Purification and characterization of a kanamycin nucleotidyltransferase from plasmid pUB110-carrying cells of Bacillus subtilis.

The nucleotidyltransferase encoded by plasmid pUB110 was purified to greater than 95% purity with a 33% yield. The enzyme is a monomeric protein with a molecular weight of 34,000. The optimum pH for activity is 5, and the optimum MgCl2 concentration for activity is 18 mM. The enzyme, which is synthesized constitutively, is stable for several weeks at 4 degrees C. This enzyme would appear to be a good model gene product for the development of a pUB110 deoxyribonucleic acid-dependent in vitro protein-synthesizing system from Bacillus subtilis.     

Repair deficiency, mutator activity, and thermal prophage inducibility in dna-8132 strains of Bacillus subtilis.

A ts mutation, dna-8132 (Hara and Yoshikawa, 1973), in the region of chromosome replication origin of Bacillus subtilis was found to cause pleiotropic effects at a permissive temperature (30 C). Strains carrying this mutation were lethan at 48 C but exhibited higher spontaneous mutation frequency and a lower capacity for repairing radiation damages at 30C. Introduction of the polA59 (Gass et al., 1971) mutation further enhanced the repair deficiency and the mutator activity. These results suggest that the dna-8132 gene product may be directly involved in chromosome replication and repair. SPO2 lysogens carrying this mutation produced mature phages upon a temperature shift from 30 to 48 C. Phage production at nonpermissive temperature suggests that there are few defects in the precursors of deoxyribonucleic acid synthesis in the mutant.     

A Particle Model for Prediction of Cement Infiltration of Cancellous Bone in Osteoporotic Bone Augmentation

Femoroplasty is a potential preventive treatment for osteoporotic hip fractures. It involves augmenting mechanical properties of the femur by injecting Polymethylmethacrylate (PMMA) bone cement. To reduce the risks involved and maximize the outcome, however, the procedure needs to be carefully planned and executed. An important part of the planning system is predicting infiltration of cement into the porous medium of cancellous bone. We used the method of Smoothed Particle Hydrodynamics (SPH) to model the flow of PMMA inside porous media. We modified the standard formulation of SPH to incorporate the extreme viscosities associated with bone cement. Darcy creeping flow of fluids through isotropic porous media was simulated and the results were compared with those reported in the literature. Further validation involved injecting PMMA cement inside porous foam blocks — osteoporotic cancellous bone surrogates — and simulating the injections using our proposed SPH model. Millimeter accuracy was obtained in comparing the simulated and actual cement shapes. Also, strong correlations were found between the simulated and the experimental data of spreading distance (R² = 0.86) and normalized pressure (R² = 0.90). Results suggest that the proposed model is suitable for use in an osteoporotic femoral augmentation planning framework.     

Bio-functional pickles that reduce blood pressure of rats

Addition of γ-aminobutyric acid (GABA) and angiotensin converting enzyme (ACE)-inhibitory peptides to the pickles was studied in order to develop a new type of pickles that reduce blood pressure. Based on the outcome of these studies, a new type of fermentation bed composed of rice bran and white miso has been successfully developed. The advantage of such pickles is that they not only contain both GABA and ACE-inhibitory peptides, but also that their taste and flavor are excellent, with colors close to the original ones. The new type of pickles could temporarily reduce blood pressure in two types of rats, spontaneously hypertensive rats and NaCl-sensitive model rats. Thus, the newly developed pickles appear to be beneficial for pickle business.     

A low trophic position of Japanese eel larvae indicates feeding on marine snow

What eel larvae feed on in the surface layer of the ocean has remained mysterious. Gut contents and bulk nitrogen stable isotope studies suggested that these unusual larvae, called leptocephali, feed at a low level in the oceanic food web, whereas other types of evidence have suggested that small zooplankton are eaten. In this study, we determined the nitrogen isotopic composition of amino acids of both natural larvae and laboratory-reared larvae of the Japanese eel to estimate the trophic position (TP) of leptocephali. We observed a mean TP of 2.4 for natural leptocephali, which is consistent with feeding on particulate organic matter (POM) such as marine snow and discarded appendicularian houses containing bacteria, protozoans and other biological materials. The nitrogen isotope enrichment values of the reared larvae confirm that the primary food source of natural larvae is consistent only with POM. This shows that leptocephali feed on readily available particulate material originating from various sources closely linked to ocean primary production and that leptocephali are a previously unrecognized part of oceanic POM cycling.     

Effect of Water Content on the Glass Transition Temperature of Calcium Maltobionate and its Application to the Characterization of Non-Arrhenius Viscosity Behavior

To understand the fundamental physical properties of calcium maltobionate (MBCa), its water sorption isotherm, glass transition temperature (T _g), and viscosity (η) were investigated and compared with those of maltobionic acid (MBH) and maltose. Although amorphous maltose crystalized at water activity (a _w) higher than 0.43, MBCa and MBH maintained an amorphous state over the whole a _w range. In addition, MBCa had a higher T _g and greater resistance to water plasticizing than MBH and maltose. These properties of MBCa likely originate from the strong interaction between MBCa and water induced by electrostatic interactions. Moreover, the effects of temperature and water content on η of an aqueous MBCa solution were evaluated, and its behavior was described using a semi-empirical approach based on a combination of T _g extrapolated by the Gordon-Taylor equation and a non-Arrhenius formula known as the Vogel–Fulcher–Tammann equation. This result will be useful for understating the effect of MBCa addition on the solution’s properties.     

Glutathione maintenance is crucial for survival of melanocytes after exposure to rhododendrol

Rhododendrol is a phenolic compound that shows a tyrosinase‐dependent toxicity for melanocytes and occasionally induces a vitiligo‐like skin depigmentation. The post‐tyrosinase mechanisms determining melanocyte death or survival, however, are far from clear. Here, we find that rhododendrol treatment leads to a reduction in the levels of cellular glutathione but also induces a cellular antioxidant response that eventually increases glutathione levels. We further find that rhododendrol toxicity is enhanced when glutathione levels are experimentally reduced and alleviated when glutathione levels are increased. Hence, it appears that the size of the preexisting glutathione pool along with the capacity to supply glutathione via the antioxidant response determines whether melanocytes survive or die after rhododendrol exposure. It is conceivable, therefore, that rhododendrol‐induced leukoderma depends on the capacity to maintain appropriate glutathione levels and that enhancement of glutathione levels may preserve a patient's melanocytes and potentially help in repigmentation.