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151.
We studied a recently described method for the determination of total nitrogen in natural waters involving sample oxidation with persulfate and subsequent determination of nitrate-nitrogen with second derivative spectroscopy and compared it to the USEPA approved method involving the sum of nitrate-nitrogen and Kjeldahl-nitrogen as measured with an automated analyzer. The overall objective was to determine if the two methods gave the same estimates of total nitrogen and if the detection limits, precision and accuracy of the new method were as good as those of the USEPA method. The new method was used to make measurements on replicated blanks, standards and lake water samples covering a range of concentrations. We also collaborated with certified laboratories to make comparative measurements on 5 standards and 21 lake water samples that were run by us with the new method and by them with the USEPA method. The new method had an instrument detection limit of 0.07 mg 1–1, and the standard deviation of 20 sets of lake water samples averaged 0.03 mg 1–1. The new method gave concentrations equivalent to those found with the USEPA method, was more sensitive, and had a higher degree of precision. We concluded that the new method is suitable as a substitute for the USEPA method. We also found that the addition of acid to lake water samples stored under refrigeration was not necessary to preserve them for later determinations of nitrate-nitrogen and total nitrogen and that freezing was an effective means of sample preservation for 90 days. 相似文献
152.
Changes in the species composition, photosynthesis, calcification and size-fractionated carbon metabolism by natural phytoplankton assemblages were monitored in three mesocosms under different nutrient conditions during May 1993. In the 3 enclosures, the decline of the diatom-dominated assemblages was followed by the development of a bloom of the coccolithoporid Emiliania huxleyi. Highest growth of E. huxleyi was observed in the mesocosm with a high N : P ratio, suggesting this species is a good competitor at low phosphate concentrations. The transition from diatom- to E. huxleyi-dominated assemblages brought about a sharp reduction of the phytoplankton standing stock and carbon-specific photosynthetic rate. The relative contribution of the smaller size fraction to total photosynthesis increased as the succession progressed. Calcification rate and E. huxleyi cell-specified calcite production were highest during the early stages of development of the E. huxleyi bloom. Distinct changes in the patterns of 14C allocation into biomolecules were noticed during the diatom-E. huxleyi succession. The diatom-dominated assemblage showed high relative 14C incorporation into low molecular weight metabolites (LMWM), whereas proteins and, specially, lipids accounted for the largest proportion of carbon incorporation in the E. huxleyi bloom. The patterns of photoassimilated carbon metabolism proved to be strongly dependent on cellular size, as protein relative synthesis was significantly higher in the smaller than in the larger size fraction, irrespective of the nutrient regime and the successional stage. These results are discussed in relation to the ecological and physiological features of small phytoplankton. 相似文献
153.
Substituted enzyme (or ping-pong) mechanisms usually involve enzymes that exist in two forms that alternate during the catalytic
reaction. A method is described here for determining the position of the equilibrium of a half reaction in a ping-pong enzyme
mechanism that is based on the kinetics of the burst reaction which occurs upon addition of reactants that recycle the enzyme
from one form to another. The theoretical basis for the analysis is developed, and the method is applied to the half reaction
of the aldimine form of aspartate transaminase with difluoro-oxaloacetate.
Special issue dedicated to Herman Bachelard 相似文献
154.
Elumalai Sivamani Ping Shen Natacha Opalka Roger N. Beachy Claude M. Fauquet 《Plant cell reports》1996,15(5):322-327
The microprojectile bombardment of immature embryos has proven to be effective in transforming many indica rice varieties. One of the drawbacks of using immature embryos is the requirement of a large number of high quality immature embryos, which itself is a tedious and laborious process. To circumvent these problems, we have developed a procedure, using indica variety TN1 as a model that generates highly homogenous populations of embryogenic subcultured calli by selectively propagating a small number of regeneration-proficient calli derived from seeds. Thousands of embryogenic calli were produced from 50 seeds within 10 weeks. Ten to 20 independent R0 transgenic lines were regenerated per 500 embryogenic calli bombarded. The convenience and reliability offered by this transformation system has made transformation of indica rice a routine procedure.Abbreviations 2,4-D
2,4-dichlorophenoxy acetic acid
- NAA
naphthalene acetic acid
- BAP
6-benzylaminopurine
- kb
kilobase
- GUS
-glucuronidase
- X-gluc
5-bromo-4-chloro-3-indolyl--D-glucuronide
- HPH
hygromycin B phosphotransferase 相似文献
155.
Regeneration of fertile transgenic indica (group 1) rice plants following microprojectile transformation of embryogenic suspension culture cells 总被引:8,自引:0,他引:8
Shiping Zhang Lili Chen Rongda Qu Philippe Marmey Roger Beachy Claude Fauquet 《Plant cell reports》1996,15(7):465-469
Regenerable embryogenic suspensions of elite Indica (group 1) rice varieties IR24, IR64, IR72 and an advanced Indica rice breeding line IR57311-95-2-3 were established within 6–8 weeks from 3–4 week old calli derived from mature seeds. Transgenic rice plants were obtained by introducing a plasmid carrying genes encoding hygromycin phosphotransferase (hph, conferring resistance to hygromycin B) and ß-glucuronidase (uidA), both driven by the CaMV 35S promoter, via particle bombardment of embryogenic suspensions. The effect of osmotic conditioning on transformation was evaluated. Regenerated plants were resistant to hygromycin B and expressed the uidA (GUS) gene. The growth of mother plants (R0) was normal and seeds were produced. Southern blot analysis of R0 and R1 plants showed that hygromycin resistant plants contained intact hph genes that were inherited in a Mendelian fashion. A protocol for a simple, efficient, repeatable, genotype- and environment-independent Indica rice transformation system is described.Abbreviations 2,4-D
2,4-dichlorophenoxy acetic acid
- NAA
-naphthalene acetic acid
- kb
kilobase
- GUS
ß-glucuronidase
- hph
hygromycin B phosphotransferase 相似文献
156.
Kichoon Lee Jeff Buhr Gary J. Hausman Thomas Wright Roger Dean 《Molecular and cellular biochemistry》1996,156(1):31-35
Calcium oxalate crystal growth and aggregation leads to the formation of renal calculi. It is known to be inhibited by several compounds both in vitro and in vivo conditions. The present study highlights the inhibitory potential of sodium pentosan polysulphate (SPP), a semi-synthetic glycosaminoglycan (GAG) on calcium oxalate crystal growth in vitro. Its efficacy was compared with those of known inhibitors like pyrophosphate, heparin and chondroitin-4-sulphate. Of the above compounds pyrophosphate was found to be the most potent inhibitor. Among the GAGs, SPP exhibited 80% inhibitory activity as compared to heparin. A lesser degree of inhibition was observed with chondroitin-4-sulphate. 相似文献
157.
Roger F. Castilho Paulo C. Carvalho-Alves Anibal E. Vercesi Sérgio T. Ferreira 《Molecular and cellular biochemistry》1996,159(2):105-114
The major protein in the sarcoplasmic reticulum (SR) membrane is the Ca2+ transporting ATPase which carries out active Ca2+ pumping at the expense of ATP hydrolysis. The aim of this work was to elucidate the mechanisms by which oxidative stress induced by Fenton's reaction (Fe2+ + H2O2 HO· + OH–+ Fe3+) alters the function of SR. ATP hydrolysis by both SR vesicles (SRV) and purified ATPase was inhibited in a dose-dependent manner in the presence of 0–1.5 MM H2O2 plus 50 M Fe2+ and 6 mM ascorbate. Ca2+ uptake carried out by the Ca2+-ATPase in SRV was also inhibited in parallel. The inhibition of hydrolysis and Ca2+ uptake was not prevented by butylhydroxytoluene (BHT) at concentrations which significantly blocked formation of thiobarbituric acid-reactive substances (TBARS), suggesting that inhibition of the ATPase was not due to lipid peroxidation of the SR membrane. In addition, dithiothreitol (DTT) did not prevent inhibition of either ATPase activity or Ca2+ uptake, suggesting that inhibition was not related to oxidation of ATPase thiols. The passive efflux of 45Ca2+ from pre-loaded SR vesicles was greatly increased by oxidative stress and this effect could be only partially prevented (ca 20%) by addition of BHT or DTT. Trifluoperazine (which specifically binds to the Ca2+-ATPase, causing conformational changes in the enzyme) fully protected the ATPase activity against oxidative damage. These results suggest that the alterations in function observed upon oxidation of SRV are mainly due to direct effects on the Ca2+-ATPase. Electrophoretic analysis of oxidized Ca2+-ATPase revealed a decrease in intensity of the silver-stained 110 kDa Ca2+-ATPase band and the appearance of low molecular weight peptides (MW < 100 kDa) and high molecular weight protein aggregates. Presence of DTT during oxidation prevented the appearance of protein aggregates and caused a simultaneous increase in the amount of low molecular weight peptides. We propose that impairment of function of the Ca2+-pump may be related to aminoacid oxidation and fragmentation of the protein.Abbreviations AcP
acetylphosphate
- BHT
butylhydroxytoluene
- DTT
dithiothreitol
- Hepes
4-(2-hydroxyethyl)-1-piperazine-ethanesulfonic acid
- SDS
sodium dodecyl sulfate
- SDS-PAGE
polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate
- SR
sarcoplasmic reticulum
- SRV
sarcoplasmic reticulum vesicles
- TBA
thiobarbituric acid
- TBARS
thiobarbituric acid-reactive substances
- TFP
trifluoperazine 相似文献
158.
159.
It is unlikely that the intact or commonly preserved varieties of Ordovician-Silurian nautiloid shells were able to drift for any distance at the surface of the sea even if they died there. Their cameral capacity was much larger than the volume of the extracted or decayed body, and it would have contained a partial vacuum and cameral liquid when they were alive. The closely spaced and thin septa of the shallow-water adapted species were liable to buckle in compression and then implode in local tension during reverse hydrostatic loading by water pressure. This reverse loading and internal implosion of the septa was probably initiated by the sudden cameral refilling of an apical chamber caused by the depositional rupture of the apical siphuncle at or near the maximum habitat depth of these species. The instantaneous buckling of the more adorai septa was potentially terminated by variations in the septum thickness and cameral fill-fractions at that time, and they imply that some of the Silurian nautiloids from Bohemia were deposited at a minimum depth of about 65 m. Alternative interpretations involving the breakage of the same septa in tension, or buckling due to the difference in pressure between adjacent flooded chambers, set a maximum depth limit of about 160 m for the same facies. Many of the smaller Silurian nautiloids were unlikely to buckle during refilling, and they were potentially flooded faster than they could sink, below a depth of 100–300 m. 相似文献
160.
For the first time a complete ostracode and trilobite fauna is described from the Dobrovitá Formation (Ordovician) of Perunica. In contrast to the trilobite fauna the ostracode fauna evidences close relations to both Armorica and Baltica. The trilobite fauna comprises 11 species out of 11 genera, the ostracode fauna comprises 15 (5 new) species out of 15 (1 new) genera. 相似文献