Circulating small extracellular vesicles mediate vascular hyperpermeability in diabetes.

AIMS/HYPOTHESIS: A hallmark chronic complication of type 2 diabetes mellitus is vascular hyperpermeability, which encompasses dysfunction of the cerebrovascular endothelium and the subsequent development of associated cognitive impairment. The present study tested the hypothesis that during type 2 diabetes circulating small extracellular vesicles (sEVs) exhibit phenotypic changes that facilitate pathogenic disruption of the vascular barrier. METHODS: sEVs isolated from the plasma of a mouse model of type 2 diabetes and from diabetic human individuals were characterised for their ability to disrupt the endothelial cell (EC) barrier. The contents of sEVs and their effect on recipient ECs were assessed by proteomics and identified pathways were functionally interrogated with small molecule inhibitors. RESULTS: Using intravital imaging, we found that diabetic mice (Leprdb/db) displayed hyperpermeability of the cerebrovasculature. Enhanced vascular leakiness was recapitulated following i.v. injection of sEVs from diabetic mice into non-diabetic recipient mice. Characterisation of circulating sEV populations from the plasma of diabetic mice and humans demonstrated increased quantity and size of sEVs compared with those isolated from non-diabetic counterparts. Functional experiments revealed that sEVs from diabetic mice or humans induced the rapid and sustained disruption of the EC barrier through enhanced paracellular and transcellular leak but did not induce inflammation. Subsequent sEV proteome and recipient EC phospho-proteome analysis suggested that extracellular vesicles (sEVs) from diabetic mice and humans modulate the MAPK/MAPK kinase (MEK) and Rho-associated protein kinase (ROCK) pathways, cell-cell junctions and actin dynamics. This was confirmed experimentally. Treatment of sEVs with proteinase K or pre-treatment of recipient cells with MEK or ROCK inhibitors reduced the hyperpermeability-inducing effects of circulating sEVs in the diabetic state. CONCLUSIONS/INTERPRETATION: Diabetes is associated with marked increases in the concentration and size of circulating sEVs. The modulation of sEV-associated proteins under diabetic conditions can induce vascular leak through activation of the MEK/ROCK pathway. These data identify a new paradigm by which diabetes can induce hyperpermeability and dysfunction of the cerebrovasculature and may implicate sEVs in the pathogenesis of cognitive decline during type 2 diabetes.

Directional Endothelial Communication by Polarized Extracellular Vesicle Release.

BACKGROUND: Extracellular vesicles (EVs) contain bioactive cargo including miRNAs and proteins that are released by cells during cell-cell communication. Endothelial cells (ECs) form the innermost lining of all blood vessels, interfacing with cells in the circulation and vascular wall. It is unknown whether ECs release EVs capable of governing recipient cells within these 2 separate compartments. Given their boundary location, we propose ECs use bidirectional release of distinct EV cargo in quiescent (healthy) and activated (atheroprone) states to communicate with cells within the circulation and blood vessel wall. METHODS: EVs were isolated from primary human aortic ECs (plate and transwell grown; ±IL [interleukin]-1ß activation), quantified, visualized, and analyzed by miRNA transcriptomics and proteomics. Apical and basolateral EC-EV release was determined by miRNA transfer, total internal reflection fluorescence and electron microscopy. Vascular reprogramming (RNA sequencing) and functional assays were performed on primary human monocytes or smooth muscle cells±EC-EVs. RESULTS: Activated ECs increased EV release, with miRNA and protein cargo related to atherosclerosis. EV-treated monocytes and smooth muscle cells revealed activated EC-EV altered pathways that were proinflammatory and atherogenic. ECs released more EVs apically, which increased with activation. Apical and basolateral EV cargo contained distinct transcriptomes and proteomes that were altered by EC activation. Notably, activated basolateral EC-EVs displayed greater changes in the EV secretome, with pathways specific to atherosclerosis. In silico analysis determined compartment-specific cargo released by the apical and basolateral surfaces of ECs can reprogram monocytes and smooth muscle cells, respectively, with functional assays and in vivo imaging supporting this concept. CONCLUSIONS: Demonstrating that ECs are capable of polarized EV cargo loading and directional EV secretion reveals a novel paradigm for endothelial communication, which may ultimately enhance the design of endothelial-based therapeutics for cardiovascular diseases such as atherosclerosis where ECs are persistently activated.

Study of the Mechanism of Hydrolysis of Hemicellulose from Lignocellulose during Alkali Thermal Pretreatment by Density Functional Theory and Experiment.

The covalent bond fracture of hemicellulose leads to hemicellulose hydrolysis during lignocellulosic alkali thermal pretreatment, which has not previously been reported. Density functional theory was used to study the mechanism of hydrolysis of the hemicellulose model compounds under alkali conditions. There are four reaction paths for xylose formation, among which the reaction path with the lowest energy barrier is that in which the nucleophile captures H30 to generate water. The deprotonated hydroxyl group attacks the carbon on the glycoside bond, resulting in the cleavage of the glycoside bond and the formation of a new carbon-oxygen covalent bond, with an energy barrier of 154.2 kJ/mol. The nucleophile further attacks the glycosidic bond to form a new xylose residue with an energy barrier of 111.9 kJ/mol. When the glycosidic bond breaks, the orbital interaction with the largest proportion causes the transfer of ∼0.511 electron from the glycosidic bond oxygen to the deprotonated hydroxy oxygen. In situ Fourier transform infrared spectroscopy is used for the identification of functional groups during the alkali thermal pretreatment. As the temperature increases, the feasibility of the reaction increases. This study lays a theoretical foundation for the development of the alkali thermal pretreatment of lignocellulose.

Brain network integration underpins differential susceptibility of adolescent anxiety.

BACKGROUND: Parenting is a common and potent environmental factor influencing adolescent anxiety. Yet, the underlying neurobiological susceptibility signatures remain elusive. Here, we used a longitudinal twin neuroimaging study to investigate the brain network integration and its heritable relation to underpin the neural differential susceptibility of adolescent anxiety to parenting environments. METHODS: 216 twins from the Beijing Twin Study completed the parenting and anxiety assessments and fMRI scanning. We first identified the brain network integration involved in the influences of parenting at age 12 on anxiety symptoms at age 15. We then estimated to what extent heritable sensitive factors are responsible for the susceptibility of brain network integration. RESULTS: Consistent with the differential susceptibility theory, the results showed that hypo-connectivity within the central executive network amplified the impact of maternal hostility on anxiety symptoms. A high anti-correlation between the anterior salience and default mode networks played a similar modulatory role in the susceptibility of adolescent anxiety to paternal hostility. Genetic influences (21.18%) were observed for the connectivity pattern in the central executive network. CONCLUSIONS: Brain network integration served as a promising neurobiological signature of the differential susceptibility to adolescent anxiety. Our findings deepen the understanding of the neural sensitivity in the developing brain and can inform early identification and personalized interventions for adolescents at risk of anxiety disorders.

Endothelial cells secrete small extracellular vesicles bidirectionally containing distinct cargo to uniquely reprogram vascular cells in the circulation and vessel wall.

Rationale: Extracellular vesicles (EVs) contain bioactive cargo including microRNAs (miRNAs) and proteins that are released by cells as a form of cell-cell communication. Endothelial cells (ECs) form the innermost lining of all blood vessels and thereby interface with cells in the circulation as well as cells residing in the vascular wall. It is unknown whether ECs have the capacity to release EVs capable of governing recipient cells within two separate compartments, and how this is affected by endothelial activation commonly seen in atheroprone regions. Objective: Given their boundary location, we propose that ECs utilize bidirectional release of distinct EV cargo in quiescent and activated states to communicate with cells within the circulation and blood vessel wall. Methods and Results: EVs were isolated from primary human aortic endothelial cells (ECs) (+/-IL-1ß activation), quantified, and analysed by miRNA transcriptomics and proteomics. Compared to quiescent ECs, activated ECs increased EV release, with miRNA and protein cargo that were related to atherosclerosis. RNA sequencing of EV-treated monocytes and smooth muscle cells (SMCs) revealed that EVs from activated ECs altered pathways that were pro-inflammatory and atherogenic. Apical and basolateral EV release was assessed using ECs on transwells. ECs released more EVs apically, which increased with activation. Apical and basolateral EV cargo contained distinct transcriptomes and proteomes that were altered by EC activation. Notably, basolateral EC-EVs displayed greater changes in the EV secretome, with pathways specific to atherosclerosis. In silico analysis determined that compartment-specific cargo released by the apical and basolateral surfaces of ECs can reprogram monocytes and SMCs, respectively. Conclusions: The demonstration that ECs are capable of polarized EV cargo loading and directional EV secretion reveals a novel paradigm for endothelial communication, which may ultimately enhance our ability to design endothelial-based therapeutics for cardiovascular diseases such as atherosclerosis where ECs are persistently activated.

Synergistic antitumor activity of regorafenib and rosuvastatin in colorectal cancer.

Introduction: Colorectal cancer is one of the most prevalent life-threatening malignant tumors with high incidence and mortality. However, the efficacy of current therapeutic regimens is very limited. Regorafenib has been approved for second- or third-line treatment of patients who are refractory to standard chemotherapy diagnosed with metastatic colorectal cancer, but its clinical efficacy needs to be further improved. Accumulating evidence demonstrates that statins also possess potent anticancer activities. However, whether regorafenib and statins pose synergistic anticancer effects in colorectal cancer is still unclear. Methods: Sulforhodamine B (SRB) assays were applied to evaluate the anti-proliferative activity of regorafenib or/and rosuvastatin in vitro, and immunoblotting analysis were applied to detect the effects of regorafenib/rosuvastatin combined treatment on mitogen-activated protein kinase (MAPK) signaling and apoptosis-related proteins. MC38 tumors were applied to investigate the synergistic anticancer effects of regorafenib in combination with rosuvastatin in vivo. Results: We found that regorafenib in combination with rosuvastatin exerted significant synergistic inhibition against colorectal cancer growth in vitro and in vivo. Mechanistically, regorafenib and rosuvastatin combination synergistically suppressed MAPK signaling, a crucial signaling pathway promoting cell survival, as indicated by the reduction of phosphorylated MEK/ERK. In addition, regorafenib in combination with rosuvastatin synergistically induced the apoptosis of colorectal cancer in vitro and in vivo. Discussion: Our study demonstrated the synergistic anti-proliferative and pro-apoptotic effects of regorafenib/rosuvastatin combined treatment in colorectal cancer in vitro/vivo and might potentially be evaluated as a novel combination regimen for clinical treatment of colorectal cancer.

Field evidences of fluorescent dissolved organic matter (FDOM) as potential fingerprints for agricultural and urban sources in river environment.

Field evidences of the fluorescence differences between agricultural and urban river reaches are still lack. In this study, the middle reaches of Danhe River (DH) and Mihe River (MH) in Shouguang, China, were designed as agricultural and urban river reaches, respectively, to compare the the fluorescence differences in disparate river reaches using excitation-emission matrix coupled with parallel factor analysis (EEM-PARAFAC). Three fluorescence components were identified. C1 (Ex/Em=230,255,295 nm/420 nm) was categorized as humic-like fluorophores, C2 (Ex/Em=230,275 nm/330 nm) was recognized as tryptophan-like substances, and C3 (Ex/Em=215 nm/290 nm) was noted as tyrosine-like and phenylalanine-like compounds. The results showed that the FDOM posed significant differences between agricultural and urban river reaches (P < 0.001). The monitoring sites in DH were rich in C2 (1.90 ± 0.62 Raman Unit (RU), mean ± standard deviation), and the monitoring sites in MH were rich in C3 (1.32 ± 0.51 RU). Redundancy analysis revealed that C2 could be regarded as a fluorescence indicator of agricultural sewage in river environment, while C3 was recognized as a fluorescence indicator of domestic sewage in river environment. In conclusion, this study provided field evidences of FDOM as potential fingerprints for agricultural and urban sources in river environment.

Microstructure and Properties of Nickel-Based Gradient Coatings Prepared Using Cold Spraying Combined with Laser Cladding Methods.

A cold spray-laser cladding composite gradient coating (CLGC) was successfully formed on a Cu substrate. In comparison with traditional laser cladding gradient coatings (LGC), cold spraying the pre-set Ni-Cu alloy's intermediate transition layer not only mitigates the negative impacts due to the high reflectivity of the copper substrate but also helps to minimize the difference in the coefficients of thermal expansion (CTE) between the substrate and coating. This reduces the overall crack sensitivity and improves the cladding quality of the coating. Besides this, the uniform distribution of hard phases in CLGC, such as Ni11Si12 and Mo5Si3, greatly increases its microhardness compared to the Cu substrate, thus resulting in the value of 478.8 HV0.5 being approximately 8 times that of the Cu substrate. The friction coefficient of CLGC is lowered compared to both the Cu substrate and LGC with respective values of 0.28, 0.54, and 0.43, and its wear rate is only one-third of the Cu substrate's. These results suggest CLGC has excellent anti-wear properties. In addition, the wear mechanism was determined from the microscopic morphology and element distribution and was found to be oxidative and abrasive. This approach combines cold spraying and laser cladding to form a nickel-based gradient coating on a Cu substrate without cracks, holes, or other faults, thus improving the wear resistance of the Cu substrate and improving its usability.

The risk-taking behavioural intentions of pilots in adverse weather conditions: an application of the theory of planned behaviour.

This paper examined pilots' risk-taking behavioural intentions based on the theory of planned behaviour, as well as the impact of experience on behavioural intentions in adverse weather conditions. Two hundred and seventy-three airline pilots and flying cadets were divided into two groups and asked to complete a questionnaire based on two decision-making scenarios. This questionnaire measured pilots' intentions to take risks, along with the attitude towards the behaviour, subjective norms, perceived behavioural control (PBC), risk perception, and self-identity. The results showed that attitude, subjective norm, PBC, and risk perception explained 52% of the variance in behavioural intentions. Additionally, pilots' risk-taking decisions can be influenced by experience. Inexperienced pilots had a relatively stronger intention to take risks and a more favourable attitude towards risky behaviour. Moreover, pilots were more likely to rely on their own direct experience in the decision-making process. Practitioner summary: This study examined the pilots' risk-taking intentions under adverse weather conditions using a questionnaire based on the TPB theory. Results demonstrated that the TPB model can be applied to the risk-taking scenario and that experience can influence pilots' decisions. These findings have implications for improving flight safety and lowering accident rates.

68Ga-DOTA-FAPI-04 PET/CT Imaging in a Case of Radioactive Iodine-Induced Chronic Parotitis.

ABSTRACT: 68Ga-DOTA-FAPI-04 is a promising PET agent for tumor imaging. However, inflammatory lesions can also show increased FAPI uptake. Herein, we reported a 52-year-old woman who underwent total thyroidectomy for thyroid papillary carcinoma 1 year ago and underwent adjuvant radioiodine therapy 1 month later. After 131I therapy, she began to develop pain and swelling in bilateral cheeks, which developed into oral dryness. The patient was diagnosed with radioactive iodine-induced parotitis. 68Ga-DOTA-FAPI-04 PET/CT showed the density of bilateral parotid glands increased, and the volume decreased with intense tracer uptake. 99mTcO4- salivary gland scintigraphy showed decreased tracer uptake in the bilateral parotid glands.

Associations of Maternal Fructosamine before Delivery in Gestational Diabetes Mellitus Pregnancies with Neonatal Glucometabolic Disorders.

Background: The offspring of pregnant women with gestational diabetes mellitus (GDM) are vulnerable to be glucometabolic disorders. However, to date, few current studies focused on the associations of maternal accumulated glucose exposure before delivery with neonatal glucometabolic disorders and large for gestational age (LGA) infants. This study is aimed at exploring the associations of maternal fructosamine (FMN) before delivery in GDM pregnant women with neonatal glucometabolic disorders in the first 3 days of life and LGA infants. Methods: The study subjects were the GDM pregnant women, who gave birth in our hospital from September 1, 2018 to January 31, 2021, and their newborns. The maternal FMN adjusted by serum albumin (FMNALB) before delivery was selected as exposure factors. A multivariate logistical regression model was used to calculate the odds ratios (OR) for neonatal glucometabolic disorders, hypoglycemia needing intervention (<2.6 mmol/L), and glucose intolerance (>7.0 mmol/L) in the first 3 days and LGA infants. Results: In GDM pregnant women, the newborns in the maternal FMNALB ≥ 75th percentile (≥5.89 mmol/g) group had higher risks in neonatal glucometabolic disorders (aOR 2.50, 95% CI 1.34-4.65, P = 0.004) and hypoglycemia (aOR 2.18, 95% CI 1.16-4.10, P = 0.016). However, FMNALB ≥ 75th percentile seemed to be not predictive of the glucose intolerance (aOR 1.76, 95% CI 0.82-3.79, P = 0.149) and LGA (aOR 1.56, 95% CI 0.81-3.02, P = 0.185). Further, in the sensitivity analysis, the newborns in the maternal FMNALB ≥ 90th percentile (≥6.40 mmol/g) group also had higher risks in neonatal glucometabolic disorders (aOR 5.70, 95% CI 2.18-14.89, P < 0.001) and hypoglycemia (aOR 3.72, 95% CI 1.48-9.31, P = 0.005). Conclusions: The maternal FMNALB before delivery in GDM pregnant women was a useful biomarker to identify the offspring with high risk of neonatal glucometabolic disorders. However, the association between maternal FMNALB and the risk of LGA infants was not so strong.

Distribution and Relationships of Polycyclic Aromatic Hydrocarbons (PAHs) in Soils and Plants near Major Lakes in Eastern China.

The distributions and correlations among polycyclic aromatic hydrocarbons (PAHs) in soils and plants were analyzed. In this study, 9 soil samples and 44 plant samples were collected near major lakes (Hongze Lake, Luoma Lake, Chaohu, Changhu, Danjiangkou Reservoir, Wuhan East Lake, Longgan Lake, Qiandao Lake and Liangzi Lake) in eastern China. The following results were obtained: The total contents of PAHs in soil varied from 99.17 to 552.10 ng/g with an average of 190.35 ng/g, and the total contents of PAHs in plants varied from 122.93 to 743.44 ng/g, with an average of 274.66 ng/g. The PAHs in soil were dominated by medium- and low-molecular-weight PAHs, while the PAHs in plants were dominated by low-molecular-weight PAHs. The proportion of high-molecular-weight PAHs was the lowest in both soil and plants. Diagnostic ratios and principal component analysis (PCA) identified combustion as the main source of PAHs in soil and plants. The plant PAH monomer content was negatively correlated with Koa. Acenaphthylene, anthracene, benzo[k]fluoranthene, benzo[b]fluoranthene and dibenzo[a,h]anthracene were significantly correlated in plants and soil. In addition, no significant correlation between the total contents of the 16 PAHs and the content of high-, medium-, and low-molecular-weight PAHs in plants and soil was found. Bidens pilosa L. and Gaillardia pulchella Foug in the Compositae family and cron in the Poaceae family showed relatively stronger accumulation of PAHs, indicating their potential for phytoremediation.

A human model of arteriovenous malformation (AVM)-on-a-chip reproduces key disease hallmarks and enables drug testing in perfused human vessel networks.

Brain arteriovenous malformations (AVMs) are a disorder wherein abnormal, enlarged blood vessels connect arteries directly to veins, without an intervening capillary bed. AVMs are one of the leading causes of hemorrhagic stroke in children and young adults. Most human sporadic brain AVMs are associated with genetic activating mutations in the KRAS gene. Our goal was to develop an in vitro model that would allow for simultaneous morphological and functional phenotypic data capture in real time during AVM disease progression. By generating human endothelial cells harboring a clinically relevant mutation found in most human patients (activating mutations within the small GTPase KRAS) and seeding them in a dynamic microfluidic cell culture system that enables vessel formation and perfusion, we demonstrate that vessels formed by KRAS4AG12V mutant endothelial cells (ECs) were significantly wider and more leaky than vascular beds formed by wild-type ECs, recapitulating key structural and functional hallmarks of human AVM pathogenesis. Immunofluorescence staining revealed a breakdown of adherens junctions in mutant KRAS vessels, leading to increased vascular permeability, a hallmark of hemorrhagic stroke. Finally, pharmacological blockade of MEK kinase activity, but not PI3K inhibition, improved endothelial barrier function (decreased permeability) without affecting vessel diameter. Collectively, our studies describe the creation of human KRAS-dependent AVM-like vessels in vitro in a self-assembling microvessel platform that is amenable to phenotypic observation and drug delivery.

A Domain-General Monitoring Account of Bilingual Language Control in Recognition: The Role of Language Dominance and Bilingual Experience.

The ability of bilingual individuals to manage two competing languages is assumed to rely on both domain-specific language control and domain-general control mechanisms. However, previous studies have reported mixed findings about the extent and nature of cross-domain generality. The present study examined the role of language dominance, along with bilingual language experience, in the relationship between word recognition and domain-general cognitive control. Two single-language lexical decision tasks (one in L1 and another in L2) and a domain-general flanker task were administered to bilinguals who live in the sociolinguistic context of a minority and a majority language, namely, Uyghur (L1) and Chinese (L2), respectively. The results showed a diversity in language dominance patterns with better performance in L2 than L1 in the recognition modality, even for participants who self-identified as globally being dominant in L1. This finding reflected all bilinguals' self-evaluation that their preferred language for reading was L2, suggesting that language dominance is dynamic, depending on what language modality is measured. Furthermore, it was found that an earlier onset age of L2 acquisition (but not recent exposure) and a higher across-modality dominance in L2 were related to faster L2 word recognition. When self-reported language dominance was operationalised as a grouping variable, it was further found that both across-modality L1- and L2-dominant bilingual participants demonstrated a significant relationship between L2 word recognition and domain-general monitoring control, while only L1-dominant bilinguals additionally tapped into inhibitory control, indexed by the flanker effect during L2 word recognition. These findings suggest that language dominance has an impact on the extent and nature of the overlap in control mechanisms across specific linguistic and domain-general cognitive domains and add evidence to a domain-general monitoring account of bilingual word recognition.

Cardiovascular signatures of COVID-19 predict mortality and identify barrier stabilizing therapies.

BACKGROUND: Endothelial cell (EC) activation, endotheliitis, vascular permeability, and thrombosis have been observed in patients with severe coronavirus disease 2019 (COVID-19), indicating that the vasculature is affected during the acute stages of SARS-CoV-2 infection. It remains unknown whether circulating vascular markers are sufficient to predict clinical outcomes, are unique to COVID-19, and if vascular permeability can be therapeutically targeted. METHODS: Prospectively evaluating the prevalence of circulating inflammatory, cardiac, and EC activation markers as well as developing a microRNA atlas in 241 unvaccinated patients with suspected SARS-CoV-2 infection allowed for prognostic value assessment using a Random Forest model machine learning approach. Subsequent ex vivo experiments assessed EC permeability responses to patient plasma and were used to uncover modulated gene regulatory networks from which rational therapeutic design was inferred. FINDINGS: Multiple inflammatory and EC activation biomarkers were associated with mortality in COVID-19 patients and in severity-matched SARS-CoV-2-negative patients, while dysregulation of specific microRNAs at presentation was specific for poor COVID-19-related outcomes and revealed disease-relevant pathways. Integrating the datasets using a machine learning approach further enhanced clinical risk prediction for in-hospital mortality. Exposure of ECs to COVID-19 patient plasma resulted in severity-specific gene expression responses and EC barrier dysfunction, which was ameliorated using angiopoietin-1 mimetic or recombinant Slit2-N. INTERPRETATION: Integration of multi-omics data identified microRNA and vascular biomarkers prognostic of in-hospital mortality in COVID-19 patients and revealed that vascular stabilizing therapies should be explored as a treatment for endothelial dysfunction in COVID-19, and other severe diseases where endothelial dysfunction has a central role in pathogenesis. FUNDING: This work was directly supported by grant funding from the Ted Rogers Center for Heart Research, Toronto, Ontario, Canada and the Peter Munk Cardiac Center, Toronto, Ontario, Canada.

MiR-30 promotes fatty acid beta-oxidation and endothelial cell dysfunction and is a circulating biomarker of coronary microvascular dysfunction in pre-clinical models of diabetes.

BACKGROUND: Type 2 diabetes (T2D) is associated with coronary microvascular dysfunction, which is thought to contribute to compromised diastolic function, ultimately culminating in heart failure with preserved ejection fraction (HFpEF). The molecular mechanisms remain incompletely understood, and no early diagnostics are available. We sought to gain insight into biomarkers and potential mechanisms of microvascular dysfunction in obese mouse (db/db) and lean rat (Goto-Kakizaki) pre-clinical models of T2D-associated diastolic dysfunction. METHODS: The microRNA (miRNA) content of circulating extracellular vesicles (EVs) was assessed in T2D models to identify biomarkers of coronary microvascular dysfunction/rarefaction. The potential source of circulating EV-encapsulated miRNAs was determined, and the mechanisms of induction and the function of candidate miRNAs were assessed in endothelial cells (ECs). RESULTS: We found an increase in miR-30d-5p and miR-30e-5p in circulating EVs that coincided with indices of coronary microvascular EC dysfunction (i.e., markers of oxidative stress, DNA damage/senescence) and rarefaction, and preceded echocardiographic evidence of diastolic dysfunction. These miRNAs may serve as biomarkers of coronary microvascular dysfunction as they are upregulated in ECs of the left ventricle of the heart, but not other organs, in db/db mice. Furthermore, the miR-30 family is secreted in EVs from senescent ECs in culture, and ECs with senescent-like characteristics are present in the db/db heart. Assessment of miR-30 target pathways revealed a network of genes involved in fatty acid biosynthesis and metabolism. Over-expression of miR-30e in cultured ECs increased fatty acid ß-oxidation and the production of reactive oxygen species and lipid peroxidation, while inhibiting the miR-30 family decreased fatty acid ß-oxidation. Additionally, miR-30e over-expression synergized with fatty acid exposure to down-regulate the expression of eNOS, a key regulator of microvascular and cardiomyocyte function. Finally, knock-down of the miR-30 family in db/db mice decreased markers of oxidative stress and DNA damage/senescence in the microvascular endothelium. CONCLUSIONS: MiR-30d/e represent early biomarkers and potential therapeutic targets that are indicative of the development of diastolic dysfunction and may reflect altered EC fatty acid metabolism and microvascular dysfunction in the diabetic heart.

The Effect of Fasting on Human Metabolism and Psychological Health.

Fasting is a prevalent approach to weight loss and is a feasible method for treating some diseases, such as type 2 diabetes. Meanwhile, the effects of intermittent fasting on health, aging, and disease process are hot issues and are of concern by researchers of multiple areas, even the public. This article introduces the effects of fasting on human lipid metabolism, glucose metabolism, protein metabolism, and neuroendocrine metabolism; demonstrates the metabolic conversion caused by fasting; and describes the effects of fasting on human psychological health, the relationship between mood regulation and glucose, and the emotional enhancing effect induced by fasting.

Identification of HnRNPC as a novel Tau exon 10 splicing factor using RNA antisense purification mass spectrometry.

Alternative splicing in Tau exon 10 generates 3 R- and 4 R-Tau proteoforms, which have equal abundance in healthy adult human brain. Aberrant alternative splicing in Tau exon 10 leads to distortion of the balanced 3 R- and 4 R-Tau expression levels, which is a causal factor to trigger toxic Tau aggregation, neuron dysfunction and patient death in a group of neurodegenerative diseases known as tauopathies. Hence, identification of regulators upstream of the Tau exon 10 splicing events are crucial to understanding pathogenic mechanisms driving tauopathies. In this study, we used RNA Antisense Purification with Mass Spectrometry (RAP-MS) analysis to identify RNA-binding proteins (RBPs) that interact with the Tau pre-mRNA near exon 10. Among the newly identified RBP candidates, we show that knockdown of hnRNPC induces Tau exon 10 skipping whereas overexpression of hnRNPC promotes Tau exon 10 inclusion. In addition, we show that hnRNPC interacts with the poly-uridine (U-tract) sequences in introns 9 and 10 of Tau pre-mRNA. Mutation of these U-tract motifs abolished binding of hnRNPC with Tau pre-mRNA fragment and blocked its impact on Tau exon 10 inclusion. These findings indicate that hnRNPC binds and utilizes these U-tract motifs located in introns 9 and 10 of Tau pre-mRNA to promote Tau exon 10 inclusion. Intriguingly, high hnRNPC expression level is associated with progressive supranuclear palsy (PSP), a sporadic tauopathy with pathological accumulation of Tau species that contain exon 10, which suggests a putative therapeutic role of hnRNPC for PSP treatment. [Figure: see text].