Sleep disturbance correlated with severity of neuropathic pain in sciatic nerve crush injury model.

The association between sleep and pain has been investigated widely. However, inconsistent results from animal studies compared with human data show the need for a validated animal model in the sleep-pain association field. Our study aims to validate common neuropathic pain models as a tool for evaluating the sleep-pain association. Electrodes electroencephalogram (EEG) and electromyogram (EMG) were surgically implanted to measure sleep. The von Frey test was used to measure pain sensitivity. Following the baseline data acquisition, two pain-modelling procedures were performed: sciatic nerve crush injury (SCI) and common peroneal nerve ligation (CPL). Post-injury measurements were performed on days 1, 5, 10, and 15 post-surgery. The results presented decreased paw withdrawal thresholds and reduced NREM sleep duration in both models on the first post-surgery day. In the SCI model, NREM sleep duration was negatively correlated with paw withdrawal thresholds (p = 0.0466), but not in the CPL model. Wake alpha and theta EEG powers were also correlated with the pain threshold. The results confirm that the SCI model shows disturbed sleep patterns associated with increased pain sensitivity, suggesting it is a reliable tool for investigating sleep disturbances associated with neuropathic pain.

Effects of the etching process on the surface, friction and wear characteristics of silicone rubber coated with micro-sized ceramic particles.

This study investigates the friction and wear characteristics of silicone rubber used in hydraulic systems, focusing on surface properties achieved through coating strategies. Silicone rubber specimens with varying surface characteristics, prepared by coating with micro-sized ceramic particles and employing etching processes, were examined. Surface morphology, roughness, water droplet contact angles, and friction and wear characteristics were evaluated. The silicone rubber was coated with ceramic particles (average size: 16 µm) and subsequently etched for different durations (1, 5, 10, 30, and 60 minutes). The results revealed that longer etching times led to increased surface roughness, while shorter etching times resulted in improved wear characteristics. The friction coefficient demonstrated a discernible reduction with escalating etching durations, with Etching-60M showing approximately 50% lower friction coefficient compared to Etching-1M. Wear rates ranged from 2.47 × 10-7 to 1.43 × 10-6 mm3 N-1 mm-1, indicating an increasing trend with longer etching times. Distinct wear mechanisms were observed between non-etched and etched specimens, with the latter exhibiting more pronounced wear tracks. Finite element analysis highlighted variations in stress behavior during contact sliding, indicating that surface modifications significantly impact wear resistance. While longer etching times improved friction characteristics, shorter etching times yielded superior wear characteristics. Further research is recommended to explore optimal etching conditions considering various variables.

A preliminary study about the potential risks of the UV-weathered microplastic: The proteome-level changes in the brain in response to polystyrene derived weathered microplastics.

The growing use of plastic materials has resulted in a constant increase in the risk associated with microplastics (MPs). Ultra-violet (UV) light and wind break down modify MPs in the environment into smaller particles known as weathered MPs (WMPs) and these processes increase the risk of MP toxicity. The neurotoxicity of weathered polystyrene-MPs remains unclear. Therefore, it is important to understand the risks posed by WMPs. We evaluated the chemical changes of WMPs generated under laboratory-synchronized environmentally mimetic conditions and compared them with virgin MPs (VMPs). We found that WMP had a rough surface, slight yellow color, reduced molecular weight, and structural alteration compared with those of VMP. Next, 2 µg of ∼100 µm in size of WMP and VMP were orally administered once a day for one week to C57BL/6 male mice. Proteomic analysis revealed that the WMP group had significantly increased activation of immune and neurodegeneration-related pathways compared with that of the VMP group. Consistently, in in vitro experiments, the human brain-derived microglial cell line (HMC-3) also exhibited a more severe inflammatory response to WMP than to VMP. These results show that WMP is a more profound inflammatory factor than VMP. In summary, our findings demonstrate the toxicity of WMPs and provide theoretical insights into their potential risks to biological systems and even humans in the ecosystem.

Efficient brazzein production in yeast (Kluyveromyces lactis) using a chemically defined medium.

The sweet-tasting protein brazzein offers considerable potential as a functional sweetener with antioxidant, anti-inflammatory, and anti-allergic properties. Here, we optimized a chemically defined medium to produce secretory recombinant brazzein in Kluyveromyces lactis, with applications in mass production. Compositions of defined media were investigated for two phases of fermentation: the first phase for cell growth, and the second for maximum brazzein secretory production. Secretory brazzein expressed in the optimized defined medium exhibited higher purity than in the complex medium; purification was by ultrafiltration using a molecular weight cutoff, yielding approximately 107 mg L-1. Moreover, the total media cost in this defined medium system was approximately 11% of that in the optimized complex medium to generate equal amounts of brazzein. Therefore, the K. lactis expression system is useful for mass-producing recombinant brazzein with high purity and yield at low production cost and indicates a promising potential for applications in the food industry.

Optimized production and quantification of the tryptophan-deficient sweet-tasting protein brazzein in Kluyveromyces lactis.

The sweet-tasting protein brazzein is a candidate sugar substitute owing to its sweet, sugar-like taste and good stability. To commercialize brazzein as a sweetener, optimization of fermentation and purification procedure is necessary. Here, we report the expression conditions of brazzein in the yeast Kluyveromices lactis and purification method for maximum yield. Transformed K. lactis was cultured in YPGlu (pH 7.0) at 25 °C and induced by adding glucose:galactose at a weight ratio of 1:2 (%/%) during the stationary phase, which increased brazzein expression 2.5 fold compared to the previous conditions. Cultures were subjected to heat treatment at 80 °C for 1 h, and brazzein containing supernatant was purified using carboxymethyl-sepharose cation exchange chromatography using 50 mM NaCl in 50 mM sodium acetate buffer (pH 4.0) as a wash buffer and 400 mM NaCl (pH 7.0) for elution. The yield of purified brazzein under these conditions was 2.0-fold higher than that from previous purification methods. We also determined that the NanoOrange assay was a suitable method for quantifying tryptophan-deficient brazzein. Thus, it is possible to obtain pure recombinant brazzein with high yield in K. lactis using our optimized expression, purification, and quantification protocols, which has potential applications in the food industry.

Splenic malignant fibrous histiocytoma with concurrent hypertension and epistaxis in an Alaskan malamute dog.

BACKGROUND: Malignant fibrous histiocytoma has been uncommonly described in dogs. Several extranasal neoplasias have been reported to result hypertensive epistaxis. There are, however, no published case reports of extranasal malignant fibrous histiocytoma with concurrent hypertension and epistaxis in dogs. CASE PRESENTATION: A 10-year-old dog presented with a spontaneous massive epistaxis persisting for 5 days. The dog exhibited unstable hypertension, which was considered as a cause of epistaxis. The complete blood count, prothrombin time, and activated partial thromboplastin time were within the reference limits, and other systemic examination showed no abnormalities except for a splenic mass occupying more than one third of the abdomen. Histologic examination of the resected spleen revealed the characteristic features of a malignant fibrous histiocytoma. One week after splenectomy, the hypertension and epistaxis resolved clinically and did not recur on the 5-month follow-up. CONCLUSIONS: The dog's blood pressure and epistaxis normalized after malignant fibrous histiocytoma resection suggesting that hypertensive epistaxis may be a rare manifestation of canine malignant fibrous histiocytoma.

Rapid preparation of a noncultured skin cell suspension that promotes wound healing.

Autologous skin cell suspensions have been used for wound healing in patients with burns and against normal pigmentation in vitiligo. To separate cells and the extracellular matrix from skin tissue, most researchers use enzymatic digestion. Therefore, this process is difficult to perform during a routine surgical procedure. We aimed to prepare a suspension of noncultured autologous skin cells (NCSCs) using a tissue homogenizer as a new method instead of harsh biochemical reagents. The potential clinical applicability of NCSCs was analyzed using a nude-rat model of burn healing. After optimization of the homogenizer settings, cell viability ranged from 52 to 89%. Scanning electron microscopy showed evidence of keratinocyte-like cell morphology, and several growth factors, including epidermal growth factor and basic fibroblast growth factor, were present in the NCSCs. The rat model revealed that NCSCs accelerated skin regeneration. NCSCs could be generated using a tissue homogenizer for enhancement of wound healing in vivo. In the NCSC group of wounds, on day 7 of epithelialization, granulation was observed, whereas on day 14, there was a significant increase in skin adnexa regeneration as compared to the control group (PBS treatment; p < 0.05). This study suggests that the proposed process is rapid and does not require the use of biochemical agents. Thus, we recommend a combination of surgical treatment with the new therapy for a burn as an effective method.

Next Place Prediction Based on Spatiotemporal Pattern Mining of Mobile Device Logs.

Due to the recent explosive growth of location-aware services based on mobile devices, predicting the next places of a user is of increasing importance to enable proactive information services. In this paper, we introduce a data-driven framework that aims to predict the user's next places using his/her past visiting patterns analyzed from mobile device logs. Specifically, the notion of the spatiotemporal-periodic (STP) pattern is proposed to capture the visits with spatiotemporal periodicity by focusing on a detail level of location for each individual. Subsequently, we present algorithms that extract the STP patterns from a user's past visiting behaviors and predict the next places based on the patterns. The experiment results obtained by using a real-world dataset show that the proposed methods are more effective in predicting the user's next places than the previous approaches considered in most cases.

Comparison between cryopreserved and glycerol-preserved allografts in a partial-thickness porcine wound model.

Human skin allografts are one of the best temporary biological coverings for severely burned patients. Cryopreserved (CPA) and glycerol-preserved (GPA) allografts are the most widely used types. This study compared the allograft efficiency of both preservation methods under the same conditions. To simulate actual clinical conditions, we used a porcine wound model. In addition, we evaluated the macroscopic and microscopic scoring of graft performance for each method. Porcine cadaver skin 1 mm thick was obtained from one pig. Cryopreserved skin cell viability was 20.8 %, glycerol-preserved skin was 9.08 %, and fresh skin was 58.6 %. We made ten partial-thickness wounds each in two pigs. The take rates on day 2 were 96.23 and 82.65 % in the GPA and CPA group (both n = 9), respectively. After 1 week, the take rates of both groups were nearly equal. The removal rate at week 5 was 98.87 and 94.41 % in the GPA and CPA group, respectively. On microscopic findings at week 2, inflammation was greater in the CPA group. Other findings such as fibroblast hyperplasia and neovascularization were not significantly different between both groups. At week 5, the score of collagen fiber synthesis was 2.67 ± 0.47 and 2.33 ± 0.47 in the GPA and CPA group, respectively. The epidermal-dermal junction was 2.22 ± 0.79 and 2.00 ± 0.47 in the GPA and CPA group, respectively. These findings suggest that wound healing takes longer in the CPA group. The preservation method of allografts is not a absolute factor in the wound healing process in this wound model.

Rapid detection of 6×-histidine-labeled recombinant proteins by immunochromatography using dye-labeled cellulose nanobeads.

A rapid and easy immunochromatography assay using dye-labeled cellulose nanobeads (CNBs) was developed to detect proteins with hexa-histidine tag (His-tag) to characterize recombinant proteins during purification. Recombinant ATG8 protein was used as a His-tagged protein, and ATG8-conjugated CNBs (A-CNBs) were prepared. The original ATG8 in the sample solution competed with A-CNBs for anti-His-tag antibodies spotted on to the strip resulting in an inverse relationship between ATG8 concentration and the colorimetric signal. The usefulness of this method was shown by adding ATG8 to a 1% Escherichia coli extract. In addition, this assay can be used to detect other His-tagged proteins without protein-specific antibodies. Because the identification of fractions containing His-tagged proteins by western blotting or ELISA is labor-intensive and expensive, our method provides an efficient and cheaper alternative.

Development of an auditory emotion recognition function using psychoacoustic parameters based on the International Affective Digitized Sounds.

The purpose of this study was to develop an auditory emotion recognition function that could determine the emotion caused by sounds coming from the environment in our daily life. For this purpose, sound stimuli from the International Affective Digitized Sounds (IADS-2), a standardized database of sounds intended to evoke emotion, were selected, and four psychoacoustic parameters (i.e., loudness, sharpness, roughness, and fluctuation strength) were extracted from the sounds. Also, by using an emotion adjective scale, 140 college students were tested to measure three basic emotions (happiness, sadness, and negativity). From this discriminant analysis to predict basic emotions from the psychoacoustic parameters of sound, a discriminant function with overall discriminant accuracy of 88.9% was produced from training data. In order to validate the discriminant function, the same four psychoacoustic parameters were extracted from 46 sound stimuli collected from another database and substituted into the discriminant function. The results showed that an overall discriminant accuracy of 63.04% was confirmed. Our findings provide the possibility that daily-life sounds, beyond voice and music, can be used in a human-machine interface.

Impact of Tribological Conditions on Collagen Coating Self-Healing.

The study examined the correlation between collagen coating damage and self-healing under various tribological conditions. It confirmed that the friction coefficient and degree of damage on the collagen coating varied based on contact and sliding conditions. The friction coefficient, measured at 0.56 for a single sliding cycle under a 350 mN normal load, demonstrated a notable decrease to 0.46 for 2295 cycles under 30 mN, further reducing to 0.15 for 90 cycles under a 20 mN normal load. As the normal load increased, the friction coefficient decreased, and with repeated sliding cycles under the same load, the coefficient also decreased. Water droplets induced a self-healing effect on collagen coating, causing wear tracks to vanish as fibers absorbed water. Severe wear tracks, with broken fibers and peeled coating, showed limited self-healing. In contrast, mild wear tracks, with compressed yet connected fibers, exhibited the self-healing phenomenon, making the wear tracks disappear. Real-time observations during 90 cycles under a 20 mN normal load highlighted the formation of mild wear tracks with intact collagen fibers, providing quantitative insights into self-healing characteristics. To preserve the self-healing effect of the collagen coating, it is essential to ensure tribological conditions during contact and sliding that prevent the disconnection of collagen fibers.

Influence of surface structure on friction and wear characteristics of silicone rubber for hydraulic rod seals.

This research investigates the impact of surface structure on the friction and wear characteristics of silicone rubber used as a material for hydraulic rod seals. Various silicone rubber specimens with different surface structures were prepared, and their surface morphology, water contact angle, and surface roughness were compared. Friction tests were conducted using a reciprocating sliding method to evaluate the friction coefficient and wear characteristics. The results revealed that the silicone rubber specimens coated with silicone powder exhibited a significant increase in surface roughness. However, this increase was accompanied by a decrease in surface energy, leading to the absorption and dispersion of contact pressure and frictional stress, resulting in a friction-reducing effect. Consequently, the silicone rubber specimens coated with silicone powder demonstrated a friction coefficient more than 70% lower on average compared to bare silicone rubber, and exhibited minimal wear characteristics. The irregular microstructures formed on the surface of the silicone rubber are believed to contribute to these friction and wear improvements. Alterations in stress and contact behavior of bare silicone rubber and silicone powder-coated silicone rubber with pre-curing time during indentation and sliding movements were validated through finite element analysis. These findings provide valuable insights for enhancing the performance and durability of hydraulic rod seals made from silicone rubber. This research is expected to contribute to further studies aimed at improving hydraulic seal materials.

Self-lubrication and tribological properties of polymer composites containing lubricant.

The purpose of this study was to improve the tribological properties of polydimethylsiloxane (PDMS) by mixing lubricants into it. The chemical composition, physical/chemical bonding state, and mechanical properties of the PDMS/lubricant composites (PLCs), prepared by mixing PDMS and lubricants at different ratios, were analyzed. With increasing lubricant content, the friction coefficient initially decreased, reaching a minimum value at a PDMS/lubricant ratio of 100 : 10; however, it gradually increased with a further increase in the lubricant content. The mechanical properties of PLCs with lubricant contents of 10% and higher decreased owing to the lubricant addition, so that the contact area with the sliding counter tip increased with lubricant content, but the frictional resistance was still decreased owing to the self-lubricating effect. In addition, owing to the effect of the lubricating film, there was no direct contact between the PLC surface and counter tip, and almost no damage was done to the PLC surface. Finite element analysis of the changes in stress during indentation and sliding confirmed that the stress applied to the PLCs was lower than that for bare PDMS.

Physiological significance of elevated levels of lactate by exercise training in the brain and body.

For the past 200 years, lactate has been regarded as a metabolic waste end product that causes fatigue during exercise. However, lactate production is closely correlated with energy metabolism. The lactate dehydrogenase-catalyzed reaction uses protons to produce lactate, which delays ongoing metabolic acidosis. Of note, lactate production differs depending on exercise intensity and is not limited to muscles. Importantly, controlling physiological effect of lactate may be a solution to alleviating some chronic diseases. Released through exercise, lactate is an important biomarker for fat oxidation in skeletal muscles. During recovery after sustained strenuous exercise, most of the lactate accumulated during exercise is removed by direct oxidation. However, as the muscle respiration rate decreases, lactate becomes a desirable substrate for hepatic glucose synthesis. Furthermore, improvement in brain function by lactate, particularly, through the expression of vascular endothelial growth factor and brain-derived neurotrophic factor, is being increasingly studied. In addition, it is possible to improve stress-related symptoms, such as depression, by regulating the function of hippocampal mitochondria, and with an increasingly aging society, lactate is being investigated as a preventive agent for brain diseases such as Alzheimer's disease. Therefore, the perception that lactate is equivalent to fatigue should no longer exist. This review focuses on the new perception of lactate and how lactate acts extensively in the skeletal muscles, heart, brain, kidney, and liver. Additionally, lactate is now used to confirm exercise performance and should be further studied to assess its impact on exercise training.

Lipid-based colloidal nanoparticles for applications in targeted vaccine delivery.

Bioactive molecules and their effects have been influenced by their solubility and administration route. In many therapeutic reagents, the performance of therapeutics is dependent on physiological barriers in the human body and delivery efficacy. Therefore, an effective and stable therapeutic delivery promotes pharmaceutical advancement and suitable biological usage of drugs. In the biological and pharmacological industries, lipid nanoparticles (LNPs) have emerged as a potential carrier to deliver therapeutics. Since studies reported doxorubicin-loaded liposomes (Doxil®), LNPs have been applied to numerous clinical trials. Lipid-based nanoparticles, including liposomes, solid lipid nanoparticles (SLNs), and nanostructured lipid nanoparticles, have also been developed to deliver active ingredients in vaccines. In this review, we present the type of LNPs used to develop vaccines with attractive advantages. We then discuss messenger RNA (mRNA) delivery for the clinical application of mRNA therapeutic-loaded LNPs and recent research trend of LNP-based vaccine development.

Disease-microenvironment modulation by bare- or engineered-exosome for rheumatoid arthritis treatment.

BACKGROUND: Exosomes are extracellular vesicles secreted by eukaryotic cells and have been extensively studied for their surface markers and internal cargo with unique functions. A deeper understanding of exosomes has allowed their application in various research areas, particularly in diagnostics and therapy. MAIN BODY: Exosomes have great potential as biomarkers and delivery vehicles for encapsulating therapeutic cargo. However, the limitations of bare exosomes, such as rapid phagocytic clearance and non-specific biodistribution after injection, pose significant challenges to their application as drug delivery systems. This review focuses on exosome-based drug delivery for treating rheumatoid arthritis, emphasizing pre/post-engineering approaches to overcome these challenges. CONCLUSION: This review will serve as an essential resource for future studies to develop novel exosome-based therapeutic approaches for rheumatoid arthritis. Overall, the review highlights the potential of exosomes as a promising therapeutic approach for rheumatoid arthritis treatment.

CHOP down-regulates cFLIP(L) expression by promoting ubiquitin/proteasome-mediated cFLIP(L) degradation.

The transcription factor CHOP/GADD153 is induced during the unfolded protein response and is related to the induction of ER stress-mediated apoptosis. However, how CHOP is organized between the pro-survival and pro-apoptotic roles of ER stress remains largely undefined. In this study, we identified the apoptosis regulating protein suppressed by CHOP. We found that treatment of Caki cells with CHOP-inducing drugs including withaferin A, thapsigargin, brefeldin A, and silybin led to a strong reduction in cFLIP(L) protein levels together with a concomitant increase in the CHOP protein. Interestingly, Wit A down-regulated cFLIP(L) expression via both suppressing mRNA transcription and increasing cFLIPL protein instability. We also found that forced expression of CHOP dose-dependently led to a decrease of cFLIP(L) protein expression but did not alter cFLIP(L) mRNA levels. Additionally, we observed that siRNA-mediated CHOP silencing recovered the cFLIP(L) expression decreased by CHOP-inducing agents in Caki cells. Finally, we showed that CHOP facilitates ubiquitin/proteasome-mediated cFLIP(L) degradation, leading to down-regulation of cFLIP(L). Finally, cFLIP(L) over-expression reduced cell death induced by treatment with brefeldin A, thapsigargin, and silybin. Taken together, our results provide novel evidence that cFLIP(L) is a CHOP control target and that CHOP-induced down-regulation of cFLIP(L) is due to activation of the ubiquitin/proteasome pathways.

Friction and Wear Characteristics of Polydimethylsiloxane under Water-Based Lubrication Conditions.

In this study, the friction and wear characteristics of polydimethylsiloxane (PDMS) were evaluated when using lubricants created by adding surfactants at various ratios to deionized (DI) water. When pure DI water is used as a lubricant, the repulsion of water from the hydrophobic PDMS surface is large and the interfacial affinity is low; thus, the lubrication properties cannot be significantly improved. However, when a lubricant with a surfactant is added to DI water, the interfacial affinity with the PDMS surface increases to form a lubricating film, and the friction coefficient is greatly reduced. In this study, under dry and pure DI water conditions, severe wear tracks were formed on the PDMS surface after 10,000 cycles of reciprocating sliding motion under a vertical load of 100 mN, whereas in the case of the surfactant-based and water-based lubricant, no severe wear tracks occurred. The friction and wear characteristics of the PDMS were evaluated by increasing the normal loads and sliding cycles with a water-based lubricant containing 1 wt % surfactant. Under normal loads of 300 mN and 500 mN, only minor scratches occurred on the PDMS surface up to 10,000 and 100,000 cycles, respectively, but after 300,000 cycles, very severe pit wear tracks occurred.

Fabrication of TiO2 ̶ KH550 ̶ PEG Super-Hydrophilic Coating on Glass Surface without UV/Plasma Treatment for Self-Cleaning and Anti-Fogging Applications.

In this study, we have developed a self-cleaning transparent coating on a glass substrate by dip coating a TiO2 ̶ KH550 ̶ PEG mixed solution with super-hydrophilicity and good antifogging properties. The fabrication of the thin-film-coated glass is a one-step solution blending method that is performed by depositing only one layer of modified TiO2 nanoparticles at room temperature. X-ray diffraction (XRD) and scanning electron microscopy (SEM) were used to determine the structure and morphology of the nanoparticles and the thin-film-coated glass. The surface functional groups were investigated using Fourier-transform infrared spectroscopy (FT-IR), and the optical properties of the glass coating were measured using a UV/Vis spectrometer. The results revealed that the KH-500-modified TiO2 film coating was in an anatase crystalline form. The hydrophilicity of the coated and uncoated glass substrates was observed by measuring their water contact angle (WCA) using a contact angle instrument. The maximum transparency of the coated glass measured in the visible region (380-780 nm) was approximately 70%, and it possessed excellent super-hydrophilic properties (WCA ~0°) at an annealing temperature of 350 °C without further need of UV or plasma treatment. These results demonstrate the super-hydrophilic coated glass surface has potential for use in self-cleaning and anti-fogging applications.