Preparation of antimicrobial reduced lysozyme compatible in food applications

The structural and antimicrobial functions of lysozyme reduced with food-compatible reducing agents-cysteine (Cys) and glutathione (GSH)-were investigated. The disulfide bonds were partially reduced by thiol-disulfide exchange reactions under heat-induced denaturing conditions from 55 to 90 degrees C. The results showed that treatment of lysozyme with Cys and GSH resulted in the introduction of new half-cystine residues (2-3 residues/mol of protein). The released SH groups, in turn, rendered the lysozyme molecule more flexible, being accompanied by a dramatic increase in the surface hydrophobicity and exposure of tryptophan residues. As a consequence, the resulting reduced lysozymes were more capable of binding to lipopolysaccharides (LPS) and permeabilizing the bacterial outer membrane, as evidenced by the liposome leakage experiment, than were native or heated lysozyme. Both reduced lysozymes displayed significantly higher antimicrobial activity than native or heated lysozyme against Salmonella enteritidis (SE) in sodium phosphate buffer (10 mM, pH 7.2) at 30 degrees C for 1 h. Their minimal inhibitory concentrations (MICs) against the tested bacteria were about 150- and 25-fold lower than their respective MICs of native or heated lysozyme. The results suggest that partially reduced lysozyme could be used as a potential antimicrobial agent for prevention of SE attack.     

A method for visualizing cesium ions adsorbed on wood charcoal using SEM–EDX

A new method based on scanning electron microscopy with energy-dispersive X-ray spectroscopy (SEM–EDX) was developed for visualizing the distribution of cesium ions (Cs⁺) adsorbed by Japanese cedar charcoal. After being soaked in saturated aqueous CsCl solutions, wood charcoal and regular wood samples were embedded in resin, and thin sections of sample were prepared for the Cs⁺ observations. The Cs⁺ ions on charcoal could be seen in EDX mapping images and also in line profiles with enough contrast to identify differences between the various carbonized cell wall regions. On the other hand, Cs⁺ could not be observed in the EDX mapping images of wood samples, despite having been soaked in a saturated aqueous CsCl solution. These results indicate that Cs⁺ adsorbed on charcoal can be visualized using this method, but not Cs⁺ in wood. Additionally, oxygen detected on the charcoal by EDX can be regarded as an indicator of the presence of surface functional groups.     

Distribution of boron in wood treated with aqueous and methanolic boric acid solutions

The distributions of boron in Japanese cedar (Cryptomeria japonica D. Don) sapwood blocks treated with aqueous or methanolic boric acid [B(OH)₃] solutions were explored through Raman spectroscopy and prompt gamma-ray analysis (PGA). B(OH)₃ was the sole boron species observed in Raman spectra of the wood blocks treated with either solution. Plots of weight gain of the treated wood blocks versus boron concentration in treatment solutions were found to be linear. The results indicated that the methanolic solution makes it possible to impregnate wood with much larger amounts of boron than the aqueous solution. PGA confirmed that B(OH)₃ was highly enriched near the end grains of the treated wood blocks. Raman measurements suggested that boron content in the bulk of the wood block is not as large as expected from the weight gain of the treated wood blocks when an ordinary air-drying method is used. It was concluded that the aqueous solution impregnates the cell walls of wood with boron more easily than the methanolic solution. Part of this report was presented at the 55th (Kyoto, March 2005) and 56th (Akita, August 2006) Annual Meetings of the Japan Wood Research Society. This article follows the previous rapid communication “Analysis of boron in wood treated with boric acid solutions using Doppler broadening method of prompt gamma-rays.” J Wood Sci (2006) 52:279–281     

Effect of temperature and cell density on ethanol fermentation by a thermotolerant aquatic yeast strain isolated from a hot spring environment

ABSTRACT: A thermotolerant, fermentative yeast strain named RND13 from a hot spring drainage was evaluated for its ethanol-producing ability at elevated temperatures at a high substrate concentration [15% (w/v) glucose] close to the level reflecting industrial practice. The RND13 was capable of utilizing glucose almost completely at 40°C with increasing inoculum size, producing ethanol up to 6.6% (w/v), which is comparable to levels (7.0–7.2%) at 30°C. The maximum rate of ethanol production by the RND13 was found to be 9.0 g/L per h at 40°C in an inoculum sized 5% (w/v). At 43°C, however, the RND13 could not utilize glucose to completion and showed a slight drop in the extent of produced ethanol [6.0% (w/v)]. Thus, the culture at 40°C with a 5% cell inoculum was considered to be the optimal condition for ethanol production at higher temperatures in terms of batch fermentation. In the phylogenetic analysis based on the small-subunit rDNA sequence, the strain was grouped together with both Candida glabrata and Kluyveromyces delphensis , which are relatively close to Saccharomyces cerevisiae .     

Stem water transport of Lithocarpus edulis, an evergreen oak with radial-porous wood

The stem water conducting system of an evergreen broad-leaved oak, Lithocarpus edulis (Mak.) Nakai, was investigated. Evergreen broad-leaved oaks (Lithocarpus, Castanopsis, Cyclobalanopsis, Quercus) belonging to the Quercoideae are a major component of Asian monsoon forests, and are characterized by the possession of radial-porous wood. A characteristic of radial-porous wood is the development of aggregate rays between radially oriented files of vessels. We measured the distribution of vessel lumen diameters in a stem cross section and calculated the theoretical water conductivity of the wood. The radial profile of the heat pulse velocity (HPV) was measured for an intact whole tree under field conditions and compared with the theoretical distribution of water conductivity. Soft X-ray photographs of frozen stem sections indicated that most of the vessel lumina were filled with water, including those of vessels more than 20 years old. Even when vessels were relatively wide (lumen diameters > 100 microm), cavitation was negligible. The rate of water uptake from the cut stem base correlated closely with HPV (r = 0.96), and HPV closely reflected the mean volume flow per stem sectional area (SFVS) around the sensor probes. However, the ray tissue sharply inhibited heat transfer, and the positioning of the probes strongly affected the absolute value of HPV. It was also found that HPV more closely reflected the mean sap flow velocity in the vessels than did SFVS.     

Comparison of two different nutritive conditions against the changes in peripheral blood mononuclear cells of periparturient dairy cows

To clarify the relationship between cellular immune status and nutritive condition in periparturient dairy cows, feeding content, blood profiles, and immune condition were observed in cows from two dairy herds with different types of feed content. Immunological analyses such as leukocyte population and peripheral blood mononuclear cell (PBMC) mRNA of IFN-gamma, TNF-alpha, IL-4, and IL-10, quantified by real-time RT-PCR were performed. With regard to feed content during dry periods, there were six cows in the herd with insufficient non-structural carbohydrate (NFC) intake (group I) and six cows in the herd with sufficient NFC intake (group II). Significantly lower levels of blood glucose were observed in group I between weeks -12 and 16 compared with group II. Serum cholesterol level was significantly lower in group I between weeks 2 and 10 than in group II. The numbers of CD3+ and CD4+ T cells in group I were significantly lower than those in group II in weeks 6 and 14. The numbers of CD21+ B cells were significantly lower in group I than in group II in weeks -16, -12, 2, and 10. On the other hand, the CD4+/CD8+ ratio in group II was significantly higher than group I between weeks 2 and 14. The IFNgamma/IL-4 mRNA rate in group I was significantly lower than group II in week 6. We concluded that cellular immune depression occurrs after calving in dairy cows with low nutritional status in the periparturient period.     

Assessment of criteria for improvement in Theileria orientalis sergenti infection tolerance

We examined the relationships between calf productivity, anemia and Theileria orientalis sergenti infection from records of 109 Japanese shorthorn calves during a grazing period and we quantified the relative importance of the T. orientalis sergenti infection‐resistance criteria used. A decrease in average parasitemia (AvePara) and an increase in average packed cell volume (AvePCV) would cause a decrease in the number of drug treatments. An increase in AvePCV would decrease the number of drug treatments more efficiently than a decrease in AvePara. An increase in AvePCV would cause an increase in dairy gain, while a decrease in AvePara would cause a decrease in dairy gain. Therefore, an improvement in the capacity to control anemia development would improve T. orientalis sergenti infection resistance more efficiently than an improvement in the capacity to control parasitemia development.     

Structure-physicochemical function relationships of soybean beta-conglycinin heterotrimers

We purified four single molecular species of beta-conglycinin heterotrimers consisting of the alpha and beta subunits or the alpha' and beta subunits from mutant soybean cultivars lacking the alpha or alpha' subunit, respectively, and examined their structural features and physicochemical functions. The extent of the hydrophobicities of the heterotrimers was related to the number of the alpha or alpha' subunit. The thermal stabilities of the heterotrimers were mainly conferred by the subunit which had lower thermal stability. Solubilities at low ionic strength (mu = 0.08) of the heterotrimers containing the alpha or alpha' subunit were very similar to those of the alpha and alpha' homotrimers, respectively. Emulsifying abilities and heat-induced associations of the heterotrimers containing one beta subunit were similar to those of the alpha or alpha' homotrimer, whereas those of the heterotrimers containing two beta subunits were similar to those of the beta homotrimer.