Synthesis and properties of polymeric latex particles and their conjugates with human immunoglobulin G

Polymeric latex particles of uniform size in the range of 750 to 10 000 Å in diameter were prepared by emulsion copolymerization of methyl methacrylate, methacrylic acid, 2-hydroxyethyl methacrylate and ethylene glycol dimethacrylate, using sodium dodecylbenzene sulphonate as an emulsifier and ammonium persulphate as an initiator. With a fixed composition of monomers the carboxyl groups on the latex particles ranged from 5 × 10³ to 180 × 10³ per particle, but the average number per unit surface area remained within a limited variation. Human immunoglobulin G (IgG) was covalently bonded to the latex particles by either the carbodiimide or the cyanogen bromide method, taking advantage of carboxyl and hydroxyl groups, respectively. The resulting conjugates exhibited a specific agglutination with rabbit anti-human IgG and sera from rheumatoid arthritis (RA) patients. The immunolatex particles were found to be superior to those derived from nylon-6, which had been used in our previous work, and are expected to offer useful means in various immunological studies.     

Product design logic for an intelligent product modelling system

Recent research in CAD systems has been conducted to realize intelligent processing. Several CAD systems and product modelling systems have been developed using AI techniques. However, in order to develop more intelligent CAD systems, the design logic which connects the functional requirement to the geometric and the technological information of the designed product must be evaluated.

A product model used in such intelligent CAD systems has to include not only the geometric and the technological information of the product but also the designer's thought process which explains the design logic.

Design logic is generally divided into two parts. One is the generalized design logic which is commonly used in the conceptual design of mechanical products. The other is the product specified design logic which is used in the fundamental and detailed design phase. Different logic is applied to each product. This type of design logic is often used in modification design and compilation design, where the dimensions of parts have to be modified according to different functional requirements. When the dimensions and accuracies of the products are defined in connection with the functional requirements through design logic, the CAD system can automatically make decisions according to the given requirements. In this paper, suitable presentation formats and processing functions for these two types of design logic are discussed.

The importance of design logic in product modelling is proven through several case studies in this paper. As a conclusion, the intelligent product modelling system is developed, which should expedite the progress of design automation in the near future. In conceptual design, the design logic is processed in the modelling system and the product structure, with the technological information decided automatically from the functional requirement. Automation in the detailed design phase is also facilitated by the modelling system using the product specified design logic in the product model.     

Production of d-glucose from pseudo paper sludge with hydrothermal treatment

To convert cellulosic organics contained in industrial paper sludge into glucose, reaction of pseudo paper sludge composed of cellulose and inorganic compounds (calcium carbonate (CaCO₃), talc (Mg₃(Si₄O₁₀)(OH)₂), kaolin (Al₂(Si₂O₅)(OH)₄)) under hydrothermal conditions was studied. Significant amounts of glucose (ca. 23%) could be produced from cellulose in the absence of CaCO₃ for reaction in water at 523 K and 12 min reaction time. On the other hand, in the presence of CaCO3, most glucose decomposed over all conditions, whereas the addition of talc and kaolin to the mixtures increased the glucose yield to about 30%. For the case of chemical recycle of paper sludge with hydrothermal treatment to obtain d-glucose, it can be concluded that it is preferable to separate the calcium carbonate from the paper sludge before hydrothermal treatment.     

Analysis of a change in bacterial community in different environments with addition of chitin or chitosan

The temporal changes of a bacterial community in soil with chitin or chitosan added were analyzed by PCR-denaturing gradient gel electrophoresis (DGGE) targeting the 16S rRNA gene using total DNAs prepared from the community. Band patterns of PCR-DGGE confirmed that 31 species become predominant after the addition of chitin or chitosan. The determination of the nucleotide sequences of the bands of the 31 species indicated that 20 species belonged to the division Proteobacteria, and that the genus Cellvibrio was apparently predominant among them (7/20). The 16S rRNA sequences of the 16 deduced species (16/31) showed less than 98% similarities to those of previously identified bacteria, indicating that the species were derived from unidentified bacteria. The total community DNAs extracted from bacterial cells adsorbed on the surface of flakes of chitin and chitosan placed in a river, a moat, or soil were subjected to PCR-DGGE to examine the extent of diversity of chitinolytic bacteria among different environments. The predominant species significantly differed between the chitin and chitosan placed in the river and moat, but not so much between those placed in the soil. The large difference between the diversities of the three bacterial communities indicated that a wide variety of bacteria including unidentified ones are involved in the degradation of chitin and chitosan in the above-mentioned natural environments.     

Application of microalgae hydrolysate as a fermentation medium for microbial production of 2-pyrone 4,6-dicarboxylic acid

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Molecular properties of the two-component cell lysis system encoded by prophage phigaY of Lactobacillus gasseri JCM 1131T: cloning, sequencing, and expression in Escherichia coli

Shotgun cloning of the Lactobacillus gasseri JCM 1131T whole DNA yielded two recombinant plasmids, p118gaY1 and p118gaY2, which directed cell lysis activity. Sequencing analysis revealed that the two plasmids carried almost identical inserted genes in following orders (truncated genes, in parentheses): in p118gaY1, (orf149)-orf92-holgaY-lysgaY-orf35-attL-(mnaAgaY1); in p118gaY2, (orfXgaY1)-orf169-orf149-orf92-holgaY-lysgaY-orf35-attP-(intgaY). The lysgaY-encoded protein (designated as LysgaY, 33.7 kDa) showed significant homology with putative muramidases (peptidoglycan-degrading enzyme) of the Lactobacillus phage phiadh, Lj965, Lj928, LL-H, mv4, and mv1. By zymogram analysis, LysgaY overproduced in Escherichia coli exhibited lytic activity towards 17 Gram-positive bacterial strains, including lactobacilli, lactococci, and staphylococci. The holgaY-encoded protein (15.7 kDa) contained three potential transmembrane helices, resembling putative holins (cytoplasmic membrane-disrupting protein) of Lj928 and Lj965. On the other hand, another clone p118gaYR obtained by EcoRI-shotgun cloning carried the (ptsCgaY1)-attR-(intgaY) genes. Three sequences, attL, attP, and attR, had a 47-bp common (core) sequence, and the core of attR was located in 3' region of a potential tRNA(Arg) gene. These results suggested that (i) attL and attR are phage-host junctions, left- and right-arms, respectively, (ii) attP is a phage attachment site, and (iii) intgaY is an integrase gene for phage integration and/or excision. After mitomycin C-induction, phage particles were demonstrated by electron microscopy. The prophage (phigaY) is somewhat leaky in the host, and has the two-component lysis system (HolgaY-LysgaY), closely resembling that of Lj928 as well as Lj965.     

Evaluation of drug toxicity with hepatocytes cultured in a micro-space cell culture system

A micro-space cell culture system was recently developed in which cells such as hepatocytes can be cultured and formed into a multicellular three-dimensional (3D) architecture. In this study, we assessed the performance of HepG2 cells cultured in this micro-space cell culture system in a drug toxicity test, and evaluated the effects of micro-space culture on their hepatocyte-specific functions. The micro-space cell culture facilitated the formation of 3D HepG2 cell architecture. HepG2 cells cultured in a micro-space culture plate exhibited increased albumin secretion and enhanced mRNA expression levels of cytochrome P450 (CYP) enzyme compared to those cultured in a monolayer culture. When the cells were exposed to acetaminophen, a hepatotoxic drug, the damage to the HepG2 cells grown in micro-space culture was greater than the damage to the HepG2 cells grown in monolayer culture. In addition, human primary hepatocytes grown in micro-space culture also exhibited increased albumin secretion, enhanced CYP mRNA expression levels and increased sensitivity to acetaminophen compared to those grown in monolayer culture. These results suggest that this micro-space culture method enhances the hepatocyte-specific functions of hepatocytes, including drug-metabolizing enzyme activities, making hepatocytes grown in the micro-space culture system a useful tool for evaluating drug toxicity in vitro.     

Expression of aldehyde dehydrogenase gene increases hydrogen production from low concentration of acetate by Rhodobacter sphaeroides

Rhodobacter sphaeroides RV (RV) is a hydrogen-producing bacterium exhibiting the highest yield of hydrogen production from organic acids such as lactate and acetate, which are the byproducts of hydrogen fermentation by hydrogen-producing anaerobic bacteria. Co-fermentation of the RV strain with anaerobic bacteria is an efficient method of hydrogen production. However, less than 21 mM acetate is produced by the anaerobic bacteria, which is too low for efficient hydrogen production by the RV strain; it requires approximately 75 mM acetate. In this study, 2 distinct isozymes of aldehyde dehydrogenase from Rhodospirillum rubrum were separately overexpressed in the RV strain. The recombinant RV strains that were designated as RVAD1 and RVAD2, exhibited 13-fold higher ALDH activities than the wild-type RV strain. Hydrogen yields of both of the recombinant strains were 1.4-fold higher than that of the RV strain in 21 mM acetate. In 43 mM acetate, the RVAD1 strain showed higher yield, though the RVAD2 strain showed lower yield as compared to that of the RV strain. In 64 mM acetate and all concentrations of lactate tested (21, 43 and 64 mM), the yields of the recombinant strains were lower than those of the RV strain. The intact (empty) expression plasmid increased the ALDH activity and had little effect on the hydrogen production in acetate, however, it decreased the production in lactate. At the beginning of the fermentation process, when very little hydrogen had been produced, the recombinant strains expressing the ALDH gene consumed smaller amounts of acetate compared to the wild-type strain. We have discussed the effects of ALDH on hydrogen production in this report.     

Sensitivity of charged particle activation analysis for long-lived radioactive nuclide determination

ABSTRACT

Charged Particle Activation Analysis (CPAA) utilizing an 8-MeV proton beam has been studied for determination of 35 long-lived radioactive nuclides. We accumulated the reaction cross section and nuclear decay data by referring to nuclear database supplied by National Nuclear Data Center in Brookhaven National Laboratory. We also calculated the reaction cross sections by using statistical model code ALICE. By using the nuclear data, we have derived determination sensitivity of the radioactive nuclides relative to unit weight and specific radioactivity. The result indicates that several hardly measurable nuclides with long half-lives such as ¹³⁵Cs, ²⁴⁴Pu, ¹²⁹I, ¹²⁶Sn, ⁹³Mo, ¹⁰⁷Pd, ²³⁶U, ²⁴⁸Cm, and ²³⁷Np have high sensitivity. It may be concluded that CPAA can be applied to determination of several long-lived nuclei and will provide a quick and non-destructive analysis method.