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Abstract Background Insulin-degrading enzyme (IDE) is a widely studied zinc-metalloprotease implicated in the pathogenesis of type 2 diabetes mellitus, Alzheimer disease (AD) and varicella zoster virus infection. Despite more than six decades of research on IDE, progress has been hampered by the lack of well-characterized reagents targeting this biomedically important protease. To address this important need, we generated and characterized new mouse monoclonal antibodies (mAbs) targeting natively folded human and rodent IDE. Results Eight monoclonal hybridoma cell lines were derived in house from mice immunized with full-length, natively folded, recombinant human IDE. The mAbs derived from these lines were shown to detect IDE selectively and sensitively by a wide range of methods. Two mAbs in particular—designated 6A1 and 6H9—proved especially selective for IDE in immunocytochemical and immunohistochemical applications. Using a variety of methods, we show that 6A1 selectively detects both human and rodent IDE, while 6H9 selectively detects human, but not rodent, IDE, with both mAbs showing essentially no cross reactivity with other proteins in these applications. Using these novel anti-IDE mAbs, we also developed sensitive and quantitative sandwich ELISAs capable of quantifying IDE levels present in human brain extracts. Conclusion We succeeded in developing novel mAbs that selectively detect rodent and/or human IDE, which we have shown to be suitable for a wide range of applications, including western blotting, immunoprecipitation, immunocytochemistry, immunohistochemistry, and quantitative sandwich ELISAs. These novel anti-IDE mAbs and the assays derived from them constitute important new tools for addressing many unresolved questions about the basic biology of IDE and its role in multiple highly prevalent human diseases.

Abstract Insulin-degrading enzyme (IDE) is a ubiquitously expressed zinc-metalloprotease that degrades several pathophysiologically significant extracellular substrates, including insulin and the amyloid β-protein (Aβ), and accumulating evidence suggests that IDE dysfunction may be operative in both type 2 diabetes mellitus and Alzheimer disease (AD). Although IDE is well known to be secreted by a variety of cell types, the underlying trafficking pathway(s) remain poorly understood. To address this topic, we investigated the effects of known inhibitors or stimulators of protein secretion on the secretion of IDE from murine hepatocytes and HeLa cells. IDE secretion was found to be unaffected by the classical secretion inhibitors brefeldin A (BFA), monensin, or nocodazole, treatments that readily inhibited the secretion of α1-antitrypsin (AAT) overexpressed in the same cells. Using a novel cell-based Aβ-degradation assay, we show further that IDE secretion was similarly unaffected by multiple stimulators of protein secretion, including glyburide and 3'-O-(4-benzoyl)benzoyl-ATP (Bz-ATP). The calcium ionophore, A23187, increased extracellular IDE activity, but only under conditions that also elicited cytotoxicity. Our results provide the first biochemical evidence that IDE export is not dependent upon the classical secretion pathway, thereby identifying IDE as a novel member of the select class of unconventionally secreted proteins. Further elucidation of the mechanisms underlying IDE secretion, which would be facilitated by the assays described herein, promises to uncover processes that might be defective in disease or manipulated for therapeutic benefit.

Predicting the strength parameters of multi-type sediments containing hydrates is the basis and precondition for the safe and efficient development of natural gas hydrates. However, studies on the shear mechanical behavior and morphology of multi-type hydrate-bearing sediments (HBS) are still insufficient. Herein, this study presents an integrated test system that can be used to measure the interfacial strength and morphology of multi-type sediments containing hydrates. This device integrates specimen preparation, shear test, morphology observation, and data analysis, which is helpful to comprehensively evaluate interfacial strength, roughness, and morphology. The propagation and development characteristics of microfractures of HBS during shearing can be obtained, which is favorable for identifying the damage and failure modes. Preliminary validation experiments have been conducted on massive pure hydrate, hydrate–sediment interface, and homogenous HBS to verify the applicability of the device for multi-type HBS. The device and corresponding analysis method are expected to support the evaluation of interfacial strength and morphology, thereby promoting a deeper understanding of hydrate–sediment interactions and failure mechanisms of hydrate reservoirs. [ABSTRACT FROM AUTHOR]

The relationship between molybdenum and kidney-related disease outcomes, including hyperuricemia, is not well investigated. This study aims to determine whether molybdenum and its antioxidative property are associated with systemic inflammation and kidney-related disease parameters including hyperuricemia. Urinary molybdenum's epidemiological relationship to hyperuricemia and kidney-disease related outcomes was evaluated in 15,370 adult participants in the National Health and Nutrition Examination Survey (NHANES) collected between 1999 and 2016. Individuals' urinary molybdenum levels were corrected to their urinary creatinine concentrations. The association between urinary molybdenum-to-creatinine ratio and kidney-disease related outcomes were assessed by multivariable linear and logistic regression analyses, adjusting for covariates including age, sex, ethnicity, diabetes mellitus, hypertension, body mass index, and estimated glomerular filtration rate. Antimony and tungsten were used as control trace metals. Experimentally, HK-2 cell was used to assess molybdenum's antioxidative properties. HK-2 cells were challenged with H2O2-induced oxidative stress. Oxidative stress was measured using a fluorescent microplate assay for reactive oxygen species (ROS) and antioxidation levels were assessed by measuring the expression of manganese superoxide dismutase. In the adult NHANES population, urinary molybdenum-to-creatinine ratio was significantly associated with decreased serum uric acid (β, -0.119; 95% CI, -0.148 to -0.090) concentrations, and decreased prevalence of hyperuricemia (OR, 0.73; 95% CI, 0.64–0.83) and gout (OR, 0.71; 95% CI, 0.52–0.94). Higher urinary molybdenum levels were associated with lower levels of systemic oxidative stress (gamma-glutamyltransferase levels; β, -0.052; 95% CI, -0.067 to -0.037) and inflammation (C-reactive protein levels; β, -0.184; 95% CI, -0.220 to -0.148). In HK-2 cells under H2O2-induced oxidative stress, molybdenum upregulated manganese superoxide dismutase expression and decreased oxidative stress. Urinary molybdenum levels are associated with decreased prevalence of hyperuricemia and gout in adult population. Molybdenum's antioxidative properties might have acted as an important mechanism for the reduction of systemic inflammation, ROS, and uric acid levels. [ABSTRACT FROM AUTHOR]

In this paper, the effect of micro-dimple textures (produced by a laser) on the tribological properties of bearings is investigated. This study offers guidelines to reduce the friction torque of the bearing pair and addresses the problem of difficult start-ups after shutdowns. Micro-pits with different texture diameters and depths were machined on the surface of journal bearings. Then, the impact of several different texture parameters on the tribological performance of the bearing pairs was studied using an orthogonal experimental design. Subsequently, the surface morphology of the bearings before and after the friction and wear test was observed using scanning electron microscopy (SEM) and energy-dispersive spectrometry (EDS). These observations were then used to determine the type/state of friction and wear, which also improves our understanding of how texture affects the service life of bearings. The results indicate that the bearings' micro-pit surface hardness follows an approximate parabolic spatial distribution that decreases along the micro-pit wall. Furthermore, the laser processing of surface textures was found to cause hardening in certain areas, and the chemical composition of elemental carbon and oxygen at the inner surface of processed bearings increased by 31.1% and 7.9%, respectively. Moreover, abrasive wear was identified as the primary form of wear. The textured surface's antifriction mechanism primarily functioned to trap particles, which acted as a secondary lubrication source and altered the lubrication states by serving as a medium for supplied lubricants. The results confirm that a suitable selection of texture parameters can not only effectively reduce the friction coefficient without shortening the service life of the bearing pair but also facilitate the smooth start-up of the rotor–bearing system. [ABSTRACT FROM AUTHOR]

In order to investigate the turbulent lubrication performance of a textured hydrodynamic journal bearing (THJB), a model of turbulent lubrication was established in this paper. The variations in the Reynolds number, oil film thickness, oil film pressure, bearing capacity, attitude angle, and side leakage flow with structural and working parameters were studied, and the axis whirl orbit was further analyzed. The results show that turbulent lubrication is suitable for the actual operating conditions of THJBs. The Reynolds number decreases with the eccentricity ratio in the pressure-bearing zone but increases with rotational speeds, whereas the variation in the maximum oil film pressure increases and the minimum oil film thickness decreases with the eccentricity ratio under various Reynolds numbers. The bearing capacity decreases with the dimple diameter, depth, oil film thickness, and clearance ratio but increases with the length/diameter ratio and dimple spacing. As the eccentricity ratio increases, the attitude angle decreases, but the side leakage flow increases. In addition, the system tends to be unstable as the rotational speed and length/diameter ratio increase, and the friction and wear on the surface are three-body friction. This work not only helps in analyzing the characteristics of a THJB under actual operating conditions but also provides support for research on the simulation of THJB's lubrication mechanism of THJB via computational fluid dynamics. [ABSTRACT FROM AUTHOR]

Introduction: This prospective cohort study investigated the clinical role of circulating tumor necrosis factor receptor (cTNFR) levels as prognostic biomarkers in severe acute kidney injury (AKI) patients requiring continuous renal replacement therapy (CRRT). Methods: We enrolled 136 patients from 7 hospitals participating in the VENUS (VolumE maNagement Under body composition monitoring in critically ill patientS on CRRT) trial from July 2017 to October 2019. The levels of cTNFR1 and cTNFR2 were measured using plasma samples collected on days 0 (D0), 2 (D2), and 7 (D7). Patients were divided into high- and low-cTNFR groups based on their receptor concentrations. Results: D0 concentrations of cTNFR1 and cTNFR2 were positively correlated with one another (R2 = 0.37, p < 0.001). The high-cTNFR1 group displayed a higher in-hospital mortality rate than the low-TNFR1 group (p = 0.002). Moreover, the mortality rate was significantly higher in the high-TNFR1 group than in the low-TNFR1 group after adjusting for age, sex, and acute physiology, and chronic health evaluation II scores (hazard ratio 1.82, 95% confidence interval 1.09–3.03, p = 0.025). D2 and D7 cTNFR1 levels were also associated with in-hospital mortality; contrastingly, cTNFR2 levels were not associated with this outcome. Additionally, patients were divided into three groups according to the change in cTNFR levels from D0 to D2 (ΔcTNFR). Those in the highest ΔcTNFR tertile had a higher mortality rate than the remaining patients (p = 0.033 for ΔcTNFR1; p = 0.025 for ΔcTNFR2). Patients who underwent AKI-to-chronic kidney disease transition had higher concentrations of cTNFR1 (p = 0.014). Discussion/Conclusion: Plasma cTNFR1 concentrations at CRRT initiation and changes in cTNFR1 and 2 levels immediately following CRRT initiation are significant biomarkers for predicting the outcomes of patients with severe AKI. [ABSTRACT FROM AUTHOR]