Physicochemical characterization and biological effects of inulin enzymatically synthesized from sucrose

We first developed the method to produce inulin from sucrose using an enzyme from Bacillus sp. 217C-1. Synthesized inulin consists of a linear polymer having beta(2-1) linkages of d-fructose with one terminal glucose. The synthesized inulin has similar properties (pH and thermal stability, Maillard reaction, and in vitro fermentation) to plant-derived inulin. The marked difference is the polydispersity of the inulin chain length. Synthesized inulin with a narrow degree range of fructose polymerization shows better solubility in water than plant-derived inulin. Synthesized inulin (5%, w/w) treatment for 12 weeks reduced the elevation in body weight and serum and liver lipids in rats fed high fat- and high sucrose-supplemented diets, and blood glucose in rats fed a standard diet. Synthesized inulin (15%, w/w) significantly suppressed the elevation in blood glucose of human healthy subjects after dextrin loading. These results suggest that daily intake of synthesized inulin modulates carbohydrate and lipid metabolism.     

Insect antifeedants from tropical plants: structures of dumnin and dumsenin

Two novel A-seco limonoids, dumnin and dumsenin, were isolated from the methanolic extract of Croton jatrophoides by bioassay-guided fractionation, and the structures were determined by nuclear magnetic resonance, circular dichroism, and mass spectrometry experiments. These compounds showed potent antifeedant activity (PC(50) 相似文献   

Pharmacokinetics of cycloalliin, an organosulfur compound found in garlic and onion, in rats

Cycloalliin, an organosulfur compound found in garlic and onion, has been reported to exert several biological activities and also to remain stable during storage and processing. In this study, we investigated the pharmacokinetics of cycloalliin in rats after intravenous or oral administration. Cycloalliin and its metabolite, (3R,5S)-5-methyl-1,4-thiazane-3-carboxylic acid, in plasma, urine, feces, and organs was determined by a validated liquid chromatography-mass spectrometry method. When administered intravenously at 50 mg/kg, cycloalliin was rapidly eliminated from blood and excreted into urine, and its total recovery in urine was 97.8% +/- 1.3% in 48 h. After oral administration, cycloalliin appeared rapidly in plasma, with a tmax of 0.47 +/- 0.03 h at 25 mg/kg and 0.67 +/- 0.14 h at 50 mg/kg. Orally administered cycloalliin was distributed in heart, lung, liver, spleen, and especially kidney. The Cmax and AUC0-inf values of cycloalliin at 50 mg/kg were approximately 5 times those at 25 mg/kg. When administered orally at 50 mg/kg, cycloalliin was excreted into urine (17.6% +/- 4.2%) but not feces. However, the total fecal excretion of (3R,5S)-5-methyl-1,4-thiazane-3-carboxylic acid was 67.3% +/- 5.9% (value corrected for cycloalliin equivalents). In addition, no (3R,5S)-5-methyl-1,4-thiazane-3-carboxylic acid was detected in plasma (<0.1 microg/mL), and negligible amounts (1.0% +/- 0.3%) were excreted into urine. In in vitro experiments, cycloalliin was reduced to (3R,5S)-5-methyl-1,4-thiazane-3-carboxylic acid during anaerobic incubation with cecal contents of rats. These data indicated that the low bioavailability (3.73% and 9.65% at 25 and 50 mg/kg, respectively) of cycloalliin was due mainly to reduction to (3R,5S)-5-methyl-1,4-thiazane-3-carboxylic acid by the intestinal flora and also poor absorption in the upper gastrointestinal tract. These findings are helpful for understanding the biological effects of cycloalliin.     

Comparative study of anti-oncogenic microRNA-145 in canine and human malignant melanoma

MicroRNA-145 (miRNA-145; miR-145) is aberrantly expressed in most of human cancers and plays a significant role in carcinogenesis and cancer progression. In the current study, we focused on how miR-145 plays a role in canine and human malignant melanomas. MiR-145 was significantly downregulated in canine malignant melanoma tissues and canine melanoma cell lines, as well as human melanoma cell lines tested. The ectopic expression of miR-145 showed a significant growth inhibition in both canine and human melanoma cells tested, and the effect was achieved partly by suppressing c-MYC in canine melanoma LMeC and in human melanoma A2058 and Mewo cells. At the same time, a suppressive tendency on cell migration in canine melanoma KMeC cells and significant suppression of cell migration in human melanoma A2058 cells by suppressing FASCIN1 were also found. These findings suggest that miR-145 acts as a tumor suppressor in both canine and human malignant melanomas.     

Rapid Identification of Mycoplasma pulmonis Isolated from Laboratory Mice and Rats Using Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry

Mycoplasma species identification is based on biochemical, immunological, and molecular methods that require several days for accurate identification. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a novel method for identification of bacteria and has recently been introduced into the clinical microbiology laboratory as a rapid and accurate technique. This method allows a characteristic mass spectral fingerprint to be obtained from whole inactivated mycoplasmal cells. In this study, we evaluated the performance of the MALDI-TOF MS for the identification of Mycoplasma by comparison with standard sequence analysis of 16S rRNA. We developed the first database of MALDI-TOF MS profiles of Mycoplasma species, containing Mycoplasma pulmonis, M. arthritidis, and M. neurolyticum, which are the most common pathogens in mice and/or rats, and species-specific spectra were recorded. Using the database, 6 clinical isolates were identified. Six tracheal swabs from 4 mice and 2 rats were cultured on PPLO agar for 4 to 7 days, and the colonies were directly applied to analyze the protein profiles. Five strains were identified as M. pulmonis, and 1 strain from a mouse was identified as M. neurolyticum (spectral scores were >2.00); the results were consistent with the results of the 16S rRNA gene sequence analysis (homologies>97.0%). These data indicate that MALDI-TOF MS can be used as a clearly rapid, accurate, and cost-effective method for the identification of M. pulmonis isolates, and this system may represent a serious alternative for clinical laboratories to identify Mycoplasma species.     

Occurrence of stilbene oligomers in Cyperus rhizomes

Investigation of the chemical constituents of Rhizoma Cyperi (Cyperus rotundus Linneus) resulted in the isolation of novel enantiomeric and meso-stilbene trimers [i.e., (+)- and (−)-(E)-cyperusphenol A (1, 2 respectively) and (E)-mesocyperusphenol A (3)], a trimer bearing a novel hexacyclic ring system [cyperusphenol B (5)], as well as known stilbenoids (cyperusphenols C (4) and D (6), scirpusins A (7) and B (8), and piceid (9)) and luteolin. HPLC was used for the optical resolution of 1 and 2 as well as for the identification of cooccurrence of enantiomers of 7. The structures of the isolates were established by spectroscopic analyses, including a detailed NMR spectroscopic investigation. The isolates were evaluated in terms of their antiproliferative activity employing the Jurkat cell line (human T-cell leukemia cells), while the IC₅₀ potencies of a racemate of 1 and 2, 3, 5, and 6 were estimated as 27.4, 40.5, 26.4, and 26.3 μM, respectively. The suppression of cell growth by 6 was due to the induction of apoptosis, which was characterized by nuclear changes and PARP-1 cleavage determined by western blotting. We also evaluated the free radical scavenging activity of the isolates.     

Fasting and refeeding lead to more efficient growth in lean pacu (Piaractus mesopotamicus)

We evaluated whether body fat content affects the energetic metabolism and growth in pacu submitted to daily feeding, fasting and refeeding. For 15 days, fish were fed different diets to obtain lean and fat conditions, and then subjected, for 20 days to: (1) continuously feeding (control), or (2) fasting for 15 days and refeeding for 5 days. Blood (glucose, triglycerides, cholesterol, non‐esterified fatty acids and total protein) and tissue (liver lipid and glycogen, muscle lipid and mesenteric fat) metabolic indicators, and growth performance parameters (weight gain, specific growth rate, daily feed intake and feed conversion ratio) were measured. Fasting led both lean and fat pacu to make notable use of their energy reserves, through glycogenolysis and lipolysis, reflected in reduced blood glucose and triglycerides, liver glycogen and muscle lipid levels. Lipolysis was confirmed by the high levels of non‐esterified fatty acids, especially in fat pacu. Refeeding led to higher plasma glucose and liver lipid in lean fish. Muscle fat increased in fat fish but was not restored in lean fish, while mesenteric fat index (MFI) remained the same in fat fish and increased in lean fish. Although refeeding occurred only for 5 days, lean fish grew more and were more efficient at utilizing food (higher weight gain and better feed conversion ratio). In conclusion, our results suggest that fat pacu have higher glycogenic and lipogenic abilities, and the higher deposition of lipids in fish does not mean higher availability of energy for growth when compensatory growth is stimulated by refeeding after fasting.     

Prophylactic efficacy of desmopressin acetate for diabetes insipidus after hypophysectomy in the dog

The prophylactic efficacy of desmopressin acetate (DDAVP) on diabetes insipidus (DI) after hypophysectomy was investigated in the dog. In the control group, hypernatremia with a plasma level of 155 mEq/l or higher persisted for 12 hr from the 4th to the 16th hour after hypophysectomy, and symptoms of DI developed within five days after surgery. In the DDAVP treatment group, these changes were not observed, showing that administration of DDAVP (4 microg, installation, twice daily) effectively prevented hypernatremia that develops immediately after surgery and DI-like symptoms that persists for about one week after surgery.     

Relationship between seminal plasma lactoferrin and gonadal function in horses

Total 78 semen samples were obtained from 27 Thoroughbred stallions (aged 6 to 27 years), and were subjected to quantification of lactoferrin (Lf) in seminal plasma and examination of the seminal properties. The seminal plasma Lf concentration varied from 21 to 689 microg/ml, with a mean value of 244 +/- 151 microg/ml (S.D.). The seminal plasma Lf concentration and total seminal plasma Lf positively correlated with the sperm concentration (r=0.5938, P<0.001) and with the total sperm number (r=0.6959, P<0.001), respectively. There was no correlation between seminal plasma Lf and sperm motility. These results suggest that seminal plasma Lf reflects gonadal function.     

Evaluation of enzyme-linked immunosorbent assay (ELISA) and immunofluorescent antibody (IFA) test for the detection of porcine reproductive and respiratory syndrome virus (PRRSV) antibody in pigs from conventional farms

To evaluate the immunofluorescent antibody (IFA) test and enzyme-linked immunosorbent assay (ELISA) for detecting the porcine reproductive and respiratory syndrome virus (PRRSV) antibody, conventional pigs in PRRSV-positive and -negative commercial farms were examined. Antibody development patterns in ELISA and IFA tests were compared in 3 week old piglets experimentally infected with the PRRSV. The virus was detected from 2 days post infection (PI) and then the antibody titers and S/P ratios rose by both methods. A total of 208 serum samples were collected from 4 PRRSV-negative farms and 210 samples from PRRSV-positive farms, and were tested for the PRRSV antibody by IFA and ELISA. The titer of 64 should be set as the cut-off point in IFA for field sera. Similarly, the cut-off S/P ratio should be set at 0.4 in ELISA. A high degree of correlation was observed between antibody titers by the two methods in these 418 samples, with a correlation coefficient of 0.84. The coincidence rate between the two tests was 84.7% (354/418). In non-coincident cases, ELISA was able to detect the antibody with a low titer in the serum samples which were negative in IFA but from PRRSV positive farms. ELISA was more sensitive than IFA to detect PRRSV infected animals or farms.