Evaluation of Intestinal Absorption of Dietary Halocynthiaxanthin,a Carotenoid from the Sea Squirt Halocynthia roretzi

Halocynthiaxanthin is an acetylenic carotenoid mainly found in Halocynthia roretzi. To date, several bioactivities of halocynthiaxanthin have been reported, but its mechanism of digestion and absorption in mammals has not been studied yet. In this study, we evaluated the intestinal absorption of halocynthiaxanthin in mice. The halocynthiaxanthin-rich fraction was prepared from the tunicate Halocynthia roretzi. Mice were orally administered the fraction at a dose of 5 mg/kg body weight. The halocynthiaxanthin levels in the plasma, liver, and small intestine, were quantified using HPLC-PDA, 1, 3, 6, and 9 h after ingestion. The halocynthiaxanthin-rich fraction mainly consisted of the all-trans form and a small amount of cis forms. These three isomers were detected in the plasma of mice 3 h after ingestion. Time-course changes after the ingestion of this fraction were found, with cis isomers being more abundant than the all-trans isomer in the mouse plasma and liver. In the small intestine, however, the all-trans isomer was primarily detected. The possibility that cis isomers might be absorbed rapidly from the small intestine cannot be denied, but our results suggest that dietary all-trans-halocynthiaxanthin might be isomerized to the cis isomer after intestinal absorption.     

Evaluation of the drug sensitivity and expression of 16 drug resistance-related genes in canine histiocytic sarcoma cell lines

Canine histiocytic sarcoma (HS) is an aggressive tumor type originating from histiocytic cell lineages. This disease is characterized by poor response to chemotherapy and short survival time. Therefore, it is of critical importance to identify and develop effective antitumor drugs against HS. The objectives of this study were to examine the drug sensitivities of 10 antitumor drugs. Using a real-time RT-PCR system, the mRNA expression levels of 16 genes related to drug resistance in 4 canine HS cell lines established from dogs with disseminated HS were determined and compared to 2 canine lymphoma cell lines (B-cell and T-cell). These 4 canine HS cell lines showed sensitivities toward microtubule inhibitors (vincristine, vinblastine and paclitaxel), comparable to those in the canine B-cell lymphoma cell line. Moreover, it was shown that P-gp in the HS cell lines used in this study did not have enough function to efflux its substrate. Sensitivities to melphalan, nimustine, methotrexate, cytarabine, doxorubicin and etoposide were lower in the 4 HS cell lines than in the 2 canine lymphoma cell lines. The data obtained in this study using cultured cell lines could prove helpful in the developing of advanced and effective chemotherapies for treating dogs that are suffering from HS.     

Sodium bicarbonate protects uranium-induced acute nephrotoxicity through uranium-decorporation by urinary alkalinization in rats

To evaluate the effectiveness of sodium bicarbonate (SB) in removing uranium and protecting animals from uranium toxicity, we intramuscularly administered 1 mg/kg of uranyl nitrate to 8-wk-old male SD rats, and 20 min after administration of uranyl nitrate, the animals were given a single oral administration of SB at 0.1, 0.3 or 1 g/kg. The SB treatment at a dose of 0.3 g/kg or more raised the pH of the rats’ urine until 4 h after treatment, and it significantly reduced the uranium amounts in the kidneys at 1 day after treatment. In another experiment, rats were intramuscularly administered 1 mg/kg of uranyl nitrate, and 20 min later, the animals were treated with sodium bicarbonate (0.1 or 1 g/kg). The rats were autopsied at 1, 3 and 7 days after uranium treatment. High-dose SB resulted in a significant increase in urinary uranium excretion in the first 24 h and a reduction of uranium deposition in the kidneys and femurs, and it also significantly suppressed uranium-induced renal toxicity, as shown by both histopathology and clinical chemistry at 3 days after uranium treatment. Low-dose SB did not show such marked effects. Our findings demonstrated that the uranium decorporation effect of sodium bicarbonate was observed at the dosage showing urine alkalinization in rats and that decorporation effect of sodium bicarbonate might be beneficial if it is administered immediately after incorporation of soluble uranium.     

Effects of utilization of local food by‐products as total mixed ration silage materials on fermentation quality and intake,digestibility, rumen condition and nitrogen availability in sheep

Four wethers were used in a 4 × 4 Latin square design experiment to evaluate in vivo digestibility of total mixed ration (TMR) silage with food by‐products for dairy cows, and the ruminal condition and nitrogen (N) balance were examined. Five by‐products (i.e. potato waste, noodle waste, soybean curd residue, soy sauce cake and green tea waste) were obtained. Four types of TMR silage were used: control (C) containing roughage and commercial concentrate, T1:20% and T1:40% containing the five by‐products replacing 20% and 40% of the commercial concentrate on a dry matter (DM) basis, respectively, and T2:40% containing three by‐products (potato waste, noodle waste and soybean curd residue) replacing 40% of the commercial concentrate on a DM basis. The ingredients were mixed and preserved in oil drum silos for 4 months. The TMR silages showed 4.02–4.44% and 1.75–2.19% for pH and lactic acid contents, respectively. The digestibility of DM and neutral detergent fiber, and total digestible nutrient content were higher (P < 0.05) for T2:40% feeding than for C feeding. Urinary nitrogen excretion tended to be lower (P = 0.07) for T2:40% than for C. The results suggested 40% replacing of commercial concentrate by using the three food by‐products can be most suitable for TMR silage.     

Intramammary infusion of an Enterococcus faecium SF68 preparation promoted the involution of drying off Holstein cows partly related to neutrophil‐associated matrix metalloproteinase 9

A problem for dairy cows following milk stasis is to cope with a high risk of intramammary infection and there is a need to initiate an extensive renewal of secretory modules in mammary glands so that milk production in next lactation may be optimized. We recently reported that ultrasonicated Enterococcus faecium SF68 (SF68) is compatible with cow mammary glands and an enhancer of innate immunity during the immediate post‐milk stasis period. The current study further examines the concomitant effect of ultrasonicated SF68 on mammary tissue remodeling. Four Holstein cows each received intramammary infusions of regular antibiotic dry‐cow formula (positive control) and two different doses of SF68 in different quarters. Analyses of individual quarter secretion samples showed faster neutrophil infiltration, earlier modifications in protein composition, including caseins and lactoferrins, as well as more prompt elevation of the specific unit of 92‐kDa matrix metalloproteinase 9 (MMP9) in SF68‐infused quarters compared to the positive controls. Intramammary infusion of ultrasonicated SF68 seems able to accelerate the regression of mammary synthetic capacity and potentiate the breakdown of glandular extracellular matrix, indicating a more efficient mammary gland involution. Correlation analyses imply that the ability of ultrasonicated SF68 to induce faster neutrophil chemotaxis and the associated MMP9 release is partly responsible.     

Reactivity of secondary hydroxyl groups in methylβ-d-xylopyranoside toward aβ-o-4-type quinone methide

Methyl-d-xylopyranoside was allowed to react with-O-4-type quinone methide without a catalyst to elucidate the reactivities of secondary hydroxyl groups at the C2, C3, and C4 positions. Benzyl ether-type lignin-carbohydrate complex (LCC) compounds linked at the C2 and C4 positions were predominant, at a ratio of 23. However, the reactivity of the hydroxyl group at the C3 position was quite low. These results strongly suggest that the reactivity of the C2 hydroxyl group in xylan toward quinone methide intermediate is higher than that of the C3 hydroxyl group during biosynthesis of LCCs.     

Simultaneous measurements of quantum yield and CO2 uptake for the assessment of non-assimilative electron flow in tree leaves

Using attached and detached leaves ofAcer palmatum Thunb. andRhaphiolepsis umbellata Makino, pulse-modulated chlorophyll fluorescence and CO₂ exchange were measured. Quantum yield of photosynthesis was determined from the fluorescence parameter(Fm′−Fs)/Fm′, where (Fm′−Fs) was defined as the difference between steady state chlorophyll fluorescence (Fs) and maximum fluorescence (Fm′) elicited by a saturating light pulse. The rate of electron transport through photosystem II (total electron flow) was calculated from the product of quantum yield andA (PFD), whereA is the rate of absorbed photons as given by leaf absorptance, and PFD is the photon flux density at the leaf surface. The rate of electron transport dependant on CO₂ uptake (assimilative electron flow) was calculated from the gross photosynthetic rate in a leaf. The difference between the rates of total and assimilative electron transport was denoted as the rate of non-assimilative electron transport which depends on photorespiration and oxygen reduction. Available data provided quantitative information on the rate of non-assimilative electron flow in intact leaves. When leaf photosynthesis ofA. palmatum was measured under sunlight, the rates of total and assimilative electron transport were determined to be approximately 900 and 150 μmol equiv. e/mg Chl·h, respectively. The difference (750 μmol equiv. e/mg Chl·h) was attributed to the activity of non-assimilative electron flow. The ratio of total to assimilative electron flow was found to increase gradually with rising in irradiance. The results suggest that non-assimilative electron flow occurred at much higher rate than assimilative electron flow at high irradiance. Implications of the results are briefly discussed in relation to photosynthesis limitation in tree leaves.     

Quantitative nested real-time PCR detection of <Emphasis Type="Italic">Verticillium longisporum</Emphasis> and <Emphasis Type="Italic">V. dahliae</Emphasis> in the soil of cabbage fields

Verticillium longisporum and V. dahliae, causal agents of Verticillium wilt, are spreading through the cabbage fields of Gunma Prefecture. Using the V. longisporum-specific intron within the 18S rDNA and differences between ITS 5.8S rDNA sequences in Japanese isolates of V. longisporum and V. dahliae, we developed three quantitative nested real-time (QNRT) PCR assays. The QNRT-PCR quantification of V. longisporum or V. dahliae in cabbage field soil was consistent with the severity of Verticillium wilt disease in those fields. In field trials of resistant cultivar YR Ranpo grown for three seasons in soil infested with the pathogen, disease severity and pathogen density in the soil were significantly reduced in a field moderately contaminated by V. dahliae, but only slightly reduced in a highly contaminated field. These results suggest that continuous cultivation of a resistant cultivar is an effective way to reduce the pathogen population. QNRT-PCR assays provide a powerful analytical tool to evaluate the soil population dynamics of V. longisporum and V. dahliae for disease management.