Antioxidative effects of lactic acid bacteria on the colonic mucosa of iron-overloaded mice

The antioxidative effects of lactic acid bacteria on lipid peroxidation in the colonic mucosa were investigated. Among 49 strains of lactic acid bacteria, Streptococcus thermophilus YIT 2001 showed the highest inhibitory activity against lipid peroxidation in liposomes induced by ferrous iron. Feeding a diet containing 0.4% St. thermophilus YIT 2001 (2 x 10(8) colony-forming units per mouse per day) for 2 weeks caused a significant decrease of lipid peroxide (thiobarbituric acid reactive substance) in the colonic mucosa of iron-overloaded mice (0.07% Fe in the diet). The mucosal lipid peroxide level did not correlate with the soluble iron concentration of the cecal contents. Therefore, it is suggested that the antioxidative effect of St. thermophilus YIT 2001 in the colonic mucosa was not due to the removal of ferrous iron from the reaction system of lipid peroxidation.     

Identification of reciprocal translocations observed in several Melilotus species (subgenus Eumelilotus) by interspecific triple crossings

Summary Melilotus alba differs by a reciprocal translocation from 7 other species which are categorized into M. officinalis and M. dentata groups. The two species groups, however, remained to be studied in relation to their cytological relations because of early degeneration of hybrid embryo. Interspecific triple crosses were successfully made in order to examine whether or not the reciprocal translocations observed were of the same origin. When F₁ hybrids between M. alba and M. officinalis group were crossed with M. dentata group, about a half of hybrids plants were heterozygous for a reciprocal translocation while the remaining plants were normal chromosome pairing, showing a segregation ratio of 1:1. This result indicates that reciprocal translocations observed among the three groups are of the same origin. Accordingly, it is expected that M. officinalis group and M. dentata group have the same chromosomal constitutions.     

Effect of floral morphology on pollination in Brassica rapa L

Summary Flower structure, especially the anther–stigma separation (ASS), is well known to affect pollination efficiency, and thus to potentially increase or decrease seed production in crops. Therefore, investigating the relationship between flower characteristics and pollination ability is crucial to a full understanding of mechanisms to improve F₁ seed production in Brassica rapa. We used image analysis to measure three flower characteristics: short stamen height (SSH); long stamen height (LSH); and pistil height (PH) in seven cultivars. We calculated the ratio of PH to LSH as an index of anther–stigma separation (ASS). We investigated the number of pollen grains (NPG) deposited on the stigma and the seed-set percentage (SSP) under open-pollination and self-pollination conditions (with and without insects, respectively). Nested ANOVA indicated significant differences between the seven cultivars in the floral characteristics except for PH. Moreover, much larger variation was observed in NPG and SSP than in floral characteristics. Although stepwise multiple regression analysis indicated that plants with relatively high PHs produced more seed under self-pollination, the number of seeds that resulted from self-pollination did not affect seed production because of incompatibility. Therefore, the effect of the spatial position of pistils on the F₁ seed production was low. Possibly other factors such as the total pollen production and visiting times of pollinators were important factors in the low yields observed in some cultivars     

Use of an automatic cell-counting system with LED illumination for enumeration of marine bacteria

ABSTRACT:   The concentration of aquatic bacteria is basic information required to evaluate the status of environments and to assess bacterial contribution to material cycles. However, the standard direct counting method using epifluorescence microscopy (EFM) is tedious and there is variation in the counts among workers. Here an automatic counting system that consists of Bioplorer (BP) and image analysis has been applied to marine bacteria. BP is composed of a light-emitting diode (LED) illuminant, an optical unit, a driving stage and a charge-coupled device camera. In combination with fluorescent labeling and simplified membrane filtration, bacteria are enumerated automatically. The reproducibility, sensitivity and accuracy of the system were tested for natural marine bacteria, in comparison with EFM and flow cytometry (FCM). The counts obtained by BP showed good correlation with those obtained by EFM and FCM methods. The counts were significantly higher in inshore and oceanic samples, indicating high sensitivity with low background noise. Considering its reproducibility, objectivity, ease of use and compact size, BP can be used as a routine tool for counting aquatic bacteria in substitution for EFM or FCM.     

Characterization of depth-related changes and site-specific differences of microbial communities in marine sediments using quinone profiles

ABSTRACT:   Respiratory quinone compositions were analyzed by a high-performance liquid chromatography to characterize the depth-related changes and site-specific differences of microbial communities in marine sediments. Two deep-sea sediment samples and one coastal sediment sample were investigated from three sites on the coast of Japan, Sagami Bay, Suruga Bay and Tokyo Bay. Although depth-related changes in microbial community structures were observed, site-specific differences appeared to have greater influence on overall community structures. A variety of quinone homologs was commonly identified at all sampling sites and depths examined, but a few minor quinone fractions, mainly derived from Actinobacteria , were detected only at specific sampling sites. Methylmenaquinone-7 (MMK-7) was the major component throughout the examined depths in Sagami Bay and Tokyo Bay. Most abundant quinone homologs changed with each depth in Suruga Bay. Menaquinone-6 (MK-6) predominated in the 0 to 2-cm layer (19%). Below 0–2 cm, the most abundant homolog in each sampling depth was phylloquinone (K₁; 13%, 4–6 cm), MK-8 (19%, 8–10 cm) and MK-7 (13%, 16–18 cm). The microbial respiratory quinone profiling method shown here, successfully demonstrates the usefulness of this approach to characterize microbial communities in marine sediments.     

Bending resistance of repaired column members and shear resistance of opening frames with repaired columns of conventional Japanese wooden houses

When it is necessary to repair conventional Japanese wooden houses, the decayed lower parts of columns should be replaced with new wood material. The bending resistance of columns repaired by four methods and the shear resistance of opening frames with those repaired columns were investigated in this study. Bending tests of the repaired columns showed differences in initial bending stiffness and maximum bending moment related to the repair methods and loading direction. Racking tests were conducted on door opening frames with conventional door head members or upper partial walls sheathed with 12-mmthick plywood. The conventional frame specimens broke at door head-column joints with no obvious bending deformation of the columns, resulting in little difference in load-shear deformation curves among the repair methods. The columns of plywood-sheathed specimens, on the other hand, clearly were bent after the nails at the plywood-to-wood frame joints started to pull off. The load-shear deformation curves of the plywood-sheathed specimens did not vary regardless of the repair methods when shear deformations were small but were affected by repair methods as shear deformation increased.     

Metabolism of the herbicides chlorotoluron, diuron, linuron, simazine, and atrazine by CYP1A9 and CYP1C1 from Japanese eel (Anguilla japonica)

Through the use of a number of bioconversion experiments we demonstrated that P450 proteins (CYP1A9 and CYP1C1) from Japanese eel (Anguilla japonica) metabolized a number of herbicides and the drug phenacetin. We performed bioconversion experiments in which substrates were added directly to incubation medium. The resulting metabolites were extracted and analyzed by high-performance liquid chromatography. Proteins CYP1A9 and CYP1C1 metabolized 50 nmol of the drug phenacetin to yield 12.1 and 1.1 nmol of product (acetaminophen), respectively. Further incubation of CYP1A9 with 50 nmol of the herbicides chlorotoluron, diuron, linuron, simazine, or atrazine yielded 16.5, 18.5, 7.3, 1.6, or 0.8 nmol of product, respectively. CYP1C1 also metabolized linuron, diuron, and simazine yield 5.4, 4.6, or 0.7 nmol of product, respectively. Next, polyclonal antibody was isolated by immunizing with two conjugated-peptides (amino acid residues 272–290 and 294–310) of CYP1A9. This antibody did not recognize human CYP1A2 or CYP1C1. Western blotting using the antibody revealed one band in the livers of Japanese eel and tilapia (Oreochromis niloticus). Theses results suggest that CYP1A9 and CYP1C1 metabolize herbicides, and that CYP1A9 is an useful biomarker of contamination when detected with this antibody.     

Multiplex PCR and Genescan analysis to detect immunoglobulin heavy chain gene rearrangement in feline B-cell neoplasms

Lymphoid neoplasms are usually diagnosed on the basis of cytological and histopathological findings. However, in some cases, discrimination of lymphoid neoplasms from reactive lymphoid proliferation is difficult. PCR amplification of complementarity-determining region 3 (CDR3) of the immunoglobulin heavy-chain variable region (IGHV) gene can be used to assess clonality of B-cell populations as a supportive diagnostic tool for B-cell neoplasms. Because of the sequence variation and possible somatic hypermutation of the IGHV gene, sensitivity of the PCR-based assay to detect clonal IGHV gene rearrangement largely depends on the sequences and numbers of primer sets. Prior to the development of an efficient assay, we cloned and sequenced 97 IGHV complementary DNAs (48 IGHV-1 and 49 IGHV-3 clones) from normal cat spleens. On the basis of these sequences, we designed 6 forward primers at the variable region and 5 reverse primers at the joining region. Using each of 6 forward primers and a mixture of 5 reverse primers, we amplified CDR3 of IGHV genes and analyzed the PCR products by conventional PAGE and Genescan analyses using fluorescence-labeled primers. Twenty-six feline B-cell neoplasms diagnosed by histopathological and immunohistochemical examinations were subjected to the newly developed analysis of IGHV gene rearrangement. Clonal IGHV gene rearrangement was detected in 22 of 26 (84%) samples by both PAGE and Genescan analyses. To reduce the number of PCR reactions, we constructed a multiplex PCR analysis system using a mixture of IGHV-1- and IGHV-3-specific primers as forward primers and a mixture of 5 joining region reverse primers. Results of the multiplex PCR were 100% concordant with those obtained by each of the singleplex PCRs. The multiplex PCR-based assay and Genescan analysis developed in the present study would be useful and practical tools to detect clonal IGHV gene rearrangement in feline B-cell neoplasms.