Preparation and partial characterization of monoclonal antibodies against the protective protein antigen of Erysipelothrix rhusiopathiae

Thirteen mouse monoclonal antibodies (Mabs) against the protective protein antigen (P64) of Erysipelothrix rhusiopathiae were prepared and partially characterized. The titres of the Mabs varied from 200 to 1,638,400 as determined by enzyme-linked immunosorbent assay (ELISA). Of the 13 Mabs 10, two and one belonged to the IgG2a, IgG1 and IgM subclasses, respectively. All Mabs reacted strongly with the 64 kDa protein and weakly with the 43 kDa protein upon Western blotting of the alkaline extract (AE) of E. rhusiopathiae. The protective activity (PD50/ml) of the 13 Mabs against E. rhusiopathiae infection in mice varied from < 50 to > 50,000. These Mabs were classified into three groups, highly protective Mabs, moderately protective Mabs and Mabs which did not possess protective activity, based on the protective index (ratio of the PD50/ml to the antibody titre). These results suggest that the 64 kDa protein is an effective protective antigen, which is easily cleaved into many small proteins, including the 43 kDa protein, and possesses at least two epitopes related to its protective activity and at least one epitope which is not related to protection of mice against E. rhusiopathiae infection.     

Malignant basal cell tumor in a Djungarian hamster

A malignant basal cell tumor was found in the skin of the abdomen of a female Djungarian hamster of unknown age and weighing 40 g. Histologically, the tumor mass was composed of cells resembling the basal cells of the epidermis, and these cells exhibited solid type proliferation. In the tumor tissue and necrotic foci, horn cysts were observed. Immunohistochemically, cytokeratin was present in the tumor cells and horncysts. By electron microscopic examination, the tumor cells had scanty cell organelles and a few desmozomes. This paper describes a rare malignant basal cell tumor in a Djungarian hamster.     

Degradation of paternal mitochondria by fertilization-triggered autophagy in C. elegans embryos

The mitochondrial genome is believed to be maternally inherited in many eukaryotes. Sperm-derived paternal mitochondria enter the oocyte cytoplasm upon fertilization and then normally disappear during early embryogenesis. However, the mechanism responsible for this clearance has been unknown. Here, we show that autophagy, which delivers cytosolic components to lysosomes for degradation, is required for the elimination of paternal mitochondria in Caenorhabditis elegans. Immediately after fertilization, sperm-derived components trigger the localized induction of autophagy around sperm mitochondria. Autophagosomes engulf paternal mitochondria, resulting in their lysosomal degradation during early embryogenesis. In autophagy-defective zygotes, paternal mitochondria and their genome remain even in the first larval stage. Thus, fertilization-triggered autophagy is required for selective degradation of paternal mitochondria and thereby maternal inheritance of mitochondrial DNA.     

The protective effect of stevia extract on the gastric mucosa of rainbow trout Oncorhynchus mykiss (Walbaum) fed dietary histamine

Stevia extract was made from stevia stem by extraction with boiling water and fermentation. The protective effect of a Stevia extract on rainbow trout (average weight, 12 g) fed 1% of dietary histamine for 4 weeks was investigated. Administration of dietary histamine to trout did not result in a reduction in growth rate or feed consumption but caused a gastric abnormality, e.g. exfoliation of the mucosal epithelium and atrophy of mucosal lamina propria. Stevia extract protected gastric tissue from histamine‐induced damage. Pepsin activity in gastric fluid increased and liver α‐tocopherol content was reduced after histamine treatment, but Stevia treatment prevented these abnormalities. The results suggest that the stevia extract might protect the rainbow trout stomach from histamine toxicity.     

Survival of inoculant Bradyrhizobium japonicum in an Andosol

The survival of an antibiotic-resistant mutant of a commercial inoculant Bradyrhizobium japonicum strain, A1017ks, was studied in a volcanic ash soil (Andosol) in comparison with a non-volcanic ash soil (Fluvisol) over a period of 84 days. In a non-sterile soil system, the population decline in the Andosol (15% or 1.2 log units) was larger than in the Fluvisol (6% or 0.54 log units). In both soils, however, the inoculant bradyrhizobium survived at fairly high population levels after the period of incubation [10⁶ and 10⁷ colony-forming units (CFU) g^-1 dry soil in the Andosol and Fluvisol, respectively]. In sterile control soil, viable bradyrhizobium cells could not be detected after 1 week of incubation in the Andosol, whereas in the Fluvisol population of introduced bradyrhizobium was maintained throughout the period of incubation. Overall changes in the population of indigenous bacteria and fungi were also monitored. However, no clear pattern of interaction between the inoculant Bradyrhizobium japonicum and the indigenous microbes could be identified. The antibiotic-resistant mutant maintained its resistance in the Fluvisol throughout the 3-month period of incubation, making it a useful model for conducting ecological studies in the soil.     

Determination of p-hydroxybenzoic acid esters in cosmetics by liquid chromatography with ultraviolet and fluorescence detection

A simple, precise, and accurate liquid chromatographic method with both ultraviolet (UV) and fluorescence detection is described for the determination of methyl, ethyl, propyl, and butyl p-hydroxybenzoates (PHBA-esters) in cosmetics. sec-Butyl p-hydroxybenzoate is added to the sample as an internal standard. Then the PHBA-esters are extracted with ether, the ether is evaporated to dryness, and the residue is dissolved in 60% (v/v) acetonitrile. The acetonitrile solution is passed through a Sep-Pak C18 cartridge to remove co-extracted lipids. PHBA-esters are determined by reverse-phase liquid chromatography with UV detection at 254 nm and fluorescence detection at ex 280 nm, em 305 nm. The mobile phase is acetonitrile-water (35 + 65). The method was linear over the concentration range of 0.005-0.15 mg/mL. Mean recoveries of each PHBA-ester were 98.9-102.7% (coefficients of variation less than or equal to 2.0%).     

Polymeric procyanidins as radical-scavenging components in red-hulled rice

The extracts from white-, black-, and red-hulled rice were prepared by sequential extraction with six different polar solvents, and their radical-scavenging activities were measured by methods using 1,1-diphenyl-2-picrylhydrazyl radical (DPPH*) and tert-butyl hydroperoxyl radical (t-BuOO*). The extracts prepared with highly polar solvents, methanol and deionized water, exhibited higher DPPH* and t-BuOO* scavenging activities in all three cultivars. In addition, the acetone extract from red-hulled rice exhibited a high DPPH* and t-BuOO* scavenging activity, while no such activity was detected for the acetone extracts from white- and black-hulled rice. The major components responsible for the radical scavenging in the acetone extract from red-hulled rice were identified as procyanidins by acidic hydrolysis, vanillin assay, and Sephadex LH-20 chromatography. GPC analysis of the acetylated procyanidins revealed that the average molecular weight is about 5000, in a range of about 500-18,000.     

Genetic engineering of maize (Zea mays) for high-level tolerance to treatment with the herbicide dicamba

Herbicide-tolerant crops have been widely and rapidly adopted by farmers in several countries due to enhanced weed control, lower labor and production costs, increased environmental benefits, and gains in profitability. Soon to be introduced transgenic soybean and cotton varieties tolerant to treatments with the herbicide dicamba offer prospects for excellent broadleaf weed control in these broadleaf crops. Because monocots such as maize (Zea mays) can be treated with dicamba only during a limited window of crop development and because crop injury is sometimes observed when conditions are unfavorable, transgenic maize plants have been produced and tested for higher levels of tolerance to treatment with dicamba. Maize plants expressing the gene encoding dicamba monooxygenase (DMO) linked with an upstream chloroplast transit peptide (CTP) display greatly enhanced tolerance to dicamba applied either pre-emergence or postemergence. Comparisons of DMO coupled to CTPs derived from the Rubisco small subunit from either Arabidopsis thaliana or Z. mays showed that both allowed production of transgenic maize plants tolerant to treatment with levels of dicamba (i.e., 27 kg/ha) greatly exceeding the highest recommended rate of 0.56 kg/ha.