Small-Molecule Ferritin Degrader as a Pyroptosis Inducer.

Exploring the response of malignant cells to intracellular metabolic stress is critical for understanding pathologic processes and developing anticancer therapies. Herein, we developed ferritin-targeting proteolysis targeting chimeras (PROTACs) to establish the iron excess stress inside cancer cells and investigated subsequent cellular behaviors. We conjugated oleic acid that binds to the ferritin dimer to the ligand of von Hippel-Lindau (VHL) E3 ligase through an alkyl linker. The screened chimera, DeFer-2, degraded ferritin and then rapidly elevated the free iron content, thereby initiating the caspase 3-GSDME-mediated pyroptosis in cancer cells rather than typical ferroptosis that is always associated with iron ion overload. According to its structural and physicochemical characteristics, DeFer-2 was loaded into a tailored albumin-based nano-formulation, which substantially inhibited tumor growth and prolonged the survival time of mice bearing B16F10 subcutaneous tumors with negligible adverse effects. This study developed a ferritin-targeting PROTAC for iron overload stress, revealed iron metabolic dysregulation-mediated pyroptosis, and provided a PROTAC-based pyroptosis inducer for anticancer treatment.

Greater Plasma Protein Adsorption on Mesoporous Silica Nanoparticles Aggravates Atopic Dermatitis.

Purpose: The protein corona surrounding nanoparticles has attracted considerable attention as it induces subsequent inflammatory responses. Although mesoporous silica nanoparticles (MSN) are commonly used in medicines, cosmetics, and packaging, the inflammatory effects of the MSN protein corona on the cutaneous system have not been investigated till date. Methods: We examined the greater plasma protein adsorption on MSN leads to serious inflammatory reactions in Dermatophagoides farinae extract (DFE)-induced mouse atopic dermatitis (AD)-like skin inflammation because of increased uptake by keratinocytes. Results: We compare the AD lesions induced by MSN and colloidal (non-porous) silica nanoparticles (CSN), which exhibit different pore architectures but similar dimensions and surface chemistry. MSN-corona treatment of severe skin inflammation in a DFE-induced in vivo AD model greatly increases mouse ear epidermal thickness and infiltration of immune cells compared with the CSN-corona treatment. Moreover, MSN-corona significantly increase AD-specific immunoglobulins, serum histamine, and Th1/Th2/Th17 cytokines in the ear and lymph nodes. MSN-corona induce more severe cutaneous inflammation than CSN by significantly decreasing claudin-1 expression. Conclusion: This study demonstrates the novel impact of the MSN protein corona in inducing inflammatory responses through claudin-1 downregulation and suggests useful clinical guidelines for MSN application in cosmetics and drug delivery systems.

An improved Raphanus sativus cv. WK10039 genome localizes centromeres, uncovers variation of DNA methylation and resolves arrangement of the ancestral Brassica genome blocks in radish chromosomes.

KEY MESSAGE: This study presents an improved genome of Raphanus sativus cv. WK10039 uncovering centromeres and differentially methylated regions of radish chromosomes. Comprehensive genome comparison of radish and diploid Brassica species of U's triangle reveals that R. sativus arose from the Brassica B genome lineage and is a sibling species of B. nigra. Radish (Raphanus sativus L.) is a key root vegetable crop closely related to the Brassica crop species of the family Brassicaceae. We reported a draft genome of R. sativus cv. WK10039 (Rs1.0), which had 54.6 Mb gaps. To study the radish genome and explore previously unknown regions, we generated an improved genome assembly (Rs2.0) by long-read sequencing and high-resolution genome-wide mapping of chromatin interactions. Rs2.0 was 434.9 Mb in size with 0.27 Mb gaps, and the N50 scaffold length was 37.3 Mb (40-fold larger assembly compared to Rs1.0). Approximately 38% of Rs2.0 was comprised of repetitive sequences, and 52,768 protein-coding genes and 4845 non-protein-coding genes were predicted and annotated. The improved contiguity and coverage of Rs2.0, along with the detection of highly methylated regions, enabled localization of centromeres where R. sativus-specific centromere-associated repeats, full-length OTA and CRM LTR-Gypsy retrotransposons, hAT-Ac, CMC-EnSpm and Helitron DNA transposons, and sequences highly homologous to B. nigra centromere-specific CENH3-associated CL sequences were enriched. Whole-genome bisulfite sequencing combined with mRNA sequencing identified differential epigenetic marks in the radish genome related to tissue development. Synteny comparison and genomic distance analysis of radish and three diploid Brassica species of U's triangle suggested that the radish genome arose from the Brassica B genome lineage through unique rearrangement of the triplicated ancestral Brassica genome after splitting of the Brassica A/C and B genomes.

Study on the Viability of Canine Nose Pattern as a Unique Biometric Marker.

The uniqueness of the canine nose pattern was studied. A total of 180 nose images of 60 dogs of diverse age, gender, and breed were collected. The canine nose patterns in these images were examined visually and by a biometric algorithm. It was found that the canine nose pattern remains invariant regardless of when the image is taken; and that the canine nose pattern is indeed unique to each dog. The same study was also performed on an enlarged dataset of 278 nose images of 70 dogs of 19 breeds. The study of the enlarged dataset also leads to the same conclusion. The result of this paper confirms and enhances the claims of earlier works by others that the canine nose pattern is indeed unique to each animal and serves as a unique biometric marker.

The Formation and Invariance of Canine Nose Pattern of Beagle Dogs from Early Puppy to Young Adult Periods.

The formation and invariance of the canine nose pattern is studied. Nose images of ten beagle dogs were collected for ten months from month two to month eleven. The nose patterns in these images are examined visually and by a biometric algorithm. It is found that the canine nose pattern is fully formed at the end of the second month since birth and remains invariant until the end of the eleventh month. This study also strongly indicates that the canine nose pattern can be used as a unique biometric marker for each individual dog.

Seeded Ising Model and Distributed Biometric Template Storage and Matching.

It is known that a variant of Ising model, called Seeded Ising Model, can be used to recover the information content of a biometric template from a fraction of information therein. The method consists in reconstructing the whole template, which is called the intruder template in this paper, using only a small portion of the given template, a partial template. This reconstruction method may pose a security threat to the integrity of a biometric identity management system. In this paper, based on the Seeded Ising Model, we present a systematic analysis of the possible security breach and its probability of accepting the intruder templates as genuine. Detailed statistical experiments on the intruder match rate are also conducted under various scenarios. In particular, we study (1) how best a template is divided into several small pieces called partial templates, each of which is to be stored in a separate silo; (2) how to do the matching by comparing partial templates in the locked-up silos, and letting only the results of these intra-silo comparisons be sent to the central tallying server for final scoring without requiring the whole templates in one location at any time.

Unfolded Protein Corona Surrounding Nanotubes Influence the Innate and Adaptive Immune System.

The plasma proteins around nanoparticles (NPs) form an outer protein corona, significantly influencing the subsequent immune response. However, it was uncertain whether the protein corona around NPs influences immune response. This study clarified that the immune response mediated by the protein corona is greatly dependent on the type of plasma proteins surrounding the NPs. Structural changes in the unfolded protein corona elevated reactive oxygen species (ROS) levels and induced major proinflammatory cytokine release in both murine and human macrophage cell lines. In contrast, negligible structural changes in the protein corona provoke neither ROS production nor proinflammatory cytokine release. Furthermore, in vivo analysis confirms that a stimulated immune response by an unfolded protein corona triggers selective activation of innate and adaptive immunity in the spleen. Specifically, neutrophils, natural killer cells, and CD8+ T cells are overpopulated by unfolded protein corona structures surrounding nanotubes, whereas innate and adaptive immunologic responses are not triggered by a normal protein corona. In conclusion, highly unfolded protein corona structures are strongly correlated with subsequent activation of proinflammatory cytokines and innate immune responses; thus, the protein corona can be used in immune-enhancing therapy.

Dual-Mode Radar Sensor for Indoor Environment Mapping.

In this paper, we introduce mapping results in an indoor environment based on our own developed dual-mode radar sensor. Our radar system uses a frequency-modulated continuous wave (FMCW) with a center frequency of 62 GHz and a multiple-input multiple-output antenna system. In addition, the FMCW radar sensor we designed is capable of dual-mode detection, which alternately transmits two waveforms using different bandwidths within one frame. The first waveform is for long-range detection, and the second waveform is for short-range detection. This radar system is mounted on a small robot that moves in indoor environments such as rooms or hallways, and the radar and the robot send and receive necessary information to each other. The radar estimates the distance, velocity, and angle information of targets around the radar-equipped robot. Then, the radar receives information about the robot's motion from the robot, such as its speed and rotation angle. Finally, by combining the motion information and the detection results, the radar-equipped robot maps the indoor environment while finding its own position. Compared to the actual map data, the radar-based mapping is effectively achieved through the radar system we developed.

Potential Therapeutic Usage of Nanomedicine for Glaucoma Treatment.

Glaucoma is a group of diseases characterized by progressive degeneration of retinal ganglion cells, leading to irreversible blindness. Currently, intraocular pressure reduction is the only established treatment available for glaucoma. With this treatment, the progression of the disease can only be delayed and there is no recovery. In addition, the commercially available eye drops have the disadvantage of low compliance and short therapeutic time, while glaucoma surgery always has the risk of failure due to wound fibrosis. Nanotechnology can overcome the limitations of the current treatment through the encapsulation and conjugation of drugs used for lowering intraocular pressure and antifibrotic agents using biodegradable or biocompatible nanoparticles for the sustained release of the drugs to protect the damaged ocular cells. Furthermore, using nanotechnology, treatment can be administered in various forms, including eye drops, contact lens, and ocular inserts, according to the convenience of the patients. Despite the promising results of delaying the progression of glaucoma, the regeneration of damaged ocular cells, including trabecular meshwork and retinal ganglion cells, is another critical hurdle to overcome. Bone marrow-derived mesenchymal stem cells and Müller glia cells can secrete neurogenic factors that trigger the regeneration of associated cells, including trabecular meshwork and retinal ganglion cells. In conclusion, this review highlights the potential therapeutic applications of nanotechnology- and stem cell-based methods that can be employed for the protection and regeneration of ocular cells.

Triamcinolone-Gold Nanoparticles Repolarize Synoviocytes and Macrophages in an Inflamed Synovium.

Understanding the crosstalk between synoviocytes and macrophages is very important for the development of strategies to regulate inflammatory responses in an inflamed synovium. Simultaneous regulation of the pro- and anti-inflammatory responses of synoviocytes and macrophages (repolarization) is critical for the treatment of arthritis. Thus, the immune regulatory functions of an ideal nanodrug should not only decrease the pro-inflammatory response but also effectively increase the anti-inflammatory response. In this study, crosstalk between synoviocytes and macrophages was found to be significantly involved in the activation and deactivation of inflammatory responses in the synovium. Interestingly, a developed triamcinolone-gold nanoparticle (Triam-AuNP) complex both decreased the pro-inflammatory responses and increased the anti-inflammatory responses of fibroblast-like synoviocytes (FLSs) and macrophages via repolarization of macrophages from the M1 to the M2 phenotype. In contrast, triamcinolone alone only decreased the pro-inflammatory responses of FLSs and macrophages without upregulating their anti-inflammatory responses. In vitro (human), ex vivo (human), and in vivo (mouse) analyses clearly indicated that Triam-AuNPs effectively regulated the expression of both pro- and anti-inflammatory cytokines in FLSs and effectively repolarized activity of macrophages in the inflamed synovium. Furthermore, Triam-AuNPs significantly promoted cartilage regeneration, whereas triamcinolone alone did not induce either FLS anti-inflammatory activity or macrophage repolarization.

Ectopic transient overexpression of OCT-4 facilitates BMP4-induced osteogenic transdifferentiation of human umbilical vein endothelial cells.

Limitation in cell sources for autologous cell therapy has been a recent focus in stem cell therapy and tissue engineering. Among various research advances, direct conversion, or transdifferentiation, is a notable and feasible strategy for the generation and acquirement of wanted cell source. So far, utilizing cell transdifferentiation technology in tissue engineering was mainly restricted at achieving single wanted cell type from diverse cell types with high efficiency. However, regeneration of a complete tissue always requires multiple cell types which poses an intrinsic complexity. In this study, enhanced osteogenic differentiation was achieved by transient ectopic expression of octamer-binding transcription factor 4 (OCT-4) gene followed by bone morphogenetic protein 4 treatment on human umbilical vein endothelial cells. OCT-4 transfection and bone morphogenetic protein 4 treatment resulted in enhanced expression of osteogenic markers such as core-binding factor alpha 1, alkaline phosphatase, and collagen 1 compared with bone morphogenetic protein 4 treatment alone. Furthermore, we employed gelatin-heparin cryogel in cranial defect model for in vivo bone formation. Micro-computed tomography and histological analysis of in vivo samples showed that OCT-4 transfection followed by bone morphogenetic protein 4 treatment resulted in efficient transdifferentiation of endothelial cells to osteogenic cells. These results suggest that the combination of OCT-4 and bone morphogenetic protein 4 on endothelial cells would be a reliable multicellular transdifferentiation model which could be applied for bone tissue engineering.

Apoptotic lysosomal proton sponge effect in tumor tissue by cationic gold nanorods.

Despite the lysosomal "proton sponge hypothesis" being considered to be an additional factor for stimulating the cellular toxicity of nanoparticle-based drug delivery systems, a clear relationship between the massive influx of calcium ions and the proton sponge effect, both of which are associated with cancer cell apoptosis, has still not been elucidated. Cetrimonium bromide (CTAB: cationic quaternary amino group based) gold nanorods possessed a more effective electric surface charge for inducing the lysosomal proton sponge effect than anionic gold nanoparticles. In this aspect, identifying released cytoplasmic Cl-, arising from the ruptured lysosomal compartment, in the cytoplasm is critical for supporting the "proton sponge hypothesis". This study clarified that the burst release of Cl-, as a result of lysosomal swelling by CTAB gold nanorods, stimulates the transient receptor potential channels melastatin 2 (TRPM2) channels, and subsequently induces a massive Ca2+ influx, which independently increases apoptosis of cancer cells. Although the previous concept of elevated cancer apoptosis acting through the proton sponge effect is unclear, this study supports the evidence that a massive Ca2+ influx mediated in response to a burst release of Cl- significantly influenced cytotoxicity of cancer cells in tumor tissues.

Mesenchymal stem cell therapy assisted by nanotechnology: a possible combinational treatment for brain tumor and central nerve regeneration.

Mesenchymal stem cells (MSCs) intrinsically possess unique features that not only help in their migration towards the tumor-rich environment but they also secrete versatile types of secretomes to induce nerve regeneration and analgesic effects at inflammatory sites. As a matter of course, engineering MSCs to enhance their intrinsic abilities is growing in interest in the oncology and regenerative field. However, the concern of possible tumorigenesis of genetically modified MSCs prompted the development of non-viral transfected MSCs armed with nanotechnology for more effective cancer and regenerative treatment. Despite the fact that a large number of successful studies have expanded our current knowledge in tumor-specific targeting, targeting damaged brain site remains enigmatic due to the presence of a blood-brain barrier (BBB). A BBB is a barrier that separates blood from brain, but MSCs with intrinsic features of transmigration across the BBB can efficiently deliver desired drugs to target sites. Importantly, MSCs, when mediated by nanoparticles, can further enhance tumor tropism and can regenerate the damaged neurons in the central nervous system through the promotion of axon growth. This review highlights the homing and nerve regenerative abilities of MSCs in order to provide a better understanding of potential cell therapeutic applications of non-genetically engineered MSCs with the aid of nanotechnology.

Comparative Analysis of the Developmental Toxicity in Xenopus laevis and Danio rerio Induced by Al2 O3 Nanoparticle Exposure.

Engineered aluminum oxide nanoparticles (Al2 O3 NPs) having high-grade thermal stability and water-dispersion properties are extensively used in different industries and personal care products. Toxicological response evaluation of these NPs is indispensable in assessing the health risks and exposure limits because of their industrial disposal into the aquatic environment. We assessed and compared the developmental toxicity of Al2 O3 NPs in Xenopus laevis and Danio rerio over a period of 96 h using the frog embryo teratogenic assay Xenopus and a fish embryo toxicity assay. Engineered Al2 O3 NP exposure produced dose-dependent embryonic mortality and decreased the embryo length, indicating a negative effect on growth. Moreover, Al2 O3 NPs induced various malformations, such as small head size, a bent/deformed axis, edema, and gut malformation, dose-dependently and altered the expression of heart- and liver-specific genes in both X. laevis and D. rerio, as revealed by whole-mount in-situ hybridization and reverse transcriptase polymerase chain reaction. In conclusion, the toxicological data suggest that Al2 O3 NPs are developmentally toxic and teratogenic and negatively affect the embryonic development of X. laevis and D. rerio. Our study can serve as a model for the toxicological evaluation of nanomaterial exposure on vertebrate development that is critical to ensure human and environmental safety. Environ Toxicol Chem 2019;38:2672-2681. © 2019 SETAC.

Clinical Application of Bone Morphogenetic Protein-2 Microcarriers Fabricated by the Cryopolymerization of Gelatin Methacrylate for the Treatment of Radial Fracture in Two Dogs.

Bone morphogenetic protein-2 (BMP-2) effectively induces bone healing. However, the efficacy of BMP-2 relies heavily on its delivery vehicle because of its short half-life. We utilized a microcarrier fabricated by the cryopolymerization of gelatin methacrylate (cryoGelMA) infused with bone morphogenetic protein-2 (cryoGelMA-BMP-2) for the sustained and localized release of growth factors. Two dogs with radius and ulnar fractures were treated with implanted cryoGelMA-BMP-2 to accelerate bone healing. The cases were followed up for 6 months and 2 months after surgery, respectively. Distinctive healing processes were observed. The operated limb regained its premorbid function, the fracture line disappeared, and the gait was functionally stable. Implantation of cryoGelMA-BMP-2 resulted in the successful healing of bone fractures.

Bioglass-Incorporated Methacrylated Gelatin Cryogel for Regeneration of Bone Defects.

Cryogels have recently gained interest in the field of tissue engineering as they inherently possess an interconnected macroporous structure. Considered to be suitable for scaffold cryogel fabrication, methacrylated gelatin (GelMA) is a modified form of gelatin valued for its ability to retain cell adhesion site. Bioglass nanoparticles have also attracted attention in the field due to their osteoinductive and osteoconductive behavior. Here, we prepare methacrylated gelatin cryogel with varying concentration of bioglass nanoparticles to study its potential for bone regeneration. We demonstrate that an increase in bioglass concentration in cryogel leads to improved mechanical property and augmented osteogenic differentiation of mesenchymal cells during in vitro testing. Furthermore, in vivo testing in mice cranial defect model shows that highest concentration of bioglass nanoparticles (2.5 w/w %) incorporated in GelMA cryogel induces the most bone formation compared to the other tested groups, as studied by micro-CT and histology. The in vitro and in vivo results highlight the potential of bioglass nanoparticles incorporated in GelMA cryogel for bone regeneration.

Difference in food and nutrient intakes in Korean elderly people according to chewing difficulty: using data from the Korea National Health and Nutrition Examination Survey 2013 (6th).

BACKGROUND/OBJECTIVES: Chewing difficulty is a factor contributing to a poor nutritional status in the elderly. The aim of this study was to examine disparities in food and nutrition intakes among Korean elderly people with and without chewing difficulty. SUBJECTS/METHODS: This study utilized data from the sixth Korea National Health and Nutrition Examination Survey conducted in 2013. The study subjects included males and females over 65 years of age who were not required to adhere to a special diet due to disease or sickness. They were divided into groups according to their chewing ability. Those who found chewing "very difficult" or "difficult", were combined to form the chewing difficulty group. Similarly, those who found chewing "moderately difficult", "easy", and "very easy" were combined to form the normal chewing group. RESULTS: Of the 999 subjects, 47.7% had chewing difficulties and the prevalence of chewing difficulty was higher in females than in males (P = 0.03) and higher in those 75 years of age and over than in younger individuals (P < 0.001). The chewing difficulty group had a significantly lower intake of fruits and vegetables (P < 0.05) and lower vitamin C and potassium intake than those in the normal group. Comparison of the percentages of Dietary Reference Intakes for Koreans (KDRIs) in the two groups indicated that the intake of most nutrients (energy, vitamin C, thiamin, riboflavin, niacin, calcium, phosphorus, sodium, potassium, and iron) were significantly lower in the chewing difficulty group than in the normal group. In particular, calcium intake was inadequate (51% of KDRIs) in the chewing difficulty group. CONCLUSIONS: The results indicate that chewing difficulty is closely related to food and nutrient intake in the elderly and can result in vitamin and mineral intake deficiencies. It is evident that the care of elderly subjects with chewing difficulty is essential for maintaining a healthy lifestyle.

Clinical Manifestations of Chemical Eye Injury

PURPOSE: The purpose of this study is to investigate the clinical characteristics of patients admitted to emergency rooms with ocular chemical injuries, the substances responsible for such injuries, in order to improve their prevention and treatment. METHODS: The subjects of this study were 73 patients admitted to the hospital with ocular chemical injuries from January 1, 2009 to December 31, 2010. The subjects were divided into groups based on the PSS (poisoning severity score) grade of their injuries, where grades 0 and 1 were classified as the low group (low PSS) and grades 2 and 3 were classified as the high group (high PSS). There were 28 subjects in the low PSS group (38.3%) and 45 subjects in the high PSS group (61.6%). The results were retrospectively reviewed by examining the medical records of the patients. RESULTS: Upon their arrival to the hospital, the most common symptoms were red eye (50.7%), pain (39.7%), and eye irritation (39.7%). Red eyes were more frequently observed from 27 persons (60%) in the high PSS group (p=0.04). The most common causative substances were chemicals and adhesives/glues for both groups (21 cases each). Among the 15 patients exposed to acids or alkalis, 13 patients belonged to the high PSS group. Only two patients visited the hospital after having washed their eyes. CONCLUSION: Worse damage was observed from work-related exposure (including alkali or acid exposure), causing red eye and ocular chemical injuries. Onsite first aid was lacking.

Compensation Methods for Non-uniform and Incomplete Data Sampling in High Resolution PET with Multiple Scintillation Crystal Layers / 핵의학분자영상

PURPOSE: To establish the methods for sinogram formation and correction in order to appropriately apply the filtered backprojection (FBP) reconstruction algorithm to the data acquired using PET scanner with multiple scintillation crystal layers. MATERIAL AND METHODS: Formation for raw PET data storage and conversion methods from listmode data to histogram and sinogram were optimized. To solve the various problems occurred while the raw histogram was converted into sinogram, optimal sampling strategy and sampling efficiency correction method were investigated. Gap compensation methods that is unique in this system were also investigated. All the sinogram data were reconstructed using 2D filtered backprojection algorithm and compared to estimate the improvements by the correction algorithms. RESULTS: Optimal radial sampling interval and number of angular samples in terms of the sampling theorem and sampling efficiency correction algorithm were pitch/2 and 120, respectively. By applying the sampling efficiency correction and gap compensation, artifacts and background noise on the reconstructed image could be reduced. CONCLUSION: Conversion method from the histogram to sinogram was investigated for the FBP reconstruction of data acquired using multiple scintillation crystal layers. This method will be useful for the fast 2D reconstruction of multiple crystal layer PET data.