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Experimental Culture (experimental + culture)
Selected AbstractsNitric oxide regulates cell survival in purified cultures of avian retinal neurons: involvement of multiple transduction pathwaysJOURNAL OF NEUROCHEMISTRY, Issue 2 2007T. A. Mejía-García Abstract Nitric oxide (NO) is an important signaling molecule in the CNS, regulating neuronal survival, proliferation and differentiation. Here, we explored the mechanism by which NO, produced from the NO donor S -nitroso-acetyl- d - l -penicillamine (SNAP), exerts its neuroprotective effect in purified cultures of chick retinal neurons. Cultures prepared from 8-day-old chick embryo retinas and incubated for 24 h (1 day in culture, C1) were treated or not with SNAP, incubated for a further 72 h (up to 4 days in culture, C4), fixed, and the number of cells estimated, or processed for cell death estimation, by measuring the reduction of the metabolic dye 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). Experimental cultures were run in parallel but were re-fed with fresh medium in the absence or presence of SNAP at culture day 3 (C3), incubated for a further 24 h up to C4, then fixed or processed for the MTT assay. Previous studies showed that the re-feeding procedure promotes extensive cell death. SNAP prevented this death in a concentration- and time-dependent manner through the activation of soluble guanylate cyclase; this protection was significantly reversed by the enzyme inhibitors 1H-[1,2,4]oxadiazolo-[4,3-a]quinoxalin-1-one (ODQ) or LY83583, and mimicked by 8-bromo cyclic guanosine 5,-phosphate (8Br-cGMP) (GMP) or 3-(5,-hydroxymethyl-2,-furyl)-1-benzyl indazole (YC-1), guanylate cyclase activators. The effect was blocked by the NO scavenger 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (PTIO). The effect of NO was also suppressed by LY294002, Wortmannin, PD98059, KN93 or H89, indicating the involvement, respectively, of phosphatidylinositol-3 kinase, extracellular-regulated kinases, calmodulin-dependent kinases and protein kinase A signaling pathways. NO also induced a significant increase of neurite outgrowth, indicative of neuronal differentiation, and blocked cell death induced by hydrogen peroxide. Cyclosporin A, an inhibitor of the mitochondrial permeability transition pore considered an important mediator of apoptosis and necrosis, as well as boc-aspartyl (OMe) fluoromethylketone (BAF), a caspase inhibitor, also blocked cell death induced by re-feeding the cultures. These findings demonstrate that NO inhibits apoptosis of retinal neurons in a cGMP/protein kinase G (PKG)-dependent way, and strengthens the notion that NO plays an important role during CNS development. [source] The response of Aeromonas hydrophila to oxidative stress induced by exposure to hydrogen peroxideJOURNAL OF APPLIED MICROBIOLOGY, Issue 1 2000J.P.B. Landre Aeromonas hydrophila, an opportunist human pathogen of low virulence, was shown to display a high degree of sensitivity upon exposure to hydrogen peroxide. As with other species, Aer. hydrophila is able to develop the capacity to resist loss of viability induced by such oxidative stress. Development of stress resistance follows the archetypal profile where pre-exposure of a population to sub-lethal levels of H2O2 stimulates onset of tolerance to further exposure. Acquisition of tolerance critically requires nascent protein synthesis. Further analysis demonstrated population growth phase influences the degree of sensitivity of the organism. Late stationary phase cultures demonstrate a decreased sensitivity compared with younger populations. Significantly, it was also determined that stock culture age influenced the level of sensitivity of the derived experimental culture, where an increased stock culture age corresponded with enhanced resistance to H2O2. These data show that Aer. hydrophila population phenotype is influenced by the phenotype of the donor stock culture. [source] Weaning Chinese perch Siniperca chuatsi (Basilewsky) onto artificial diets based upon its specific sensory modality in feedingAQUACULTURE RESEARCH, Issue 2001X F Liang Abstract Chinese perch are one of the most valuable food fish in China, but the sole source of feed for intensive culture is live prey fish. Our previous studies on systematic sensory physiology revealed that this species have a mechanism for this peculiar feeding habit. In the present study, a specific training procedure was designed, and both experimental (initial body weight 171.0 g; 120 days) and commercial (initial body weight 52.4 g; 240 days) net-cage cultures were conducted to investigate the training success, growth performance and survival of the trained yearlings fed with nonlive or Oregon-type moist diet. The training successes of minced prey fish and the Oregon moist diet were 100 and 89.9%, respectively, in experimental culture, and 92.2 and 83.5% in commercial culture. In an experimental trial, the fish fed minced prey fish or the Oregon moist diet attained final body weights of 472.7 g or 344.7 g, although the specific growth rates of these groups were significantly lower than that of the fish fed live prey fish (final body weight 560.0 g). Mortality was not significantly related to dietary treatment. In commercial culture, the final body weights were as follows: 750 g on live prey fish, 705 g on minced prey fish and 651 g on the Oregon moist diet. Feed costs to produce 1 kg fish were estimated to be US$6.59 for live prey fish, US$1.76 for minced prey fish and US$2.07 for the Oregon moist diet. The results of the present study confirmed that sensory modality and associative learning appear to be critical factors in determining food discrimination of Chinese perch, indicating that both minced trash fish and Oregon-type moist diet can be substituted for live prey fish in intensive commercial production. [source] Translational and transcriptional analysis of Sulfolobus solfataricus P2 to provide insights into alcohol and ketone utilisationPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 3 2007Poh Kuan Chong Abstract The potential of Sulfolobus solfataricus P2 for alcohol or ketone bioconversion was explored in this study. S. solfataricus was grown in different concentrations (0.1,0.8% w/v) of alcohols or ketones (ethanol, iso-propanol, n -propanol, acetone, phenol and hexanol) in the presence of 0.4% w/v glucose. Consequently, the addition of these alcohols or ketones into the growth media had an inhibitory effect on biomass production, whereby lag times increased and specific growth rates decreased when compared to a glucose control. Complete glucose utilisation was observed in all cultures, although slower rates of glucose consumption were observed in experimental cultures (average of 14.9,mg/L/h compared to 18.9,mg/L/h in the control). On the other hand, incomplete solvent utilisation was observed, with the highest solvent consumption being approximately 51% of the initial concentration in acetone cultures. Translational responses of S. solfataricus towards these alcohols or ketones were then investigated using the isobaric tags for relative and absolute quantitation (iTRAQ) technique. The majority (>80%) of proteins identified and quantified showed no discernable changes in regulation compared to the control. These results, along with those obtained from transcriptional analysis of key genes involved within this catabolic process using quantitative RT-PCR and metabolite analysis, demonstrate successful alcohol or ketone conversion in S. solfataricus. [source] Effect of antibiotic treatment on the growth and survival of juvenile northern Chilean scallop, Argopecten purpuratus Lamarck (1819), and associated microflora in experimental culturesAQUACULTURE RESEARCH, Issue 12 2009Jorge Fierro Abstract Juvenile northern Chilean scallops of 937±55 ,m shell height were exposed to five different concentrations of chloramphenicol (CHL) (5, 10, 25, 50 and 100 ,g mL,1), plus a control without antibiotics. To determine the effect of CHL on the accompanying microflora, the number of colony-forming units (CFU) that grew on TGE culture medium was counted in the seawater of containers with juveniles, and in containers with microalgae used as food. Both were exposed to the same concentrations of CHL. The growth rates of juveniles treated with CHL and the control without antibiotic showed highly significant differences (P=0.0001). The growth rate was inversely proportional to the CHL concentration. The control sample presented the highest growth rate (84.4±14.3 ,m day,1), followed by the sample treated with 5 ,g mL,1 (64.2±14.3 ,m day,1). The survival in the control and in the treated samples with 5,50 ,g mL,1 was rather high, with a mean value of 95%. Only the sample treated with 100 ,g mL,1 had a low survival (36.7%). The CFU count was larger in the containers with juveniles, when compared with the ones with food. The CFU count tended to decrease with increasing CHL concentration in the juveniles. [source] | ||||||||||