In this study, the aggregation mechanism of polyion complex (PIC) particles from chitosan (CHI) and chondroitin sulfate C (CS) in phosphate-buffered saline (PBS) was analyzed, and a novel method for ...the fabrication of hydrogels via aggregation was developed. The PBS induced a decrease in the ζ-potential of the CS/CHI PIC particles, increase in their diameter, and aggregation in a concentration-dependent manner. The hydrogels prepared by mixing CS/CHI PIC particle dispersion and PBS showed the PIC components, with porous structure, high swelling ratio (161.4 ± 13.3%), and high storage moduli (26.2 ± 1.4 kPa). By mixing PBS with suspended adhesive cells and CS/CHI PIC particle dispersion, hydrogels with high cell-loading efficiency were successfully synthesized. The loaded cells within the hydrogels exhibited high viability, uniform distribution, and formation of cell aggregates. These results indicate that CS/CHI-based hydrogels have a potential application as three-dimensional scaffolds for cell culture in tissue engineering.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
Silica nonwoven fabrics (SNFs) with high mechanical strength and porosity are known to exhibit high cell proliferation and osteogenic differentiation potential of mesenchymal stem cells (MSCs) by ...morphologically mimicking the extracellular matrix (ECM). To further improve the osteoinductive ability of SNFs, it could be effective to increase the interaction between MSCs and ECM components because exogenous ECM components seem to modulate the fate of MSCs differentiation. In this study, we developed immobilization methods for ECM components, such as collagen, fibronectin, and chondroitin sulphate C on SNFs, to improve cell–matrix interactions and examined their suitability for bone tissue regeneration. Collagen and fibronectin were immobilized via physical adsorption and chondroitin sulphate C was also immobilized by the layer-by-layer method combined with chitosan on SNF surfaces to maintain the high porosity of SNFs. The treated SNFs were characterized using scanning electron microscopy, Fourier transform infrared spectroscopy, and X-ray photoelectron spectroscopy. In osteogenic differentiation culture, modified SNFs showed significantly increased expression of osteogenic differentiation marker genes compared to unmodified SNFs. These results suggest that the present methods improve cell–matrix interactions and enhance the cellular functions of MSCs. We are convinced that these simple modification techniques for ECM components are effective in functionalizing various 3D fabric scaffolds possessing hydrophilic groups.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
Polysaccharides have been extensively studied as biomaterials in various fields for their biocompatibility, biodegradability, and biological functions. To obtain water-insoluble materials from ...polysaccharides, polyion complexes (PICs) formed between cationic and anionic polysaccharides have been widely used in drug delivery systems and tissue engineering. Understanding the atomic interaction mechanism of oppositely charged polysaccharides is important in the design and application of PIC-based materials. In this work, the interaction between single-stranded chitosan and four kinds of anionic oligosaccharides was systematically investigated to elucidate the effects of the functional groups of chitosan and chemical species of anionic oligosaccharides on complex formation using molecular dynamics (MD) simulation with atomic detail. We verified that chitosan and anionic oligosaccharides form complexes, regardless of the functional groups of chitosan. For ‒NH3+ chitosan, due to the strong electrostatic interaction, a higher number of hydrogen bonds between ‒NH3+ in chitosan and anionic charged groups of anionic oligosaccharides were formed. Our results also suggested that ‒NH2 and ‒NHAc chitosan could form complexes with anionic oligosaccharides due to hydrogen bonds. These findings might be important for the design and stabilization of PICs based on polysaccharides.Molecular dynamics simulation of chitosan and anionic oligosaccharides was conduced to elucidate the formation mechanism and structure of polyion complex (PIC) of polysaccharides. The association of oligosaccharides with scrambled structure indicated that PIC of polysaccharides takes on the structure of “Scrambled egg model”. The formation of hydrogen bond between deprotonated chitosan and anionic oligosaccharides suggested that hydrogen bonds contribute to the stabilization of PIC under neutral condition.
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EMUNI, FIS, FZAB, GEOZS, GIS, IJS, IMTLJ, KILJ, KISLJ, MFDPS, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, SBMB, SBNM, UKNU, UL, UM, UPUK, VKSCE, ZAGLJ
Biomaterials made of natural polysaccharides have attracted much attention due to the fact of their excellent properties, such as high biocompatibility and biodegradability, and their specific ...biological functions based on their chemical structures. This study demonstrates that polysaccharide composite films can be fabricated from polyion complexes (PICs) with their particles used as building components. Dispersion of PIC particles prepared by mixing, centrifugation, and re-dispersion of dilute solutions of cationic and anionic polysaccharides were cast, dried, and formed into films several micrometers thick. These films were homogenous and water insoluble. It was revealed that the component anionic polysaccharides affected the film's properties such as the swelling behavior and mechanical characteristics. Adhesion of NIH3T3 cells (integrin: high, CD44: lack or weak) and A549 cells (integrin: high, CD44: high) to the composite films were examined. Both NIH3T3 and A549 cells adhered to heparin/chitosan (HEP/CHI) film because HEP has an affinity for integrin through fibronectin. However, A549 cells adhered to chondroitin sulfate (CS)/CHI and hyaluronic acid (HYA)/CHI films, whereas NIH3T3 cells did not, because both CS and HYA have affinity for CD44. These results indicated that the biological functions of anionic polysaccharides were maintained on the surface of the composite films. It was also possible to fabricate films composed of three kinds of polysaccharides: one cationic polysaccharide and two kinds of anionic polysaccharides. These results show that the properties of films composed of three kinds of polysaccharides may be controllable depending on the anionic polysaccharide composition rates.
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IZUM, KILJ, NUK, PILJ, PNG, SAZU, UL, UM, UPUK
The balanced corpus of contemporary written Japanese (BCCWJ) is Japan's first 100 million words balanced corpus. It consists of three subcorpora (publication subcorpus, library subcorpus, and ...special-purpose subcorpus) and covers a wide range of text registers including books in general, magazines, newspapers, governmental white papers, best-selling books, an internet bulletin-board, a blog, school textbooks, minutes of the national diet, publicity newsletters of local governments, laws, and poetry verses. A random sampling technique is utilized whenever possible in order to maximize the representativeness of the corpus. The corpus is annotated in terms of dual POS analysis, document structure, and bibliographical information. The BCCWJ is currently accessible in three different ways including Chunagon a web-based interface to the dual POS analysis data. Lastly, results of some pilot evaluation of the corpus with respect to the textual diversity are reported. The analyses include POS distribution, word-class distribution, entropy of orthography, sentence length, and variation of the adjective predicate. High textual diversity is observed in all these analyses.
Heat stress has negative effects on biological defense mechanisms such as the immune response in chickens, and organs of the immune system, such as the spleen, are atrophied by heat stress in broiler ...chickens. In order to assess heat stress-induced functional changes of immune mechanisms in the spleen of broiler chickens, expression of genes encoding splenic cytokines (Th1 type, Th2 type, and pro-inflammatory cytokines) was analyzed. Heat exposure at 34°C for 15 days significantly induced the spleen involution, increased interleukin (IL)-4 and IL-12 expression and decreased interferon (IFN)-γ. However, expression of IL-6, 10, 13, and 18 was not affected. Heat stress reduced feed intake, which may affect the spleen weight and cytokine expression. Therefore, a pair-fed group at 24°C (24PF) was included in which chickens received the same amount of feed as those in the group at 34°C. Spleen weight was not affected by the reduction in feed intake. IL-4 expression in the 24PF group was higher than that in the control group. Further, IFN-γ expression increased and IL-12 expression was not affected by the reduction of feed intake, suggesting that the feed intake reduction induced by heat stress does not modulate splenic cytokine expression in broiler chickens. These data suggest that heat stress induces spleen involution and affects the expression of splenic cytokines such as IL-12 and IFN-γ in broiler chickens, but not through the feed intake reduction.
Heat stress in laying hens negatively affects egg production and shell quality by disrupting the homeostasis of plasma calcium and phosphorus levels. Although the kidney plays an important role in ...calcium and phosphorus homeostasis, evidence regarding the effect of heat stress on renal injury in laying hens is yet to be elucidated. Therefore, the aim of this study was to evaluate the effects of chronic heat stress on renal damage in hens during laying periods.
A total of 16 white-leghorn laying hens (32 weeks old) were randomly assigned to two groups (n = 8). One group was exposed to chronic heat stress (33 °C for 4 weeks), whereas the other group was maintained at 24 °C.
Chronic heat exposure significantly increased plasma creatinine and decreased plasma albumin levels (P < 0.05). Heat exposure also increased renal fibrosis and the transcription levels of fibrosis-related genes (COLA1A1, αSMA, and TGF-β) in the kidney. These results suggest that renal failure and fibrosis were induced by chronic heat exposure in laying hens. In addition, chronic heat exposure decreased ATP levels and mitochondrial DNA copy number (mtDNA-CN) in renal tissue, suggesting that renal mitochondrial dysfunction occurs under conditions of heat stress. Damaged mitochondria leak mtDNAs into the cytosol and mtDNA leakage may activate the cyclic GMP-AMP synthase (cGAS) stimulator of interferon genes (STING) signaling pathway. Our results showed that chronic heat exposure activated the cGAS-STING pathway as indicated by increased expression of MDA5, STING, IRF7, MAVS, and NF-κB levels. Furthermore, the expression of pro-inflammatory cytokines (IL-12) and chemokines (CCL4 and CCL20) was upregulated in heat-stressed hens.
These results suggest that chronic heat exposure induces renal fibrosis and mitochondrial damage in laying hens. Mitochondrial damage by heat stress may activate the mtDNA-cGAS-STING signaling and cause subsequent inflammation, which contributes to the progression of renal fibrosis and dysfunction.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
The aim of this study was to evaluate the effects of dietary brown rice on the growth performance, systemic oxidative status, and splenic inflammatory responses of broiler chickens under both ...thermo-neutral and chronic heat stress conditions. Forty 12-day-old male broiler chickens (ROSS 308) were randomly assigned to two groups and fed either a control diet (corn-based) or a brown rice-based diet. After seven days (19 days old), both groups were randomly divided into two sub-groups (n=10), one of which was exposed to heat stress (33°C for 14 days), while the other was maintained at 24°C. Heat exposure reduced the body weight gain and feed intake (p<0.01) of both groups. In terms of oxidative plasma states, heat exposure reduced the glutathione peroxidase activity and increased the ceruloplasmin content, while the 2-thiobarbituric acid reactive substance and reduced glutathione levels were not affected adversely. Heat exposure activated the immune responses, as evidenced by increased plasma immunoglobin levels, and altered splenic immune-related gene expressions including heat shock proteins, toll-like receptor 4, and interleukin-12. Under both thermo-neutral and heat stress conditions, dietary brown rice improved the growth performance, decreased the immunoglobulin levels, and down-regulated the expression of splenic immune-related genes of broilers, although their systemic oxidative status was not affected. Dietary brown rice should be considered as a valuable component of broiler chicken feeds subjected to both thermo-neutral and heat stress conditions. The positive effects of brown rice on bird performance may be associated with the modulation of the immune responses, as reflected by the decreased production of immunoglobulins and altered splenic immune-related gene expression.
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