PNPLA1 knockdown inhibits esterification of γ-linolenic acid to ceramide 1 in differentiated keratinocytes.

Patatin-like phospholipase domain-containing 1 (PNPLA1) is crucial in the esterification of linoleic acid (LA; 18:2n-6) to ω-hydroxy fatty acids (FA) of ceramide 1 (Cer1), the major barrier lipid of the differentiated epidermis. We previously reported that γ-linolenic acid (GLA; 18:3n-6) as well as LA is esterified to Cer1 subspecies with sphingosine (d18:1) or eicosasphingosine (d20:1) amide-linked to two different ω-hydroxy FA (30wh:0; 32wh:1). Here, we further investigated whether PNPLA1 is also responsible for esterification of GLA to these Cer1 subspecies in normal human keratinocytes (NHK). As late/terminal differentiation was induced in NHK, PNPLA1 and differentiation markers were expressed, and LA-esterified Cer1 subspecies (18:2n-6/C30wh:0 or C32wh:0/d18:1; 18:2n-6/C32wh:0/d20:1) were detected, which were further increased with LA treatment. GLA-esterified Cer1 subspecies (18:3n-6/C30wh:0 or C32wh:0/d18:1; 18:3n-6/C32wh:0/d20:1) were detected only with GLA treatment. Specific small interfering RNA-mediated knockdown of PNPLA1 (KDP) in differentiated NHK decreased levels of these LA-esterified Cer1 subspecies overall and of involucrin (IVL), a terminal differentiation marker. Moreover, KDP resulted in lesser LA/GLA responses as characterized by more significant decreases in IVL and LA/GLA-esterified Cer1 subspecies overall and an accumulation of non-esterified ω-hydroxy ceramides, their putative precursors; the decrease of 18:3n-6/C32wh:0/d18:1, the predominant GLA-esterified Cer1 subspecies, specifically paralleled the increase of C32wh:0/d18:1, its corresponding precursor. PNPLA1 is responsible for NHK terminal differentiation and also for esterification of GLA to the ω-hydroxy FA of Cer1.

A comparative study of the electromyographic activities of lower extremity muscles during level walking and Pedalo riding.

[Purpose] To analyze the electromyographic (EMG) activities of several lower extremity muscles during ground walking and pedaling using the Pedalo Reha-Bar device. [Subjects and Methods] Fifteen healthy adults aged 20-29 year participated in this study. The subjects' surface EMG signals while walking and Pedalo Reha-Bar riding were recorded. The subjects performed 20 steps on flat ground and 20 cycles on the Pedalo Reha-Bar. During the tasks, EMG signals of the rectus femoris, biceps femoris, tibialis anterior, soleus, and gastrocnemius within a 20-second period were recorded. The mean EMG signals within the 10 seconds from 6 to 15 seconds were used for the data analysis. [Results] There was a significant increase in the bilateral use of the rectus femoris and a significant decrease in the use of the left tibialis anterior and left soleus in pedaling using the Pedalo Reha-Bar device compared to ground walking. [Conclusion] Level walking and the Pedalo Reha-Bar riding utilize different types of muscles activities. These results suggest that Pedalo Reha-Bar riding may be used for neuromuscular activation, especially of the rectus femoris.

Laboratory Evaluation of Storage Stability for Asphalt Binder Modified with Crumb Rubber and Styrene-Isoprene-Styrene Depending on Evaluation Factors and Blending Condition.

Modified asphalt binders are still considered important in asphalt pavement. However, the comprehensive use of various modifiers is limited due to storage stability issues. Moreover, there is a scarcity of detailed analyses regarding the degree of separation for asphalt binders among each method despite the utilization of various methods to assess the storage stability of binders. Therefore, a comprehensive analysis was conducted to assess the storage stability of asphalt binder modified with a crumb rubber modifier (CRM) and styrene-isoprene-styrene (SIS), utilizing five evaluation factors following the ASTM D7173 guidelines based on four mixing methods (A: high-shear mixing method, B: low-speed agitating method, C: high-shear mixing method + low mixing method, D: low-speed agitating method + low mixing method). To produce the modified asphalt binder, the proportions of the CRM were 5% and 10% for each binder, and 10% SIS was added to all binders. The results in this study convey that (1) the addition of the modifier led to an increase in G*/sin δ with different mixing methods, but using mixing methods (C and D) for a relatively long time resulted in a lower G*/sin δ, indicating suboptimal performance; (2) through the multiple stress creep recovery (MSCR), rheological properties of Jnr and % rec exhibited trends similar to G*/sin δ evaluation, highlighting an improved elastic recovery with a higher modifier content; (3) storage stability assessment revealed consistent trends in high-shear mixing groups (A and C), while low-speed mixing groups (B and D) exhibited an elevated separation index (SI), suggesting a sensitivity to modification conditions; (4) evaluation using the MSCR method indicated that % rec with a 3.2 kPa load is effective for the sensitive assessment of binder storage stability and Jnr showed a limited sensitivity across varying loads, advocating for % rec for precise evaluation; and (5) despite permitting various tests, achieving consistent results remains challenging. Future research should explore diverse modifiers and optimal evaluation methods to enhance knowledge of binder behavior and separation dynamics.

Characterization of Base Oil Effects on Aged Asphalt Binders Considering Bicycle Road.

Demand for various bicycles and sharing systems has constantly been growing worldwide as they improve the quality of life and promote eco-friendly transportation. Accordingly, it is inevitable that bicycle roads should be expanded. As bicycle roads have a relatively lower load applied than automobile roads, adopting a design method that uses a high reclaimed asphalt pavement (RAP) content can be beneficial. However, much uncertainty still exists about the relation between the mixing method and application in field sites, without appropriately considering the quality control of the rejuvenator. Therefore, this study aims to demonstrate the effect of base oil as a rejuvenator on aged binders, considering the use of a high RAP content for bicycle roads. To prepare the aged binder, a rolling thin-film oven (RTFO) and pressure aging vessel (PAV) were used to imitate the life cycle of asphalt pavement from production to service life, and then three contents of aged binder (0%, 50%, and 100%) were added and mixed with fresh PG 64-22 base binder. Finally, each type of prepared aged asphalt binder was blended with three different base oil contents (0%, 5%, and 10%). The results indicated that (1) the addition of base oil effectively decreased the viscosity of aged binders, (2) aged binders containing base oil showed less G*/sin δ compared to originally aged binders, and (3) the application of base oil improves the cracking properties of the aged binder by decreasing stiffness. In conclusion, the most striking observation from the data analysis from the Superpave test and statistical results was the effect of reducing the asphaltene portion based on the use of base oil in the aged binder. Therefore, using base oil in RAP can enable the application of a high RAP content to the bicycle road.

Effect of Blending and Curing Conditions on the Storage Stability of Rubberized Asphalt Binders.

Crumb rubber modifier (CRM) binders easily suffer from instability at high temperatures, with many suggestions being developed to evaluate their storage stability. However, much uncertainty around CRM binders still exists regarding the relationship between mixing methods and experiments in order to calculate the separation index. In this study, a laboratory investigation into how CRM binders behave regarding storage stability using different mixing methods and experiments was conducted. The CRM asphalt binder in this study was prepared through a wet mixing process with the addition of 5% and 10% crumb rubber modifier (by weight) at 200 °C. The three main modification methods were method A: high-shear mixing (8000 rpm) for 2 h, method B: low mixing (300 rpm) for 8 h, and method C: high-shear mixing (8000 rpm) for 2 h + low mixing (300 rpm) for 6 h. In addition, the effect of separation index (SI) on storage stability was assessed, measuring viscosity, G*/sin δ, and percentage recovery. In general, the results of this study revealed that method C appeared to have the most prominent effect on decreasing the difference between the values of the top and bottom parts; the results for 5% CRM using method C showed that there were no differences among the values for the top, middle, and bottom parts; from the MSCR test, method C was effective in decreasing the difference between the values of the top and bottom parts. It was observed that method C improved storage stability. However, the results for percentage recovery were relatively higher than the separation index when using rotational viscosity and G*/sin δ.

Effect of Kaolinite and Cloisite Na+ on Storage Stability of Rubberized Binders.

This study aimed to evaluate the impact of a two-step modification process involving kaolinite and cloisite Na+ on the storage stability of rubberized binders. The process involved the manual combination of virgin binder PG 64-22 with crumb rubber modifier (CRM), which was heated to condition it. The preconditioned rubberized binder was then modified for two hours at a high speed of 8000 rpm using wet mixing. The second stage modification was performed in two parts, with part 1 using only crumb rubber as the modifier and part 2 involving the use of kaolinite and montmorillonite nano clays at a replacement percentage of 3% to the original weight of the binder along with the crumb rubber modifier. The Superpave and multiple shear creep recovery (MSCR) test methods were used to calculate the performance characteristics and separation index % of each modified binder. The results showed that the viscosity properties of kaolinite and montmorillonite improved the performance class of the binder, with montmorillonite demonstrating greater viscosity values than kaolinite even at high temperatures. Additionally, kaolinite with rubberized binders showed higher resistance to rutting, and the % recovery value from multiple shear creep recovery testing indicated that kaolinite with rubberized binders was more effective than montmorillonite with rubberized binders, even at higher load cycles. The use of kaolinite and montmorillonite reduced phase separation between the asphaltene phase and rubber-rich phase at higher temperatures, but the performance of the rubber binder was affected by higher temperatures. Overall, kaolinite with the rubber binder generally demonstrated greater binder performance.

Fabrication of Pressure Conductive Silicone Rubber Socket Device by Shape-Controlled Nickel Powders Produced by High-Energy Ball Milling.

The pressure conductive silicone rubber socket (PCR) is one of the promising test socket devices in high-speed testing environments. In this study, we report highly dense PCR device channels comprised of high aspect-ratio flake-shaped Ni powders. The shape-controlled Ni powders are prepared by the high-energy milling process. The scanning electron microscopy (SEM) and particle size analyzer (PSA) results of the synthesized powder samples showed well-defined flake type Ni powder morphology, and the powder sizes are distributed in the range of ~24-49 µm. The cross-sectional SEM study of the fabricated PCR revealed that the channels consisting of flake Ni powder are uniformly, densely distributed, and connected as face-to-face contact. The resistance of the PCR channels comprised of flake-shaped Ni powders showed ~23% lower resistance values than the spherical-shaped Ni powders-based channels, which could be due to the face-to-face contact of the powders in the channels. The magnetic properties study for the flake-type Ni powder showed a high remanence (~2.2 emu/g) and coercivity (~5.24 mT), owing to the shape anisotropy factor. Finally, the fabricated highly dense and conductive channels of the silicone rubber socket device by shape-controlled Ni powder could be a potential test socket device.

Effect of Processed Oil on Asphalt Binder Properties.

This study investigates the effectiveness of processed oil in the modification of PG 64-22 and PG 76-22 by assessing their physical and rheological properties, and multiple comparison was conducted between the two binders. The base binders PG 64-22 and PG 76-22 were blended with processed oil at four different percentages of contents (3%, 6%, 9% and 12% by the weight of the binder) and compared with the control binder in each test. The base and modified binders were artificially short-term and long-term aged using a rolling thin film oven (RTFO) and pressure aging vessel (PAV) procedures. Superpave binder tests were performed on the modified binders by applying a rotational viscometer (RV), dynamic shear rheometer (DSR), and bending beam rheometer (BBR). The comparisons and results presented in this study indicate that (1) the processed oil has a significant effect on the binders' viscosity, which changes with respect to the increment of processed oil content. The viscosity of both modified binders decreased with the addition of 3, 6, 9 and 12% processed oil; (2) the performed DSR test showed that the addition of processed oil had a negative effect on the rutting resistance for both binders, since in PG 64-22, G*/Sin δ values decreased by 55, 65, 75 and 83% with the addition of 3, 6, 9 and 12% processed oil, respectively, while a decrement of G*/Sin δ of 24, 45, 58 and 65% with the addition of 3, 6, 9 and 12% processed oil was observed in PG 76-22; meanwhile, the fatigue cracking performance was improved and was found to be effective, while G* Sin δ in PG76-22 decreased by 9, 30, 36, and 52% and in PG 64-22 by 27, 44, 53, and 67% with the addition of 3, 6, 9 and 12% processed oil; (3) the results from the BBR test indicate significant improvement in the thermal cracking properties of the binders. The addition of 3, 6, 9 and 12% processed oil resulted in a decrease in the stiffness of both the PG 64-22 and PG 76-22 binders, with a positive effect consequently being observed on the m-values of the binders.

Evaluation of Effect of Thermoplastic Polyurethane (TPU) on Crumb Rubber Modified (CRM) Asphalt Binder.

Crumb rubber binder with thermoplastic polyurethane (TPU) has been experimented with to characterize the performance properties considering the workability, rutting, fatigue cracking and cracking resistance at low temperatures depending on the temperatures and aging states. Physical and rheological properties were evaluated to proceed with the study by applying Superpave asphalt binder testing and multi-stress creep recovery (MSCR). Based on the targeted experiments, the binder samples were produced at three aging states (original, short-term aged and long-term aged) using a rolling thin film oven (RTFO) and pressure aging vessel (PAV). The results revealed that (i) the addition of TPU into CRM binders has a potential effect on increasing viscoelasticity at the original condition, (ii) CRM binders containing TPU showed improved anti-aging performance based on results of RTFO residues and (iii) the inclusion of TPU made it possible for CRM asphalt binder to improve its fatigue and cracking resistance at low temperature.

Laboratory Characterization of Asphalt Binders Modified with Styrene Butadiene Rubber (SBR).

The study describes the laboratory assessment (physical and rheological properties) of the binders (PG 64-22 and PG 76-22) modified with Styrene Butadiene Rubber (SBR), and a comprehensive comparison between these two modified binder types. PG 64-22 and PG 76-22 were used as base binders. Both of the base binders were blended with SBR at four different percentages of content (0%, 4%, 6%, and 8% by the weight of the binder). The base and modified binders were artificially short-term and long-term aged using a rolling thin film oven (RTFO) and pressure aging vessel (PAV) procedures. Superpave binder tests were conducted on the SBR modified binder using rotational viscometer (RV), dynamic shear rheometer (DSR), and bending beam rheometer (BBR). In depth rutting performance was investigated using Multiple Stress Creep Recovery (MSCR). The results of this study indicated that (1) the addition of SBR into both binders increased the viscosity and polymer modified asphalt (PMA) binders observed to have more significant effect on its viscosity property; (2) the higher the SBR content, the better the rutting resistance of the binder and it is observed that the effect is prominent on the control binder; (3) MSCR test results showed that the SBR modified binders improved the binder percentage recovery and found to have a more significant effect on the PG 76-22 binder compared to PG 64-22; and (4) both the control PG 64-22 and PMA PG 76-22 binders resulted in similar trends on the cracking properties and were found to have insignificant effects due to the addition of an SBR modifier.

Chemical proteomic identification of T-plastin as a novel host cell response factor in HCV infection.

Hepatitis C virus (HCV) infection is the leading cause of chronic liver disease that currently affects at least 170 million people worldwide. Although significant efforts have been focused on discovering inhibitors of a viral polymerase (NS5B) or protease (NS3), strategies to cure HCV infection have been hampered by the limited therapeutic target proteins. Thus, discovery of a novel target remains a major challenge. Here, we report a method that combines transcriptome expression analysis with unbiased proteome reactivity profiling to identify novel host cell response factors in HCV infection. A chemical probe for non-directed proteomic profiling was selected based on genome-wide transcriptome expression analysis after HCV infection, which revealed noticeable alterations related to disulfide bond metabolism. On the basis of this result, we screened the proteome reactivity using chemical probes containing thiol-reactive functional groups and discovered a unique labeling profile in HCV-infected cells. A subsequent quantitative chemical proteomic mapping study led to the identification of a target protein, T-plastin (PLST), and its regulation of HCV replication. Our approach demonstrates both a straightforward strategy for selecting chemical probes to discriminate disease states using a model system and its application for proteome reactivity profiling for novel biomarker discovery.

Rational design of a photo-crosslinking BODIPY for in situ protein labeling.

Photo-crosslinking agents have emerged as critical tools to investigate protein-protein interactions in complex proteomes, but there are few photocrosslinkers available at the moment. Here, we report the first rational design of a photo-crosslinking BODIPY fluorophore (pcBD) and its biological application for biomolecule labeling. As a photosensitizing functional motif, an aryl ketone group was incorporated into the BODIPY fluorophore, and a series of proteins were labeled by pcBD compounds upon UV irradiation. In order to investigate protein-protein interactions in a protein mixture, amino-functionalized pcBD was prepared and covalently attached to a ubiquitin ligase binding peptide. Upon UV irradiation, we could successfully visualize the substrates in the total lysate. These results provided a proof of concept for spatially controllable tagging via photo-activation of the pcBD scaffold, and demonstrated its potential usage for in situ labeling applications.

The novel resveratrol analogue HS-1793 induces apoptosis via the mitochondrial pathway in murine breast cancer cells.

Resveratrol (3,4',5 tri-hydroxystilbene), a natural plant polyphenol, has gained interest as a non-toxic chemopreventive agent capable of inducing tumor cell death in a variety of cancer types. Several studies were undertaken to obtain synthetic analogues of resveratrol with potent anticancer activity. The aim of the present study was to investigate the effect of HS-1793 as a new resveratrol analog on apoptosis via the mitochondrial pathway in murine breast cancer cells. A pharmacological dose (1.3-20 µM) of HS-1793 exerted a cytotoxic effect on murine breast cancer cells resulting in apoptosis. HS-1793-mediated cytotoxicity in FM3A cells by several apoptotic events including mitochondrial cytochrome c release, activation of caspase-3 and PARP occurred. In addition, HS-1793 induced collapse of ∆Ψm and enhanced AIF and Endo G release from mitochondria while undergoing apoptosis. These results demonstrate that the cytotoxicity by HS-1793 in FM3A cells can mainly be attributed to apoptosis via a mitochondrial pathway by caspase activation or contributions of AIF and Endo G.