Guewha Steven Huang

Assessment of gold nanoparticles as a size-dependent vaccine carrier for enhancing the antibody response against synthetic foot-and-mouth disease virus peptide

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.

Crack-free GaN∕AlN distributed Bragg reflectors incorporated with GaN∕AlN superlattices grown by metalorganic chemical vapor deposition

A crack-free GaN∕AlN distributed Bragg reflector (DBR) incorporated with GaN∕AlN superlattice (SL) layers was grown on a c-plane sapphire substrate by metalorganic chemical vapor deposition. Three sets of half-wave layers consisting of 5.5 periods of GaN∕AlN SL layers and GaN layer were inserted in every five pairs of the 20 pair GaN∕AlN DBR structure to suppress the crack generation. The grown GaN∕AlN DBRs with SL insertion layers showed no observable cracks in the structure and achieved high peak reflectivity of 97% at 399nm with a stop band width of 14nm. Based on the x-ray analysis, the reduction in the in-plane tensile stress in the DBR structure with insertion of SL layers could be responsible for the suppression of crack formation and achievement of high reflectivity.

A novel melanin-like pigment derived from black tea leaves with immuno-stimulating activity

Abstract Melanin-like pigment (MLP) was isolated from black tea leaves by alkaline extraction, acid hydrolysis, and repeated precipitation, and a yield of 2% (dry weight basis) was obtained. The isolated MLP possessed physical and chemical properties comparable to melanins extracted from other sources. The immuno-modulating activity of MLP was measured using plaque-forming cells assay. Oral administration of MLP significantly stimulated the spleen lymphoid tissue in mice at doses ranging from 50 to 200 mg/kg body weight, with a maximal antibody response at 75 mg/kg. The immuno-stimulating activity of tea MLP may serve as a platform for drug discovery, and the abundance of MLP from tea suggests an economical source for health food application.

Isolation and characterization of melanic pigments derived from tea and tea polyphenols

The dark brown pigments derived from tea and tea polyphenols were studied. Physical and chemical properties revealed that pigments directly extracted from tea leaves and derived from tea polyphenols were similar to typical melanins. Further investigation showed that both melanic pigments possessed similar antioxidant capability, due to their chelating and scavenging properties. The inhibitory effect of melanic pigments, either from tea or tea polyphenols, was significantly stronger than that of non-treated tea polyphenols. According to these properties, we have extracted melanin from tea. In addition, oxidation of tea polyphenols also provided an alternative method to maximize the yields. The extracted melanin is an antioxidant, which interrupted free radical reactions at a step in the development chain by its scavenging properties and, at the step of initiation, by its ability to chelate metals.

Aluminum incorporation into AlGaN grown by low-pressure metal organic vapor phase epitaxy

Aluminum (Al) incorporation in AlxGa1−xN films grown by low-pressure metal organic vapor phase epitaxy using trimethylaluminum (TMAl) and trimethylgallium as group III precursors has been systematically studied. The solid phase Al composition of the AlxGa1−xN films varied nonlinearly with the Al gas phase composition. The incorporation kinetics of AlxGa1−xN alloy has been analyzed by using an adsorption-trapping model. Two parameters were used to characterize the properties of Al incorporation, i.e., the capture radius and the adsorption time of Al atoms. An exponential function of the Al composition of the AlxGa1−xN films versus the TMAl gas flow rate was obtained. It was demonstrated that the adsorption time of the Al atom was larger than the growth time of one atomic layer. The effects of ammonia flow rate, crystal growth rate, and growth temperature on the adsorption parameters were also discussed.

Microarray Analysis of Ovarian Cancer

Microarray experiments were performed to normal ovaries, benign ovarian tumor and stage I and III of ovarian cancer. By treating benign ovarian tumor as pre-cancerous stage and comparing to later stages of ovarian cancer we identified 46 genes overly expressed in early-staged cancer. We also identified over-expressed genes at all stages of ovarian tumor/cancer. The over-expressed genes might be alternative candidates for developing early diagnostic markers. JUN and nuclear receptor subfamily genes were repeatedly identified in the group of down-regulated genes indicating their important regulatory roles in the oncogenesis of ovary

Effects of static magnetic fields on the development and aging of Caenorhabditis elegans.

SUMMARY 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.

Spatial Control of Cell-Nanosurface Interactions by Tantalum Oxide Nanodots for Improved Implant Geometry.

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.

Detection of foot and mouth disease and porcine reproductive and respiratory syndrome viral genes using microarray chip.

Two viral pathogens, namely, porcine reproductive and respiratory syndrome virus (PRRSV) and foot and mouth disease virus (FMDV), were selected as models for multiple pathogen detection in a cDNA microarray. Two signature regions selected from ORF2 (around 500 bp) and ORF5 (around 600 bp) of PRRVS (America serotype), and one signature region from structural genes VP1 (around 500 bp) of FMDV type O were designed and spotted on a nylon membrane. For PCR sensitivity study, the cloned FMDV–VP1 template could be diluted to near one copy and its PCR product was still detectable in gel electrophoresis. In the microarray detection, the labelling FMDV probes (3 mg/ml) could be diluted 320 times and still maintained a visible colour when hybridized with the chip. Using the mixing primers, the microarray chip demonstrated rapid and accurate detection of the specific genes. To our knowledge, this preliminary study is the first example reported applying the long signature sequences to the multiple pathogen detection in cDNA microarray.

Nanochips of Tantalum Oxide Nanodots as artificial-microenvironments for monitoring Ovarian cancer progressiveness.

Nanotopography modulates cell characteristics and cell behavior. Nanotopological cues can be exploited to investigate the in-vivo modulation of cell characteristics by the cellular microenvironment. However, the studies explaining the modulation of tumor cell characteristics and identifying the transition step in cancer progressiveness are scarce. Here, we engineered nanochips comprising of Tantalum oxide nanodot arrays of 10, 50, 100 and 200 nm as artificial microenvironments to study the modulation of cancer cell behavior. Clinical samples of different types of Ovarian cancer at different stages were obtained, primary cultures were established and then seeded on different nanochips. Immunofluorescence (IF) was performed to compare the morphologies and cell characteristics. Indices corresponding to cell characteristics were defined. A statistical comparison of the cell characteristics in response to the nanochips was performed. The cells displayed differential growth parameters. Morphology, Viability, focal adhesions, microfilament bundles and cell area were modulated by the nanochips which can be used as a measure to study the cancer progressiveness. The ease of fabrication of nanochips ensures mass-production. The ability of the nanochips to act as artificial microenvironments and modulate cell behavior may lead to further prospects in the markerless monitoring of the progressiveness and ultimately, improving the prognosis of Ovarian cancer.