Nanoscale modification of porous gelatin scaffolds with chondroitin sulfate for corneal stromal tissue engineering

Recent studies reflect the importance of using naturally occurring biopolymers as three-dimensional corneal keratocyte scaffolds and suggest that the porous structure of gelatin materials may play an important role in controlling nutrient uptake. In the current study, the authors further consider the application of carbodiimide cross-linked porous gelatin as an alternative to collagen for corneal stromal tissue engineering. The authors developed corneal keratocyte scaffolds by nanoscale modification of porous gelatin materials with chondroitin sulfate (CS) using carbodiimide chemistry. Scanning electron microscopy/energy dispersive X-ray spectroscopy and Fourier transform infrared spectroscopy showed that the amount of covalently incorporated polysaccharide was significantly increased when the CS concentration was increased from 0% to 1.25% (w/v). In addition, as demonstrated by dimethylmethylene blue assays, the CS content in these samples was in the range of 0.078–0.149 nmol per 10 mg scaffold. When compared with their counterparts without CS treatment, various CS-modified porous gelatin membranes exhibited higher levels of water content, light transmittance, and amount of permeated nutrients but possessed lower Young’s modulus and resistance against protease digestion. The hydrophilic and mechanical properties of scaffolds modified with 0.25% CS were comparable with those of native corneas. The samples from this group were biocompatible with the rabbit corneal keratocytes and showed enhanced proliferative and biosynthetic capacity of cultured cells. In summary, the authors found that the nanoscale-level modification has influence on the characteristics and cell-material interactions of CS-containing gelatin hydrogels. Porous membranes with a CS content of 0.112 ± 0.003 nmol per 10 mg scaffold may hold potential for use in corneal stromal tissue engineering

In Vitro Response of Retinal Pigment Epithelial Cells Exposed to Chitosan Materials Prepared with Different Cross-Linkers

The interaction between cells and biopolymers is the evaluation indicator of the biocompatibility of materials. The purpose of this work was to examine the responses of retinal pigment epithelial (RPE) cells to genipin (GP) or glutaraldehyde (GTA) cross-linked chitosan by means of cell viability assays, cytokine expression analyses, and apoptosis assays. Evaluations of non-cross-linked chitosan were conducted simultaneously for comparison. Both GP and GTA treated samples with the same extent of cross-linking (around 80%) were prepared by varying cross-linking time. Our results showed that GP cross-linking was carried out by either radical polymerization of the monomers or SN2 nucleophilic substitution reaction involving the replacement of the ester group on the monomer with a secondary amide linkage. On the other hand, GTA could react with free amino groups of chitosan, leading to the formation of either the Schiff bases or the Michael-type adducts with terminal aldehydes. The biocompatibility of non-cross-linked chitosan membranes was demonstrated by the absence of any signs of toxicity or inflammation reaction. The present study showed that the ARPE-19 cells exposed to GTA cross-linked chitosan membranes had significantly higher cytotoxicity, interleukin-6 levels, and number of TUNEL-positive nuclei than did those exposed to GP treated samples. In addition, the materials modified with GTA trigger apoptosis at an early stage and may induce toxicity in the RPE cells later. The findings suggest that while the chitosan molecules bridged by GP are satisfactorily cytocompatible, the counterparts treated by GTA do not seem to be tolerated. In terms of material safety, the GP cross-linked chitosan may be compatible with human RPE cells and may have a potential application as delivery carriers in the treatment of posterior segment diseases

Evaluation of an Epitypified Ophiocordyceps formosana

The substantial merit of Cordyceps s.l. spp. in terms of medicinal benefits is largely appreciated. Nevertheless, only few studies have characterized and examined the clinical complications of the use of health tonics containing these species. Here, we epitypified C. formosana isolates that were collected and characterized as Ophiocordyceps formosana based on morphological characteristics, molecular phylogenetic analyses, and metabolite profiling. Thus, we renamed and transferred C. formosana to the new protologue Ophiocordyceps formosana (Kobayasi & Shimizu) Wang, Tsai, Tzean & Shen comb. nov. Additionally, the pharmacological potential of O. formosana was evaluated based on the hot-water extract from its mycelium. The relative amounts of the known bioactive ingredients that are unique to Cordyceps s.l. species in O. formosana were found to be similar to the amounts in O. sinensis and C. militaris, indicating the potential applicability of O. formosana for pharmacological uses. Additionally, we found that O. formosana exhibited antioxidation activities in vitro and in vivo that were similar to those of O. sinensis and C. militaris. Furthermore, O. formosana also displayed conspicuously effective antitumor activity compared with the tested Cordyceps s.l. species. Intrinsically, O. formosana exhibited less toxicity than the other Cordyceps species. Together, our data suggest that the metabolites of O. formosana may play active roles in complementary medicine

[[alternative]]Gazing at the Life of a Flower--A Study on the Thoughts of Beauty in Ikenbono Ikebana

[[abstract]]台灣自1970年代後經濟起飛生活日漸優渥，追逐時尚喜愛插花和從事插花教學的人口也日益增多，「花」也不知不覺融入日常生活之中。不論民間宗教祭拜神明的儀式或婚喪禮俗，乃至一般的生活裝飾、怡情養性的休閒娛樂皆有花藝的呈現。「花道」日文漢字書寫為「華道」（kadou），又稱「生花」（ikebana），都是透過插花傳達內在思想，追求瞬間美感的藝術；台灣日式插花始於日據時代，上流社會以此文化交流聯誼，國民政府撤台後，則由早年留日返台者或少數台日通婚的日籍夫人教授插花，上流社會婦女插花因此蔚為風尚，其中以教授及研習池坊花道（Ikenobo ikebana ）者最多，歷程最久，而台灣學習池坊花道及喜愛池坊插花藝術之花友，礙於與日本語言與文化上之隔閡，對日本池坊花道藝術之涉獵大部份停留於雪泥鴻爪之片段，對於花形、樣式之插作行為，往往只知其然不知其所以然，而淪為純技術性的形式操作，然，日本池坊花道在外顯的形式樣貌中，乃蘊含深刻的生命哲學，從初期先民的依代信仰到佛教傳入，對其傳統文化影響頗深，面對其無常的生命、與「空寂」、「閑寂」、「幽玄」的美感思維與我們所習慣的審美經驗與思維模式相去甚遠，若未充分理解其獨特的審美意識，便有如隔紗看花，未能一窺其堂奧之美，實有遺珠之憾；惟插花行為以非主流形式依附於文化生活的脈絡發展，目前台灣以「插花藝術」所發表的學術論文或研究，在大中華文化背景薰陶下，多半著墨於「中華插花藝術」研究，面對多元社會文化的發展型態，「日本花道」這塊曾經風及一時又再度黯然的園地，則有待積極開創發掘，為研究瞭解日本池坊花道之生命美學，本文將從二個面向予以探討，第一：從日本歷史文化層面著手，探討依代信仰及佛教東傳對池坊花道歷史淵源之影響，第二：從美學面向著手，探討池坊花道之美感生命與哲學思維。《凝視一朵花的生命》乃筆者論文研究的主要方向，因涉及範圍較廣、層次較深，有待逐步論述之。[[abstract]]Since 1970s in Taiwan, economy has been growing and people have become richer and richer, so there are more and more people devoted to the teaching of flower arrangement. Who love flower arrangement and follow this trend get more. Flowers become a part of life, such as in offering to gods or ancestors, decorating in funerals, wedding, people’s normal life, or even people’s leisure life. “Flower Arrangement” written as “Kadou” in Japanese, called as “Ikebana,” implies that people convey their thoughts and present the transient sense of beauty by arranging flowers. In Taiwan, Kadou, which became a vital way, for high social class, to communicate with the others in the same social class, begins during Japanese colonial period. After the Nationalist government settled in Taiwan, only few who had ever studied in Japan or Japanese women who married Taiwanese men taught women in high social class to arrange flowers. This becomes a fashion. What they teach and learn most as well as what lasts longest is Ikenbo. However, those who learn and love Ikenbo are not good at Japanese language and culture, so “Ikenobo Ikebana” they learn is fragmented. Not only in flower patterns but styles, they know how to arrange flowers but they don’t know why. Aside from caring about external images, “Ikenobo ikebana” has specific sense of beauty. This sense of beauty contains more profound life philosophy, influenced by both their original belief and Buddhism, which includes the thoughts that are quite different from people’s traditional thoughts in Taiwan, such as “impermanence,” “kongji-empty silence,” leisurely quietness,” and “youxuan.” If we don’t understand the unique sense of beauty hidden in Japanese humanity spirit, we can’t fully know “Ikenobo ikebana.” However, flower arrangement is a pat of our culture in Taiwan, yet not domain in fashion. Until now, influenced by our traditional culture, the studies on the art of ikebana (i. e., the art of flower arrangement) mostly focus on Chinese art of ikebana. In order to understand and promote multi-cultural development in our own society, the study on the life beauty hidden and presented in Japanese Ikenobo Kadou is necessary. For the sake of achieving this goal, two aspects in this study are explored. Firstly, concerning Japanese history and culture, how its original belief and Buddhism influence the formation of Ikenobo Kadou is revealed. Secondly, as for aesthetics, the thoughts of beauty and vitality hidden and shown in Ikenobo Kado are explored

[[alternative]]Gazing at the Life of a Flower--A Study on the Thoughts of Beauty in Ikenobo Ikebana

[[abstract]]　　台灣自1970年代後經濟起飛帶動生活品質日漸優渥，追逐時尚喜愛插花和從事插花教學的人口也日益增多，「花」也不知不覺融入日常生活之中。不論民間宗教祭拜神明的儀式或婚喪禮俗，乃至一般的生活裝飾、怡情養性的休閒娛樂皆有花藝的呈現，做為追逐時尚生活的表徵。台灣日式插花始於日據時代，上流社會以此文化交流聯誼，國民政府撤台後，則由早年留日返台者或少數台日通婚的日籍夫人教授插花，上流社會婦女插花因此蔚為風尚，其中以教授及研習池坊花道（Ikenobo Ikebana）者最多，歷程亦最久，日本「花道」為中文稱呼，日文漢字書寫為「華道」（kadou），又稱「生花」（ikebana），「華」字為花的古字，兩者音相通，皆有花開之義，「道」是行走的路，也引申有方向、方法、技能、思想、知覺、真實生命之義，為人內心深處所建立通向理想的途徑，「花道」是透過學習插花技能，發展自覺的意志力量，以尋求或傳達內在精神思想，進而追求瞬間美感的藝術。而台灣學習池坊花道及喜愛池坊插花藝術的花友，礙於與日本語言與文化上之隔閡，對日本池坊花道藝術之涉獵大部份停留於雪泥鴻爪之片段，對於花形、樣式之插作行為，往往只知其然不知其所以然，而淪為純技術性的形式操作；然，日本池坊花道在外顯的形式樣貌中，乃蘊含深刻的生命哲學，從初期先民的依代信仰到佛教供花傳入，對日本傳統文化影響頗深，面對無常的生命、與「幽玄」、「空寂」、「閑寂」的美感思維與我們所習慣的審美經驗與思維模式相去甚遠，若未充分理解其獨特的審美意識，便有如隔紗看花，未能一窺其堂奧之美，實有遺珠之憾。　　惟插花行為以非主流形式依附於文化生活的脈絡發展，受限於生活條件優渥與否影響，使得插花人口侷限於少數之數，目前台灣以「插花藝術」所發表的學術論文或研究，在大中華文化背景薰陶下，多半著墨於「中華插花藝術」研究，面對多元社會文化的發展型態，「日本花道」這塊曾經風及一時又再度黯然的園地，則有待積極開創發掘，為研究瞭解日本池坊花道之生命美學，本文將從三個面向予以探討，第一：從日本歷史文化層面著手，探討依代信仰及佛教東傳對池坊花道歷史淵源的演繹發展，其次探討日本佛教天台本覺思想對池坊花道精神內涵的影響，第三：從美學面向著手，探討池坊花道的生命美感與哲學思維。[[abstract]]　　Since 1970s in Taiwan, economy has been growing and people have become richer and richer, so there are more and more people devoted to the teaching of flower arrangement. Who love flower arrangement and follow this trend get more. Flowers become a part of life, such as in offering to gods or ancestors, decorating in funerals, wedding, people’s normal life, or even people’s leisure life. “Flower Arrangement” written as “Kadou” in Japanese, called as “Ikebana,” implies that people convey their thoughts and present the transient sense of beauty by arranging flowers. In Taiwan, Kadou, which became a vital way, for high social class, to communicate with the others in the same social class, begins during Japanese colonial period. After the Nationalist government settled in Taiwan, only few who had ever studied in Japan or Japanese women who married Taiwanese men taught women in high social class to arrange flowers. This becomes a fashion. What they teach and learn most as well as what lasts longest is Ikenobo. However, those who learn and love Ikenobo are not good at Japanese language and culture, so “Ikenobo Ikebana” they learn is fragmented. Not only in flower patterns but styles, they know how to arrange flowers but they don’t know why. Aside from caring about external images, “Ikenobo ikebana” has specific sense of beauty. This sense of beauty contains more profound life philosophy, influenced by both their original belief and Buddhism, which includes the thoughts that are quite different from people’s traditional thoughts in Taiwan, such as “impermanence,” “kongji-empty silence,” leisurely quietness,” and “youxuan.” If we don’t understand the unique sense of beauty hidden in Japanese humanity spirit, we can’t fully know “Ikenobo ikebana.” However, flower arrangement is a pat of our culture in Taiwan, yet not domain in fashion. Until now, influenced by our traditional culture, the studies on the art of ikebana (i. e., the art of flower arrangement) mostly focus on Chinese art of ikebana. In order to understand and promote multi-cultural development in our own society, the study on the life beauty hidden and presented in Japanese Ikenobo Kadou is necessary. For the sake of achieving this goal, three aspects in this study are explored. Firstly, concerning Japanese history and culture, how its original belief and Buddhism influence the formation of Ikenobo Kadou is revealed. Secondly, explore the impact of the idea of Tian tai Japanese Buddhism on the spiritual connotation of Ikenobo Ikebana. Finally, as for aesthetics, the thoughts of beauty and vitality hidden and shown in Ikenobo Kado are explored

High Rainfall Inhibited Soil Respiration in an Asian Monsoon Forest in Taiwan

Soil respiration represents the second largest carbon flux, next to photosynthesis of the terrestrial biosphere, and thus plays a dual role in regional and global carbon cycles. However, soil respiration in Asian monsoon forests with high rainfall has rarely been studied. In this study, we continuously measured soil respiration using a 12-channel automated chamber system in a 61-year-old Japanese cedar forest in central Taiwan with annual rainfall greater than 2500 mm. A 4-year (2011–2014) continuous half-hourly dataset was used to quantify the influences of soil temperature and moisture, especially rainfall events, on both total soil respiration (Rs) and heterotrophic respiration (Rh). The annual mean Rs was approximately 10.8 t C ha−1 (ranging from 10.7 to 10.9) t C ha−1, with Rh contributing approximately 74.6% (ranging from 71.7% to 80.2%). Large seasonal variations in both Rs and Rh were primarily controlled by soil temperature. Over 45.8% of total annual rainfall amounts were provided by strong rainfall events (over 50 mm), and over 40% of rainfall events occurred during summers between 2012 and 2014. These strong rainfall events caused rainwater to enter soil pores and cover the soil surface, which resulted in limited soil microorganism activity and, consequently, restricted CO2 production. The mean Q10 values were 2.38 (ranging from 1.77 to 2.65) and 2.02 (ranging from 1.71 to 2.34) for Rs and Rh, respectively. The Q10 values in this study, which were lower than in global forest ecosystems, may imply that the interannual Rs values observed in this study that were caused by high rainfall were less temperature-dependent than the Rs levels in global forest ecosystems. Both Rs and Rh were negatively correlated with soil moisture, which indicated that the soil moisture levels in the studied forest were usually under saturated conditions. These results also provide the lack of data for respiration in the Asian monsoon region under high-rainfall conditions

The Influence of Mg-Based Inclusions on the Grain Boundary Mobility of Austenite in SS400 Steel

In this study, the effects of the addition of Mg to the grain growth of austenite and the magnesium-based inclusions to mobility were investigated in SS400 steel at high temperatures. A high-temperature confocal scanning laser microscope (HT-CSLM) was employed to directly observe, in situ, the grain structure of austenite under 25 torr Ar at high temperatures. The grain size distribution of austenite showed the log-normal distribution. The results of the grain growth curves using 3D surface fitting showed that the n and Q values of the growth equation parameters ranged from 0.2 to 0.26 and from 405 kJ/mole to 752 kJ/mole, respectively, when adding 5.6–22 ppm of Mg. Increasing the temperature from 1150 to 1250 °C for 20 min and increasing the addition of Mg by 5.6, 11, and 22 ppm resulted in increases in the grain boundary velocity. The effects of solute drag and Zener pinning on grain boundary mobility were also calculated in this study

Antimicrobial activity of Antrodia camphorata extracts against oral bacteria.

Antrodia camphorata (A. camphorata) is a unique, endemic and extremely rare mushroom species native to Taiwan, and both crude extracts of and purified chemical compounds from A. camphorata have been reported to have a variety of significant beneficial effects, such as anti-tumor and anti-inflammatory activity. However, reports on the effects of A. camphorata against dental pathogens have been limited. Oral health is now recognized as important for overall general health, including conditions such as dental caries, periodontal disease and rheumatoid arthritis. Streptococcus mutans (S. mutans) and Porphyromonas gingivalis (P. gingivalis) are the most common bacteria associated with dental plaque and periodontopathic diseases, respectively. Thus, our study examined the ability of five various crude extracts of A. camphorata to inhibit the growth of dental bacteria and anti-adherence in vitro. Among the extracts, the ethanol, ethyl acetate and chloroform extracts exhibited the lowest MICs against P. gingivalis and S. mutans (MIC = 4∼16 µg/mL). The MIC of the aqueous extract was greater than 2048 µg/mL against both P. gingivalis and S. mutans. In vitro adherence of S. mutans was significantly inhibited by the addition of either the ethyl acetate extract or chloroform extract (MIC = 16∼24 µg/mL), while the ethanol extract (MIC = 32∼64 µg/mL) exhibited moderate inhibitory activity. Based on the result of this study, the ethyl acetate and chloroform extracts of A. camphorata may be good candidates for oral hygiene agents to control dental caries and periodontopathic conditions

Characterization of Cross-Linked Porous Gelatin Carriers and Their Interaction with Corneal Endothelium: Biopolymer Concentration Effect

<div><p>Cell sheet-mediated tissue regeneration is a promising approach for corneal reconstruction. However, the fragility of bioengineered corneal endothelial cell (CEC) monolayers allows us to take advantage of cross-linked porous gelatin hydrogels as cell sheet carriers for intraocular delivery. The aim of this study was to further investigate the effects of biopolymer concentrations (5–15 wt%) on the characteristic and safety of hydrogel discs fabricated by a simple stirring process combined with freeze-drying method. Results of scanning electron microscopy, porosity measurements, and ninhydrin assays showed that, with increasing solid content, the pore size, porosity, and cross-linking index of carbodiimide treated samples significantly decreased from 508±30 to 292±42 µm, 59.8±1.1 to 33.2±1.9%, and 56.2±1.6 to 34.3±1.8%, respectively. The variation in biopolymer concentrations and degrees of cross-linking greatly affects the Young’s modulus and swelling ratio of the gelatin carriers. Differential scanning calorimetry measurements and glucose permeation studies indicated that for the samples with a highest solid content, the highest pore wall thickness and the lowest fraction of mobile water may inhibit solute transport. When the biopolymer concentration is in the range of 5–10 wt%, the hydrogels have high freezable water content (0.89–0.93) and concentration of permeated glucose (591.3–615.5 µg/ml). These features are beneficial to the in vitro cultivation of CECs without limiting proliferation and changing expression of ion channel and pump genes such as ATP1A1, VDAC2, and AQP1. In vivo studies by analyzing the rabbit CEC morphology and count also demonstrate that the implanted gelatin discs with the highest solid content may cause unfavorable tissue-material interactions. It is concluded that the characteristics of cross-linked porous gelatin hydrogel carriers and their triggered biological responses are in relation to biopolymer concentration effects.</p></div

Swelling tests.

<p>Time course of swelling ratio of various gelatin discs after incubation in BSS at 34°C. An asterisk indicates statistically significant differences (*<i>P</i><0.05; <i>n</i> = 5) for the mean value of swelling ratio compared with the value at the previous time point.</p