Comparisons of ELISA and Western blot assays for detection of autophagy flux

We analyzed autophagy/mitophagy flux in vitro (C2C12 myotubes) and in vivo (mouse skeletal muscle) following the treatments of autophagy inducers (starvation, rapamycin) and a mitophagy inducer (carbonyl cyanide m-chlorophenylhydrazone, CCCP) using two immunodetection methods, ELISA and Western blotting, and compared their working range, accuracy, and reliability. The ELISAs showed a broader working range than that of the LC3 Western blots (Table 1). Table 2 showed that data value distribution was tighter and the average standard error from the ELISA was much smaller than those of the Western blot, directly relating to the accuracy of the assay. Test-retest reliability analysis showed good reliability for three individual ELISAs (interclass correlation, ≥ 0.7), but poor reliability for three individual Western blots (interclass correlation, ≤ 0.4) (Table 3). Keywords: Autophagy, Mitophagy, ELISA, Western blot, Skeletal muscl

Carbon Monoxide Protects against Hepatic Ischemia/Reperfusion Injury via ROS-Dependent Akt Signaling and Inhibition of Glycogen Synthase Kinase 3β

Carbon monoxide (CO) may exert important roles in physiological and pathophysiological states through the regulation of cellular signaling pathways. CO can protect organ tissues from ischemia/reperfusion (I/R) injury by modulating intracellular redox status and by inhibiting inflammatory, apoptotic, and proliferative responses. However, the cellular mechanisms underlying the protective effects of CO in organ I/R injury remain incompletely understood. In this study, a murine model of hepatic warm I/R injury was employed to assess the role of glycogen synthase kinase-3 (GSK3) and phosphatidylinositol 3-kinase (PI3K)-dependent signaling pathways in the protective effects of CO against inflammation and injury. Inhibition of GSK3 through the PI3K/Akt pathway played a crucial role in CO-mediated protection. CO treatment increased the phosphorylation of Akt and GSK3-beta (GSK3β) in the liver after I/R injury. Furthermore, administration of LY294002, an inhibitor of PI3K, compromised the protective effect of CO and decreased the level of phospho-GSK3β after I/R injury. These results suggest that CO protects against liver damage by maintaining GSK3β phosphorylation, which may be mediated by the PI3K/Akt signaling pathway. Our study provides additional support for the therapeutic potential of CO in organ injury and identifies GSK3β as a therapeutic target for CO in the amelioration of hepatic injury

Islands stretch test for measuring the interfacial fracture energy between a hard film and a soft substrate

We present a technique for measuring the interfacial fracture energy, $\Gamma_i$, between a hard thin film and a soft substrate. A periodic array of hard thin islands is fabricated on a soft substrate, which is then subjected to uniaxial tension under an optical microscope. When the applied strain reaches a critical value, delamination between the islands and the substrate starts from the edge of the islands. As the strain is increased, the interfacial cracks grow in a stable fashion. At a given applied strain, the width of the delaminated region is a unique function of the interfacial fracture energy. We have calculated the energy release rate driving the delamination as a function of delamination width, island size, island thickness, and applied strain. For a given materials system, this relationship allows determination of the interfacial fracture energy from a measurement of the delamination width. The technique is demonstrated by measuring the interfacial fracture energy of plasma-enhanced chemical vapor deposition SiNx islands on a polyimide substrate. We anticipate that this technique will find application in the flexible electronics industry where hard islands on soft substrates are a common architecture to protect active devices from fracture.Engineering and Applied Science

Effects of education on low-phosphate diet and phosphate binder intake to control serum phosphate among maintenance hemodialysis patients: A randomized controlled trial.

Background:For phosphate control, patient education is essential due to the limited clearance of phosphate by dialysis. However, well-designed randomized controlled trials about dietary and phosphate binder education have been scarce. Methods:We enrolled maintenance hemodialysis patients and randomized them into an education group (n = 48) or a control group (n = 22). We assessed the patients' drug compliance and their knowledge about the phosphate binder using a questionnaire. Results:The primary goal was to increase the number of patients who reached a calcium-phosphorus product of lower than 55. In the education group, 36 (75.0%) patients achieved the primary goal, as compared with 16 (72.7%) in the control group (P = 0.430). The education increased the proportion of patients who properly took the phosphate binder (22.9% vs. 3.5%, P = 0.087), but not to statistical significance. Education did not affect the amount of dietary phosphate intake per body weight (education vs. control: -1.18 ± 3.54 vs. -0.88 ± 2.04 mg/kg, P = 0.851). However, the dietary phosphate-to-protein ratio tended to be lower in the education group (-0.64 ± 2.04 vs. 0.65 ± 3.55, P = 0.193). The education on phosphate restriction affected neither the Patient-Generated Subjective Global Assessment score (0.17 ± 4.58 vs. -0.86 ± 3.86, P = 0.363) nor the level of dietary protein intake (-0.03 ± 0.33 vs. -0.09 ± 0.18, P = 0.569). Conclusion:Education did not affect the calcium-phosphate product. Education on the proper timing of phosphate binder intake and the dietary phosphate-to-protein ratio showed marginal efficacy

Inorganic Islands on a Highly Stretchable Polyimide Substrate

For a flexible electronic device integrating inorganic materials on a polymer substrate, the polymer can deform substantially, but the inorganic materials usually fracture at small strains. This paper describes an approach to make such a device highly stretchable. A polyimide substrate is first coated with a thin layer of an elastomer, on top of which SiNx islands are fabricated. When the substrate is stretched to a large strain, the SiNx islands remain intact. Calculations confirm that the elastomer reduces the strain in the SiNx islands by orders of magnitude.Engineering and Applied Science

Implant selection for successful reverse total shoulder arthroplasty

Reverse total shoulder arthroplasty (RTSA) emerged as a new concept of arthroplasty that does not restore normal anatomy but does restore function. It enables the function of the torn rotator cuff to be performed by the deltoid and shows encouraging clinical outcomes. Since its introduction, various modifications have been designed to improve the outcome of the RTSA. From the original cemented baseplate with peg or keel, a cementless baseplate was designed that could be fixed with central and peripheral screws. In addition, a modular-type glenoid component enabled easier revision options. For the humeral component, the initial design was an inlay type of long stem with cemented fixation. However, loss of bone stock from the cemented stem hindered revision surgery. Therefore, a cementless design was introduced with a firm metaphyseal fixation. Furthermore, to prevent complications such as scapular notching, the concept of lateralization emerged. Lateralization helped to maintain normal shoulder contour and better rotator cuff function for improved external/internal rotation power, but excessive lateralization yielded problems such as subacromial notching. Therefore, for patients with pseudoparalysis or with risk of subacromial notching, a medial eccentric tray option can be used for distalization and reduced lateralization of the center of rotation. In summary, it is important that surgeons understand the characteristics of each implant in the various options for RTSA. Furthermore, through preoperative evaluation of patients, surgeons can choose the implant option that will lead to the best outcomes after RTSA.Level of evidence: IV

Neural Correlates of Transient Mal de Debarquement Syndrome: Activation of Prefrontal and Deactivation of Cerebellar Networks Correlate With Neuropsychological Assessment

Background: Mal de debarquement syndrome (MdDS) is characterized by a subjective perception of self-motion after exposure to passive motion, mostly after sea travel. A transient form of MdDS (t-MdDS) is common in healthy individuals without pathophysiological certainty. In the present cross-sectional study, the possible neuropsychiatric and functional neuroimaging changes in local fishermen with t-MdDS were evaluated. Methods: The present study included 28 fishermen from Buan County in South Korea; 15 (15/28, 53.6%) participants experienced t-MdDS for 1–6 h, and 13 were asymptomatic (13/28, 46.4%). Vestibular function tests were performed using video-oculography, the video head impulse test, and ocular and cervical vestibular-evoked myogenic potentials. Visuospatial function was also assessed by the Corsi block test. Brain imaging comprised structural MRI, resting-state functional MRI, and [18F]FDG PET scans. Results: The results of vestibular function tests did not differ between the fishermen with and those without t-MdDS. However, participants with t-MdDS showed better performance in visuospatial memory function than those without t-MdDS (6.40 vs. 5.31, p-value = 0.016) as determined by the Corsi block test. Structural brain MRIs were normal in both groups. [18F]FDG PET showed a relative hypermetabolism in the bilateral occipital and prefrontal cortices and hypometabolism in the vestibulocerebellum (nodulus and uvula) in participants with t-MdDS compared to those without t-MdDS. Resting-state functional connectivities were significantly decreased between the vestibular regions of the flocculus, superior temporal gyrus, and parietal operculum and the visual association areas of the middle occipital gyrus, fusiform gyrus, and cuneus in participants with t-MdDS. Analysis of functional connectivity of the significant regions in the PET scans revealed decreased connectivity between the prefrontal cortex and visual processing areas in the t-MdDS group. Conclusion: Increased visuospatial memory, altered metabolism in the prefrontal cortex, visual cognition cortices, and the vestibulocerebellum, and decreased functional connectivity between these two functional areas might indicate reductions in the integration of vestibular input and enhancement of visuospatial attention in subjects with t-MdDS. Current functional neuroimaging similarities from transient MdDS via chronic MdDS to functional dizziness and anxiety disorders suggest a shared mechanism of enhanced self-awareness as a kind of continuum or as overlap disorders

Sensitivity of censored data analysis to determine the characteristic value of structural timber

In structural timber tests, unintended failure mechanisms occur frequently in specimens and their results are called censored data. There are two censored data analysis: censored maximum likelihood estimation (CMLE) and Kaplan–Meier (KM) method. In this study, the precision of the censored data analysis was investigated to determine the characteristic value, 5th percentile value, of the structural timber. The results show that (1) the 5th percentile value was underestimated by ordinary data analysis methods; maximum likelihood estimation (MLE) and Order statistics. (2) CMLE with 30% lower tail censored data and KM method provided much more precise 5th percentile value. (3) The amount of under-measurement (5 MPa, 10 MPa, and 15 MPa in this simulation study) did not show significant effect on the 5th percentile determination in CMLE and KM method, but the proportion of censored data (percentage of unintended failure specimen; 10%, 20%, 30%, and 40%) affected the determination of 5th percentile value. (4) CMLE with 30% lower tail censored data and KM method showed good agreement in case that the data included unintended failure data up to 20%.(1) This work was supported by Research Resettlement Fund for the new fac‑ulty of Seoul National University. (2) This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIT) (No. 2017R1A2B1010748)

TSLP Induces Mast Cell Development and Aggravates Allergic Reactions through the Activation of MDM2 and STAT6

Thymic stromal lymphopoietin (TSLP) is known to promote T helper type 2 cell–associated inflammation. Mast cells are major effector cells in allergic inflammatory responses. We noted that the population and maturation of mast cells were reduced in TSLP-deficient mice (TSLP-/-). Thus, we hypothesized that TSLP might affect mast cell development. We found that TSLP induced the proliferation and differentiation of mast cells from bone marrow progenitors. TSLP-induced mast cell proliferation was abolished by depletion of mouse double minute 2 (MDM2) and signal transducers and activators of transcription 6 (STAT6), as an upstream activator of MDM2. TSLP-/-, in particular, had a considerable deficit in the expression of MDM2 and STAT6. Also, the TSLP deficiency attenuated mast cell–mediated allergic reactions through the downregulation of STAT6 and MDM2. In an antibody microarray chip analysis, MDM2 expression was increased in atopic dermatitis patients. These observations indicate that TSLP is a factor for mast cell development, and that it aggravates mast cell–mediated immune responses