Differentiation of canine bone marrow stromal cells into voltage- and glutamate-responsive neuron-like cells by basic fibroblast growth factor

We investigated the in vitro differentiation of canine bone marrow stromal cells (BMSCs) into voltage- and glutamate-responsive neuron-like cells. BMSCs were obtained from the bone marrow of healthy beagle dogs. Canine BMSCs were incubated with the basal medium for neurons containing recombinant human basic fibroblast growth factor (bFGF; 100 ng/ml). The viability of the bFGF-treated cells was assessed by a trypan blue exclusion assay, and the morphology was monitored. Real-time RT-PCR was performed to evaluate mRNA expression of neuronal, neural stem cell and glial markers. Western blotting and immunocytochemical analysis for the neuronal markers were performed to evaluate the protein expression and localization. The Ca²⁺ mobilization of the cells was evaluated using the Ca²⁺ indicator Fluo3 to monitor Ca²⁺ influx. To investigate the mechanism of bFGF-induced neuronal differentiation, the fibroblast growth factor receptor inhibitor, the phosphoinositide 3-kinase inhibitor or the Akt inhibitor was tested. The bFGF treatment resulted in the maintenance of the viability of canine BMSCs for 10 days, in the expression of neuronal marker mRNAs and proteins and in the manifestation of neuron-like morphology. Furthermore, in the bFGF-treated BMSCs, a high concentration of KCl and L-glutamate induced an increase in intracellular Ca²⁺ levels. Each inhibitor significantly attenuated the bFGF-induced increase in neuronal marker mRNA expression. These results suggest that bFGF contributes to the differentiation of canine BMSCs into voltage- and glutamate-responsive neuron-like cells and may lead to the development of new cell-based treatments for neuronal diseases.     

浅谈英语课堂教学活动的设计

教学活动的设计和使用成为英语课堂一个十分重要的环节,对最终的教学效果起不可或缺的作用.本文试从英语课堂教学活动的设计和活动方法两个方面,从教师的角度对该问题进行了初步探讨,对英语教师的课堂活动设计有一定参考作用.     

Anthocyanin composition and antioxidant activity of the Crowberry (Empetrum nigrum) and other berries

The anthocyanin composition and antioxidant activity of the crowberry ( Empetrum nigrum) were studied. High-performance liquid chromatography (HPLC) combined with a diode array detector and electrospray ionization mass spectrometry were used for identification and quantification of individual anthocyanins. Freeze-dried crowberry powder was extracted with 80% methanol containing 0.5% acetic acid and subjected to HPLC. Thirteen kinds of anthocyanins were identified. The major anthocyanins were cyanidin-3-galactoside and delphinidin-3-galactoside, at 8.04 and 8.62 mg/g extract, respectively. The HPLC profile of crowberry extract was similar to bilberry and blueberry. The total content of anthocyanins in crowberry was 41.8 mg/g extract, higher than the other nine major berry species (2.5-38.8 mg/g extract). The antioxidant activity was also evaluated using the 1,1-diphenyl-2-picrylhydrazyl and 2,2'-azinobis(3-ethybenzothiazoline-6-sulfonic acid) radical quenching assays and the ferric reducing activity power assy. Crowberry extract exerted the strongest antioxidant activity. In conclusion, individual anthocyanins in crowberry were identified and then quantified in this study. Additionally, crowberry is suggested to be associated with a reduction in the risk of developing chronic diseases because of its strong antioxidant activity.     

新入侵害虫螺旋粉虱的田间药效试验及防效

螺旋粉虱(Aleurodicus disperses Russell)是新入侵我国海南省的l种外来有害生物,对农业生产和生态环境具有较大危害性.田间防治研究结果表明:10%(质量分数,以下各农药浓度均为质量分数)联苯菊酯乳油、10%高效灭百可乳油、4.5%高效氯氰菊酯乳油等3种菊酯类杀虫剂对螺旋粉虱均表现出良好的速效性和持效性,其中又以10%高效灭百可乳油防效最高,为98.32%～100%;40%乐果乳油、30%敌百虫乳油和40%乙酰甲胺磷乳油等3种有机磷类农药的防效以40%乙酰甲胺磷乳油较好,但都低于上述3种菊酯类农药的防效;98%巴丹粉剂速效性较好,持效性较低;18%杀虫双水剂的速效性和持效性均较差.     

Characterization of the digestive tract of wild ayu

ABSTRACT: The gut and gut contents of ayu Plecoglossus altivelis caught in the Ohta River in Hiroshima prefecture, Japan were examined. Relative intestine length (intestine length/body length) was almost constant. Except for relative intestine length, characteristics of the gut of wild ayu were not significantly different from those of cultured ayu. The height of intestinal folds increased and their width declined from stomach to anus. Cyanobacteria ( Homoeathrix sp. and Calothrix sp.) and diatoms ( Cymbella sp., Gomphonema sp., Melosira sp., Navicula sp., and Synedra sp.) were well digested during passage through the gut despite a lack of apparent destruction. In contrast, green algae ( Dictyosphearium sp., Coelastrum sp., and Pediastrum sp.) exhibited little degradation. The pH of stomach contents ranged from 2.8 to 7.4 (mean ± SD = 4.1 ± 1.0), whereas that of the posterior intestine was 7.0–8.5 (7.9 ± 0.4). Gut contents of wild ayu increased with bodyweight, and were more than threefold greater than those of cultured ayu, suggesting that wild ayu compensate for low nutrient content of algae through the continuous ingestion of large quantities of feed organisms.     

Production of bacteriocin by Leuconostoc mesenteroides 406 isolated from Mongolian fermented mare's milk,airag

The purification and characterization of a bacteriocin produced by Leuconostoc mesenteroides strain 406 that was isolated from traditional Mongolian fermented mare's milk, airag, were carried out. Leuconostoc mesenteroides strain 406 was identified on the basis of its morphological and biochemical characteristics and carbohydrate fermentation profile and by API 50 CH kit and 16S ribosomal DNA analyses. The neutral‐pH cell‐free supernatant of this bacterium inhibited the growth of several lactic acid bacteria and food spoilage and pathogenic organisms, including Listeria monocytogenes and Clostridium botulinum. The bacteriocin was heat‐stable and not sensitive to acid and alkaline conditions, but was sensitive to several proteolytic enzymes such as pepsin, pronase E, proteinase K, trypsin, and α‐chymotrypsin, but not catalase. Optimum bacteriocin production (4000 activity units/mL) was achieved when the strain was cultured at 25°C for 24–36 h in Man Rogosa Sharpe medium. The bacteriocin was partially purified by ammonium sulfate precipitation (80% saturation), dialysis (cut‐off MW: 1000), and gel filtration chromatography. Sodium dodecyl sulfate‐polyacrylamide gel electrophoresis revealed that the bacteriocin had a molecular weight of approximately 3.3 kDa. To our knowledge, this is the first report of the isolation of a bacteriocin‐producing Leuconostoc strain from airag. An application to fermented milks would be desired.     

Proliferation capacity, neuronal differentiation potency and microstructures after the differentiation of canine bone marrow stromal cells into neurons

We examined the proliferation capacity and neuronal differentiation potency of canine bone marrow stromal cells (BMSCs). In addition, the microstructures of neuron-like cells after neuronal differentiation were observed under a scanning electron microscope. Canine BMSCs grew to confluency at 10.0 ± 2.5 days, and 3.8 ± 2.1 × 10(6) BMSCs were collected in one passage. Approximately 65% of canine BMSCs changed to neuron-like morphology after neuronal differentiation, and nearly all neuron-like cells stained positive against neuron-specific enolase. In addition, microstructures such as the cellular organelles, filaments and growth cones of these cells bore a close resemblance to those of the original mature neurons. These results suggested that canine BMSCs might be capable of differentiating into neurons.