Phase Behavior of a Carbon Dioxide/Methyl Trimethoxy Silane/Polystyrene Ternary System.

Recently, polymeric foams filled with a silica aerogel have been developed. The phase behavior of CO2/silicon alkoxide binary systems and CO2/silicon alkoxide/polymer ternary systems is an important factor that affects the design of novel processes. The phase behavior of a carbon dioxide (CO2)/methyl trimethoxy silane (MTMS)/polystyrene (PS) ternary system was measured using a synthetic method involving the observation of the bubble and cloud point. The phase boundaries were measured at temperatures ranging from 313.2 to 393.2 K and CO2 weight fractions between 0.01 and 0.08. The CO2/MTMS/PS system showed a similar CO2 mass fraction dependence of the phase behavior to that observed for the CO2/tetramethyl orthosilicate (TMOS)/PS system. When the phase boundaries of these systems were compared, the vapor-liquid (VL) and vapor-liquid-liquid (VLL) lines were found to be nearly identical, while the liquid-liquid (LL) lines were different. These results indicate that the affinity between the silicon alkoxide and polymer greatly influences the liquid-liquid phase separation.

Two Family B DNA Polymerases From Aeropyrum pernix, Based on Revised Translational Frames.

Living organisms are divided into three domains, Bacteria, Eukarya, and Archaea. Comparative studies in the three domains have provided useful information to understand the evolution of the DNA replication machinery. DNA polymerase is the central enzyme of DNA replication. The presence of multiple family B DNA polymerases is unique in Crenarchaeota, as compared with other archaeal phyla, which have a single enzyme each for family B (PolB) and family D (PolD). We analyzed PolB1 and PolB3 in the hyperthermophilic crenarchaeon, Aeropyrum pernix, and found that they are larger proteins than those predicted from the coding regions in our previous study and from public database annotations. The recombinant larger PolBs exhibited the same DNA polymerase activities as previously reported. However, the larger PolB3 showed remarkably higher thermostability, which made this enzyme applicable to PCR. In addition, the high tolerance to salt and heparin suggests that PolB3 will be useful for amplification from the samples with contaminants, and therefore it has a great potential for diagnostic use in the medical and environmental field.

A longer finger-subdomain of family A DNA polymerases found by metagenomic analysis strengthens DNA binding and primer extension abilities.

The family A DNA polymerases from thermophilic bacteria are useful for PCR. The DNA polymerase from Thermus aquaticus (Taq polymerase) was the original enzyme used when practical PCR was developed, and it has remained the standard enzyme for PCR to date. Knowledge gained from structure-function relationship studies of Taq polymerase is applicable to create PCR enzymes with enhanced performance. We collected the deduced amino acid sequences of the regions from motif A to motif C in the family A DNA polymerases from metagenomic sequence data, obtained by sequencing DNAs from microorganisms isolated from various hot spring areas in Japan. The corresponding regions of the polA gene for Taq polymerase were substituted with the metagenomic DNA gene fragments, and various chimeric DNA polymerases were prepared. Based on the properties of these chimeric enzymes and their sequences, we found an insertion sequence that affects the primer extension ability of the family A DNA polymerases. The insertion sequence is located in the finger subdomain, and it may enhance the affinity of the enzyme to DNA. Mutant Taq polymerases with the corresponding 9 amino acid insertion displayed enhanced PCR performance.

A useful strategy to construct DNA polymerases with different properties by using genetic resources from environmental DNA.

DNA polymerases synthesize new DNA strands according to the template DNA, using deoxynucleotide triphosphates during DNA replication and repair, and are essential to maintain genome integrity in DNA metabolism. In addition, these enzymes are widely used for genetic engineering techniques, including dideoxy-sequencing, PCR, DNA labeling, mutagenesis, and other in vitro experiments. Thermostable DNA polymerases are especially useful for PCR and cycle-sequencing. We propose a powerful strategy using environmental DNA as a genetic resource to investigate the structure-function relationships of the family B DNA polymerases. The region corresponding to the active center of the DNA polymerizing reaction in the structural gene of P. furiosus DNA polymerase I (PolBI) was substituted by PCR fragments amplified from DNAs within soil samples from various locations in Japan. The chimeric pol genes were constructed within the PolBI expression plasmid. The chimeric enzymes thus produced revealed DNA polymerase activities with different properties.

Technical evaluation of dialysate flow in a newly designed dialyzer.

Rexeed was developed by Asahi Kasei Medical using wave-shaped hollow fibers, a full baffle, and a short taper housing to improve dialysate flow. The present study is clarifies improvement in dialysate flow with Rexeed-15 compared with that of a conventional dialyzer. Dialysate flow was evaluated by the pulse-response method. Dialysate pressure and tracer concentration were measured at a blood-side flow rate (QBeta) of 200 ml/min, a dialysate-side flow rate (QD) of 500 ml/min, and a net filtration rate (QF) of 0 ml/min using needles placed in the test dialyzer. Dialyzer performance was evaluated by measuring urea and vitamin B12 clearance at QB = 200 and 400 ml/min, QD = 300-800 ml/min, and QF = 0 ml/min. In the conventional dialyzer, dialysate channeling was observed. In contrast, Rexeed-15 had a uniform dialysate flow. Urea and vitamin B12 clearance with Rexeed-15 was slightly sensitive to QD. The overall mass transfer coefficient for urea with Rexeed-15 was more than 50% higher than that of the conventional dialyzer, indicating the possibility of reduced dialysate usage with Rexeed. Rexeed has a highly optimal dialysate flow, due to the wave-shaped hollow fibers and the new housing, and gives increased clearance for lower-molecular-weight substances.