Roles of Protein Post-Translational Modifications During Adipocyte Senescence.

Adipocytes are adipose tissues that supply energy to the body through lipids. The two main types of adipocytes comprise white adipocytes (WAT) that store energy, and brown adipocytes (BAT), which generate heat by burning stored fat (thermogenesis). Emerging evidence indicates that dysregulated adipocyte senescence may disrupt metabolic homeostasis, leading to various diseases and aging. Adipocytes undergo senescence via irreversible cell-cycle arrest in response to DNA damage, oxidative stress, telomere dysfunction, or adipocyte over-expansion upon chronic lipid accumulation. The amount of detectable BAT decreases with age. Activation of cell cycle regulators and dysregulation of adipogenesis-regulating factors may constitute a molecular mechanism that accelerates adipocyte senescence. To better understand the regulation of adipocyte senescence, the effects of post-translational modifications (PTMs), is essential for clarifying the activity and stability of these proteins. PTMs are covalent enzymatic protein modifications introduced following protein biosynthesis, such as phosphorylation, acetylation, ubiquitination, or glycosylation. Determining the contribution of PTMs to adipocyte senescence may identify new therapeutic targets for the regulation of adipocyte senescence. In this review, we discuss a conceptual case in which PTMs regulate adipocyte senescence and explain the mechanisms underlying protein regulation, which may lead to the development of effective strategies to combat metabolic diseases.

Tschimganidine reduces lipid accumulation through AMPK activation and alleviates high-fat diet-induced metabolic diseases.

Obesity increases the risk of mortality and morbidity because it results in hypertension, heart disease, and type 2 diabetes. Therefore, there is an urgent need for pharmacotherapeutic drugs to treat obesity. We performed a screening assay using natural products with anti-adipogenic properties in 3T3-L1 cells and determined that tschimganidine, a terpenoid from the Umbelliferae family, inhibited adipogenesis. To evaluate the anti-obesity effects of tschimganidine in vivo. Mice were fed either a normal chow diet (NFD) or a high-fat chow diet (HFD) with or without tschimganidine for 12 weeks. Treatment with tschimganidine decreased lipid accumulation and adipogenesis, accompanied by reduced expression of adipogenesis and lipid accumulation-related factors. Tschimganidine significantly increased the phosphorylation of AMP-activated protein kinase (AMPK) and decreased that of AKT. Depletion of AMPK relieved the reduction in lipid accumulation resulting from tschimganidine treatment. Moreover, tschimganidine administration drastically reduced the weight and size of both gonadal white adipose tissue (WAT) and blood glucose levels in high-fat diet-induced obese mice. We suggest that tschimganidine is a potent antiobesity agent, which impedes adipogenesis and improves glucose homeostasis. Tschimganidine can then be evaluated for clinical application as a therapeutic agent. [BMB Reports 2023; 56(4): 246-251].

Psychological Benefits of Purchasing Home Meal Replacement in the Context of Eco-Friendly TV Home Shopping Broadcast: The Moderating Role of Personal Norm.

This study observed the relationship between psychological benefits and the theory of planned behavior (TPB) in the context of an eco-friendly TV home shopping broadcasts. The theoretical framework was enhanced even further by examining the moderating role of personal norm on proenvironmental attitudes in the TV home shopping context. An online survey was conducted with Korean customers who had purchased home meal replacement (HMR) products from a TV home shopping broadcast within the past 6 months. A total of 305 samples were collected and used for the data analysis. All six of the hypotheses in the psychological benefits and TPB model were supported, meaning all constructs of psychological benefits, including warm glow, self-expressive benefits, and nature experiences, impacted TPB and behavioral intentions. Furthermore, personal norm had a moderating role in the relationship between warm glow and attitude. This research provides significant theoretical and managerial implications for the home shopping industry.

Depletion of Adipocyte Becn1 Leads to Lipodystrophy and Metabolic Dysregulation.

Becn1/Beclin-1 is a core component of the class III phosphatidylinositol 3-kinase required for autophagosome formation and vesicular trafficking. Although Becn1 has been implicated in numerous diseases such as cancer, aging, and neurodegenerative disease, the role of Becn1 in white adipose tissue and related metabolic diseases remains elusive. In this study, we show that adipocyte-specific Becn1 knockout mice develop severe lipodystrophy, leading to adipose tissue inflammation, hepatic steatosis, and insulin resistance. Ablation of Becn1 in adipocytes stimulates programmed cell death in a cell-autonomous manner, accompanied by elevated endoplasmic reticulum (ER) stress gene expression. Furthermore, we observed that Becn1 depletion sensitized mature adipocytes to ER stress, leading to accelerated cell death. Taken together, these data suggest that adipocyte Becn1 would serve as a crucial player for adipocyte survival and adipose tissue homeostasis.

De novo assembly and characterization of transcriptome in the medicinal plant Euphorbia jolkini.

BACKGROUND: Euphorbia jolkini, a medicinal herb that grows on the warm beaches in Japan and South Korea, is known to be used for traditional medicines to treat a variety of ailments, including bruises, stiffness, indigestion, toothache, and diabetes. OBJECTIVE: It is to analyze the whole transcriptome and identify the genes related to the phenylpropanoid biosynthesis in the medicinally important herb E jolkini. METHODS: Paired-end Illumina HiSeq™ 2500 sequencing technology was employed for cDNA library construction and Illumina sequencing. Public databases like TAIR (The Arabidopsis Information Resource), Swissprot and KEGG (Kyoto Encyclopedia of Genes and Genomes) were used for annotations of unigenes obtained. RESULTS: The transcriptome of E. jolkini generated 139,215 assembled transcripts with an average length of 868 bp and an N50 value of 1460 bp that were further clustered using CD-HIT into 93,801 unigenes with an average length of 847 bp (N50-1410 bp). Sixty-three percent of the coding sequences (CDS) were annotated from the longest open reading frame (ORF). A remarkable percentage of unigenes were annotated against various databases. The differentially expressed gene analysis revealed that the expression of genes related to the terpenoid backbone biosynthesis pathway was higher in the flowers, whereas that of genes related to the phenylpropanoid biosynthesis pathway was both up- and downregulated in flowers and leaves. A search of against the transcription factor domain found 1023 transcription factors (TFs) that were from 54 TF families. CONCLUSION: Assembled sequences of the E. jolkini transcriptome are made available for the first time in this study E. jolkini and lay a foundation for the investigation of secondary metabolite biosynthesis.

Ginsenoside Rp1, A Ginsenoside Derivative, Augments Anti-Cancer Effects of Actinomycin D via Downregulation of an AKT-SIRT1 Pathway.

Novel strategies for overcoming multidrug resistance are urgently needed to improve chemotherapy success and reduce side effects. Ginsenosides, the main active components of Panax ginseng, display anti-cancer properties and reverse drug resistance; however, the biological pathways mediating this phenomenon remain incompletely understood. This study aimed to evaluate the anti-cancer effects of ginsenoside Rp1, actinomycin D (ActD), and their co-administration in drug-resistant cells and murine xenograft model of colon cancer, and explore the underlying mechanisms. ActD increased expression and activity of SIRT1 in drug-resistant LS513 colon cancer, OVCAR8-DXR ovarian cancer, and A549-DXR lung cancer cells, but not in ActD-sensitive SW620 colon cancer cells. Inhibition of SIRT1, either pharmacologically, with EX527 or through siRNA, stimulated p53 acetylation and apoptosis in LS513 cells when treated with ActD. ActD also increased AKT activation in drug-resistant cells. Inhibition of AKT abrogated ActD-induced upregulation of SIRT1, suggesting that the AKT-SIRT1 pathway is important in ActD resistance. Rp1 inhibited both ActD-induced AKT activation and SIRT1 upregulation and re-sensitized the cells to ActD. Synergistic antitumor effects of Rp1 with ActD were also observed in vivo. Our results suggest that combining Rp1 with chemotherapeutic agents could circumvent drug resistance and improve treatment efficacy.

SIRT1 modulates cell cycle progression by regulating CHK2 acetylation-phosphorylation.

Both the stress-response protein, SIRT1, and the cell cycle checkpoint kinase, CHK2, play critical roles in aging and cancer via the modulation of cellular homeostasis and the maintenance of genomic integrity. However, the underlying mechanism linking the two pathways remains elusive. Here, we show that SIRT1 functions as a modifier of CHK2 in cell cycle control. Specifically, SIRT1 interacts with CHK2 and deacetylates it at lysine 520 residue, which suppresses CHK2 phosphorylation, dimerization, and thus activation. SIRT1 depletion induces CHK2 hyperactivation-mediated cell cycle arrest and subsequent cell death. In vivo, genetic deletion of Chk2 rescues the neonatal lethality of Sirt1-/- mice, consistent with the role of SIRT1 in preventing CHK2 hyperactivation. Together, these results suggest that CHK2 mediates the function of SIRT1 in cell cycle progression, and may provide new insights into modulating cellular homeostasis and maintaining genomic integrity in the prevention of aging and cancer.

Diverse therapeutic efficacies and more diverse mechanisms of nicotinamide.

BACKGROUND: Nicotinamide (NAM) is a form of vitamin B3 that, when administered at near-gram doses, has been shown or suggested to be therapeutically effective against many diseases and conditions. The target conditions are incredibly diverse ranging from skin disorders such as bullous pemphigoid to schizophrenia and depression and even AIDS. Similar diversity is expected for the underlying mechanisms. In a large portion of the conditions, NAM conversion to nicotinamide adenine dinucleotide (NAD+) may be a major factor in its efficacy. The augmentation of cellular NAD+ level not only modulates mitochondrial production of ATP and superoxide, but also activates many enzymes. Activated sirtuin proteins, a family of NAD+-dependent deacetylases, play important roles in many of NAM's effects such as an increase in mitochondrial quality and cell viability countering neuronal damages and metabolic diseases. Meanwhile, certain observed effects are mediated by NAM itself. However, our understanding on the mechanisms of NAM's effects is limited to those involving certain key proteins and may even be inaccurate in some proposed cases. AIM OF REVIEW: This review details the conditions that NAM has been shown to or is expected to effectively treat in humans and animals and evaluates the proposed underlying molecular mechanisms, with the intention of promoting wider, safe therapeutic application of NAM. KEY SCIENTIFIC CONCEPTS OF REVIEW: NAM, by itself or through altering metabolic balance of NAD+ and tryptophan, modulates mitochondrial function and activities of many molecules and thereby positively affects cell viability and metabolic functions. And, NAM administration appears to be quite safe with limited possibility of side effects which are related to NAM's metabolites.

Mechanisms and disease implications of sirtuin-mediated autophagic regulation.

Accumulating evidence has indicated that sirtuins are key components of diverse physiological processes, including metabolism and aging. Sirtuins confer protection from a wide array of metabolic and age-related diseases, such as cancer, cardiovascular and neurodegenerative diseases. Recent studies have also suggested that sirtuins regulate autophagy, a protective cellular process for homeostatic maintenance in response to environmental stresses. Here, we describe various biological and pathophysiological processes regulated by sirtuin-mediated autophagy, focusing on cancer, heart, and liver diseases, as well as stem cell biology. This review also emphasizes key molecular mechanisms by which sirtuins regulate autophagy. Finally, we discuss novel insights into how new therapeutics targeting sirtuin and autophagy may potentially lead to effective strategies to combat aging and aging-related diseases.

Deep-Blue Thermally Activated Delayed Fluorescence Emitters Containing Diphenyl Sulfone Group for Organic Light Emitting Diodes.

Novel blue thermally activated delayed fluorescence (TADF) emitters, D1-DPS and D2-DPS, were designed and synthesized. Diphenyl sulfone (DPS) group functioned as a common acceptor, and it combined with each of two different spiro-acridine groups, D1 and D2. The calculated energy differences (ΔEST) of the singlet and triplet excited states of D1-DPS (0.062 eV) and D2-DPS (0.128 eV) had sufficiently small ΔEST values, which is favorable in the thermally activated reverse intersystem crossing (RISC) process from the T1 state to the S1 state. A device doped 10 wt% of D2-DPS with ADN host material, obtained 5.05% of external quantum efficiency with deep-blue emission having CIExy coordinates of (0.152, 0.065). The results showed that these molecules are promising host-free TADF deep-blue emitters by inhibiting concentration quenching.

Deacetylation of CHK2 by SIRT1 protects cells from oxidative stress-dependent DNA damage response.

Growing evidence indicates that metabolic signaling pathways are interconnected to DNA damage response (DDR). However, factors that link metabolism to DDR remain incompletely understood. SIRT1, an NAD+-dependent deacetylase that regulates metabolism and aging, has been shown to protect cells from DDR. Here, we demonstrate that SIRT1 protects cells from oxidative stress-dependent DDR by binding and deacetylating checkpoint kinase 2 (CHK2). We first showed that essential proteins in DDR were hyperacetylated in Sirt1-deficient cells and that among them, the level of acetylated CHK2 was highly increased. We found that Sirt1 formed molecular complexes with CHK2, BRCA1/BRCA2-associated helicase 1 (BACH1), tumor suppressor p53-binding protein 1 (53BP1), and H2AX, all of which are key factors in response to DNA damage. We then demonstrated that CHK2 was normally inhibited by SIRT1 via deacetylation but dissociated with SIRT1 under oxidative stress conditions. This led to acetylation and activation of CHK2, which increased cell death under oxidative stress conditions. Our data also indicated that SIRT1 deacetylated the K235 and K249 residues of CHK2, whose acetylation increased cell death in response to oxidative stress. Thus, SIRT1, a metabolic sensor, protects cells from oxidative stress-dependent DDR by the deacetylation of CHK2. Our findings suggest a crucial function of SIRT1 in inhibiting CHK2 as a potential therapeutic target for cancer treatment.

Anti-cancer effect of doxorubicin is mediated by downregulation of HMG-Co A reductase via inhibition of EGFR/Src pathway.

Doxorubicin is a widely used DNA damage-inducing anti-cancer drug. However, its use is limited by its dose-dependent side effects, such as cardiac toxicity. Cholesterol-lowering statin drugs increase the efficacy of some anti-cancer drugs. Cholesterol is important for cell growth and a critical component of lipid rafts, which are plasma membrane microdomains important for cell signaling. 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase (HMG-CR) is a critical enzyme in cholesterol synthesis. Here, we show that doxorubicin downregulated HMG-CR protein levels and thus reduced levels of cholesterol and lipid rafts. Cholesterol addition attenuated doxorubicin-induced cell death, and cholesterol depletion enhanced it. Reduction of HMG-CR activity by simvastatin, a statin that acts as an HMG-CR inhibitor, or by siRNA-mediated HMG-CR knockdown enhanced doxorubicin cytotoxicity. Doxorubicin-induced HMG-CR downregulation was associated with inactivation of the EGFR-Src pathway. Furthermore, a high-cholesterol-diet attenuated the anti-cancer activity of doxorubicin in a tumor xenograft mouse model. In a multivulva model of Caenorhabditis elegans expressing an active-EGFR mutant, doxorubicin decreased hyperplasia more efficiently in the absence than in the presence of cholesterol. These data indicate that EGFR/Src/HMG-CR is a new pathway mediating doxorubicin-induced cell death and that cholesterol control could be combined with doxorubicin treatment to enhance efficacy and thus reduce side effects.

Ahnak promotes tumor metastasis through transforming growth factor-ß-mediated epithelial-mesenchymal transition.

Previously, we reported a molecular mechanism by which Ahnak potentiates transforming growth factor-ß (TGFß) signaling during cell growth. Here, we show that Ahnak induces epithelial-mesenchymal transition (EMT) in response to TGFß. EMT phenotypes, including altered in cell morphology, and expression patterns of various EMT marker genes were detected in HaCaT keratinocytes transfected with Ahnak-specific siRNA. Knockdown of Ahnak expression in HaCaT keratinocytes resulted in attenuated cell migration and invasion. We found that Ahnak activates TGFß signaling via Smad3 phosphorylation, leading to enhanced Smad3 transcriptional activity. To validate function of Ahnak in EMT of B16F10 cells having high metastatic and tumorigenic properties, we established B16F10 cells with stable knockdown of Ahnak. N-cadherin expression and Smad3 phosphorylation were significantly decreased in B16F10-shAhnak cells, compared to B16F10-shControl cells after treatment of TGFß. Moreover, TGFß failed to induce cell migration and cell invasion in B16F10-shAhnak cells. To determine whether Ahnak regulates the metastatic activity of B16F10 cells, we established a lung metastasis model in C57BL/6 mice via tail vein injection of B16F10-shAhnak cells. Lung metastasis was significantly suppressed in mice injected with B16F10-shAhnak cells, compared to those injected with B16F10-shControl cells. Taken together, we propose that TGFß-Ahnak signaling axis regulates EMT during tumor metastasis.

Improvement of Liquid Crystal Alignment Using Novel Photo-Activators.

Photo-activator is a kind of additive that can improve the anchoring energy by attacking some of the bonds of polyimide (PI). Photo-activators were synthesized from the reaction of cyclohexanone oxime with three different anhydrides, respectively. Each activator generates different active radicals when irradiated. These fragmented and activated radicals are responsible for the liquid crystal (LC) alignment of PI film. The reactivity was confirmed through UV-Visible spectroscopy. All the three photo-activators had characteristic bimodal-shaped absorption peaks at 270∼280 nm. The photofragmentation reactions were completed within 1 min of UV irradiation, which implies that the activators are highly reactive to UV light. The short reaction time is very useful for liquid crystal display (LCD) factory applications. The photo-activator using crotonic anhydride (CAP) showed the highest surface anchoring energy, of 6.92 × 10-5 J/m2, compared to that of the other activators and that obtained by rubbing methods; (1.11 × 10-5 J/m2). This result was obtained due to resonance stabilization from the allyl radicals of CAP. The photo-activator using acetic anhydride (AAP) reached its maximum anchoring energy in less than 3 min of irradiation, which is the shortest optimum irradiation time. Considering the fact that this process does not require additional procedure and time, the photo-activators can be considered an innovate additive.

Emergency preparedness for the accidental release of radionuclides from the Uljin Nuclear Power Plant in Korea.

Site specific radionuclide dispersion databases were archived for the emergency response to the hypothetical releases of 137Cs from the Uljin nuclear power plant in Korea. These databases were obtained with the horizontal resolution of 1.5 km in the local domain centered the power plant site by simulations of the Lagrangian Particle Dispersion Model (LPDM) with the Unified Model (UM)-Local Data Assimilation Prediction System (LDAPS). The Eulerian Dispersion Model-East Asia (EDM-EA) with the UM-Global Data Assimilation Prediction System (UM-GDAPS) meteorological models was used to get dispersion databases in the regional domain. The LPDM model was performed for a year with a 5-day interval yielding 72 synoptic time-scale cases in a year. For each case hourly mean near surface concentrations, hourly mean column integrated concentrations, hourly total depositions for 5 consecutive days were archived by the LPDM model in the local domain and by the EDM-EA model in the regional domain of Asia. Among 72 synoptic cases in a year the worst synoptic case that showed the highest mean surface concentration averaged for 5 days in the LPDM model domain was chosen to illustrate the emergency preparedness to the hypothetical accident at the site. The simulated results by the LPDM model with the 137Cs emission rate of the Fukushima nuclear power plant accident for the first 5-day period were found to be able to provide prerequisite information for the emergency response to the early phase of the accident whereas those of the EDM-EA model could provide information required for the environmental impact assessment of the accident in the regional domain. The archived site-specific database of 72 synoptic cases in a year could have a great potential to be used as a prognostic information on the emergency preparedness for the early phase of accident.

SP-LL-37, human antimicrobial peptide, enhances disease resistance in transgenic rice.

Human LL-37 is a multifunctional antimicrobial peptide of cathelicidin family. It has been shown in recent studies that it can serve as a host's defense against influenza A virus. We now demonstrate in this study how signal peptide LL-37 (SP-LL-37) can be used in rice resistance against bacterial leaf blight and blast. We synthesized LL-37 peptide and subcloned in a recombinant pPZP vector with pGD1 as promoter. SP-LL-37 was introduced into rice plants by Agrobacterium mediated transformation. Stable expression of SP-LL-37 in transgenic rice plants was confirmed by RT-PCR and ELISA analyses. Subcellular localization of SP-LL-37-GFP fusion protein showed evidently in intercellular space. Our data on testing for resistance to bacterial leaf blight and blast revealed that the transgenic lines are highly resistant compared to its wildtype. Our results suggest that LL-37 can be further explored to improve wide-spectrum resistance to biotic stress in rice.

Estimation of radionuclide (137Cs) emission rates from a nuclear power plant accident using the Lagrangian Particle Dispersion Model (LPDM).

A methodology for the estimation of the emission rate of 137Cs by the Lagrangian Particle Dispersion Model (LPDM) with the use of monitored 137Cs concentrations around a nuclear power plant has been developed. This method has been employed with the MM5 meteorological model in the 600 km × 600 km model domain with the horizontal grid scale of 3 km × 3 km centered at the Fukushima nuclear power plant to estimate 137Cs emission rate for the accidental period from 00 UTC 12 March to 00 UTC 6 April 2011. The Lagrangian Particles are released continuously with the rate of one particle per minute at the first level modelled, about 15 m above the power plant site. The presently developed method was able to simulate quite reasonably the estimated 137Cs emission rate compared with other studies, suggesting the potential usefulness of the present method for the estimation of the emission rate from the accidental power plant without detailed inventories of reactors and fuel assemblies and spent fuels. The advantage of this method is not so complicated but can be applied only based on one-time forward LPDM simulation with monitored concentrations around the power plant, in contrast to other inverse models. It was also found that continuously monitored radionuclides concentrations from possibly many sites located in all directions around the power plant are required to get accurate continuous emission rates from the accident power plant. The current methodology can also be used to verify the previous version of radionuclides emissions used among other modeling groups for the cases of intermittent or discontinuous samplings.

Spatial and temporal distributions of aerosol concentrations and depositions in Asia during the year 2010.

Aerosol Modeling System (AMS) that is consisted of the Asian Dust Aerosol Model2 (ADAM2) and the Community Multi-scale Air Quality (CMAQ) modeling system has been employed to document the spatial distributions of the monthly and the annual averaged concentration of both the Asian dust (AD) aerosol and the anthropogenic aerosol (AA), and their total depositions in the Asian region for the year 2010. It is found that the annual mean surface aerosol (PM10) concentrations in the Asian region affect in a wide region as a complex mixture of AA and AD aerosols; they are predominated by the AD aerosol in the AD source region of northern China and Mongolia with a maximum concentration exceeding 300 µg m(-3); AAs are predominated in the high pollutant emission regions of southern and eastern China and northern India with a maximum concentration exceeding 110 µg m(-3); while the mixture of AA and AD aerosols is dominated in the downwind regions extending from the Yellow Sea to the Northwest Pacific Ocean. It is also found that the annual total deposition of aerosols in the model domain is found to be 485 Tg (372 Tg by AD aerosol and 113 Tg by AA), of which 66% (319 Tg) is contributed by the dry deposition (305 Tg by AD aerosol and 14 Tg by AA) and 34% (166 Tg) by the wet deposition (66 Tg by AD aerosol and 100 Tg by AA), suggesting about 77% of the annual total deposition being contributed by the AD aerosol mainly through the dry deposition process and 24% of it by AA through the wet deposition process. The monthly mean aerosol concentration and the monthly total deposition show a significant seasonal variation with high in winter and spring, and low in summer.

Incidence and levels of deoxynivalenol, fumonisins and zearalenone contaminants in animal feeds used in Korea in 2012.

The objective of this study was to evaluate the occurrence and levels of deoxynivalenol (DON), fumonisins B1 and B2 (FBs), and zearalenone (ZEN) contaminants in animal feeds used in Korea in 2012. Contamination with DON was observed in 91.33% and 53.33% in compound feeds and feed ingredients, respectively. Among compound feeds, poultry layer feed (laying) exhibited the highest contaminant level of 1.492 mg/kg. FBs contaminants were present in compound feeds and feed ingredients at 93.33% and 83.33%, respectively. Most poultry broiler (early) feeds were highly contaminated with FBs, and one of these feeds detected the level as 12.823 mg/kg as the highest level. The levels of ZEN in compound feeds and feed ingredients were 71.33% and 47%, respectively. Ninety-eight percent of compound feeds for cattle were contaminated with ZEN, and the highest contamination level of 0.405 mg/kg was observed in cattle fatting feeds.

Autophagy regulates endothelial cell processing, maturation and secretion of von Willebrand factor.

Endothelial secretion of von Willebrand factor (VWF) from intracellular organelles known as Weibel-Palade bodies (WPBs) is required for platelet adhesion to the injured vessel wall. Here we demonstrate that WPBs are often found near or within autophagosomes and that endothelial autophagosomes contain abundant VWF protein. Pharmacological inhibitors of autophagy or knockdown of the essential autophagy genes Atg5 or Atg7 inhibits the in vitro secretion of VWF. Furthermore, although mice with endothelial-specific deletion of Atg7 have normal vessel architecture and capillary density, they exhibit impaired epinephrine-stimulated VWF release, reduced levels of high-molecular weight VWF multimers and a corresponding prolongation of bleeding times. Endothelial-specific deletion of Atg5 or pharmacological inhibition of autophagic flux results in a similar in vivo alteration of hemostasis. Thus, autophagy regulates endothelial VWF secretion, and transient pharmacological inhibition of autophagic flux may be a useful strategy to prevent thrombotic events.