The Synergistic Effect of Carbon Black/Carbon Nanotube Hybrid Fillers on the Physical and Mechanical Properties of EPDM Composites after Exposure to High-Pressure Hydrogen Gas.

This study investigated the synergistic effect of carbon black/multi-wall carbon nanotube (CB/MWCNT) hybrid fillers on the physical and mechanical properties of Ethylene propylene diene rubber (EPDM) composites after exposure to high-pressure hydrogen gas. The EPDM/CB/CNT hybrid composites were prepared by using the EPDM/MWCNT master batch (MB) with 10 phr CNTs to enhance the dispersion of CNTs in hybrid composites. The investigation included a detailed analysis of cure characteristics, crosslink density, Payne effect, mechanical properties, and hydrogen permeation properties. After exposure to 96.3 MPa hydrogen gas, the hydrogen uptake and the change in volume and mechanical properties of the composites were assessed. We found that as the MWCNT volume fraction in fillers increased, the crosslink density, filler-filler interaction, and modulus of hybrid composites increased. The hydrogen uptake and the solubility of the composites decreased with an increasing MWCNT volume fraction in fillers. Moreover, after exposure to hydrogen gas, the change in volume and mechanical properties exhibited a diminishing trend with a higher MWCNT volume fraction. We conclude that the hybridization of CB and CNTs formed strong filler-filler networks in hybrid composites, consequently reinforcing the EPDM composites and enhancing the barrier properties of hydrogen gas.

High-Resolution Spatial Transcriptomic Atlas of Mouse Soleus Muscle: Unveiling Single Cell and Subcellular Heterogeneity in Health and Denervation.

Skeletal muscle is essential for both movement and metabolic processes, characterized by a complex and ordered structure. Despite its importance, a detailed spatial map of gene expression within muscle tissue has been challenging to achieve due to the limitations of existing technologies, which struggle to provide high-resolution views. In this study, we leverage the Seq-Scope technique, an innovative method that allows for the observation of the entire transcriptome at an unprecedented submicron spatial resolution. By applying this technique to the mouse soleus muscle, we analyze and compare the gene expression profiles in both healthy conditions and following denervation, a process that mimics aspects of muscle aging. Our approach reveals detailed characteristics of muscle fibers, other cell types present within the muscle, and specific subcellular structures such as the postsynaptic nuclei at neuromuscular junctions, hybrid muscle fibers, and areas of localized expression of genes responsive to muscle injury, along with their histological context. The findings of this research significantly enhance our understanding of the diversity within the muscle cell transcriptome and its variation in response to denervation, a key factor in the decline of muscle function with age. This breakthrough in spatial transcriptomics not only deepens our knowledge of muscle biology but also sets the stage for the development of new therapeutic strategies aimed at mitigating the effects of aging on muscle health, thereby offering a more comprehensive insight into the mechanisms of muscle maintenance and degeneration in the context of aging and disease.

Effect of a looming visual cue on situation awareness and perceived urgency in response to a takeover request.

This study aimed to investigate the effect of a looming visual cue on situation awareness and perceived urgency in response to a takeover request (TOR), and to explore the underlying mechanisms of this effect through three experiments. In Experiment 1, the optimal size and speed of a red disk were determined, which were effective in capturing looming motion and conveying the urgency of the situation. The results indicated that both looming speed and size ratio had significant effects on situation awareness and perceived urgency. In Experiment 2, the effects of looming stimuli were compared with dimming stimuli, and the results showed that the looming visual cue was more effective in promoting perceived urgency and situation awareness. The results also indicated that the looming visual cue attracted more visual attention than the dimming visual cue, in line with previous studies. Experiment 3 utilized a driving simulator to test the effectiveness of the looming visual cue in promoting fast and appropriate responses to TORs in complex driving scenarios. The results showed that the looming visual cue was more effective in promoting perceived urgency and enhancing situation awareness, especially in highly complex driving situations. Overall, the findings suggest that the looming visual cue is a powerful tool for promoting fast and appropriate responses to TORs and enhancing situation awareness, particularly in complex driving scenarios. These results have important implications for designing effective TOR systems and improving driver safety on the road.

Evaluation of circulating IGF-I and IGFBP-3 as biomarkers for tumors in dogs.

BACKGROUND: Serum-based parameters are considered non-invasive biomarkers for cancer detection. In human studies, insulin-like growth factor-I and II (IGF-I and IGF-II) and insulin-like growth factor binding protein-3 (IGFBP-3) are useful as diagnostic or prognostic markers and potential therapeutic targets. OBJECTIVES: This study examined the diagnostic utility of circulating IGF-I, IGF-II, and IGFBP-3 levels in healthy dogs and dogs with tumors. METHODS: The serum concentrations of these biomarkers in 86 dogs with tumors were compared with those in 30 healthy dogs using an enzyme-linked immunosorbent assay (ELISA). RESULTS: The ELISA results showed no difference between healthy dogs and dogs with tumors in the serum IGF-II concentrations. On the other hand, there was a significant difference in the circulating IGF-I and IGFBP-3 levels between healthy dogs and dogs with tumors. The concentrations of serum IGF-I (median [interquartile range], 103.4 [59.5-175] ng/mL) in dogs with epithelial tumors were higher than those (58.4 ng/mL [43.5-79.9]) in healthy dogs. Thus, the concentrations of serum IGFBP-3 (43.4 ng/mL [33.2-57.2]) in dogs with malignant mesenchymal tumors were lower than those (60.8 ng/mL [47.6-70.5]) in healthy dogs. CONCLUSIONS: The serum IGF-I and IGFBP-3 levels can be used as diagnostic biomarkers in dogs with tumors.

Estimating Bar Graph Averages: Overcoming Within-the-Bar Bias.

Although most people are not aware of it, bias can occur when interpreting graphs. Within-the-bar bias describes a misinterpretation of the distribution of data underlying bar graphs that indicate an average or where the average estimation point moves inside the bar when the average of several graphs is estimated. This study proposes and tests two methods based on information processing to reduce within-the-bar bias. The first method facilitates bottom-up processing by changing various graph features, such as presenting confidence intervals, placing boundaries around the graph, and showing cumulative bars with different tones. The second method facilitates top-down processing by instructing participants to estimate the mean based on a dot at the end of each bar. Testing of the first method showed that cumulative bars reduced bias, but the other methods did not. The second method was found to reduce bias. Overall, our results demonstrate that the accurate interpretation of bar graphs can be facilitated through the manipulation of specific graph features and instruction.

What Makes Consumers' Intention to Purchase Paid Stickers in Personal Messenger? The Role of Personality and Motivational Factors.

Many messengers and social networking services (SNSs) use emojis and stickers as a means of communication. Stickers express individual emotions well, allowing long texts to be replaced with small pictures. As the use of stickers increased, stickers were commercialized on a few platforms and showed remarkable growth as people bought and used stickers with their favorite characters, products, or entertainers online. Depending on their personality, individuals have different motivations for using stickers that determine the usefulness and enjoyment of stickers, affecting their purchase decisions. In the present study, participants (n = 302) who were randomly recruited from a university completed an online questionnaire assessing the Big Five personality characteristics, motivations for using stickers, and the technology acceptance model (TAM). Results using partial least squares structural equation modeling (PLS-SEM) revealed that each personality trait affected different motivations for using stickers. Moreover, motivations for using stickers also influenced different technology acceptance variables. Finally, perceived usefulness, enjoyment, and ease of use had a positive effect on the intention to purchase stickers. This study has implications in that it is an exploratory approach to the intention to purchase stickers, which has been investigated by few prior studies, and it sheds light on the relationship between personality, motivation, and TAM in purchasing stickers. It also suggests that personality and motivation factors can be considered in personalized recommendation services.

Geometric calibration of a camera and a turntable system using two views.

A calibration algorithm for a system comprising a camera and turntable is provided, in which the camera captures three-dimensional point clouds while a target object rotates on the turntable to three-dimensionally scan the entire object. A mathematical approach is used to obtain the geometry between the coordinate systems of the camera and the turntable. As a minimal setup, the camera captures two views of a chessboard calibration pattern placed on the turntable. The coordinate transformation from the camera to the turntable is then calculated through geometric reasoning. First, two camera poses are computed with respect to the chessboard images taken before and after rotation. The direction of the rotation axis of the turntable coordinate system can be calculated without any ambiguity. However, there is one degree of freedom in the choice of the other two directions and one degree of freedom in choosing the location of the origin of the turntable coordinate system. We provide a practical method for these choices. Experimental results with a stereo-vision system and a turntable validate the proposed method.

Two-dimensional semiconducting and single-crystalline antimony trioxide directly-grown on monolayer graphene.

The first successful synthesis and characterization of single-crystalline two-dimensional (2D) semiconducting antimony tri oxide (Sb2O3) by direct chemical vapor deposition (CVD) growth on monolayer graphene is presented herein. The Sb2O3 flakes on graphene are regular triangles with smooth edges and a thickness of ∼1.45 nm. The thickness dependence of the Raman spectra and scanning Kelvin probe microscopy (SKPM) were systematically studied.

Social relationships enhance the time spent eating and intake of a novel diet in pregnant Hanwoo (Bos taurus coreanae) heifers.

The objective of this study was to evaluate the effects of social relationships on the feed intake, eating behavior, and growth, upon exposure to a novel diet, in Hanwoo (Bos taurus coreanae) heifers during pregnancy. Twenty-four pregnant Hanwoo heifers, averaging 438 ± 27.8 kg in weight, 21 months in age, and 194 ± 8.5 days in pregnancy, were involved in a two-month (eight weeks) experiment. The heifers were randomly assigned to either the single housing group (SG; one individual per pen, n = 12), or the paired housing group (PG; two individuals per pen, n = 12). All pens were of the same size (5 × 5 m) and provided with one feed bin, which automatically recorded the individual feed intake and eating behavior. As the experiment began, the diet of the heifers was switched from a total mixed ration (TMR; 250 g/kg ryegrass straw and 750 g/kg concentrate mix) to a forage-only diet (mixed hay cubes composed of 500 g/kg alfalfa, 250 g/kg timothy, and 250 g/kg blue grass hay). The heifers were fed ad libitum twice a day. The individual feed intake and eating behavior were recorded daily throughout the experiment, and body weights (BWs) were measured every four weeks before the morning feeding. PG animals visited the feed bin 22% less often than SG. PG, however, stayed 39% longer in the feed bin and consumed 40% more feed per visit, compared with SG. Consequently, PG heifers spent 23% more time in eating and had 16% more daily dry matter intake than SG during the experiment. Average daily gain during the experimental period tended to be greater in PG than in SG. When pregnant Hanwoo heifers encountered a novel diet, social relationships (i.e., presence of a pen-mate) enhanced their time spent eating and feed intake. Social interactions, even with an unfamiliar individual, may be helpful for pregnant Hanwoo heifers cope with a diet challenge compared to solitary situation.

Quantitative Evaluation of the Dispersion of Graphene Sheets With and Without Functional Groups Using Molecular Dynamics Simulations.

Nanofluids with enhanced thermal properties are candidates for thermal management in automotive systems, with scope for improving energy efficiency. In particular, many studies have reported on dispersions of nanoparticles with long-term stability in the base fluid, with qualitative evaluations of the dispersion stability via either the naked eye or optical instruments. Additives such as surfactants can be used to enhance the dispersion of nanoparticles; however, this may diminish their intrinsic thermal properties. Here, we describe molecular dynamics simulations of nanofluids containing graphene sheets dispersed in ethylene glycol and water. We go on to suggest a quantitative evaluation method for the degree of dispersion, based on the ratio of the total number of nanoparticles to the number of clustered nanoparticles. Moreover, we investigate the effects of functional groups on the surface of graphene, which are expected to improve the dispersion without requiring additives such as surfactants due to steric hindrance and chemical affinity for the surrounding fluid. We find that, for pure graphene, the degree of dispersion decreased as the quantity of graphene sheets increased, which is attributed to an increased probability of aggregation at higher loadings; however, the presence of functional groups inhibited the graphene sheets from forming aggregates.

Study on a 4-week recovery test of sweet bee venom after a 13-week, repeated, intramuscular dose toxicity test in sprague-dawley rats.

OBJECTIVES: This study was performed to check for reversibility in the changes induced by a 13-week, repeated, dose toxicity test of Sweet Bee Venom (SBV) in Sprague-Dawley (SD) rats. METHODS: Fifteen male and 15 female SD rats were treated with 0.28 mg/kg of SBV (high-dosage group) and the same numbers of male and female SD rats were treated with 0.2 mL/kg of normal saline (control group) for 13 weeks. We selected five male and five female SD rats from the high-dosage group and the same numbers of male and female SD rats from the control group, and we observed these rats for four weeks. We conducted body-weight measurements, ophthalmic examinations, urinalyses and hematology, biochemistry, histology tests. RESULTS: (1) Hyperemia and movement disorder were observed in the 13-week, repeated, dose toxicity test, but these symptoms were not observed during the recovery period. (2) The rats in the high-dose group showed no significant changes in weight compared to the control group. (3) No significant differences in the ophthalmic parameters, urine analyses, complete blood cell counts (CBCs), and biochemistry were observed among the recovery groups. (4) No changes in organ weights were observed during the recovery period. (5) Histological examination of the thigh muscle indicated cell infiltration, inflammation, degeneration, necrosis of muscle fiber, and fibrosis during the treatment period, but these changes were not observed during the recovery period. The fatty liver change that was observed during the toxicity test was not observed during the recovery period. No other organ abnormalities were observed. CONCLUSION: The changes that occurred during the 13-week, repeated, dose toxicity test are reversible, and SBV can be safely used as a treatment modality.

Study of a 13-weeks, Repeated, Intramuscular Dose, Toxicity Test of Sweet Bee Venom in Sprague-Dawley Rats.

OBJECTIVES: This study was performed to analyze a 13-week repeated dose toxicity test of Sweet Bee Venom (SBV) extracted from bee venom and administered in Sprague-Dawley (SD) rats. METHODS: Male and female 5-week-old SD rats were treated once daily with SBV (high-dosage group: 0.28 mg/kg; medium-dosage group: 0.14 mg/kg; or low-dosage group: 0.07 mg/kg) for 13 weeks. Normal saline was administered to the control group in a similar manner (0.2 mL/kg). We conducted clinical observations, body weight measurements, ophthalmic examinations, urinalyses, hematology and biochemistry tests, and histological observations using hematoxylin and eosin (H&E) staining to identify any abnormalities caused by the SBV treatment. RESULTS: During this study, no mortality was observed in any of the experimental groups. Hyperemia and a movement disorder were observed around the area of in all groups that received SBV treatment, with a higher occurrence in rats treated with a higher dosage. Male rats receiving in the high-dosage group showed a significant decrease in weight during the treatment period. Compared to the control group, no significant changes in the ophthalmic parameters, the urine analyses, the complete blood cell count (CBC), and the biochemistry in the groups treated with SBV. Compared to the control group, some changes in organ weights were observed in the medium-and the high-dosage groups, but the low-dosage group showed no significant changes. Histological examination of thigh muscle indicated cell infiltration, inflammation, degeneration, and necrosis of muscle fiber, as well as fibrosis, in both the medium- and the high-dosage groups. Fatty liver change was observed in the periportal area of rats receiving medium and high dosages of SBV. No other organ abnormalities were observed. CONCLUSION: Our findings suggest that the No Observed Adverse Effect Level (NOAEL) of SBV is approximately 0.07 mg/kg in male and female SD rats.

Cellular delivery and biological activity of antisense oligonucleotides conjugated to a targeted protein carrier.

Targeted delivery can potentially improve the pharmacological effects of antisense and siRNA oligonucleotides. Here, we describe a novel bioconjugation approach to the delivery of splice-shifting antisense oligonucleotides (SSOs). The SSOs are linked to albumin via reversible S-S bonds. The albumin is also conjugated with poly(ethylene glycol) (PEG) chains that terminate in an RGD ligand that selectively binds the alphavbeta3 integrin. As a test system, we utilized human melanoma cells that express the alphavbeta3 integrin and that also contain a luciferase reporter gene that can be induced by delivery of SSOs to the cell nucleus. The RGD-PEG-SSO-albumin conjugates were endocytosed by the cells in an RGD-dependent manner; using confocal fluorescence microscopy, evidence was obtained that the SSOs accumulate in the nucleus. The conjugates were able to robustly induce luciferase expression at concentrations in the 25-200 nM range. At these levels, little short-term or long-term toxicity was observed. Thus, the RGD-PEG-albumin conjugates may provide an effective tool for targeted delivery of oligonucleotides to certain cells and tissues.

Intracellular delivery of an anionic antisense oligonucleotide via receptor-mediated endocytosis.

We describe the synthesis and characterization of a 5' conjugate between a 2'-O-Me phosphorothioate antisense oligonucleotide and a bivalent RGD (arginine-glycine-aspartic acid) peptide that is a high-affinity ligand for the alphavbeta3 integrin. We used alphavbeta3-positive melanoma cells transfected with a reporter comprised of the firefly luciferase gene interrupted by an abnormally spliced intron. Intranuclear delivery of a specific antisense oligonucleotide (termed 623) corrects splicing and allows luciferase expression in these cells. The RGD-623 conjugate or a cationic lipid-623 complex produced significant increases in luciferase expression, while 'free' 623 did not. However, the kinetics of luciferase expression was distinct; the RGD-623 conjugate produced a gradual increase followed by a gradual decline, while the cationic lipid-623 complex caused a rapid increase followed by a monotonic decline. The subcellular distribution of the oligonucleotide delivered using cationic lipids included both cytoplasmic vesicles and the nucleus, while the RGD-623 conjugate was primarily found in cytoplasmic vesicles that partially co-localized with a marker for caveolae. Both the cellular uptake and the biological effect of the RGD-623 conjugate were blocked by excess RGD peptide. These observations suggest that the bivalent RGD peptide-oligonucleotide conjugate enters cells via a process of receptor-mediated endocytosis mediated by the alphavbeta3 integrin.

Remediation of heavy metal contaminated groundwater originated from abandoned mine using lime and calcium carbonate.

Column and pilot scale experiments for a chemical treatment involving the use of coagulants to remediate heavy metal contaminated groundwater were performed. Granulated lime (Ca(OH)(2)) and calcium carbonate (CaCO(3)) were used as coagulants and contaminated groundwater obtained at an abandoned Fe-mine in Korea was used for the experiments. The main removal mechanism of heavy metals in the experiments was "sweep precipitation" by coagulation. Using granulated lime as a coagulant in the column experiment, more than 98% of As and Ni were removed from artificially contaminated water. When granulated calcium carbonate was used in the artificially contaminated water, the removal efficiencies of Ni and Zn were more than 97%, but As removal efficiency was lower than 50%. For the continuous column experiment with mixed lime and calcium carbonate at a 1:1 (v/v) ratio, almost all As was removed and more than 98 % of Ni was removed. For pilot scale experiments (acryl tank: 34 cm in length and 24 cm in diameter), the removal efficiencies of As and Cd were above 96% for 150l groundwater treatment and their accumulated removal capacities linearly maintained. This suggests that coagulants could treat more than 22 times greater groundwater volume compared with the volume of coagulants used.

Tat-conjugated PAMAM dendrimers as delivery agents for antisense and siRNA oligonucleotides.

PURPOSE: PAMAM G5 dendrimer (P) was conjugated to Tat peptide (T), a cell penetrating peptide, in search of an efficient cellular delivery vehicle for antisense and siRNA oligonucleotides. METHODS: PAMAM G5 dendrimer was reacted with 4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-propionic acid, sulfosuccinimidyl ester, sodium salt (BODIPY) for visualization to yield the conjugate BP. Bifunctional sulfosuccinimidyl 6-[alpha-methyl-alpha-(2-pyridyldithio)toluamido]hexanoate (sulfo-LC-SMPT) was then used to conjugate primary amino groups of BP to cysteine derivatized Tat peptide to give the designed conjugate, BPT. This conjugate was complexed with antisense and siRNA oligonucleotides designed to inhibit MDR1 gene expression. NIH 3T3 MDR cells were used for the evaluation of biological activity of the conjugate. RESULTS: Both antisense and siRNA readily formed complexes with the synthesized BPT, introduced into NIH 3T3 MDR cells, and primarily accumulated in intracellular vesicles. MDR1 gene expression was partially inhibited by the antisense-BPT complex and weakly inhibited by the siRNA-BPT complex when both were tested at nontoxic levels of dendrimer. Conjugation with Tat peptide did not improve the delivery efficiency of the dendrimer. CONCLUSIONS: Dendrimer-oligonucleotide complexes were moderately effective for delivery of antisense and only poorly effective for delivery of siRNA. Conjugation of the dendrimer with the Tat cell penetrating peptide failed to further enhance the effectiveness of the dendrimer.

Delivery of antisense oligonucleotides using cholesterol-modified sense dendrimers and cationic lipids.

Cholesterol modified mono-, di-, and tetrameric oligonucleotides were synthesized and hybridized with antisense oligonucleotides to study their incorporation in cationic liposomes together with the influence of this dendrimeric delivery system on biological activity. Electrostatic interactions seem to play the most important role during complexation with cationic lipids. This oligonucleotide formulation gives a small but significant increase in the inhibition of P-glycoprotein expression in a cellular system.

Epigenetic manipulation of gene expression: a toolkit for cell biologists.

Cell biologists have been afforded extraordinary new opportunities for experimentation by the emergence of powerful technologies that allow the selective manipulation of gene expression. Currently, RNA interference is very much in the limelight; however, significant progress has also been made with two other approaches. Thus, antisense oligonucleotide technology is undergoing a resurgence as a result of improvements in the chemistry of these molecules, whereas designed transcription factors offer a powerful and increasingly convenient strategy for either up- or down-regulation of targeted genes. This mini-review will highlight some of the key features of these three approaches to gene regulation, as well as provide pragmatic guidance concerning their use in cell biological experimentation based on our direct experience with each of these technologies. The approaches discussed here are being intensely pursued in terms of possible therapeutic applications. However, we will restrict our comments primarily to the cell culture situation, only briefly alluding to fundamental differences between utilization in animals versus cells.

Application of nonionic surfactant-enhanced in situ flushing to a diesel contaminated site.

Surfactant-enhanced in situ flushing was performed to remediate soil and groundwater at a diesel contaminated area, which had been used as a military vehicle repair area in Korea for 45 years. A pilot-scale site (4 m x 4 m x 4 m) was selected within the contaminated area for in situ flushing; the selected site was composed of heterogeneous sandy and silt-sandy soils, with an average hydraulic conductivity (K) of 2.0 x 10(-4)cm/s. Two percent of sorbitan monooleate (POE 20) was mixed with uncontaminated groundwater and five pore volumes of solution (three pore volumes of surfactant solution and two pore volumes of groundwater alone) were flushed to remove diesel from the site. The effluent TPH (total petroleum hydrocarbon) concentration with surfactant solution flushing increased to 1761 mg/L, which was over 200 times higher than the average concentration with only groundwater flushing. A total of 48 kg of TPH (about 88% of the initial TPH) was removed from the pilot site with five pore volumes of 2% sorbitan monooleate solution flushing; this total was more than 75 times the amount that was removed when flushing with water alone (less than 640 g). All of the extracted solution was treated by means of a chemical treatment process, which included the use of a dissolved air flotation system to lower the concentration of solution below 5mg/L and the treated solution was then disposed of in a nearby sewage drain.

Inhibition of MDR1 gene expression by chimeric HNA antisense oligonucleotides.

Hexitol nucleic acids (HNAs) are nuclease resistant and provide strong hybridization to RNA. However, there is relatively little information on the biological properties of HNA antisense oligonucleotides. In this study, we compared the antisense effects of a chimeric HNA 'gapmer' oligonucleotide comprising a phosphorothioate central sequence flanked by 5' and 3' HNA sequences to conventional phosphorothioate oligonucleotides and to a 2'-O-methoxyethyl (2'-O-ME) phosphorothioate 'gapmer'. The antisense oligomers each targeted a sequence bracketing the start codon of the message of MDR1, a gene involved in multi-drug resistance in cancer cells. Antisense and control oligonucleotides were delivered to MDR1-expressing cells using transfection with the cationic lipid Lipofectamine 2000. The anti-MDR1 HNA gapmer was substantially more potent than a phosphorothioate oligonucleotide of the same sequence in reducing expression of P-glycoprotein, the MDR1 gene product. HNA and 2'-O-ME gapmers displayed similar potency, but a pure HNA antisense oligonucleotide (lacking the phosphorothioate 'gap') was ineffective, indicating that RNase H activity was likely required. Treatment with anti-MDR1 HNA gapmer resulted in increased cellular accumulation of the drug surrogate Rhodamine 123 that correlated well with the reduced cell surface expression of P-glycoprotein. Thus, HNA gapmers may provide a valuable additional tool for antisense-based investigations and therapeutic approaches.