Polymeric microneedles fabricated by microinjection molding techniques have been demonstrated using Topas Ò COC as the molding plastic material. Open-channel microneedles with cross-sectional area of 100 lm · 100 lm were designed and fabricated on top of a shank of 4.7 mm in length, 0.6 mm in width, and 0.5 mm in depth. The tip of the microneedle has a round shape with a radius of 125 lm as limited by the drill used in fabricating the mold insert. The injection molding parameters including clamping force, shot size, injection velocity, packing pressure, and temperature were characterized in order to achieve best reproducibility. Experimentally, a fabricated microneedle was successfully injected into a chicken leg and a beef liver freshly bought from a local supermarket and about 0.04 lL of liquid was drawn from these tissues immediately. This new technology allows mass production of microneedles at a low cost for potential biomedical applications.

The whole genome sequences of Helicobacter pylori strain 26695 have been reported. Whole cell proteins of H. pylori strain 26695 cells were obtained and analyzed by two-dimensional electrophoresis, using immobilized pH gradient strips. The most abundant proteins were shown in the region of pI 4.0-9.5 with molecular masses from 10 to 100 kDa. Soluble proteins were precipitated by the use of 0-80% saturated solutions of ammonium sulfate. Soluble proteins precipitated by the 0-40% saturations of ammonium sulfate produced similar spot profiles and their abundant protein spots had acidic pI regions. However, a number of soluble proteins precipitated by more than 60% saturation of ammonium sulfate were placed in the alkaline pI regions, compared to those precipitated by 40% saturation. In addition, we have performed an extensive proteome analysis of the strain utilizing peptide MALDI-TOF-MS. Among the 345 protein spots processed, 175 proteins were identified. The identified spots represented 137 genes. One-hundred and fifteen proteins were newly identified in this study, including DNA polymerase III beta-subunit. These results might provide guidance for the enrichment of H. pylori proteins and contribute to construct a master protein map of H. pylori.

To examine the proteomes of 2 important causative agents of fish streptococcosis, Streptococcus iniae ATCC29178 and Lactococcus garvieae KG9408, we used 2-dimensional gel electrophoresis (2-DE) followed by mass spectrometry to generate 2-DE maps of these type strains. Silver-stained 2-DE gels of S. iniae ATCC29178 and L. garvieae KG9408 revealed ~320 and 300 spots, respectively, and immobilized pH gradient strips (13 cm, pH 4 to 7) revealed that the majority of the detected spots were concentrated in the pH range of 4.5 to 5.5. The spots were randomly selected from the 2-DE profiles and identified by peptide mass fingerprinting using matrix-assisted laser desorption/ionization time of flight mass spectrometry. The majority of the identified proteins were functionally related to energy and carbohydrate metabolism (e.g. enolase ATPase, glyceraldehyde-3-phosphate dehydrogenase) or translation and translocation (e.g. elongation factor G, elongation factor Tu, DNA-directed RNA polymerase alpha chain). These data, along with our partial 2-DE maps of S. iniae ATCC29178 and L. garvieae KG9408, may help suggest antigenic proteins for the development of effective diagnostic tools and vaccines against S. iniae and L. garvieae. KEY WORDS: Streptococcus iniae · Lactococcus garvieae · Two-dimensional gel electrophoresis · Mass spectrometry · Proteome Resale or republication not permitted without written consent of the publisherDis Aquat Org 70 :-71-79, 2006 amide gel electrophoresis (SDS-PAGE) are used to separate proteins according to their isoelectric point (pI) and molecular weight (MW), respectively (O'Farrell 1975). In spite of available promising technologies for studying proteins, such as multidimensional protein identification, stable isotope labeling, and protein or antibody arrays, the 2-DE is the only technique that can be routinely applied for quantitative expression profiling of a complex mixture of proteins according to pI, MW, solubility, and relative abundance (Görg et al. 2004). The proteins separated by 2-DE are then identified by immunoblotting with specific antibodies (Winterhoff et al. 2002), Edman sequencing with N-terminal sequencing (Anglade et al. 2000, Lei et al. 2000, or peptide mass fingerprinting (PMF) using matrixassisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) (Cash et al. 1999, Thongboonkerd et al. 2002, Lee et al. 2003, Len et al. 2003, Lin et al. 2003, Mortz et al. 2003). The identified proteins may then be assembled into a 2-DE map or reference map of the sampled cell, tissue, or organism (Cash 2000, Cordwell et al. 2001, Champomier-Verges et al. 2002, Hughes et al. 2002, Lee et al. 2003, Len et al. 2003. Such maps have been widely used to compare differences in protein expression among strains of the same species or among the same strains under different experimental conditions. In this way, researchers have been able to study the mechanisms involved in pathogenicity, antibiotic resistance, environmental adaptation and responses to host i...

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BackgroundAllium sativum., commonly known as garlic, is a species in the onion genus (Allium), which is a large and diverse one containing over 1,250 species. Its close relatives include chives, onion, leek and shallot. Garlic has been used throughout recorded history for culinary, medicinal use and health benefits. Currently, the interest in garlic is highly increasing due to nutritional and pharmaceutical value including high blood pressure and cholesterol, atherosclerosis and cancer. For all that, there are no comprehensive databases available for Expressed Sequence Tags(EST) of garlic for gene discovery and future efforts of genome annotation. That is why we developed a new garlic database and applications to enable comprehensive analysis of garlic gene expression.DescriptionGarlicESTdb is an integrated database and mining tool for large-scale garlic (Allium sativum) EST sequencing. A total of 21,595 ESTs collected from an in-house cDNA library were used to construct the database. The analysis pipeline is an automated system written in JAVA and consists of the following components: automatic preprocessing of EST reads, assembly of raw sequences, annotation of the assembled sequences, storage of the analyzed information into MySQL databases, and graphic display of all processed data. A web application was implemented with the latest J2EE (Java 2 Platform Enterprise Edition) software technology (JSP/EJB/JavaServlet) for browsing and querying the database, for creation of dynamic web pages on the client side, and for mapping annotated enzymes to KEGG pathways, the AJAX framework was also used partially. The online resources, such as putative annotation, single nucleotide polymorphisms (SNP) and tandem repeat data sets, can be searched by text, explored on the website, searched using BLAST, and downloaded. To archive more significant BLAST results, a curation system was introduced with which biologists can easily edit best-hit annotation information for others to view. The GarlicESTdb web application is freely available at .ConclusionGarlicESTdb is the first incorporated online information database of EST sequences isolated from garlic that can be freely accessed and downloaded. It has many useful features for interactive mining of EST contigs and datasets from each library, including curation of annotated information, expression profiling, information retrieval, and summary of statistics of functional annotation. Consequently, the development of GarlicESTdb will provide a crucial contribution to biologists for data-mining and more efficient experimental studies.