PCR amplification from single DNA molecules on magnetic beads in emulsion: application for high-throughput screening of transcription factor targets

We have developed a novel method of genetic library construction on magnetic microbeads based on solid-phase single-molecule PCR in a fine and robust water-phase compartment formed in water-in-oil (w/o) emulsions. In this method, critically diluted DNA fragments were distributed over the emulsion as templates, where beads crosslinked with multiple primers and other PCR components were encapsulated to form multiple reaction compartments. The delivered DNA was then amplified and covalently immobilized on the beads in parallel, within individual compartments, to construct a genetic library on beads (GLOBE), which was readily applicable to a genomewide global scanning of genetic elements recognized by a defined DNA-binding protein. We constructed a GLOBE of Paracoccus denitrificans and selected gene beads that were bound to the His-tagged transcription factor PhaR by flow cytometry. As a result of flow cytometry screening with an anti-His fluorescent antibody, the PhaR target fragments were enriched 1200-fold from this library with this system. Therefore, this system is a powerful tool for analyzing the transcription network on a genomewide scale

Sequence-specific cleavage of RNA by a hybrid ribonuclease H

AbstractSite-specific cleavage of the 22-, 132- and 534-base RNAs by the DNA/protein hybrid R Nase H were examined. The 22-base RNA was chemically synthesized, and 132- and 534-base RNAs were prepared by run-off transcription. The hybrid enzyme cleaves these RNAs, which contain a single target sequence, primarily at the unique phosphodiester bond within the target sequence. The hybrid enzyme performs multiple turnovers, and at a substrate/enzyme ratio of 10:1 the RNAs are almost completely cleaved by the hybrid enzyme at 37°C within 1 h. We propose that hybrid RNase H molecules with various oligodeoxyribonucleotides function as RNA restriction enzymes and are useful for structural and functional studies of RNA

Cystic Lymphangioma of the Pancreas with Spontaneous Rupture: Report of a Case

Lymphangioma is a benign and congenital malformation of the lymphatic system. Most lymphangiomas are preferentially located in the head and neck region. The abdominal organs are uncommon sites of origin. Several cases of lymphangioma in abdominal organs were reported, however, the pancreas is one of the rarest origins. Generally, intra-abdominal lymphangioma is asymptomatic and found incidentally, but in some cases, the patient complains of abdominal distension or a palpable mass. We describe the case of a 38-year-old male who presented with sudden-onset upper abdominal pain. Rupture of a cystic tumor of the pancreatic head was suspected, based on the findings of computed tomography, magnetic resonance imaging and endoscopic ultrasonography. Subtotal stomach-preserving pancreaticoduodenectomy was undertaken. The tumor, which was 4 × 4.5 × 8 cm in size, was pathologically diagnosed as a cystic lymphangioma. In conclusion, pancreatic lymphangioma is mostly asymptomatic, a ruptured case causing ‘acute abdomen’ has never been reported. Since lymphangioma is benign, it could be observed with accurate diagnosis. The surgical indication would be limited to cases of symptomatic lymphangiomas

Diel patterns of zooplankton community structure in nearshore waters of different substrates off Tinggi and Sibu Islands, Malaysia, with special reference to copepods

The relationship between zooplankton community structure and selected environmental conditions was investigated by simultaneous two-day consecutive sampling in the waters overlying coral and sand sites off Sibu Island and a seagrass site off Tinggi Island, Johor, Malaysia. A total of 129 taxa were identified, 60 non-copepod taxa and 69 copepod species in all samples. Uni- and multi-variate analyses reveal distinct coral, sand, seagrass copepod assemblages with indicator copepod species and attributes of their size fractions. Small fraction (100–335 μm) samples contained greatest number of individuals, few rare species and were densest at onshore depths, and the opposite for large fraction (>335 μm) samples but were densest at nighttime and most species rich in the coral site at night. Higher species diversity at offshore stations is due likely to ecotonal effects of overlapping oceanic and nearshore communities. This study demonstrates the usefulness of uni- and multi-variate analyses in identifying patterns in zooplankton community structure in representative shallow tropical habitats, and the need for accurate zooplankton taxonomy, nighttime and daytime and onshore and offshore sampling, and size fractionation of samples

Density and surface tension measurements of molten Al–Si based alloys

This study is part of a series of studies aimed at measuring the thermophysical properties of molten phase change material-type metallic thermal energy storage materials near 873 K (600°C). The target material is Al–Si based alloys. First, as a feasibility study, density measurements of the molten state of three Al–Si binary alloys (Al–12.2Si, Al–50Si and Al–90Si in atomic%) were performed. A highly accurate non-contact density measurement method based on the static magnetic field superposition electromagnetic levitation (EML) method was employed as the density measurement method. The validity of this experimental method was confirmed, and density of molten Al–Si base alloys (ADC12 and Al–5.9mass%Si–1.6mass%Fe) were measured as a function of temperature with an expanded uncertainty of 1.2%. In addition, the surface tension of the alloys was measured by the droplet oscillation method using the EML technique. The surface tension was successfully obtained as a function of temperature with expanded uncertainty of 2.3%