Gastroprotective effect of red pigments in black chokeberry fruit (Aronia melanocarpa Elliot) on acute gastric hemorrhagic lesions in rats

It has been reported that the fruits and leaves of berries such as the blackberry, raspberry, and strawberry contain a high level of scavenging activity for chemically generated active oxygen species. This study investigated the antioxidative activities of black chokeberry fruit (Aronia melanocarpa Elliot) both in vitro and in vivo using the DPPH stable radical and rats with ethanol-induced gastric injury, respectively. The red pigment fraction of the black chokeberry contained three main components, one of which was identified as cyanidin 3-O-beta-glucoside by HPLC analysis and (1)H NMR. The black chokeberry red pigment fraction scavenged >44% of DPPH radicals at a concentration of 25 microg/mL compared to the control solution. The black chokeberry extract and its hydrolysate administrated at 2 g/kg of body weight each had nearly the same protective effect as quercetin administrated at 100 mg/kg of body weight in suppressing the area of gastric mucosal damage caused by the subsequent application of ethanol to <30% compared to the control group. The black chokeberry red pigment fraction had a similarly significant protective effect on gastric mucosa in a dose-dependent manner when administered at 30-300 mg/kg of body weight, and the administration of 30 mg/kg of body weight could suppress ethanol-induced gastric mucosal damage by approximately 50% (ID(50) = 30 mg/kg of body weight).     

Purification and characterization of lipoxygenase from Pleurotus ostreatus

Lipoxygenase was purified homogeneously from cups of Pleurotus ostreatus by Sephacryl S-400 HR gel filtration, Dyematrex Green A affinity, and DEAE-Toyopearl 650M ion-exchange chromatographies. The molecular weight of the enzyme was estimated to be 67,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 66,000 by gel filtration; the isoelectric point was pH 5.1. The optimum pH and temperature of the enzymatic activity were 8.0 and 25 degrees C, respectively. The enzyme contained non-heme iron, and a thiol group seemed to be involved in its activity. The K(m), V(max), and k(cat) values of the enzyme for linoleic acid were 0.13 mM, 23.4 micromol.min(-1).mg(-1), and 25.7 s(-1), respectively. The enzyme showed high specificity toward linoleic acid. When linoleic acid was incubated with the enzyme, 13-hydroperoxy-9Z,11E-octadecadienoic acid was found to be the main oxidative product.     

Protein characterizations and immunological properties of the low-molecular-mass protein A isolated from Staphylococcus aureus KS 1034.

The extracellular low-molecular-mass protein A isolated from Staphylococcus aureus KS 1034 was analyzed for its amino acid contents and terminal amino acid sequences. The C-terminal and the N-terminal amino acid sequences of the low-molecular-mass protein A were -Asn-Ala-Phe and Ala-Gln-His-Asp-Glu-Ala-Gln-, respectively. For immunochemical properties, the low-molecular-mass protein A was similar to protein A isolated from S. aureus Cowan I. Extracellular protease activity of KS 1034 strain was considerably lower than S. aureus C-30 isolated from chicken that produced the extracellular low-molecular-mass protein A.     

儿茶素抗氧化作用的协同增效效应

用电子自旋共振(ESR)和化学发光技术,研究了(-)-EGCG、(-)-ECG、(-)-EC、(-)-EGC 4种儿茶素及其不同组合,对超氧阴离子自由基(O(?))的清除作用。结果表明,单一儿茶素以(-)-EGCG 的清除能力最强,其纳摩尔清除率为316.3,其次为(-)-ECG,纳摩尔清除率为262.9;两两不同组合对O(?)的清除率以(-)-EGCG+(-)-ECG 效果最佳,比各自单独存在时要增效,三三组合时以(-)-EGCG+(-)-ECG+(-)-EC 组合效果最佳,增效效果明显,且强于两两组合。这些组合的增效效果与体系中存在的儿茶素的摩尔浓度呈高度的线性正相关。4种儿茶素组成不同的比例,以茶叶天然组成增效效果最明显。用伏安法测得的各儿茶素第一还原电位由高到低的次序为:(-)-EGCG>(-)-ECG>(-)-EGC≈(-)-EC,这一电位次序是儿茶素构成抗氧化循环的根本所在。     

Procedure for bovine ICSI, not sperm freeze-drying, impairs the function of the microtubule-organizing center

This study was designed to investigate whether freeze-dried (FD) bull spermatozoa maintained the function of the microtubule-organizing center (MTOC) after rehydration and intracytoplasmic sperm injection (ICSI). In a preliminary attempt, the cleavage and blastocyst formation rates in FD-ICSI zygotes (36 and 1%, respectively) were found to be considerably lower than those in control ICSI zygotes (67 and 21%, respectively) or in IVF zygotes (78 and 43%, respectively). An alkaline comet assay indicated that the DNA fragmentation index (length of comet tail % DNA liberated) was not significantly different between fresh and FD spermatozoa. In the main experiment, formation of sperm-asters in the FD-ICSI oocytes 7 h postinsemination occurred at a similar rate when compared with the control ICSI oocytes (41 vs. 49%). Among the oocytes exhibiting sperm aster formation, the extent of microtubule network assembly was comparable between the FD-ICSI and control ICSI groups. However, the MTOC of the ICSI oocytes was not as functional as that of IVF oocytes in terms of the aster formation rate (97%) and the fluorescent intensity of the microtubule network (2.0 folds). These results suggest that the freeze-drying process per se had no adverse effect on maintaining the MTOC function in bull spermatozoa.     

Challenging endeavour for preservation of freeze-dried mammalian spermatozoa

Freeze-drying (lyophilization) has been proposed as an alternative method for sperm preservation to overcome the disadvantages of the current cryopreservation method such as the high maintenance cost of frozen stocks, the problems associated with transportation of frozen materials and the potential risk of total loss of the frozen stock. Since freeze-dried spermatozoa after rehydration lose their motility, which is an essential requirement to complete physiological fertilization, a relatively difficult microinsemination technique must be applied to rehydrated spermatozoa. Theoretically, it has been supposed that freeze-dried spermatozoa could maintain their functions and abilities to interact with the oocyte cytoplasm after prolonged storage at refrigerator temperature. However, sufficient yield of transferable blastocysts and production of live offspring derived from freeze-dried sperm samples are still subjects to be challenged and overcome in large domestic species.     

Detection and genotyping of canine coronavirus RNA in diarrheic dogs in Japan

To clarify the prevalence of canine coronavirus (CCoV) infection in Japan, faecal samples from 109 dogs with diarrhoea were examined for CCoV RNA together with canine parvovirus type 2 (CPV-2) DNA. The detection rates of CCoV and CPV-2 for dogs aged less than 1 year were 66.3% and 43.8%, while those for dogs aged 1 year or older were 6.9% and 10.3%, respectively, which were significantly different (p < 0.0001 and p = 0.0003, respectively), indicating not CPV-2 but CCoV is an important diarrhoea-causing organism in juvenile dogs. Among the CCoV-positive dogs, 65.5% and 72.7% showed to be positive for CCoV types I and II, respectively, and simultaneous detection rate of both types was high at 40.0%. Furthermore, transmissible gastroenteritis virus (TGEV)-like CCoV RNA was detected from 8 dogs. These findings indicate that CCoV type I and TGEV-like CCoV are already circulating in Japan, though no reports have been presented to date.     

Potato extract (Potein) suppresses food intake in rats through inhibition of luminal trypsin activity and direct stimulation of cholecystokinin secretion from enteroendocrine cells

Dietary proteins and trypsin inhibitors are known to stimulate the secretion of the satiety hormone cholecystokinin (CCK). A potato extract (Potein) contains 60% carbohydrate and 20% protein including trypsin inhibitory proteins. In this study, we examined whether Potein suppresses food intake in rats and whether it directly stimulates CCK secretion in enteroendocrine cells. In fasted rats, food consumption was measured up to 6 h after the oral administration of Potein or soybean trypsin inhibitor (SBTI). CCK-releasing activities of Potein and SBTI were examined in the murine CCK-producing cell line STC-1. Potein inhibited the trypsin activity in vitro with a potency 20-fold lower than that of SBTI. Oral administration of Potein dose-dependently suppressed food intake for 1-6 h. Potein, but not the SBTI, dose-dependently induced CCK secretion in STC-1 cells. These results suggest that Potein suppresses food intake through the CCK secretion induced by direct stimulation on enteroendocrine cells and through inhibition of luminal trypsin.     

Plasma growth hormone (GH) responses after administration of the peptidergic GH secretagogue KP102 into the oral cavity, rumen, abomasum and duodenum in adult goats

The study was performed to determine whether orally administered KP102 (also known as GHRP-2) stimulates GH release in adult goats, and how the orally administered KP102 passes through the digestive tract and stimulates GH release in ruminant animals. Five mg/kg body weight (BW) of KP102 dissolved in 9 ml of saline were administered into the oral cavity, rumen, omasum and duodenum of adult goats, and GH release after administration of KP102 was examined. The GH levels were significantly elevated at 20 min after administration of KP102 into the oral cavity, and plasma concentrations of GH remained significantly elevated until 60 min (P < 0.05). The GH levels after administration of KP102 into the abomasum were variable. However, the GH level tended to increase within 30 min after administration, and were significantly higher than those of controls after 120 to 150 min (P < 0.05). The GH levels after administration of KP102 into the duodenum were significantly elevated at 40 min after administration, and plasma concentrations of GH remained significantly elevated until 140 min (P < 0.05). The administration of KP102 into the rumen failed to stimulate GH release. The GH response curves (AUC) produced after administration of KP102 into the abomasum or duodenum were 2.2-fold greater than those for after administration into the oral cavity (P < 0.05). The oral administration of 5 mg/kg BW of KP102 in the powder state, not dissolved in 9 ml of saline, failed to stimulate GH release. These results suggested that orally administered KP102 dissolved in saline transiently stimulates GH release in adult goats, and this phenomenon might be due to small amounts of the peptides entering directly into the abomasum with liquid bypassing the rumen.