Retention of the developmental pluripotency in medaka embryonic stem cells after gene transfer and long-term drug selection for gene targeting in fish

Embryonic stem (ES) cells provide a unique tool for introducing random or targeted genetic alterations, because it is possible that the desired, but extremely rare recombinant genotypes can be screened by drug selection. ES cell-mediated transgenesis has so far been limited to the mouse. In the fish medaka (Oryzias latipes) several ES cell lines have been made available. Here we report the optimized conditions for gene transfer and drug selection in the medaka ES cell line MES1 as a prelude for gene targeting in fish. MES1 cells gave rise to a moderate to high transfection efficiency by the calcium phosphate co-precipitation (5%), commercial reagents Fugene (11%), GeneJuice (21%) and electroporation (>30%). Transient gene transfer and CAT reporter assay revealed that several enhancers/promoters and their combinations including CMV, RSV and ST (the SV40 virus early gene enhancer linked to the thymidine kinase promoter) were suitable regulatory sequences to drive transgene expression in the MES1 cells. We show that neo, hyg or pac conferred resistance to G418, hygromycin or puromycin for positive selection, while the HSV-tk generated sensitivity to ganciclovir for negative selection. The positive-negative selection procedure that is widely used for gene targeting in mouse ES cells was found to be effective also in MES1 cells. Importantly, we demonstrate that MES1 cells after gene transfer and long-term drug selection retained the developmental pluripotency, as they were able to undergo induced differentiation in vitro and to contribute to various tissues and organs during chimeric embryogenesis.

Temporal and spatial variations in kinetics of alkaline phosphatase in sediments of a shallow Chinese eutrophic lake (Lake Donghu)

Monthly sediment and interstitial water samples were collected in a shallow Chinese freshwater lake (Lake Donghu) from three areas to determine if alkaline phosphatase activity (APA) plays an important role in phosphorus cycling in sediment. The seasonal variability in the kinetics of APA and other relevant parameters were investigated from 1995-1996. The phosphatase hydrolyzable phosphorus (PHP) fluctuated seasonally in interstitial water, peaking in the spring. A synchronous pattern was observed in chlorophyll a contents in surface water in general. The orthophosphate (o-P) concentrations in the interstitial water increased during the spring. An expected negative relationship between PHP and V-max of APA is not evident in interstitial water. The most striking feature of the two variables is their co-occurring, which can be explained in terms of an induction mechanism. It is argued that phosphatase activity mainly contributes to the driving force of o-P regeneration from PHP in interstitial water, supporting the development of phytoplankton biomass in spring. The V-max values in sediment increased during the summer, in Conjunction with lower K-m values in interstitial water that suggest a higher affinity for the substrate. The accumulation of organic matter in the sediment could be traced back to the breakdown of the algal spring bloom, which may stimulate APA with higher kinetic efficiency, by a combination of the higher V-max in sediments plus lower K-m values in interstitial water, in Summer. In summary, a focus On phosphatase and its substrate in annual scale may provide a useful framework for the development of novel P cycling, possible explanations for the absence of a clear relationship between PHP and APA were PHP released from the sediment which induced APA, and the presence of kinetically higher APA both in sediment and interstitial water which permitted summer mineralization of organic matter derived from the spring bloom to occur. The study highlighted the need for distinguishing functionally distinct extracellular enzymes between the sediment and interstitial water of lakes. (C) 2002 Elsevier Science Ltd. All rights reserved.

Compensatory growth in the gibel carp following feed deprivation: temporal patterns in growth, nutrient deposition, feed intake and body composition

To investigate the nature of compenstory growth in fish, an 8 week study at 28 degreesC was performed on juvenile gibel carp Carassius auratus gibelio weighing 6.6 g. Fish were starved for 0 (control), 1 (Sl)or 2 (S2) weeks and then re-fed to satiation For 5 weeks. Weekly changes in weight gain, feed intake and body composition were monitored during re-feeding. No significant difference was found in final body weight between the three groups, indicating complete compensation in the deprived fish, The deprived groups caught up in body weight with that of the control after 2 weeks of re-feeding. Body fat:lean body mass ratio was restored to the control level within 1 week of re-feeding. In the re-feeding period, weekly gains in body weight, protein. lipid, ash and energy in the S1 group were significantly higher than in the controls for 1 week. For the S2 group, weekly gains in body weight. lipid. ash and energy were higher than in the controls for 2 weeks, and gain in protein was higher than in the controls for 3 weeks, though gain in body energy became elevated again during the last 2 weeks of the experiment. Feed intake remained higher than the control level for 3 weeks in the S1 group and 3 weeks in the SZ group. Growth efficiency was not significantly different among the three groups in any of the weeks during re-feeding. Compensatory responses in growth and especially feed intake tended to last longer than the recovery of body composition. (C) 2001 The Fisheries Society of the British Isles.

FREQUENCY AND TEMPORAL COHERENCE PROPERTIES OF DISTRIBUTED BRAGG REFLECTOR LASER

This article presents the investigation of frequency and temporal coherence properties of distributed Bragg reflector laser. In this scheme, a square-wavefrom voltage is applied to the phase section of the laser to little optical wavelength, and delayed optical heterodyne technique is used for the analysis of spectral characteristics. Experiments show that lightwaves emitted from the same active region asynchronously are partially frequency and temporal coherent. When the two wavelengths are closer, the two waves are strong v coherent, and the coherence properties get weak as the delay v time increases. (C) 2010 Wiley Periodicals, Inc. Microwave Opt Technol Left 52: 822-825, 2010 Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/mop.25031

Electro-optical properties and temporal stability of the guest-host DCNP/PEK-c polymer thin film

The polyetherketone (PEK-c) guest-host system thin films in which the range of the weight percent of 3-(1,1-dicyanothenyl)-1-phenyl-4, 5- dihydro-1H-pryazole (DCNP) is from 20% to 50% were prepared. The predicted high value of electro-optical (EO) coefficient gamma(33) = 48.8 pm/V by using two-level model was obtained when the weight percent of DCNP in the polymer system is 40%, whereas EO coefficients are attenuated at higher chromophore loading then 40%. The temporal stability of the EO activity of the guest-host polymer was evaluated by probing the decay of the orientational order of the chromophores in the polymer system.