Studies on absorption and hydrolysis of ethyl alpha-D-glucoside in rat intestine

Ethyl alpha-D-glucoside (alpha-EG) is normally contained in Sake, which has been taken by Japanese people since ancient times. In this study, the intestinal absorption of alpha-EG was investigated using rat everted intestinal sac. Furthermore, the alpha-EG hydrolytic activity in rat intestine was compared with disaccharides hydrolytic activities, and the effects of alpha-EG on disaccharides hydrolysis were examined using crude enzyme preparation from rat intestinal acetone powder. Glucose liberated from alpha-EG was detected in a serosal solution of everted rat intestinal sac, but it was only less than 4% of absorbed intact alpha-EG. alpha-EG absorption into small intestinal tissue was reduced by elimination of sodium ion from the mucosal solution or under the presence of phlorizin. The hydrolytic activity for alpha-EG was detected in crude enzyme preparation from rat intestinal acetone powder, but it showed a low value as compared to those for disaccharides. alpha-EG showed mixed type inhibition for maltose and sucrose hydrolysis, but inhibitory concentrations of alpha-EG required for 50% inhibition for the maltose and sucrose hydrolysis were higher than those of arabinose and acarbose. In conclusion, a small amount of alpha-EG was hydrolyzed and most of it was absorbed via SGLT1 as an intact form in the rat small intestine, and the inhibitory effect of alpha-EG on disaccharides hydrolysis was weak.     

Identification and sensory evaluation of volatile compounds in oxidized porcine liver

Headspace solid phase microextraction (HS-SPME) was used to isolate the off-flavor volatile compounds, which are formed during the oxidation of porcine liver induced by iron. Poly(dimethylsiloxane)/divinylbenzene fiber was used in the HS-SPME. Changes in the volatile compounds of oxidized porcine liver and unsaturated fatty acids induced by iron were examined. Results showed that 1-octen-3-one (metallic), hexanol (weak metallic), 1-octen-3-ol (mushroomlike), (E)-2-nonenal (cardboardlike), and (E,E)-2,4-decadienal (fatty, oily) were the main contributors to the overall off-flavor of porcine liver. The results of the sensory evaluation revealed that oxidized arachidonic acid has a major impact on metallic and liverlike off-flavor and that when liverlike off-flavor is perceived, metallic is also included. Oxidized linolenic acid was the most important contributor to the objectionable fishy off-flavor. Oxidized porcine liver exhibited distinct metallic, liverlike, and weak fishy background notes. Liverlike flavor had a high correlation coefficient with odor characteristics such as metallic (0.839) and fishy (0.777). In this study, it was clearly observed that the stronger the metallic and fishy off-flavor the higher the perception of liverlike off-flavor.     

Identification of a novel cytotoxic protein, Cry45Aa, from Bacillus thuringiensis A1470 and its selective cytotoxic activity against various mammalian cell lines

Parasporal inclusion proteins produced by Bacillus thuringiensis strain A1470 exhibit strong cytotoxicity against human leukemic T cells when activated by protease treatment. One of the cytotoxic proteins was separated by anion exchange chromatography and gel filtration chromatography and designated Cry45Aa. Its gene was then expressed in recombinant Escherichia coli, in which the Cry45Aa precursor was accumulated in an inclusion body. It was solubilized in sodium carbonate buffer and processed with proteinase K, and cytotoxic activities of the protein against various mammalian cell lines were evaluated using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide assay. The protein exhibited high cytotoxic activity against CACO-2, Sawano, MOLT-4, TCS, and HL60 cells and moderate activity against U-937 DE-4, PC12, and HepG2 cells. On the other hand, the EC50 values against Jurkat, K562, HeLa, A549, Vero, COS-7, NIH3T3, CHO, and four normal tissue cells (human primary hepatocyte cells, UtSMC, MRC-5, and normal T cells) were >2 microg/mL.     

Histamine behavior during the fermentation process in the manufacture of fish sauce.

The behavior of histamine in fish sauce making was investigated using fresh and spoiled fish with or without histidine added during fermentation. The histamine content in the 2% histidine added fresh fish mixture did not change significantly even after a lapse of 4 months incubation. However, when histidine was added to spoiled fish, the histamine content rose to a high level but decreased continuously with incubation time. This decrease may suggest the presence of histamine-decomposing bacteria in the samples. Eight of the 10 commercial fish sauces analyzed contained histamine levels below the "decomposition level" of 50 mg/kg set by the FDA. The increase in histamine at the initial stage and the decrease in histidine might suggest that histidine was converted to histamine by a microorganism possessing the enzyme histidine decarboxylase.     

CO_2施肥时树脂膜培养容器在文心兰试管苗培养中的应用

采用在培养室内CO2 施肥的方法 ,以C3植物文心兰 (Oncidium)品种‘AlohaIwanaga’试管苗为实验植物材料 ,分别以氟乙烯树脂膜容器 (CP)和三角瓶作为培养容器进行培养 .结果表明 :CO2 施肥时 ,树脂膜培养容器 (CP)内生长的试管苗在株高、叶数、叶长、叶幅、根数、根长、鲜重、干物重、干物率、叶绿素指数 (SPAD值 )等生长指标上高于无CO2施肥处理 .采用SSR法进行比较 (显著水平P≤ 0 0 5 ) ,其中在株高、叶长、叶幅、根数、鲜重、干物重等重要生长指标之间存在显著性差异 .以三角瓶为培养容器时其生长虽优于无CO2 施肥处理 ,但多数指标差异并不显著 .实验证明 ,使用CP作为培养容器 ,培养室内CO2 施肥对促进文心兰试管苗的生长和提高其品质效果明显 .     

Effects of permanent magnetic fields on the proliferation of Phalaenopsis protocorm-like bodies using liquid medium

Magnetic fields (MFs) have been applied for the first time in orchid micropropagation. Protocorm-like bodies (PLBs) – approximately 3 mm in diameter – first derived from leaf segment culture of Phalaenopsis Gallant Beau ‘George Vazquez’, were subcultured every 2 months, and served as initial explants. The proliferation of Phalaenopsis PLBs in liquid medium in the Miracle Pack^® culture system was affected by the action of different intensities and polarities of MFs: 0.1, 0.15 and 0.2 Tesla (T) at North (N) and South (S) poles. The MF of 0.1 T – S resulted in the greatest fresh weight of regenerated PLBs. The average number of neo-PLBs formed per clusters in the PLB treated by MF: 0.1–0.2 T was decreased compared to the control exposed to natural MF (5 × 10⁻⁶ T). The proliferation of PLBs under 0.15 T – MF at both N and S poles for 2 and 7 weeks demonstrated that a longer duration of exposure to an MF of 0.15 T, regardless of the polarity, resulted in greater biomass of newly formed PLBs and smaller average number of newly formed PLBs. The S pole of MF had stronger effects on Phalaenopsis PLBs proliferation than the N pole did in all treatments.     

Effect of cold cathode fluorescent lamps on growth of Gerbera jamesonii plantlets in vitro

The goal of this study was to evaluate the effect of cold cathode fluorescent lamps (CCFLs) on the growth of Gerbera jamesonii var. ‘Rui Kou’ plantlets in vitro in six different light quality ratios: 100% red CCFL (R), 80% R + 20% blue CCFL (B), 70% R + 30% B, 60% R + 40% B, 100% B and white CCFLs (W). Control radiation was provided by conventional heat-generating plant growth fluorescent lamps (PGFLs). Plantlets under CCFLs showed better plantlet height, SPAD value (i.e., leaf chlorophyll content) and root activity (as assessed by root dehydrogenase activity) than those growing under PGFLs while all other growth parameters were comparable with plants under conventional lighting systems.     

Lymphocyte blastogenic responses to inciting food allergens in dogs with food hypersensitivity

Lymphocyte blastogenic responses against food allergens in dogs with food hypersensitivity were evaluated in this study. Eleven dogs with food hypersensitivity, based on food elimination and oral food provocation tests and allergic responses to food allergens, were examined by various tests such as intradermal testing, antigen-specific IgE testing, and lymphocyte blastogenic responses. The number and kinds of food allergens identified as positive by these tests were compared with the offending food allergens that were found in an oral food provocation test. In 9 (82%) of the 11 dogs with food hypersensitivity, there was close agreement for positive allergens between the results of lymphocyte blastogenic responses and oral food provocation test; however, there was little agreement for intradermal and IgE testing of the positive allergens with those of the oral food provocation test (11% and 31%, respectively). In the 9 dogs, the stimulation indices of lymphocyte blastogenic responses increased to 2.0-10.1 upon food provocation but decreased significantly to 0.7-1.4 upon feeding the elimination diet until clinical signs disappeared. These results indicate that lymphocyte blastogenic responses may fluctuate because of exposure to offending food allergens in dogs with food hypersensitivity. Lymphocytes reactive to food allergens may play an important role in the pathogenesis of food hypersensitivity in dogs.     

Identification of CpG oligodeoxynucleotide sequences that induce IFN-gamma production in canine peripheral blood mononuclear cells

Oligodeoxynucleotides containing the cytosine-phosphate-guanine (CpG) motif (CpG-ODNs) have been shown to induce T(H)1 immune responses in animals. Since the sequences of CpG-ODNs that induce T(H)1 responses are considered to vary among animal species, it is necessary to identify effective CpG-ODNs in each animal. In order to identify the sequences of CpG-ODNs that induce T(H)1 responses in dogs, mRNA expression and protein production of IFN-gamma were examined in peripheral blood mononuclear cells (PBMCs) from healthy dogs treated with 11 kinds of synthetic CpG-ODNs. One of the 11 CpG-ODNs (No. 2 CpG-ODN, 5'-GGTGCATCGATGCAGGGGGG-3') was shown to significantly increase mRNA expression and protein production of IFN-gamma in canine PBMCs in a manner dependent on the sequence of the CpG motif. This CpG-ODN also enhanced the expression of IL-12 p40 mRNA in canine PBMCs, whereas expression of IL-12 p35, IL-18, and IL-4 mRNAs was not induced by this CpG-ODN. These results indicate that this CpG-ODN was able to produce IFN-gamma by induction of T(H)1-skewed immune response in dogs. CpG-ODNs may be useful for inducing prophylactic and therapeutic immunity against allergic diseases, viral infection, and tumors in dogs.