Meng-Yen Hong

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.

A protein transistor made of an antibody molecule and two gold nanoparticles

A major challenge in molecular electronics is to attach electrodes to single molecules in a reproducible manner to make molecular junctions that can be operated as transistors. Several attempts have been made to attach electrodes to proteins, but these devices have been unstable. Here, we show that self-assembly can be used to fabricate, in a highly reproducible manner, molecular junctions in which an antibody molecule (immunoglobulin G) binds to two gold nanoparticles, which in turn are connected to source and drain electrodes. We also demonstrate effective gating of the devices with an applied voltage, and show that the charge transport characteristics of these protein transistors are caused by conformational changes in the antibody. Moreover, by attaching CdSe quantum dots to the antibody, we show that the protein transistor can also be gated by an applied optical field. This approach offers a versatile platform for investigations of single-molecule-based biological functions and might also lead to the large-scale manufacture of integrated bioelectronic circuits.

Differential gene expression of livers from ApoE deficient mice

A genomic survey for differentially expressed genes was performed to livers of ApoE deficient mice using human cDNA microarray containing approximately 9,000 human cDNA clones. Due to the homology between mouse and human, hybridization was performed at lower stringency condition, 10 degrees below the regular hybridizing temperature. Gene expression profiles of livers corresponding to high levels of blood cholesterol were generated at genomic scale. Thirty-seven genes were randomly selected from a pool of differentially expressed genes and subjected to semi-quantitative RT-PCR, further confirmed the result from microarray hybridization. These included genes associated with atherosclerosis, and novel genes that implied novel pathways correlated to high levels of blood cholesterol. It is promising using human cDNA microarray, the most complete collection among all species, to study other mammalian systems with satisfying speed and accuracy.

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

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.

Self-assembly of an antireflective moth-like structure using antibody-functionalized nanowires and nanopore arrays

A novel framework to fabricate moth-like nanopillar arrays was proposed. In this scheme, nanowires were first cross-linked with anti-gold nanoparticle (GNP) antibodies and mixed with the nanopore array pre-deposited by GNP, which was then followed by centrifugation. An optimal success rate of 95% was finally obtained by choosing nanorods with an aspect ratio of 5:1 by modifying with 10 ng mL⁻¹ antibodies, and by inserting them into a pore array pre-deposited with 54.4 µM GNP. The nanopillar arrays thus fabricated showed high levels of antireflective efficiency across a broad wavelength. Here we demonstrate the assembly of nanowires and nanopores into nanopillar arrays by the assistance of antibody-antigen binding. The application of bio-nano-interaction provides an economic, time-saving, and throughput approach to manipulating objects on the nanoscale.

Incorporation of DNA chip technology to the simulation and validation of flux analysis in yeast diauxic growth.

We incorporated gene expression information from cDNA microarray into flux analysis to simulate yeast diauxic growth. Expression ratios of both growth phases were applied to assign the split ratio at glyoxylate shunt during simulation, in which the equation was mathematically unsolvable due to the singularity and artificial split ratios, which were traditionally introduced without biological evidence. In addition, the directionality of microarray dataset was used as a further constraint during simulation. Metabolic fluxes obtained by this modified approach are in general consistent with microarray analysis. However, discrepancies occurred when the quantity of fluxes was compared, probably due to the substantial reduction of substrates at phase II in which the increase in the enzymatic levels was not proportional to the increase of substrate flow, as would be predicted from microarray dataset. The modified flux analysis might have brought a new approach to investigate other cellular pathways.