Analyzing small RNA sequences from canine stem cell-derived extracellular vesicles primed with TNF-α and IFN-γ and exploring their potential in lung repair.

Acute lung injury is an acute inflammation disorder that disrupts the lung endothelial and epithelial barriers. In this study, we investigated the extracellular vesicles (EVs) obtained via priming inflammatory cytokines such as tumor necrosis factor (TNF)-α and interferon (IFN)-γ on canine adipose mesenchymal stem cells in improving their anti-inflammatory and/or immunosuppressive potential, and/or their ability to alleviate lipopolysaccharide-induced lung injury in vitro. We also explored the correlation between epithelial-to-mesenchymal transition and the inflammatory repressive effect of primed EVs. Using small RNA-Seq, we confirmed that miR-16 and miR-502 significantly increased in EVs from TNF-α and IFN-γ-primed canine adipose mesenchymal stem cells. The pro and anti-inflammatory cytokines were analyzed in a lipopolysaccharide-induced lung injury model and we found that the EV anti-inflammatory effect improved on priming with inflammatory cytokines. EVs obtained from primed stem cells effectively suppress endothelial-to-mesenchymal transition in a lung injury model. Our results suggest a potential therapeutic approach utilizing EVs obtained from adipose mesenchymal stem cells primed with TNF-α and IFN-γ against lung inflammation and endothelial to mesenchymal transition.

Simultaneous Estimation of Tonic Dopamine and Serotonin with High Temporal Resolution In Vitro Using Deep Learning.

Quantitative measurement of the phasic (changes in several seconds) and tonic (changes in minutes to hours) level changes of neurotransmitters is an essential technique for understanding brain functions and brain diseases regulated by the neurotransmitters. However, monitoring phasic and tonic levels of multiple neurotransmitters is still a challenging technology. Microdialysis can measure the tonic levels of multiple neurotransmitters simultaneously but has a low temporal resolution (minute) to analyze precisely. Fast-scan cyclic voltammetry (FSCV) has high temporal resolution and high sensitivity, but it was not able to simultaneously measure the tonic level of multiple neurotransmitters. The recently proposed deep learning-based FSCV method was still only capable of phasic concentration estimation of neurotransmitters. In this study, we estimate the tonic levels of dopamine and serotonin simultaneously by training a deep-learning network with the extracted tonic information from the FSCV. The proposed deep learning model was validated in vitro to simultaneously estimate tonic concentrations of two neurotransmitters with statistically significantly higher accuracy than previously proposed background subtraction-based models (p<0.001). In particular, in the case of serotonin concentration estimation error, the proposed model showed higher prediction performance than the background subtraction-based model (48 nM and 73 nM, respectively). We expect that the proposed technique will help simultaneous measurement of the phasic and tonic levels of numerous neurotransmitters in vivo soon.Clinical Relevance- This study proposes a method to simultaneously measure tonic dopamine and tonic serotonin with high temporal resolution with a single electrode in the brain.

Simultaneous hypersensitivity reactions to trimethoprim-sulfamethoxazole and amoxicillin-clavulanate in a dog.

Antibiotics are known to be able to cause hypersensitivity reactions through various mechanisms. We present a case of drug-induced immune thrombocytopenia (DITP) and anaphylactic shock occurring simultaneously in a dog after the administration of two classes of antibiotics, namely trimethoprim-sulfamethoxazole (TMP-SMX) and amoxicillin-clavulanate (AMC). The patient recovered completely from DITP on discontinuation of TMP-SMX and the anaphylactic shock caused by AMC was treated with intensive care. DITP is a rare adverse drug reaction (ADR), and anaphylactic shock is a life-threatening ADR. This is the first case report of a dog manifesting two types of hypersensitivity reactions caused by two antibiotics.

IL-31RA and TRPV1 Expression in Atopic Dermatitis Induced with Trinitrochlorobenzene in Nc/Nga Mice.

Atopic dermatitis (AD) is a common chronic inflammatory skin disease. Interleukin 31 (IL-31), a novel cytokine in AD, causes pruritus, typically characteristic of AD patients. The transient receptor potential vanilloid type 1 (TRPV1) is a cation channel activated by diverse noxious stimuli that has been studied in a variety of pruritic skin diseases. In this study, the AD animal model was generated by administering the hapten, trinitrochlorobenzene (TNCB), to Nc/Nga mice, and the degree of expression of the IL-31 receptor alpha (IL-31RA) and TRPV1 in the skin of these atopic models was evaluated. The Nc/Nga mice were divided into 3 groups: control, TNCB 2-weeks treated, and TNCB 8-weeks treated. After inducing AD, the skin lesions in each group were scored and compared, and the histology of the skin lesions and the IL-31RA and TRPV1 expression for each group were evaluated by analyzing immunohistochemistry. The results show a significant difference in the skin lesion scores between the groups. The immunohistochemistry evaluation highlighted the remarkable expression of IL-31RA and TRPV1 in the nerve fibers of the TNCB 8-weeks-treated group. We thus confirmed that the long-term application of TNCB induced chronic atopic-like dermatitis and that IL-31RA and TRPV1 were overexpressed in the peripheral nerve fibers in this AD model.

Automated Evaluation of Upper Airway Obstruction Based on Deep Learning.

Objectives: This study is aimed at developing a screening tool that could evaluate the upper airway obstruction on lateral cephalograms based on deep learning. Methods: We developed a novel and practical convolutional neural network model to automatically evaluate upper airway obstruction based on ResNet backbone using the lateral cephalogram. A total of 1219 X-ray images were collected for model training and testing. Results: In comparison with VGG16, our model showed a better performance with sensitivity of 0.86, specificity of 0.89, PPV of 0.90, NPV of 0.85, and F1-score of 0.88, respectively. The heat maps of cephalograms showed a deeper understanding of features learned by deep learning model. Conclusion: This study demonstrated that deep learning could learn effective features from cephalograms and automated evaluate upper airway obstruction according to X-ray images. Clinical Relevance. A novel and practical deep convolutional neural network model has been established to relieve dentists' workload of screening and improve accuracy in upper airway obstruction.

Second-Derivative-Based Background Drift Removal for a Tonic Dopamine Measurement in Fast-Scan Cyclic Voltammetry.

The dysregulation of dopamine, a neuromodulator, is associated with a broad spectrum of brain disorders, including Parkinson's disease, addiction, and schizophrenia. Quantitative measurements of dopamine are essential for understanding dopamine functional dynamics. Fast-scan cyclic voltammetry (FSCV) is the most popular electrochemical technique for measuring real-time in vivo dopamine level changes. Standard FSCV has only analyzed "phasic dopamine" (changes in seconds) because the gradual generation of background charging current is inevitable and is the primary noise source in the low-frequency band. Although "tonic dopamine" (changes in minutes to hours) is critical for understanding the dopamine system, an electrochemical technique capable of simultaneously measuring phasic and tonic dopamine in an in vivo environment has not been established. Several modified voltammetric techniques have been developed for measuring tonic dopamine; however, the sampling rates (0.1-0.05 Hz) are too low to be useful. Further investigation of the in vivo applicability of previously developed background drift removal methods for measuring tonic dopamine levels is required. We developed a second-derivative-based background removal (SDBR) method for simultaneously measuring phasic and tonic neurotransmitter levels in real-time. The performance of this technique was tested via in silico and in vitro tonic dopamine experiments. Furthermore, its applicability was tested in vivo. SDBR is a simple, robust, postprocessing technique that can extract tonic neurotransmitter levels from all FSCV data. As SDBR is calculated in individual-scan voltammogram units, it can be applied to any real-time closed-loop system that uses a neurotransmitter as a biomarker.

Anti-Inflammatory and Immune Modulatory Effects of Synbio-Glucan in an Atopic Dermatitis Mouse Model.

Many trials have been conducted to treat atopic dermatitis (AD), but these therapies are generally unsuccessful because of their insufficiency or side effects. This study examined the efficacy of ß-glucan derived from oats with fermented probiotics (called Synbio-glucan) on an AD-induced mouse model. For the experiment, Nc/Nga mice were exposed to a house dust mite extract (HDM) to induce AD. The mice were placed in one of four groups: positive control group, Synbio-glucan topical treatment group, Synbio-glucan dietary treatment group, and Synbio-glucan topical + dietary treatment group. The experiment revealed no significant difference in the serum IgE concentration among the groups. Serum cytokine antibody arrays showed that genes related to the immune response were enriched. A significant difference in the skin lesion scores was observed between the groups. Compared to the control group tissue, skin lesions were alleviated in the Synbio-glucan topical treatment group and Synbio-glucan dietary treatment group. Interestingly, almost normal structures were observed within the skin lesions in the Synbio-glucan topical + dietary treatment group. Overall, the ß-glucan extracted from oats and fermented probiotic mixture is effective in treating atopic dermatitis.

Serum Concentration of Inflammatory Cytokines in Dogs with Suspected Acute Pancreatitis.

Acute pancreatitis is an acute inflammatory process in the pancreas that is common in dogs. This study was designed to compare cytokines between healthy dogs and dogs with suspected acute pancreatitis. For the canine cytokine antibody array, three healthy dogs and three dogs with suspected acute pancreatitis were included. Interleukin (IL)-2, IL-6, IL-10, GM-CSF, and TNF-α were not detected in either group based on the results. Conversely, IL-8 (p = 0.035), Monocyte Chemoattractant Protein-1 (MCP)-1 (p = 0.0138), Receptor for Advanced Glycation Endproducts (RAGE) (p = 0.0079), and stem cell factor (SCF) (p = 0.034) were significantly increased in dogs with suspected acute pancreatitis. However, vascular endothelial growth factor (VEGF) (p = 0.6971) did not differ significantly between groups. For the canine serum Enzyme-Linked Immunosorbent Assay (ELISA), eight healthy dogs and eight dogs with suspected acute pancreatitis were included. ELISA revealed that IL-8 (p < 0.0001), MCP-1 (p < 0.0001), RAGE (p = 0.006), and SCF (p = 0.0002) were all significantly upregulated in the experimental group. We confirmed multiple patterns of cytokines in suspected acute pancreatitis of dogs via canine cytokine antibody array using a small quantity of serum. After this procedure, we reevaluated the cytokines, which were significantly increased in dogs with suspected acute pancreatitis, by ELISA, with more samples. Through this study, we confirmed that MCP-1, RAGE, and SCF were newly suggested factors in dogs with suspected acute pancreatitis.

Cholesterol-Functionalized Linear/Brush Block Copolymers for Metal-Incorporated Nanostructures with Modulated Core Density and Enhanced Self-Assembly Efficiency.

Metal-mediated self-assembly of chelating double-hydrophilic block copolymer has become a facile preparation strategy of great importance for the metal-chelated hybrid nanostructures. Herein, we present a delicate control over the morphology regulation of metal-chelated nanostructures by a terminal modification of polymer building blocks with mesogenic cholesterol. Such a molecular design motif at an end of chelating linear/brush-type block copolymer imparts not only additional hydrophobicity for enhanced cohesive force to facilitate the metal-mediated self-assembly, but also significant morphological alteration of a metal-chelated core that otherwise generally forms a spherical interior with cholesterol-free block copolymers. The presence of cholesterol entities localized at the central core further allows for the density modulation of the final PtII-chelated nanostructures while maintaining the colloidal stability, comparable to that of the cholesterol-free nanoparticles in physiological conditions. This metal-mediated assembly strategy with modified polymer building blocks can provide a potential platform for the delivery of inorganic agents.

Construction of Paramagnetic Manganese-Chelated Polymeric Nanoparticles Using Pyrene-End-Modified Double-Hydrophilic Block Copolymers for Enhanced Magnetic Resonance Relaxivity: A Comparative Study with Cisplatin Pharmacophore.

Cationic metal-mediated self-assembly of double-hydrophilic block copolymers (DHBCs) has been of great interest for the preparation of hybrid nanoparticles for versatile applications. Among many functional transition-metal ions, manganese (MnII) is a highly attractive element due to its paramagnetic property with a high coordination number. However, MnII does not lead to the efficient self-assembly of DHBCs because of the relatively high aqueous solubility of coordinated MnII. This article reports a facile method for direct conjugation of MnII ions inside sterically stabilized polymer assemblies, composed of pyrene-end-modified DHBCs. Nitroxide-mediated radical polymerization was used to prepare the poly(ethylene glycol)- b-poly(acrylate) DHBC precursor, followed by the end-modification with pyrene maleimide via the radical-exchange reaction. Employing the self-associated DHBC as the nanoscale template, the simple addition of MnII enables a large number of polyvalent MnII ions to be immobilized at the chelating blocks of DHBCs, which can be readily monitored by the excimeric fluorescence emission change of the terminal pyrene fluorophore. The resulting MnII-loaded polymeric nanoparticles (MnII-PNPs) possess nanogel-like scaffolds, which allow for efficient water permeation at the MnII-incorporated interior for enhanced magnetic resonance contrasting effect. Additionally, by comparing the coordination properties of MnII and cisplatin, we endeavor to understand the internal structures and the relevant physicochemical features of metal-chelated nanoparticles.

Cisplatin-Encapsulated Polymeric Nanoparticles with Molecular Geometry-Regulated Colloidal Properties and Controlled Drug Release.

Encapsulation of chemotherapeutic agents inside a nanoscale delivery platform can provide an attractive therapeutic strategy with many pharmaceutical benefits, such as increased plasma solubility, prolonged in vivo circulation, and reduced acute toxicity. Given that the biological activities of polymeric nanoparticles are highly dependent on their colloidal structures, the molecular geometry-regulated programming of self-assembled nanoscale architecture is of great interest for chemical design of an ideal delivery platform. In this report, we demonstrate that the molecular geometry of block-copolymer excipients can govern the level of drug-loading capacity and core hydrophobicity of polymeric nanoparticles, which can eventually control the pH-sensitive drug-release property. Atom-transfer radical polymerization was employed as a controlled synthetic method for the copolymer excipients, which contain the metal-chelating poly(acrylic acid) block linked to either a small mPEG-grafted poly(methacrylate) to generate a bulky brush-like chains or a simple linear mPEG segment. During the coordination of cis-diammineplatinum(II) as an active pharmacophore of cisplatin, aqueous-phase size-exclusion chromatography analyses exhibited highly different self-association kinetic regimes prompted by versatile molecular geometry of copolymer excipients, which further allows us to explore the molecular geometry-colloidal property relationship.

Colour stabilities of three types of orthodontic clear aligners exposed to staining agents.

The aim of this study was to evaluate and compare the colour stabilities of three types of orthodontic clear aligners exposed to staining agents in vitro. Sixty clear orthodontic aligners produced by three manufacturers (Invisalign, Angelalign, and Smartee) were immersed in three staining solutions (coffee, black tea, and red wine) and one control solution (distilled water). After 12-h and 7-day immersions, the aligners were washed in an ultrasonic cleaner and measured with a colourimeter. The colour changes (ΔE*) were calculated on the basis of the Commission Internationale de I'Eclairage L*a*b* colour system (CIE L*a*b*), and the results were then converted into National Bureau of Standards (NBS) units. Fourier transformation infrared (FT-IR) spectroscopy and scanning electron microscopy (SEM) were conducted to observe the molecular and morphologic alterations to the aligner surfaces, respectively. The three types of aligners exhibited slight colour changes after 12 h of staining, with the exception of the Invisalign aligners stained with coffee. The Invisalign aligners exhibited significantly higher ΔE* values (ranging from 0.30 to 27.81) than those of the Angelalign and Smartee aligners (ΔE* values ranging from 0.33 to 1.89 and 0.32 to 1.61, respectively, P<0.05). FT-IR analysis confirmed that the polymer-based structure of aligners did not exhibit significant chemical differences before and after the immersions. The SEM results revealed different surface alterations to the three types of aligner materials after the 7-day staining. The three types of aesthetic orthodontic appliances exhibited colour stability after the 12-h immersion, with the exception of the Invisalign aligners stained by coffee. The Invisalign aligners were more prone than the Angelalign and Smartee aligners to pigmentation. Aligner materials may be improved by considering aesthetic colour stability properties.

The synthesis and characterization of metal nanoparticles using a modified electrolysis method for conductive ink in ink-jet printing technology.

In the ink-jet patterning process, conductive ink composed of metal nanoparticles and solutions, is an important factor for improving properties of printed patterns and processes. In this study, metal (Cu) nanoparticles in conductive ink were synthesized using a modified electrolysis method that extracted to metal nanoparticles from bulk metal plates. The Cu nanoparticles were prepared with a narrow size distribution. The dispersion stability and oxidation properties in conductive inks were also studied. Cu nanoparticles were homogeneous and had a diameter of 15 approximately 20 nm. By addition of PVP, the dispersion and oxidation stability of the metal nanoparticles, which were not oxidized after 2 month, were improved.