Browsing by Author "Huang, Guewha Steven"

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Assessment of gold nanoparticles as a size-dependent vaccine carrier for enhancing the antibody response against synthetic foot-and-mouth disease virus peptide
(2010-05-14)
Chen, Yu-Shiun
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Hung, Yao-Ching
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Lin, Wei-Hsu
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Huang, Guewha Steven
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材料科學與工程學系
To assess the ability of gold nanoparticles (GNPs) to act as a size-dependent carrier, a synthetic peptide resembling foot-and-mouth disease virus (FMDV) protein was conjugated to GNPs ranging from 2 to 50 nm in diameter (2, 5, 8, 12, 17, 37, and 50 nm). An extra cysteine was added to the C-terminus of the FMDV peptide (pFMDV) to ensure maximal conjugation to the GNPs, which have a high affinity for sulfhydryl groups. The resultant pFMDV-GNP conjugates were then injected into BALB/c mice. Immunization with pFMDV-keyhole limpet hemocyanin (pFMDV-KLH) conjugate was also performed as a control. Blood was obtained from the mice after 4, 6, 8, and 10 weeks and antibody titers against both pFMDV and the carriers were measured. For the pFMDV-GNP immunization, specific antibodies against the synthetic peptide were detected in the sera of mice injected with 2, 5, 8, 12, and 17 nm pFMDV-GNP conjugates. Maximal antibody binding was noted for GNPs of diameter 8-17 nm. The pFMDV-GNPs induced a three-fold increase in the antibody response compared to the response to pFMDV-KLH. However, sera from either immunized mouse group did not exhibit an antibody response to GNPs, while the sera from pFMDV-KLH-immunized mice presented high levels of binding activity against KLH. Additionally, the uptake of pFMDV-GNP in the spleen was examined by inductively coupled plasma mass spectroscopy (ICP-MS) and transmission electron microscopy (TEM). The quantity of GNPs that accumulated in the spleen correlated to the magnitude of the immune response induced by pFMDV-GNP. In conclusion, we demonstrated the size-dependent immunogenic properties of pFMDV-GNP conjugates. Furthermore, we established that GNPs ranging from 8 to 17 nm in diameter may be ideal for eliciting a focused antibody response against a synthetic pFMDV peptide.
Camellia sinensis tea melanin suppresses transformation of the aryl hydrocarbon receptor and prevents against dioxin-induced toxicity in mice
(2008-02-01)
Hung, Yao-Ching
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Huang, Guewha Steven
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Sava, Vasyl M.
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Makan, Svetlana Y.
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Hong, Meng-Yen
The suppressive effects of Camellia sinensis tea melanin (CSTM) on transformation of aryl hydrocarbon receptor (AhR) induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) were disclosed for the first time. CSTM strongly inhibited TCDD-induced toxicity with IC50 equalling 20.4 mu g mL(-1). Daily administration of CSTM (40 mg kg(-1), p.o.) prevented TCDD-induced body weight loss, ameliorated TCDD-induced mortality and prevented TCDD-induced hepatomegaly and thymic atrophy. Co-administration of CSTM significantly inhibited TCDD-induced hepatic CYP1 A1 activity. CSTM retarded transformation of AhR in vitro. In animals treated with CSTM, the antibody-secreting cells produced significantly (P < 0.05) more antibodies (32-34%) than the antigen control. Administration of TCDD caused a suppression of antibody-forming cells of 29-33% against the antigen control level. Co-administration of CSTM restored immunity to the control level. We demonstrated that CSTM directly competed with TCDD during the transformation of AhR and suppressed the downstream activation of genes associated with TCDD toxicity.
Control of growth and inflammatory response of macrophages and foam cells with nanotopography
(2012-07-16)
Mohiuddin, Mohammed
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Pan, Hsu-An
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Hung, Yao-Ching
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Huang, Guewha Steven
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材料科學與工程學系
Macrophages play an important role in modulating the immune function of the human body, while foam cells differentiated from macrophages with subsequent fatty streak formation play a key role in atherosclerosis. We hypothesized that nanotopography modulates the behavior and function of macrophages and foam cells without bioactive agent. In the present study, nanodot arrays ranging from 10aEuro to 200aEuronm were used to evaluate the growth and function of macrophages and foam cells. In the quantitative analysis, the cell adhesion area in macrophages increased with 10- to 50-nm nanodot arrays compared to the flat surface, while it decreased with 100- and 200-nm nanodot arrays. A similar trend of adhesion was observed in foam cells. Immunostaining, specific to vinculin and actin filaments, indicated that a 50-nm surface promoted cell adhesion and cytoskeleton organization. On the contrary, 200-nm surfaces hindered cell adhesion and cytoskeleton organization. Further, based on quantitative real-time polymerase chain reaction data, expression of inflammatory genes was upregulated for the 100- and 200-nm surfaces in macrophages and foam cells. This suggests that nanodots of 100aEuroaEuro parts per thousand and 200aEuronm triggered immune inflammatory stress response. In summary, nanotopography controls cell morphology, adhesions, and proliferation. By adjusting the nanodot diameter, we could modulate the growth and expression of function-related genes in the macrophages and foam cell system. The nanotopography-mediated control of cell growth and morphology provides potential insight for designing cardiovascular implants.
Detection of gold nanoparticles using an immunoglobulin-coated piezoelectric sensor
(2008-12-10)
Chen, Yu-Shiun
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Hung, Yao-Ching
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Chen, Kaochao
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Huang, Guewha Steven
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材料科學與工程學系
Since the existence of nanoparticles in our environment has already attracted considerable attention due to their possible toxic impact on biological systems, the field detection of nanoparticles is becoming a technology that will be much in need. We have constructed a piezoelectric sensor with an antibody-coated electrode. The antiserum can bind gold nanoparticles with a high degree of selectivity and sensitivity. The biosensor thus constructed can detect 4, 5, or 6 nm gold nanoparticles (GNPs) depending on the coated antiserum. The sensitivity for the detection of 5 nm GNPs was 10.3 +/- 0.9 ng Hz(-1), with the low limit of detection at 5.5 ng. A quartz crystal microbalance (QCM) sensor was capable of detecting GNPs and other types of nanoparticle, such as ZnO, or Fe(3)O(4). The current study provides, for the first time, a platform for detecting nanoparticles in a convenient, economical manner.
Effects of static magnetic fields on the development and aging of Caenorhabditis elegans
(2010-06-15)
Hung, Yao-Ching
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Lee, Jia-Huey
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Chen, Huang-Meng
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Huang, Guewha Steven
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材料科學與工程學系奈米科技碩博班
The current study investigated the possible effects of static magnetic fields (SMFs) on the developmental and aging processes of Caenorhabditis elegans. Nematodes were grown in the presence of SMFs of strengths varying from 0 to 200 mT. The rate of development and the lifespan were recorded. Treatment with a 200 mT SMF reduced the development time from the L2 to the L3 stage by 20%, from L3 to L4 by 23%, and from L4 to young adult by 31%. After SMF treatment, the average lifespan was reduced from 31 days to 24 days for wild-type nematodes. The up-regulation of clk-1, lim-7, daf-2, unc-3 and age-1 after SMF treatment was verified by quantitative real-time RT-PCR. Apparently, induction of gene expression is selective and dose dependent. The total developmental time was significantly reduced for the lin-4, lin-14, lin-41 and lim-7 mutants, but not for the let-7, clk-1, unc-3 and age-1 mutants. Lifespan analyses revealed that the let-7, unc-3 and age-1 mutants were not affected by SMF treatment. Here we show that SMFs accelerate nematode development and shorten nematode lifespan through pathways associated with let-7, clk-1, unc-3 and age-1.
Prevention and amelioration of acetaminophen toxicity with tea melanin
(2006-07-20)
Huang, Guewha Steven
The present invention relates to a method for preventing and/or reducing the toxicity of acetaminophen which comprises administering to a mammal an amount of tea melanin before or simultaneous with dosage of acetaminophen, a pharmaceutical composition containing tea melanin, and a pharmaceutical composition containing tea melanin and acetaminophen for preventing and/or reducing the toxicity of acetaminophen.
Spatial Control of Cell-Nanosurface Interactions by Tantalum Oxide Nanodots for Improved Implant Geometry
(2016-06-30)
Dhawan, Udesh
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Pan, Hsu An
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Lee, Chia Hui
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Chu, Ying Hao
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Huang, Guewha Steven
Nanotopological cues can be exploited to understand the nature of interactions between cells and their microenvironment to generate superior implant geometries. Nanosurface parameters which modulate the cell behavior and characteristics such as focal adhesions, cell morphology are not clearly understood. Here, we studied the role of different nanotopographic dimensions in modulating the cell behavior, characteristics and ultimately the cell fate and accordingly, a methodology to improve implant surface geometry is proposed. Tantalum oxide nanodots of 50, 100nm dot diameter with an inter-dot spacing of 20, 70nm and heights 40, 100nm respectively, were engineered on Silicon substrates. MG63 cells were cultured for 72 hours and the modulation in morphology, focal adhesions, cell extensible area, cell viability, transcription factors and genes responsible for bone protein secretion as a function of the nanodot diameter, inter-dot distance and nanodot height were evaluated. Nanodots of 50nm diameter with a 20nm inter-dot spacing and 40nm height enhanced cell spreading area by 40%, promoted cell viability by 70% and upregulated transcription factors and genes twice as much, as compared to the 100nm nanodots with 70nm inter-dot spacing and 100nm height. Favorable interactions between cells and all dimensions of 50nm nanodot diameter were observed, determined with Scanning electron microscopy and Immunofluorescence staining. Nanodot height played a vital role in controlling the cell fate. Dimensions of nanodot features which triggered a transition in cell characteristics or behavior was also defined through statistical analysis. The findings of this study provide insights in the parameters of nanotopographic features which can vitally control the cell fate and should therefore be taken into account when designing implant geometries.
Ultrasensitive Detection of Cymbidium Mosaic Potexvirus Using a Single-Wall Carbon Nanotube-Functionalized Quartz Crystal Microbalance
(2010)
Chen, Yu-Shiun
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Hung, Yao-Ching
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Chiou, Jin-Chern
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Wang, Hui-Liang
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Huang, Hung-Shu
We have developed an ultrasensitive, convenient, real-time platform for detecting Cymbidium mosaic potexvirus (CymMV) based on single-wall carbon nanotube (SWNT)-functionalized quartz crystal microbalance (QCM) sensors. Functionalization was achieved by coating the QCM electrode with SWNTs, followed by 1,1'-carbonyldiimidazole-activated Tween 20 (CDI-Tween 20) modification and conjugation of antibodies. Sensitivity was enhanced from 2.18 to 11.5 Hz ng(-1) when 0.1 mu g mL(-1) CymMV was applied. The low limit of detection of SWNT-functionalized QCM sensors was improved from 2.08 to 0.502 ng. The SWNT-functionalized QCM sensor was successfully used to quantify the amount of CymMV contained in infected orchid leaves. Compared to enzyme-linked immunosorbent assay (ELISA), SWNT-functionalized QCM sensors are fast, economical, and ultra-sensitive, with comparable sensitivities. The current study demonstrates the application of QCM sensors as a convenient platform to detect and quantify CymMV. (C) 2010 The Japan Society of Applied Physics
以奈米技術開發茶葉黑色素的新功能
(2005)
黃國華
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Huang, Guewha Steven
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國立交通大學奈米科技研究所
黑色素是一種存在於植物、動物和微生物的獨特色素。黑色素主要功能在於保護生物免於太陽輻射的直接照射；黑色素不僅是個「顏色」。同時也具有清除自由機活性功能的抗氧化劑，亦是重金屬的螯合劑，也對有毒有機分子具有強吸收性。黑色素本身無毒且具有生物親和力；是對抗有毒化學物和重金屬的天然保護劑。由於動物性的黑色素不溶解於水、非晶態以及複雜的高分子結構，在研究化性、物性以及生物特性有很大的困難。因此，需要尋找以及研究出一種嶄新取得黑色素的來源。我們成功的在茶葉中萃取出在生理酸鹼值下可溶解的黑色素。從茶葉萃取出的黑色素無毒以及具有由其他黑色素取得來源相同的生物活性。它的強螯合力和高磁滯可以做為MRI 的顯影劑。對於蛋白質和有毒分子的強結合性使黑色素亦是種很好的抗蛇毒劑。其抗氧化以及清除自由機活性的能力使得能保護果蠅免於 benzidine 的破壞。黑色素的保護能力甚至可以保護肝細胞免於hydrazine 的直接破壞。此外，黑色素亦可以阻止acetaminophen 對於肝臟的傷害。然而，對於分子作用機制以及基因調節卻很少研究。茶葉黑色素是個具有單一分子量14 kD 的均勻高分子。我們使用掃瞄式電子顯微鏡第一次發現到茶葉黑色素自組裝的奈米結構。這個新發現的自組裝黑色素結構在生物上的特性還沒有被研究。我們同時也發現了茶葉黑色素的奈米晶體。對於黑色素的奈米粒子的物性、化性和生物特性方面的研究是非常重要的。我們可以製作兩種特殊的黑色素奈米結構型態：自組裝的奈米棒狀以及奈米結晶的黑色素。奈米粒子的黑色素可能具有的嶄新結構和生物特性還沒有被證明出來。我們提出對於茶葉黑色素奈米結構在物性、化性和生物特性的研究。我們將利用泡沫細胞研究奈米黑色素對於生物分子的影響。我們的特定目標為： 1. 研究黑色素對巨噬細胞以及泡沫細胞的分子影響。 2. 研究自組裝以及奈米結構的黑色素在生物活性具有的特性。 3. 研究奈米粒子的黑色素的物理和化學特性。 4. 研究奈米黑色素在生物科技的應用。
卵巢癌基因晶片資料前處理與分析(I)
(2004)
黃國華
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Huang, Guewha Steven
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國立交通大學奈米科技研究所
卵巢癌晶片之智慧型計算及分子演化與控制之研究子計畫一：卵巢癌基因晶片資料前處理及分析 Pre-processing and analysis of microarray datasets for ovarian cancers 中文摘要：卵巢癌是婦科癌症中存活率最低的癌症，主要是因為缺乏早期診斷的方法，發現時癌細胞大多轉移。縱然在對藥物反應良好的病人上，卵巢癌還容易復發。雖然已發現如c-erb-B2, c-myc, and p53 等基因在卵巢癌中有增強表現，然而皆無法用來診斷病變，或是用來追蹤及預測腫瘤的變化。微陣列（基因晶片）能大量平行偵測基因表現，可以同時觀察至少一萬個基因的活性，我們將以微陣列研究卵巢腫瘤，研究分析卵巢癌基因表現的差異，借此瞭解卵巢腫瘤轉化成為卵巢癌的機制，並尋找卵巢癌分子診斷標記。近年來，微陣列已漸漸普及化化，但是市面上的微陣列服務良莠不齊，無法提供高品質及一致的晶片，所獲得的資料無法使用，容易誤導，因而無法進行資料分析。我們已有三年執行國家型計畫基因晶片核心實驗室的寶貴經驗，本實驗室將致力於建立卵巢腫瘤基因晶片的標準化，提供高品質、高一致性的微陣列資料。第一年將建立包含三萬個基因的晶片標準化程序，第二年將分離卵巢腫瘤相關基因建立「卵巢癌晶片」，第三年將進行臨床篩檢，使計畫成果能於短期內應用。特定目標為：第一年： 1. 建立包含三萬個基因的晶片標準化程序 2. 提供二十套卵巢腫瘤微陣列資料第二年： 1. 提供二十套卵巢腫瘤微陣列資料 2. 分離卵巢腫瘤相關基因建立「卵巢癌晶片」第三年：1. 提供二十套卵巢腫瘤微陣列資料，含其他非卵巢的腫瘤 2. 進行臨床篩檢
智慧型系統在卵巢癌晶片之分子演化與控制---子計畫一---卵巢癌基因晶片資料前處理及分析(II)
(2005)
黃國華
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Huang, Guewha Steven
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國立交通大學奈米科技研究所