Cathartocytosis: How Cells Jettison Unwanted Material as They Reprogram.

Injury can cause differentiated cells to undergo massive reprogramming to become proliferative to repair tissue via a cellular program called paligenosis. Gastric digestive-enzyme-secreting chief cells use paligenosis to reprogram into progenitor-like Spasmolytic-Polypeptide Expressing Metaplasia (SPEM) cells. Stage 1 of paligenosis is to downscale mature cell architecture via a process involving lysosomes. Here, we noticed that sulfated glycoproteins (which are metaplasia and cancer markers in mice and humans) were not digested during paligenosis but excreted into the gland lumen. Various genetic and pharmacological approaches showed that endoplasmic reticulum membranes and secretory granule cargo were also excreted and that the process proceeded in parallel with, but was independent lysosomal activity. 3-dimensional light and electron-microscopy demonstrated that excretion occurred via unique, complex, multi-chambered invaginations of the apical plasma membrane. As this lysosome-independent cell cleansing process does not seem to have been priorly described, we termed it "cathartocytosis". Cathartocytosis allows a cell to rapidly eject excess material (likely in times of extreme stress such as are induced by paligenosis) without waiting for autophagic and lysosomal digestion. We speculate the ejection of sulfated glycoproteins (likely mucins) would aid in downscaling and might also help bind and flush pathogens (like H pylori which causes SPEM) away from tissue.

Development of Heteroatomic Constant Potential Method with Application to MXene-Based Supercapacitors.

We describe a method for modeling constant-potential charges in heteroatomic electrodes, keeping pace with the increasing complexity of electrode composition and nanostructure in electrochemical research. The proposed "heteroatomic constant potential method" (HCPM) uses minimal added parameters to handle differing electronegativities and chemical hardnesses of different elements, which we fit to density functional theory (DFT) partial charge predictions in this paper by using derivative-free optimization. To demonstrate the model, we performed molecular dynamics simulations using both HCPM and conventional constant potential method (CPM) for MXene electrodes with Li-TFSI/AN (lithium bis(trifluoromethane sulfonyl)imide/acetonitrile)-based solvent-in-salt electrolytes. Although the two methods show similar accumulated charge storage on the electrodes, the results indicated that HCPM provides a more reliable depiction of electrode atom charge distribution and charge response compared with CPM, accompanied by increased cationic attraction to the MXene surface. These results highlight the influence of elemental composition on electrode performance, and the flexibility of our HCPM opens up new avenues for studying the performance of diverse heteroatomic electrodes including other types of MXenes, two-dimensional materials, metal-organic frameworks (MOFs), and doped carbonaceous electrodes.

Wearable Triboelectric Nanogenerator with Ground-Coupled Electrode for Biomechanical Energy Harvesting and Sensing.

Harvesting biomechanical energy for electricity as well as physiological monitoring is a major development trend for wearable devices. In this article, we report a wearable triboelectric nanogenerator (TENG) with a ground-coupled electrode. It has a considerable output performance for harvesting human biomechanical energy and can also be used as a human motion sensor. The reference electrode of this device achieves a lower potential by coupling with the ground to form a coupling capacitor. Such a design can significantly improve the TENG's outputs. A maximum output voltage up to 946 V and a short-circuit current of 36.3 µA are achieved. The quantity of the charge that transfers during one step of an adult walking reaches 419.6 nC, while it is only 100.8 nC for the separate single-electrode-structured device. In addition, using the human body as a natural conductor to connect the reference electrode allows the device to drive the shoelaces with integrated LEDs. Finally, the wearable TENG is able to perform motion monitoring and sensing, such as human gait recognition, step count and movement speed calculation. These show great application prospects of the presented TENG device in wearable electronics.

Super-tough and self-healable all-cellulose-based electrolyte for fast degradable quasi-solid-state supercapacitor.

Recyclable and degradable supercapacitors have promising applications for a sustainable energy storage industry. Herein, we prepare a dual-physical crosslinking (DP) carboxymethyl cellulose (CMC) hydrogel with high-toughness, healability, and electric conductivity by integrating abundant ions into the matrix. The prepared hydrogel displays a maximum compressive fracture stress of 4.42 MPa, fast healing in five seconds, and full degradation within eight days. Moreover, the fabricated supercapacitor shows high specific capacitance (309 F g-1) and volumetric capacitance (2.60 F cm-3). The supercapacitor achieves a healing efficiency of 93.9 % after five cuttings, and exhibits a cycling stability of 84.6 % capacitance retention after 1000 cycles. These merits ensure that the all-cellulose-based supercapacitor can operate in case of sudden collision and deformation, which contribute to reducing the environmental hazards from supercapacitor's preparation to its abandonment.

Structure-Dynamics Interrelation Governing Charge Transport in Cosolvated Acetonitrile/LiTFSI Solutions.

Concentrated ionic solutions present a potential improvement for liquid electrolytes. However, their conductivity is limited by high viscosities, which can be attenuated via cosolvation. This study employs a series of experiments and molecular dynamics simulations to investigate how different cosolvents influence the local structure and charge transport in concentrated lithium bis(trifluoromethane-sulfonyl)imide (LiTFSI)/acetonitrile solutions. Regardless of whether the cosolvent's dielectric constant is low (for toluene and dichloromethane), moderate (acetone), or high (methanol and water), they preserve the structural and dynamical features of the cosolvent-free precursor. However, the dissimilar effects of each case must be individually interpreted. Toluene and dichloromethane reduce the conductivity by narrowing the distribution of Li+-TFSI- interactions and increasing the activation energies for ionic motions. Methanol and water broaden the distributions of Li+-TFSI- interactions, replace acetonitrile in the Li+ solvation, and favor short-range Li+-Li+ interactions. Still, these cosolvents strongly interact with TFSI-, leading to conductivities lower than that predicted by the Nernst-Einstein relation. Finally, acetone preserves the ion-ion interactions from the cosolvent-free solution but forms large solvation complexes by joining acetonitrile in the Li+ solvation. We demonstrate that cosolvation affects conductivity beyond simply changing viscosity and provide fairly unexplored molecular-scale perspectives regarding structure/transport phenomena relation in concentrated ionic solutions.

Biocompatible carboxymethyl cellulose-based super-elastic hierarchical sponge via a novel templating and plasticizing method.

Herein, a facile method to fabricate hierarchical super-elastic (SE) sponge using a water-soluble cellulose derivative, carboxymethyl cellulose (CMC), is reported. The method includes ice templating and porogen leaching steps which facilitate to generate macro-sized pores as well as pore wall structures that can dissipate stress effectively. By controlling the porogen content, the specific surface area and the morphology of the sponges can be tuned. Furthermore, a plasticizing method was used before vacuum drying to reduce the deformation of the inner structure. The derived hierarchical SE CMC sponges exhibit excellent fatigue resistance, fast shape recovery, high-water absorption, biosafeness, and fast degradation. Thus, our strategy provides a novel method for the construction of SE sponges which show great potential in green elastic wound dressing, tissue engineering, and absorbent materials.

Universal Ligands for Dispersion of Two-Dimensional MXene in Organic Solvents.

Ligands can control the surface chemistry, physicochemical properties, processing, and applications of nanomaterials. MXenes are the fastest growing family of two-dimensional (2D) nanomaterials, showing promise for energy, electronic, and environmental applications. However, complex oxidation states, surface terminal groups, and interaction with the environment have hindered the development of organic ligands suitable for MXenes. Here, we demonstrate a simple, fast, scalable, and universally applicable ligand chemistry for MXenes using alkylated 3,4-dihydroxy-l-phenylalanine (ADOPA). Due to the strong hydrogen-bonding and π-electron interactions between the catechol head and surface terminal groups of MXenes and the presence of a hydrophobic fluorinated alkyl tail compatible with organic solvents, the ADOPA ligands functionalize MXene surfaces under mild reaction conditions without sacrificing their properties. Stable colloidal solutions and highly concentrated liquid crystals of various MXenes, including Ti2CTx, Nb2CTx, V2CTx, Mo2CTx, Ti3C2Tx, Ti3CNTx, Mo2TiC2Tx, Mo2Ti2C3Tx, and Ti4N3Tx, have been produced in various organic solvents. Such products offer excellent electrical conductivity, improved oxidation stability, and excellent processability, enabling applications in flexible electrodes and electromagnetic interference shielding.

Controlling the Ion Transport Number in Solvent-in-Salt Solutions.

Solvent-in-salt (SIS) systems present promising materials for the next generation of energy storage applications. The ion dynamics is significantly different in these systems from that of ionic liquids and diluted salt solutions. In this study, we analyze the ion dynamics of two salts, Li-TFSI and Li-FSI, in highly concentrated aqueous and acetonitrile solutions. We performed high-frequency dielectric measurements covering the range of up to 50 GHz and molecular dynamics simulations. The analysis of the conductivity spectra provides the characteristic crossover time between individual charge rearrangements and the normal charge diffusion regime resulting in DC conductivity. Analysis revealed that the onset of normal charge diffusion occurs at the scale of ∼1.5-3.5 Å, comparable to the average distance between the ions. Based on the idea of momentum conservation, distinct ion correlations were estimated experimentally and computationally. The analysis revealed that cation-anion correlations can be suppressed by changing the solvent concentration in SIS systems, leading to an increase of the light ion (Li+ in our case) transport number. This discovery suggests a way for improving the light cation transport number in SIS systems by tuning the solvent concentration.

NMR and Theoretical Study of In-Pore Diffusivity of Ionic Liquid-Solvent Mixtures.

Despite having a lower energy density than common batteries, electric double-layer capacitors (EDLCs) offer several advantages for high-power applications, including high power density, quick charge and discharge time, and long cycle life. Room-temperature ionic liquids (RTILs) have been intensely studied as promising electrolytes for applications in ELDCs because of their wide potential window, low volatility, as well as thermal and chemical stability. The main deficiency of neat RTILs in such applications is the sluggish diffusivity, which restricts the EDLCs' power density. To alleviate the slow diffusivity, RTILs can be used in a mixture with organic solvents. In this study, we applied two-dimensional exchange nuclear magnetic resonance spectroscopy (2D EXSY NMR) and molecular dynamics (MD) simulations to investigate the diffusivity of anions of an RTIL, namely, 1-butyl-3-methyl-imidazolium bis(trifluoromethylsulfonyl)imide (BMIM+-TFSI-), dissolved in five different organic solvents, in the micropores of activated carbon. We determined that the relative concentrations of ions in solutions in the micropores were higher than those in the bulk solutions and were also solvent-dependent. The ion diffusivities in the pores were found to be almost 2 orders of magnitude slower than in the bulk solutions, with methanol showing the largest relative disparity. These results suggested that the interactions of solvents with the activated carbon are critical not only to the power density of EDLCs but also to the energy density. The comparisons of ion diffusivities between the experiments and the MD simulations suggest the need to consider also the surface functionalities of activated carbon for the simulation of ion diffusion in the micropores of activated carbon.

Effects of molecular weight of polyaspartic acid on nitrogen use efficiency and crop yield.

BACKGROUND: In the past decades, ever-increasing fertilizer use has led to a continuous increase in agricultural output. However, serious waste of resources occurs because of the low utilization of fertilizers. Polyaspartic acid (PASP) is a biodegradable polymer that can be used as a fertilizer synergist in agricultural production to improve the nutrient utilization capacity of plants. For polymers, the molecular weight (MW) often affects their effectiveness. However, little information is available on the effects of PASP MW in agriculture, especially on nitrogen leaching and plant element uptake. RESULTS: This work was conducted to identify the effect of PASPs with three different MWs - PASP-1 (MW: 5517), PASP-2 (MW: 6934), and PASP-3 (MW: 7568) - on nitrogen leaching, lettuce growth, and wheat cultivation. The results revealed that PASP favored plant growth and nitrogen accumulation in the soil, independent of crop species. PASP with a higher MW improved yields and the agronomic characteristics of lettuce and wheat. Furthermore, apparent amelioration of nitrogen use efficiency for lettuce (7.6%, 12.8%, and 15.0%) and wheat (4.6%, 8.1%, and 9.2%) was observed in the treatments with PASP addition. The effects and merits of PASPs on preventing ammonium nitrogen leaching and improving lettuce and wheat productivity were as follows: PASP-3 > PASP-2 > PASP-1. CONCLUSION: The MW of PASP is an essential factor affecting inorganic nitrogen leaching and crop productivity, and PASP with a higher MW (7568) is recommended for application in agriculture. © 2022 Society of Chemical Industry.

Direct Correlation of the Salt-Reduced Diffusivities of Organic Solvents with the Solvent's Mole Fraction.

Lithium bis(trifluoromethanesulfonyl)imide (LiTFSI) in organic solvents (especially propylene carbonate) has demonstrated extraordinary pseudocapacitive performance as an electrolyte in the supercapacitor configuration ( Nat. Energy 2019, 4, 241-248). However, the influence of the solvated ions on the diffusivity of the solvent molecules is yet to be understood. We examine the impact of LiTFSI on the diffusivity in five organic solvents: acetonitrile (ACN), tetrahydrofuran (THF), methanol (MeOH), dimethyl sulfoxide (DMSO), and propylene carbonate (PC) using a combination of neutron scattering, conductivity measurements, and molecular dynamics simulations. The extent of the diffusivity reduction in the concentration regime of ≤1 M directly correlates with the solvent mole fraction at which the solvation shells around Li+ ions are of similar size in all the solvents, resulting in a universal ∼50% reduction in the solvent diffusivity. These results provide guidance for formulation of the new electrolytes to enhance the performance of energy storage devices.

Effects of nuclear factor-κB signaling pathway on periodontal ligament stem cells under lipopolysaccharide-induced inflammation.

Lipopolysaccharide (LPS) induces inflammatory stress and apoptosis. This study focused on the effect of nuclear factor kappa B (NF-κB) signaling pathway on proliferation and osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) after LPS induction and its mechanism. We first isolated hPDLSCs from human tooth root samples in vitro. Then, flow cytometry detected positive expression of cell surface antigens CD146 and STRO-1 and negative expression of CD45, suggesting the hPDLSCs were successfully isolated. LPS significantly induced increased apoptosis and diminished proliferation of hPDLSCs. The NF-κB pathway agonist phorbol 12-myristate 13-acetate (PMA) or p65 overexpression inhibited the proliferation of LPS-treated hPDLSCs and promoted apoptosis. PMA also promoted LPS-induced up-regulation of the expression of inflammatory factors TNF-α and IL-6 and down-regulation of the expression of anti-inflammatory factor IL-10. Additionally, LPS was confirmed to lead to a reduction of alkaline phosphatase (ALP) activity, calcium nodules, and expression of osteogenic markers Runt-related transcription factor 2 (Runx2) and osteopontin. This reduction could be promoted by PMA. Western blotting further indicated that PMA could promote LPS-induced decrease of expression of p65 (cytoplasm), and total cellular proteins IKKα and IKKß in hPDLSCs, while protein expression of p-IκBα (cytoplasm) and p65 (nucleus), and p-IκBα/IκBα ratio was elevated. By contrast, inhibition of the NF-κB pathway (PDTC) or small-interfering RNA targeting NF-κB/p65 (p65 siRNA) showed the opposite results. In conclusion, activation of NF-κB signaling in LPS-induced inflammatory environment can inhibit the proliferation and osteogenic differentiation of hPDLSCs. This study provides a theory foundation for the clinical treatment of periodontitis.

Room temperature preparation of cellulose nanocrystals with high yield via a new ZnCl2 solvent system.

Here, a facile method to fabricate cellulose nanocrystals (CNCs) with high yield from microcrystalline cellulose (MCC) at room temperature (RT) is achieved by using a new solvent system of zinc chloride (ZnCl2) and a little amount of hydrochloric acid (HCl). Compared with sulphuric acid hydrolysis process, about one-fifth mole of acid is used for per gram of CNCs in our protocol. CNCs with rod-like morphology are regenerated with a maximum yield of 35.2% and high crystallinity of 73.8%. Moreover, with an additional 2 h of ball-milling, the yield of CNCs could significantly increase to 66.9% at RT. The possible formation mechanism for CNCs prepared by the solvent system of ZnCl2/HCl is proposed. As the first example of isolation of CNCs with high yield at RT using ZnCl2, this work provides a facile, energy-saving, and practical strategy for the preparation of cellulose nanomaterials.

Hydrogen-bonding-assisted toughening of hierarchical carboxymethyl cellulose hydrogels for biomechanical sensing.

Herein we present a CMC based hydrogel that can be engineered for texture by applying a soaking strategy after freeze-thaw. These two processes lead to a hierarchal structure within the CMC polymer hydrogels. Such hydrogels can be strain hardening; that is the gel modulus increases with higher strain levels. Both the failure stress and energy are high (1.54 MPa and 9.94 kJm-2 respectively). At sub-failure strains, the hydrogels show a high degree of recoverability as evidenced by their minimal hysteresis in a stress-strain test. Furthermore, these hydrogels are electrically conductive making them good candidates for tough sensors and wearable electronic devices.

Water insoluble and flexible transparent film based on carboxymethyl cellulose.

Preparation of renewable, insoluble, and transparent films is still a major challenge for the application of soft electronics and packing industry. Herein, a "green" protocol for preparation of such a film based on carboxymethyl cellulose (CMC) is presented, where acid assistant freeze-thaw method was used in combination with drying. We have shown that the resultant films displayed flexibility, high light transmittance (above 90 %), insolubility, high mechanical performances (elastic modulus of 29.6 MPa), and good thermal stability. Moreover, CMC film/filter paper was fabricated, and the waterproof and mechanical properties of which were investigated. This approach offers a promising route to the fabrication of flexible and transparent films with good waterproof properties based on soluble biomass.

Ripening of avocado fruits studied by spectroscopic techniques.

Avocados are considered very healthy due to the high content mono-unsaturated lipid, essential vitamins and minerals, minimal sugar and no cholesterol and are therefore sometimes referred to as "the perfect fruits". Avocados, mainly grown in Latin-America, are harvested unripe and sent overseas. However, the ripening process is very difficult to assess visually and tactilely. A tool for precise noninvasive judgment of the status would be valuable as the fruit is too expensive to be cut open unripe or overdue. A white-light source and a light-emitting diode unit with four excitation wavelengths (365, 385, 395, and 405 nm) were used for reflectance and fluorescence spectroscopy in a fiber-coupled set-up for noninvasive monitoring. Twelve non-ripe avocados, with approximately the same size and appearance, were studied and divided into three groups and kept at three different storage conditions; at room temperature, in a refrigerator and a combination of the two. We showed that fluorescence was useful for following the ripening process. A method, which compensates for the spatial variations in spectral properties around a fruit, is described. Remote fluorescence monitoring, intended for orchard use, was also demonstrated. A low-cost device based on fluorescence for avocado ripeness assessment is proposed.

microRNA­196b promotes cell migration and invasion by targeting FOXP2 in hepatocellular carcinoma.

Accumulating evidence indicates that microRNAs (miRNAs) play important roles in tumorigenesis and metastasis. Recent research has shown that miR­196b is implicated in metastasis by regulating the migration and invasion of cancer cells. However, the clinical significance of miR­196b and its role as well as the underlying mechanisms in hepatocellular carcinoma (HCC) remain largely unknown. Here, we detected miR­196b expression in HCC and matched non-tumor tissues with qRT­PCR. We found that miR­196b displayed higher expression in HCC patient tissues and cells. Clinical analysis revealed that high miR­196 expression was correlated with venous infiltration, advanced TNM stage and poor prognosis. Functionally, we demonstrated that miR­196b promoted the migration and invasion of HCC cells in vitro. Moreover, miR­196b knockdown restrained pulmonary metastasis in vivo. Mechanistically, we confirmed that miR­196b could directly bind to 3'UTR of forkhead box P2 (FOXP2) mRNA and repress its expression. miR­196b and FOXP2 showed a negative correlation in HCC tissues. More importantly, upregulation of FOXP2 antagonized miR­196b­mediated migration and invasion in Hep3B cells. Furthermore, FOXP2 knockdown partially reversed the anti­metastatic function of the miR­196b inhibitor on HCCLM3 cells. Taken together, we demonstrated that miR­196b may function as a prognostic biomarker and suppressed FOXP2 expression, subsequently leading to the metastasis of HCC. Our findings highlight a novel mechanism of miR­196b in the progression of HCC and identify miR­196b/FOXP2 axis as a promising target for HCC.

Vitamin D3 supplementation decreases a unique circulating monocyte cholesterol pool in patients with type 2 diabetes.

Cross-sectional studies indicate consistent associations between low 25(OH)D concentration and increased risk of cardiovascular disease (CVD), but results of randomized control trials (RCTs) are mixed. However, the majority of the RCTs do not focus on type 2 diabetics, potentially obscuring the effects of vitamin D in this population. In vitro 1,25(OH)2D3 downregulates macrophage cholesterol deposition, but the in vivo effects are unknown. To explore potential mechanisms of the effects of vitamin D on CVD risk in patients with type 2 diabetes, we isolated monocytes in a subset of 26 patients from our RCT of diabetics with baseline serum 25(OH)D <25ng/mL randomized to vitamin D3 4000 IU/day or placebo for 4 months. Upon enrollment, the mean 25(OH)D level was 17ng/mL, which increased to 36ng/mL after vitamin D and remained unchanged in the placebo group. Before randomization, groups demonstrated similar mean hemoglobin A1c and plasma lipids levels, none of which was significantly altered by vitamin D supplementation. Moreover, assessment of oxidized LDL uptake in monocytes cultured in the patient's own serum before vs. after treatment resulted in >50% reduction in the vitamin D group with no change in the placebo group. This was mediated through suppression of endoplasmic reticulum stress and scavenger receptor CD36 protein expression. The reduction in monocyte cholesterol uptake was reflected in a 19% decrease in total monocyte cholesterol content. Interestingly, cross-sectional analysis of circulating monocytes from vitamin D-deficient vs. sufficient diabetic patients revealed 8-fold higher cholesteryl ester content, confirming the capacity of these monocytes to uptake and carry cholesterol in the circulation. This study identifies a unique circulating cholesterol pool within monocytes that is modulated by vitamin D and has the potential to contribute to CVD in type 2 diabetes.

Immature colon carcinoma transcript-1 promotes cell growth of hepatocellular carcinoma via facilitating cell cycle progression and apoptosis resistance.

Immature colon carcinoma transcript-1 (ICT1) is a newly identified oncogene, which regulates proliferation, cell cycle progression and apoptosis of cancer cells. However, the clinical significance, biological function and underlying mechanisms of ICT1 in hepatocellular carcinoma (HCC) remain poorly known. In the present study, we showed that the expression of ICT1 in HCC tissues were notably overexpressed compared to corresponding non-tumor tissues. Accordingly, the relative levels of ICT1 were upregulated in HCC cell lines compared with LO2 cells. The positive expression of ICT1 was correlated with large tumor size and advanced TNM tumor stage. Kaplan-Meier plots indicated that ICT1-positive expression in HCC patients showed a prominent shorter survival. In addition, ICT1 knockdown inhibited proliferation and cell cycle progression, and induced apoptosis in HepG2 cells. While, ICT1 overexpression showed opposite effects on these cellular processes of Hep3B cells. In vivo experiments demonstrated that ICT1 deficiency reduced the growth of subcutaneous HCC in nude mice. Notably, ICT1 knockdown reduced the levels of CDK1, cyclin B1 and Bcl-2 and increased the expression of Bax in HepG2 cells. ICT1 overexpression resulted in upregulation of CDK1, cyclin B1 and Bcl-2, and downregulation of Bax in Hep3B cells. Furthermore, microRNA-134 (miR-134) was recognized as a direct upstream regulator and inversely modulated ICT1 abundance in HCC cells. Altogether, our data support that miR-134 regulation of ICT1 facilitates malignant phenotype of HCC cells probably via cell cycle and apoptosis-associated proteins including CDK1, cyclin B1, Bcl-2 and Bax.

Endogenous Cholesterol Excretion Is Negatively Associated With Carotid Intima-Media Thickness in Humans.

OBJECTIVE: Epidemiological studies strongly suggest that lipid factors independent of low-density lipoprotein cholesterol contribute significantly to cardiovascular disease risk. Because circulating lipoproteins comprise only a small fraction of total body cholesterol, the mobilization and excretion of cholesterol from plasma and tissue pools may be an important determinant of cardiovascular disease risk. Our hypothesis is that fecal excretion of endogenous cholesterol is protective against atherosclerosis. APPROACH AND RESULTS: Cholesterol metabolism and carotid intima-media thickness were quantitated in 86 nondiabetic adults. Plasma cholesterol was labeled by intravenous infusion of cholesterol-d7 solubilized in a lipid emulsion and dietary cholesterol by cholesterol-d5 and the nonabsorbable stool marker sitostanol-d4. Plasma and stool samples were collected while subjects consumed a cholesterol- and phytosterol-controlled metabolic kitchen diet and were analyzed by mass spectrometry. Carotid intima-media thickness was negatively correlated with fecal excretion of endogenous cholesterol (r=-0.426; P<0.0001), total cholesterol (r=-0.472; P≤0.0001), and daily percent excretion of cholesterol from the rapidly mixing cholesterol pool (r=-0.343; P=0.0012) and was positively correlated with percent cholesterol absorption (r=+0.279; P=0.0092). In a linear regression model controlling for age, sex, systolic blood pressure, hemoglobin A1c, low-density lipoprotein, high-density lipoprotein cholesterol, and statin drug use, fecal excretion of endogenous cholesterol remained significant (P=0.0008). CONCLUSIONS: Excretion of endogenous cholesterol is strongly, independently, and negatively associated with carotid intima-media thickness. The reverse cholesterol transport pathway comprising the intestine and the rapidly mixing plasma, and tissue cholesterol pool could be an unrecognized determinant of cardiovascular disease risk not reflected in circulating lipoproteins. Further work is needed to relate measures of reverse cholesterol transport to atherosclerotic disease. CLINICAL TRIAL REGISTRATION: URL: http://www.clinicaltrials.gov. Unique identifier: NCT01603758.