Stability to light, heat, and hydrogen peroxide at different pH values and DPPH radical scavenging activity of acylated anthocyanins from red radish extract

The stability of red radish extract to light, heat, and hydrogen peroxide at different pH values (3, 5, and 7) was examined, in which major anthocyanins were pelargonidin glycosides acylated with a combination of p-coumaric, ferulic, or caffeic acids. The light irradiation (fluorescence light, 5000 lx; at 25 degrees C) indicated that the red radish extract was more stable at lower pH than at higher pH. The HPLC analyses revealed that diacylated anthocyanins in the extract were more stable to light at pH 3 than monoacylated anthocyanins. No significant difference in degradation rates of acylated anthocyanins at pH 5 was observed, whereas anthocyanins acylated with p-coumaric or ferulic acids were more stable at pH 7 than ones with caffeic acids. The stability to heat (at 90-95 degrees C) showed a tendency similar to that for light. The number of intramolecular acyl units contributes to stability to light and heat at lower pH, whereas the characteristics of intramolecular acyl units influence the stability at higher pH. The degradation behavior of red radish extract to H2O2 were almost the same to those of light and heat, depending on the pH. However, HPLC analyses revealed that the stability of individual acylated anthocyanins were independent of the pH. These data suggest that the characteristics, the number, and the binding site of intramolecular acyl units affect the stability of anthocyanin to H2O2. DPPH radical scavenging activity of all acylated anthocyanins was higher than those of pelargonidin and perlargonidin-3-glucoside. The activity of acylated anthocyanins mostly depended on the activity of intramolecular acyl units (caffeic acid > ferulic acid > p-coumaric acid). However, the activity was highly affected by the binding site of intramolecular acyl units even if anthocyanins have common acyl units.     

Canine Bcl-xL gene and its expression in tumor cell lines

The canine Bcl-xL gene was cloned and sequenced. Canine Bcl-xL cDNA clone was 1252 bp in length, and encoded 233 deduced amino acides. The predicted canine Bcl-xL amino acid sequence shared 99.6%, 97.0%, 97.9%, 98.7% and 98.3% homology with that of human, mouse, rat, sheep and pig Bcl-xL, respectively. RT-PCR analysis revealed that canine Bcl-xL mRNA was constitutively expressed in CL-1 (canine lymphoma) and GL-1 (canine B cell leukemia) cell lines.     

Ultrastructural study on the morphological changes in indigenous bacteria of mucous layer and chyme throughout the rat intestine

Indigenous bacteria in the alimentary tract are exposed to various bactericidal peptides and digestive enzymes, but the viability status and morphological changes of indigenous bacteria are unclear. Therefore, the present study aimed to ultrastructurally clarify the degeneration and viability status of indigenous bacteria in the rat intestine. The majority of indigenous bacteria in the ileal mucous layer possessed intact cytoplasm, but the cytoplasm of a few bacteria contained vacuoles. The vacuoles were more frequently found in bacteria of ileal chyme than in those of ileal mucous layer and were found in a large majority of bacteria in both the mucous layer and chyme throughout the large intestine. In the dividing bacteria of the mucous layer and chyme throughout the intestine, the ratio of area occupied by vacuoles was almost always less than 10%. Lysis or detachment of the cell wall in the indigenous bacteria was more frequently found in the large intestine than in the ileum, whereas bacterial remnants, such as cell walls, were distributed almost evenly throughout the intestine. In an experimental control of long-time-cultured Staphylococcus epidermidis on agar, similar vacuoles were also found, but cell-wall degeneration was never observed. From these findings, indigenous bacteria in the mucous layer were ultrastructurally confirmed to be the source of indigenous bacteria in the chyme. Furthermore, the results suggested that indigenous bacteria were more severely degenerated toward the large intestine and were probably degraded in the intestine.     

从气流状态考察小麦深床干燥特性研究

利用通风过程中的气流状态变化，考察小麦在深床下的干燥特性，讨论在深床干燥中，气流与床上料间的热和水分交换以及各层小麦的平均含水率变化，说明气流在深床内连续变化的过程和小麦的干燥速率。     

Effect of Yucca schidigera with or without nisin on ruminal fermentation and microbial protein synthesis in sheep fed silage- and hay-based diets

A 4 × 6 Youden square design was used to study manipulating effect of Yucca schidigera with or without nisin on rumen fermentation characteristics and microbial protein synthesis in sheep fed silage‐ and hay‐based diets. Four ruminally canulated wethers were given isonitrogenous diets of timothy silage and concentrate (85:15) (S), and timothy hay and concentrate (60:40) (H), on DM basis. The six experimental diets were: (i) S; (ii) S + 240 ppm of Yucca schidigera (SY); (iii) S + 240 ppm of Yucca schidigera + 6 mg/kg bodyweight (BW)^0.75 of nisin (SYN); (iv) H; (v) H + 240 ppm of Yucca schidigera (HY); and (vi) H + 240 ppm of Yucca schidigera + 6 mg/kg BW^0.75 of nisin (HYN). Rumen pH was higher (P < 0.01) in sheep fed SYN diet than those fed S diet. Concentration of ammonia‐N (NH₃‐N) was significantly lower (P < 0.05) in sheep fed SY, SYN, HY, HYN than those of S and H. Total volatile fatty acids concentration was significantly lower (P < 0.05) in HYN than in H diet and tended to be higher in sheep fed H diets than those fed S diets. There was a significantly higher (P < 0.01) efficiency of microbial nitrogen synthesis in the rumen of sheep fed on HY diet than those fed on H diet.     

Effect of glyphosate on tryptamine production and Sekiguchi lesion formation in rice infected with Magnaporthe grisea

When exposed to light, the Sekiguchi lesion (sl) rice mutant has an enhanced resistance to Magnaporthe grisea infection responsible for Sekiguchi lesion formation and tryptamine accumulation. Glyphosate [N-(phosphonomethyl) glycine] pretreatment suppressed Sekiguchi lesion formation and tryptamine accumulation in the sl mutant after M. grisea infection even under light. This inhibition by glyphosate was blocked by the supply of exogenous tryptophan, but not by exogenous phenylalanine. In glyphosate-pretreated leaves, 5-enol-pyruvyl-shikimate-3-phosphate synthase gene expression and tryptophan biosynthesis were significantly suppressed. During tryptophan starvation, catalase activity was maintained at a high level even under light, leading to the suppression of H₂O₂ generation and DNA fragmentation. These results show a strong relationship between the tryptophan and tryptamine pathways in the induction of light-enhanced resistance to M. grisea infection in the sl mutant.     

Methane emission from stored dairy manure slurry and slurry after digestion by methane digester

To examine the effect of storage temperature on the emission of methane (CH₄) and carbon dioxide (CO₂) and on recovery of nutrients from raw dairy manure slurry (RS) and slurry digested in a methane digester (DS), both slurries were stored in closed 100 L steel tanks under psychrophilic conditions (5, 10, 15 and 20°C) for a 150 day period. As the storage temperature increased, total methane emission increased in both types of slurry. The amount of methane emitted per unit of volatile solids of the RS and DS was 0.19 L/g and 0.10 L/g, respectively. The respective carbon dioxide emissions were 0.20 L/g for RS and 0.12 L/g for DS at 20°C of storage temperature. At temperatures greater than 15°C, the methane concentration in the emitted gas remained more than 40% of the total gas. During the experimental period, in excess of 90% of the total Kjeldahl nitrogen in these slurries was recovered (91.4–93.7% for RS and 93.7–98.4% for DS) after storage, and ammonium nitrogen was recovered in excess of 100% (100.1–143.2% for RS and 106.7–143.2% for DS storage tanks) because of the mineralization of organic nitrogen in the influent. These results indicate that manure slurry characteristics and storage temperature have significant impacts on methane emission. It can be concluded that on typical farms located in northern Japan, methane emission from manure storage tanks during late fall, winter and early spring may be negligible, because of manure temperatures less than 10°C. During late spring, summer and early fall, methane emissions can be substantially reduced by using underground storage to maintain lower manure temperatures.     

Lymphangiosarcoma with bone formation of the auricle in a dog

A 12-year-old mixed-breed neutered female dog was referred with cutaneous tumors at the left auricle. Histologically, the cutaneous tumor located in the dermis comprised numerous clefts and cavernous channels lined by neoplastic endothelial cells with no erythrocytes. Bone tissue without direct contact with neoplastic cells was seen in the well-developed stromal connective tissue. The neoplastic endothelial cells exhibited mild to moderate atypia. Immunohistochemically, neoplastic cells were positive for vimentin and negative for cytokeratin and factor VIII-related antigen. Basement membrane around the neoplastic lumens was positive for laminin in a linear or granular pattern. Ultrastructural examination revealed discontinuous basement membrane beneath the tumor cells. Histopathological features of this case were consistent with lymphangiosarcoma, and stromal ossification was characteristic.     

Immunohistochemical study on the secretory host defense system of bactericidal peptides in rat digestive organs

To clarify the fundamental regulation mechanism against indigenous bacterial proliferation in the alimentary tract, we immunohistochemically examined the localization of 4 bactericidal peptides (BP) in the rat digestive exocrine glands. In the upper alimentary tract, lysozyme was detected in the gustatory, extraorbital lacrimal and parotid glands. Secretory phospholipase A2 (sPLA2) was detected in the extraorbital lacrimal glands. β-defensin1 was detected in the gustatory and extraorbital lacrimal glands. β-defensin2 was detected in the Harderian glands. In the stomach, β-defensins were detected in the gastric superficial epithelial cells. In the small and large intestines, only lysozyme and sPLA2 were detected in the Paneth cells. In the cecum, all 4 BP were detected in the middle to apical portions of the crypts, and only sPLA2 was detected in the basal portion. No BP were localized in other exocrine glands associated with the alimentary tract. In addition, all 4 BP were also detected in the columnar epithelial cells of the apical portions of intestinal villi. In the intestinal superficial epithelial cells, lysozyme and β-defensins were detected in the ascending colon, whereas only β-defensin1 was detected in the descending colon and rectum. These results suggest that BP are mainly secreted from exocrine tissues in the initial portion of the digestive tract and play a role in host defense against indigenous bacteria throughout the digestive tract. Part of the BP in the chyme might be absorbed by the epithelium at the most inner sites of mucosae in the small and large intestines.