) cohort in the United States, and 64 participants (average age 6.5 years, 39.1% female) from the Rhea study, a mother-child cohort in Greece. PFAS concentrations and miRNA levels were assessed in plasma samples from both studies. Associations between individual PFAS and plasma miRNA levels were examined after adjusting for covariates. Additionally, the cumulative effects of PFAS mixtures were evaluated using an exposure burden score. Ingenuity Pathways Analysis was employed to identify potential disease functions of PFAS-associated miRNAs.

Plasma PFAS concentrations were associated with alterations in 475 miRNAs in the Teen-LABs study and 5 miRNAs in the Rhea study (FDR p < 0.1). Specifically, plasma PFAS concentrations were consistently associated with decreased levels of miR-148b-3p and miR-29a-3p in both cohorts. Pathway analysis indicated that PFAS-related miRNAs were linked to numerous chronic disease pathways, including cardiovascular diseases, inflammatory conditions, and carcinogenesis.

Through miRNA screenings in two independent cohorts, this study identified both known and novel miRNAs associated with PFAS exposure in children. Pathway analysis revealed the involvement of these miRNAs in several cancer and inflammation-related pathways. Further studies are warranted to enhance our understanding of the relationships between PFAS exposure and disease risks, with miRNA emerging as potential biomarkers and/or mediators in these complex pathways.

We report the engineering and synthesis of peptide nucleic acid-functionalized Ti3C2Tx MXene nanosheets as a novel transducing material for amplification-free, nanoparticle-free, and isothermal electrochemical detection of microRNA biomarkers. Through bio-orthogonal copper-free click chemistry, azido-modified MXene nanosheets are covalently functionalized with clickable peptide nucleic acid probes targeting prostate cancer biomarker hsa-miR-141. The platform demonstrates a wide dynamic range, single-nucleotide specificity, and 40 aM detection limit outperforming more complex, amplification-based methods. Its versatility, analytical performance, and stability under serum exposure highlight the immense potential of this first example of click-conjugated MXene in the next generation of amplification-free biosensors.

MicroRNAs (miRNAs) are small, non-coding RNAs that control gene expression by degrading or repressing mRNA translation into proteins. Research recently suggested that food-derived miRNAs are bioavailable and may be absorbed in the gastrointestinal tract (GIT). Since these small RNAs may reach the circulation and organs, possible interactions with host genes will lead to epigenetic effects that alter metabolism. Therefore, from a precision nutrition standpoint, exogenous miRNAs may be essential in modulating health status. This review summarizes the process of miRNA biogenesis, the post-translational mechanisms of gene regulation, and their bioavailability in animal- and plant-derived foods.

MicroRNAs (miRNAs) have been recognized as promising diagnostic biomarkers for Diabetic Retinopathy (DR) due to their notable upregulation in individuals with the condition. However, the development of highly sensitive miRNAs assays for the rapid diagnosis of DR in clinical settings remains a challenging task.

In this study, we introduce an enhanced CRISPR/Cas12a assay, leveraging suboptimal PAM (sPAM)-mediated Cas12a trans-cleavage in conjunction with rolling circle amplification (RCA). sPAM was found to perform better than canonical PAM (cPAM) in the detection of Cas12a-mediated ssDNA detection at low concentrations and was used instead of canonical PAM (cPAM) to mediate the detection. The parameters of reactions have also been optimized.

In comparison with cPAM, sPAM has higher sensitivity in the detection of ssDNA at concentrations lower than 10 pM by Cas12a. By replacing cPAM with sPAM in the padlock template of RCA, ultra-high sensitivity for miR-183 detection is achieved, with a detection limit of 0.40 aM. within 25 min and a linear range spanning from 1 aM. to 1 pM. Our assay also exhibits exceptional specificity in detecting miR-183 from other miRNAs. Furthermore, the applicability of our assay for the sensitive detection of miR-183 in clinical serum samples is also validated. This study introduces a groundbreaking assay with excellent performance through a simple modification, which not only addresses existing diagnostic challenges, but also opens exciting new avenues for clinical diagnosis in the realm of DR.

Single-walled carbon nanotubes (SWCNTs) possess outstanding photophysical properties which has garnered interest towards utilizing these materials for biosensing and imaging applications. The near-infrared (NIR) fluorescence within the tissue transparent region along with their photostability and sizes in the nanoscale make SWCNTs valued candidates for the development of optical sensors. In this review, we discuss recent advances in the development and the applications of SWCNT-based nano-biosensors. An overview of SWCNT's structural and photophysical properties, sensor development, and sensing mechanisms are described. Examples of SWCNT-based optical nanosensors for detection of disease biomarkers, pathogens (bacteria and viruses), plant stressors, and environmental contaminants including heavy metals and disinfectants are provided. Molecular detection in biofluids, in vitro, and in vivo (small animal models and plants) are highlighted, and sensor integration into portable substrates for implantable and wearable sensing devices has been discussed. Recent advancements, which include high throughput assays and the use of machine learning models to predict more sensitive and robust sensing outcomes are discussed. Current limitations and future perspectives on translation of SWCNT optical probes into clinical practices have been provided.

Circulating maternal MicroRNA (miRNA) is a promising source of biomarkers for antenatal diagnostics. NanoString nCounter is a popular global screening tool due to its simplicity and ease of use, but there is a lack of standardisation in analysis methods. We examined the effect of user-defined variables upon reported changes in maternal blood miRNA during pregnancy.

Total RNA was prepared from the maternal blood of pregnant and control rats. miRNA expression was profiled using Nanostring nCounter. Raw count data were processed using nSolver using different combinations of normalisation and background correction methods as well as various background thresholds. A panel of 14 candidates in which changes were supported by multiple analysis workflows was selected for validation by RT-qPCR. We then reverse-engineered the nSolver analysis to gain further insight.

Thirty-one putative differentially expressed miRNAs were identified by nSolver. However, each analysis workflow produced a different set of reported biomarkers and none of them was common to all analysis methods. Four miRNAs with known roles in pregnancy (miR-183, miR-196c, miR-431, miR-450a) were validated. No single nSolver analysis workflow could successfully identify all four validated changes. Reverse engineering revealed errors in nSolver data processing which compound the inherent problems associated with background correction and normalisation.

Our results suggest that user-defined variables greatly influence the output of the assay. This highlights the need for standardised nSolver data analysis methods and detailed reporting of these methods. We suggest that investigators in the future should not rely on a single analysis method to identify changes and should always validate screening results.

This study aimed to establish efficient, cost-effective, and early predictive models for adverse pregnancy outcomes based on the combinations of a minimum number of miRNA biomarkers, whose altered expression was observed in specific pregnancy-related complications and selected maternal clinical characteristics.

This retrospective study included singleton pregnancies with gestational hypertension (GH, n = 83), preeclampsia (PE, n = 66), HELLP syndrome (n = 14), fetal growth restriction (FGR, n = 82), small for gestational age (SGA, n = 37), gestational diabetes mellitus (GDM, n = 121), preterm birth in the absence of other complications (n = 106), late miscarriage (n = 34), stillbirth (n = 24), and 80 normal term pregnancies. MiRNA gene expression profiling was performed on the whole peripheral venous blood samples collected between 10 and 13 weeks of gestation using real-time reverse transcription polymerase chain reaction (RT-PCR).

Most pregnancies with adverse outcomes were identified using the proposed approach (the combinations of selected miRNAs and appropriate maternal clinical characteristics) (GH, 69.88%; PE, 83.33%; HELLP, 92.86%; FGR, 73.17%; SGA, 81.08%; GDM on therapy, 89.47%; and late miscarriage, 84.85%). In the case of stillbirth, no addition of maternal clinical characteristics to the predictive model was necessary because a high detection rate was achieved by a combination of miRNA biomarkers only [91.67% cases at 10.0% false positive rate (FPR)].

The proposed models based on the combinations of selected cardiovascular disease-associated miRNAs and maternal clinical variables have a high predictive potential for identifying women at increased risk of adverse pregnancy outcomes; this can be incorporated into routine first-trimester screening programs. Preventive programs can be initiated based on these models to lower cardiovascular risk and prevent the development of metabolic/cardiovascular/cerebrovascular diseases because timely implementation of beneficial lifestyle strategies may reverse the dysregulation of miRNAs maintaining and controlling the cardiovascular system.

Endometrial cancer is one of the major cancers in women throughout the world. If diagnosed early, these cancers are treatable and the prognosis is usually good. However, one major problem in treating endometrial cancer is accurate diagnosis and staging. Till date, the choice method for diagnosis and staging is histopathology. Although there are few molecular markers identified, they are not always sufficient in making accurate diagnosis and deciding on therapeutic strategy. As a result, very often patients are under treated or over treated. In this study, our group has profiled microRNAs from Indian patients using NGS-based approach. We have identified 212 differentially expressed microRNAs in endometrial cancer. Among these, there are 17 novel miRNAs. Since this data represents only Indian cohort and also lacks survival data, validation across other populations is necessary before being considered as biomarkers. As one approach towards this, these microRNAs have also been compared to data from TCGA, which represent other populations and also correlated to relevance in overall survival. Using in-silico approaches, mRNA targets of the miRNAs have been predicted. After comparing with TCGA, we have identified 16 miRNA-mRNA pairs which could be potential prognostic biomarkers for endometrial cancer. This is the first miRNA profiling report from Indian cohort and one of the very few studies which have identified potential biomarkers of prognosis in endometrial cancer.

Pediatric sleep-disordered breathing disorders are a group of common conditions, from habitual snoring to obstructive sleep apnea (OSA) syndrome, affecting a significant proportion of children. The present article summarizes the current knowledge on diagnosis and treatment of pediatric OSA focusing on therapeutic and surgical advancements in the field in recent years. Advancements in OSA such as biomarkers, improving continuous pressure therapy adherence, novel pharmacotherapies, and advanced surgeries are discussed.

CF-related diabetes (CFRD) is a prevalent comorbidity in people with Cystic Fibrosis (CF), significantly impacting morbidity and mortality rates. This review article critically evaluates the current understanding of CFRD molecular mechanisms, including the role of CFTR protein, oxidative stress, unfolded protein response (UPR) and intracellular communication. CFRD manifests from a complex interplay between exocrine pancreatic damage and intrinsic endocrine dysfunction, further complicated by the deleterious effects of misfolded CFTR protein on insulin secretion and action. Studies indicate that ER stress and subsequent UPR activation play critical roles in both exocrine and endocrine pancreatic cell dysfunction, contributing to β-cell loss and insulin insufficiency. Additionally, oxidative stress and altered calcium flux, exacerbated by CFTR dysfunction, impair β-cell survival and function, highlighting the significance of antioxidant pathways in CFRD pathogenesis. Emerging evidence underscores the importance of exosomal microRNAs (miRNAs) in mediating inflammatory and stress responses, offering novel insights into CFRD's molecular landscape. Despite insulin therapy remaining the cornerstone of CFRD management, the variability in response to CFTR modulators underscores the need for personalized treatment approaches. The review advocates for further research into non-CFTR therapeutic targets, emphasizing the need to address the multifaceted pathophysiology of CFRD. Understanding the intricate mechanisms underlying CFRD will pave the way for innovative treatments, moving beyond insulin therapy to target the disease's root causes and improve the quality of life for individuals with CF.

Reproductive diseases and illnesses pose significant challenges in cattle farming, affecting fertility, milk production, and overall herd health. In recent years, the integration of various omics approaches, including transcriptomics, proteomics, metagenomics, miRNAomics, and metabolomics, has revolutionized the study of these conditions. This systematic review summarised the findings from studies that investigated reproductive disease biomarkers in both male and female cattle. After extracting 6137 studies according to exclusion and inclusion criteria, a total of 60 studies were included in this review. All studies identified were associated with female cattle and none were related to reproductive diseases in bulls. The analysis highlights specific biomarkers, metabolic pathways, and microbial compositions associated with bovine reproductive disease conditions, providing valuable insights into the underlying molecular mechanisms of disease. Pro-inflammatory cytokines such as IL-1β, IL-8, IL-4, IL-6, TNFα and acute-phase response proteins such as SAA and HP have been identified as promising biomarkers for bovine reproductive diseases. However, further research is needed to validate these markers clinically and to explore potential strategies for improving cow reproductive health. The role of bulls as carriers of venereal diseases has been underestimated in the current literature and therefore needs more attention to understand their impact on infectious reproductive diseases of female cattle.

There is an urgent need to accurately quantify microRNA (miRNA)-based Alzheimer's disease (AD) biomarkers, which have emerged as promising diagnostic biomarkers. In this study, we present a rapid and universal approach to establishing a target miRNA-triggered rolling circle amplification (RCA) detection strategy, which achieves ultrasensitive detection of several targets, including miR-let7a-5p, miR-34a-5p, miR-206-3p, miR-9-5p, miR-132-3p, miR-146a-5p, and miR-21-5p. Herein, the padlock probe contains three repeated signal strand binding regions and a target miRNA-specific region. The target miRNA-specific region captures miRNA, and then the padlock probe is circularized with the addition of T4 DNA ligase. Subsequently, an RCA reaction is triggered, and RCA products containing multiple signal strand binding regions are generated to trap abundant fluorescein-labeled signal strands. The addition of exonuclease III (Exo III) causes signal strand digestion and leads to RCA product recycling and liberation of fluorescein. Ultimately, graphene oxide (GO) does not absorb the liberated fluorescein because of poor mutual interaction. This method exhibited high specificity, sensitivity, repeatability, and stability toward let-7a, with a detection limit of 19.35 fM and a linear range of 50 fM to 5 nM. Moreover, it showed excellent applicability for recovering miRNAs in normal human serum. Our strategy was applied to detect miRNAs in the plasma of APP/PS1 mice, demonstrating its potential in the diagnosis of miRNA-associated disease and biochemical research.