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  • phosphorus  (73)
  • RAPD  (55)
  • apoptosis  (52)
  • Springer  (180)
  • American Chemical Society
  • American Meteorological Society
  • 1995-1999  (180)
  • 1990-1994
  • 1955-1959
  • 1999  (180)
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  • Springer  (180)
  • American Chemical Society
  • American Meteorological Society
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  • 1995-1999  (180)
  • 1990-1994
  • 1955-1959
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  • 1
    Electronic Resource
    Electronic Resource
    Molecular markers to differentiate two morphologically-close species of the genus Sitobion (1999)
    Springer
    Entomologia experimentalis et applicata 92 (1999), S. 217-225 
    Details
    ISSN: 1570-7458
    Keywords: Sitobion avenae ; Sitobion fragariae ; RAPD ; PCR ; microsatellites ; mtDNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A set of molecular markers to differentiate the aphid (Hemiptera: Aphidoidea) species Sitobion avenae (Fabricius) from Sitobion fragariae (Walker), is presented. These markers correspond to (1) a region of the mitochondrial DNA, (2) five species-specific RAPD banding patterns and (3) four microsatellite loci. Each of the markers was able to clearly distinguish between the species. The utility of each molecular marker is discussed. Mitochondrial DNA is best applicable to species determination and relative abundance, RAPDs to the evaluation of genetic diversity, and microsatellites to the assessment of the population genetic structure; the combined use of mtDNA with the other techniques can be of importance when the presence of hybrids is suspected, and RAPDs with microsatellites are best used together in population genetics and host preference studies.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1003855919065
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  • 2
    Electronic Resource
    Electronic Resource
    Ni–Zn–P alloy deposition from sulfate bath: inhibitory effect of zinc (1999)
    Springer
    Journal of applied electrochemistry 29 (1999), S. 1171-1176 
    Details
    ISSN: 1572-8838
    Keywords: alloys ; cyclic voltammetry ; electrodeposition ; electroless deposition ; nickel ; phosphorus ; zinc
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Electrical Engineering, Measurement and Control Technology
    Notes: Abstract Electroless Ni–Zn–P alloy deposition from a sulphate bath, containing sodium hypophosphite as reducer, was investigated. To increase the plating rate, the deposition parameters were optimized. The effect of process parameters (T, pH and [Zn2+]) on the plating rate and deposit composition was examined and it was found that the presence of zinc in the bath has an inhibitory effect on the alloy deposition. As a consequence, the percentage of zinc in the electroless Ni–Zn–P alloys never reaches high values. Using cyclic voltammetry the electrodeposition mechanism of Ni–Zn–P alloys was investigated. It was observed that the zinc deposition inhibits the nickel discharge and, as a consequence, its catalytic activity on hypophosphite oxidation. It was also found that increase in temperature or pH leads to the deposition of nickel rich alloys.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1003426904395
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  • 3
    Electronic Resource
    Electronic Resource
    Microstructural characterization and corrosion resistance of Ni–Zn–P alloys electrolessly deposited from a sulphate bath (1999)
    Springer
    Journal of applied electrochemistry 29 (1999), S. 637-645 
    Details
    ISSN: 1572-8838
    Keywords: alloys ; electroless ; microstructure ; morphology ; nickel ; phosphorus ; zinc
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Electrical Engineering, Measurement and Control Technology
    Notes: Abstract Electroless Ni–Zn–P alloy coatings were obtained on an iron substrate from a sulfate bath at various pH values. The effects of changes in bath pH on alloy composition, morphology, microstructure and corrosion resistance were studied. Scanning electron microscopy was performed to observe the morphological change of the deposits with bath pH. Coating crystallinity was investigated by grazing incidence asymmetric Bragg X-ray diffraction and transmission electron microscopy. A transition from an amorphous to polycrystalline structure was observed on increasing the bath alkalinity, and thus decreasing the phosphorus content of the alloys. A single crystalline phase corresponding to face-centred-cubic nickel was identified in the alloys obtained from a strong alkaline solution. An increase in zinc percentage up to 23% in the deposits does not change the f.c.c. nickel crystalline structure. Corrosion potential and polarization resistance measurements indicated that the corrosion resistance of electroless Ni–Zn–P alloys depends strongly on the microstructure and chemical composition. The deposits obtained at pH 9.0–9.5 and with 11.4–12.5% zinc and 11.8–11.2% phosphorous exhibited the best corrosion resistance.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1026441403282
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  • 4
    Electronic Resource
    Electronic Resource
    Apoptosis and p53 gene expression in male reproductive tissues of cadmium exposed rats (1999)
    Springer
    BioMetals 12 (1999), S. 131-139 
    Details
    ISSN: 1572-8773
    Keywords: cadmium ; apoptosis ; RT-PCR ; p53 gene expression ; testes ; rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Reverse transcription (RT) PCR technique was used to investigate the mechanism of apoptosis induced by Cd and the change of its related genes in testes and prostate of rats. Adult male rats were given a single (s.c.) injection of CdC l2 0, 2.5, 5.0, 10 μmol/kg. 48 h and 72 h after administration of Cd, animals were sacrificed. The results indicated that Cd can induce apoptosis in testes via p53-independent pathway. No apoptosis occurred in prostate in any of the Cd-exposed groups. There was a clearly negative relationship in testes between p53 gene expression and Cd exposure and this dose-response relationship was observed both at 48 h and 72 h. There was a very small increase of this gene expression in the dorsolateral lobe of the prostate in Cd exposed groups. The other apoptosis related gene, bcl-x, was not detectable in either control or Cd-exposed group in testes and dorsal prostate. Although the MT-I gene was expressed in testes or dorsal prostate both in control and exposed groups, no overexpression of MT-I gene was found after administration of Cd . The expression of MT-I in the ventral prostate was not detected in the control group, but a weak expression was found after Cd exposure. Since p53 is a tumo r suppressor gene which can inhibit tumorigenesis, the consequence of a Cd-induced decrease of p53 in testes may have a relation to the known risk of Cd tumorigenesis in this tissue.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1009273711068
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  • 5
    Electronic Resource
    Electronic Resource
    Effects of repeated manure and fertilizer phosphorus additions on soil phosphorus dynamics under a soybean-wheat rotation (1999)
    Springer
    Biology and fertility of soils 28 (1999), S. 150-155 
    Details
    ISSN: 1432-0789
    Keywords: Key words Phosphorus dynamics ; Olsen ; phosphorus ; Soil phosphorus fractions ; Manure ; Soybean-wheat rotation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract  Soil P availability and efficiency of applied P may be improved through an understanding of soil P dynamics in relation to management practices in a cropping system. Our objectives in this study were to evaluate changes in plant-available (Olsen) P and in different inorganic P (Pi) and organic P (P0) fractions in soil as related to repeated additions of manure and fertilizer P under a soybean-wheat rotation. A field experiment on a Typic Haplustert was conducted from 1992 to 1995 wherein the annual treatments included four rates of fertilizer P (0, 11, 22 and 44 kg ha–1 applied to both soybean and wheat) in the absence and presence of 16 t ha–1 of manure (applied to soybean only). With regular application of fertilizer P to each crop the level of Olsen P increased significantly and linearly through the years in both manured and unmanured plots. The mean P balance required to raise Olsen P by 1 mg kg–1 was 17.9 kg ha–1 of fertilizer P in unmanured plots and 5.6 kg ha–1 of manure plus fertilizer P in manured plots. The relative sizes of labile [NaHCO3-extractable Pi (NaHCO3-Pi) and NaHCO3-extractable P0 (NaHCO3-P0)], moderately labile [NaOH-extractable Pi (NaOH-Pi) and NaOH-extractable P0 (NaOH-P0)] and stable [HCl-extractable P (HCl-P) and H2SO4/H2O2-extractable P (resisual-P)] P pools were in a 1 : 2.9 : 7.6 ratio. Application of fertilizer P and manure significantly increased NaHCO3-Pi and -P0 and NaOH-Pi, and -P0 fractions and also total P. However, HCl-P and residual-P were not affected. The changes in NaHCO3-Pi, NaOH-Pi and NaOH-P0 fractions were significantly correlated with the apparent P balance and were thought to represent biologically dynamic soil P and act as major sources and sinks of plant-available P.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s003740050477
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  • 6
    Electronic Resource
    Electronic Resource
    The genetic heterogeneity of deer mouse populations (Peromyscus maniculatus) in an insular landscape (1999)
    Springer
    Population ecology 41 (1999), S. 263-268 
    Details
    ISSN: 1437-5613
    Keywords: Key words AMOVA ; Dispersion ; Gene flow ; Genetic distance ; HOMOVA ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A survey of the genetic variability in deer mouse populations was performed using specimens collected from six different islands on a lake covering approximately 50 km2. Random amplified polymorphic DNA (RAPD) was used to measure the extent of the genetic differences in this insular system. An analysis of molecular variance (AMOVA) revealed that populations are clearly separated at this microgeographic scale (F st = 0.13863; P 〈 0.001). The homogeneity of molecular variance test (HOMOVA) indicated that within-population levels vary greatly (B p = 0.76831; P 〈 0.001). The within-population molecular variance was found to be mainly correlated with the accessibility of the islands, computed as the inverse of the geographic distance separating an island from the lakeshore (r = 0.916; P 〈 0.003).
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s101440050030
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  • 7
    Electronic Resource
    Electronic Resource
    Rapid Construction of a Random Genomic Library from Date-Palm (Phoenix Dactylifera L.) (1999)
    Springer
    Plant molecular biology reporter 17 (1999), S. 409-409 
    Details
    ISSN: 1572-9818
    Keywords: Date-palm ; DNA library ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A random genomic library of Tunisian date-palm varieties has been built from total cellular DNA, previously amplified according to an RAPD procedure. The resultant recombinant DNA is characterised by a size ranging from 200 to 1600 bp inserts. This DNA would constitute a large number of anonymous probes useful in Southern hybridisation experiments. It would also provide potential markers aimed at the molecular characterisation of date-palm varieties, aid the search of those associated with bayoud disease and suggest a sex determination of trees.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1007631420875
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  • 8
    Electronic Resource
    Electronic Resource
    Stable isotope (δ13C and δ15N) signatures of sedimented organic matter as indicators of historic lake trophic state (1999)
    Springer
    Journal of paleolimnology 22 (1999), S. 205-221 
    Details
    ISSN: 1573-0417
    Keywords: carbon isotopes ; diatoms ; lake management ; nitrogen isotopes ; phosphorus ; radium-226 ; sediments ; trophic state
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences
    Notes: Abstract We explored the use of carbon and nitrogen isotopes (δ13C and δ15N) in sedimented organic matter (OM) as proxy indicators of trophic state change in Florida lakes. Stable isotope data from four 210Pb-dated sediment cores were compared stratigraphically with established proxies for historical trophic state (diatom-inferred limnetic total phosphorus, sediment C/N ratio) and indicators of cultural disturbance (sediment total P and 226Ra activity). Diatom-based limnetic total P inferences indicate a transition from oligo-mesotrophy to meso-eutrophy in Clear Lake, and from eutrophy to hypereutrophy in Lakes Parker, Hollingsworth and Griffin. In cores from all four lakes, the carbon isotopic signature of accumulated OM generally tracks trophic state inferences and cultural impact assessments based on other variables. Oldest sediments in the records yield lower diatom-inferred total limnetic P concentrations and display relatively low δ13C values. In the Clear, Hollingsworth and Parker records, diatom-inferred nutrient concentrations increase after ca. AD 1900, and are associated stratigraphically with higher δ13C values in sediment OM. In the Lake Griffin core, both proxies display slight increases before ~1900, but highest values occur over the last ~100 years. As Lakes Clear, Hollingsworth and Parker became increasingly nutrient-enriched over the past century, the δ15N of sedimented organic matter decreased. This reflects, in part, the increasing relative contribution of nitrogen-fixing cyanobacteria to sedimented organic matter as primary productivity increased in these waterbodies. The Lake Griffin core displays a narrow range of both δ13C and δ15N values. Despite the complexity of carbon and nitrogen cycles in lakes, stratigraphic agreement between diatom-inferred changes in limnetic total P and the stable isotope signatures of sedimented OM suggests that δ13C and δ15N reflect shifts in historic lake trophic state.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1008078222806
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  • 9
    Electronic Resource
    Electronic Resource
    Calibration of diatoms along a nutrient gradient in Florida Everglades Water Conservation Area-2A, USA (1999)
    Springer
    Journal of paleolimnology 22 (1999), S. 413-437 
    Details
    ISSN: 1573-0417
    Keywords: diatoms ; Everglades ; phosphorus ; wetland ; calibration ; multivariate ; Florida
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences
    Notes: Abstract The relationship between diatom taxa preserved in surface soils and environmental variables at 31 sites in Water Conservation Area 2A (WCA-2A) of the Florida Everglades was explored using multivariate analyses. Surface soils were collected along a phosphorus (P) gradient and analyzed for diatoms, total P, % nitrogen (N), %carbon (C), calcium (Ca), and biogenic silica (BSi). Phosphorus varied from 315-1781 μg g-1, and was not found to be correlated with the other geochemical variables. Canonical correspondence analysis (CCA) was used to examine which environmental variables correlated most closely with the distributions in diatom taxa. Canonical correspondence analysis with forward selection, constrained and partial CCA, and Monte Carlo permutation tests of significance show the most significant changes in diatom assemblages along the P gradient (p 〈 0.01), with additional species differences correlated with soil C, N, Ca, and BSi. Weighted-averaging (WA) regression and calibration models of diatom assemblages to P and BSi were developed. The diatom-based inference model for soil [P] had a high apparent r2 (0.86) with RMSEboot = 218 μg g-1. Indicator diatom species identified by assessing species WA optima and WA tolerance to [P], such as Nitzschia amphibia and N. palea for high [P] (~1300-1400 μ g-1) and Achnanthes minutissima var. scotica and Mastogloia smithii for low [P] (~400-600 μg g-1), may be useful as monitoring tools for eutrophication in WCA-2A as well as other areas of the Everglades. Diatom assemblages analyzed by cluster analysis were related to location within WCA-2A, and dominant taxa within clusters are discussed in relation to the geochemical variables measured as well as hydrology and pH. Diversity of diatom assemblages and a ‘Disturbance Index’ based on diatom data are discussed in relation to the historically P-limited Everglades ecosystem. Diatom assemblages should be very useful for reconstructions of [P] through time in the Florida Everglades, provided diatoms are well preserved in soil cores.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1008049224045
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  • 10
    Electronic Resource
    Electronic Resource
    Controlling soil water content in fertiliser dissolution experiments (1999)
    Springer
    Nutrient cycling in agroecosystems 55 (1999), S. 7-14 
    Details
    ISSN: 1573-0867
    Keywords: fertiliser formulation ; nutrients ; phosphorus ; relative humidity ; soil moisture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Phosphorus lost in runoff from agricultural land leads to the enrichment of surface waters and contributes to algal blooms. Fertilisers are one source of this P. To compare the water available P of different fertiliser formulations in the laboratory it is necessary to control environmental conditions, temperature, relative humidity and soil water content, prior to simulating rainfall. Two chambers were designed in which relative humidity and soil water content were controlled using salt solutions. An initial design comprising a sealed chamber with three layers of soil samples over a salt bath was found to be inferior to a single layer design. The changes in water content of soil samples were used to test the single layer chamber in a constant temperature environment (15 °C) using a saturated KCl solution (90% relative humidity). Based on the final soil water content of the samples, the spatial variation within the chamber was within tolerable limits. The single layer chamber was used for a simulation experiment comparing the water available P of two commercial fertilisers. Using a saturated resorcinol solution (95% relative humidity) soil samples were equilibrated at 15 °C for 21 days, fertiliser added, and the water available P measured up to 600 h after fertiliser application. The results indicate that the amount of water available P was related to the fertiliser compound and exponentially related to the time since fertiliser application. It was concluded that the single layer chamber is suitable for controlling relative humidity and soil water content in trials such as these where the water available P of fertilisers are being compared.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1009816716466
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  • 11
    Electronic Resource
    Electronic Resource
    Transfers of phosphorus within three dairy farming systems receiving varying inputs in feeds and fertilizers (1999)
    Springer
    Nutrient cycling in agroecosystems 55 (1999), S. 63-75 
    Details
    ISSN: 1573-0867
    Keywords: dairy systems ; feeds ; fertilizers ; phosphorus ; P surplus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Inputs of phosphorus (P) above requirements for production on dairy farms lead to surplus P with increased risk of P transfer in land run-off to surface waters causing eutrophication. The impact of reducing surplus P inputs in purchased feeds and fertilizers on milk and forage production was investigated in a comparison of three dairy farm systems on chalkland soils in southern England over a 3-year period. In accordance with current commercial practice, no attempt was made to regulate P inputs in system 1, which accumulated an average annual surplus of 23 kg P ha-1. Progressive reductions in purchased feed and/or fertilizer inputs into systems 2 and 3 decreased surplus P to 17 and 3 kg ha-1, respectively, without apparently limiting either milk or herbage dry matter production. The estimated reduction in faecal P output from system 3 cows fed a low P diet compared to system 1 cows fed a high P diet was 26%. Milk P concentrations significantly (P 〈0.001) increased in systems 2 and 3 which included maize in the diet. Output of P in milk and meat products, as a proportion of the total dietary P inputs, increased from 28% in system 1 to 36% in system 3. Surplus P was greatest in continuous maize fields receiving both dairy manure and starter P fertilizer. Withholding P fertilizer in system 3 did not reduce P offtake in cut herbage on soils of moderate P fertility. Total annual losses of P in storm run-off and leaching were no greater than annual inputs of P from the atmosphere (0.5 kg ha-1). The results indicate there is scope to reduce surplus P on commercial dairy farms without sacrificing production targets at least in the short term. Purchased feeds are the largest of the P inputs on intensive dairy farms, yet these are rarely quantified on commercial holdings.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1009860915462
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  • 12
    Electronic Resource
    Electronic Resource
    Applications of poultry litter and triple superphosphate fertilizer to a sandy soil: effects on soil phosphorus status and profile distribution (1999)
    Springer
    Nutrient cycling in agroecosystems 54 (1999), S. 233-242 
    Details
    ISSN: 1573-0867
    Keywords: leaching ; phosphorus ; poultry litter ; soil
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract To determine P loadings, added through poultry litter, sufficient to cause downward movement of P from the cultivated layer of a sandy soil, six rates of poultry litter were applied annually for four years to a site in central England. (total loading 0 – 1119 kg P ha-1). A single extra plot also received an extra 1000 kg ha-1 as triple superphosphate (TSP; total loading 2119 kg P ha-1) and three other treatments received 200 – 800 kg ha-1 P as TSP only. Annual soil sampling in 30-cm increments to 1.5-m depth provided information on P build-up in the topsoil and P movement to depth. There were strong linear trends between P balance (P applied – P removed in crops) and total P, Olsen bicarbonate extractable P and water-soluble P in the topsoil. Phosphorus from TSP and poultry litter fell on the same regression lines, suggesting that both would be equally effective as fertilizer sources. We calculated that 100 kg ha-1 surplus total P would increase the Olsen extractable P content by c. 6 mg kg-1 and the water-soluble P by c. 5 mg kg-1. Thus, relatively large amounts of P would need to be applied to raise soil P status. We found some evidence of P movement into the soil layers immediately below cultivation depth. However, neither soil sampling nor soil solution extracted through Teflon water samplers showed evidence of movement into the deep subsoil (1 m) despite large P loadings.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1009744706679
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  • 13
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    Electronic Resource
    Inorganic phosphorus and manure effects on bahiagrass production on a Spodosol (1999)
    Springer
    Nutrient cycling in agroecosystems 54 (1999), S. 259-266 
    Details
    ISSN: 1573-0867
    Keywords: bahiagrass ; manure ; pasture fertilization ; phosphorus ; phosphorus cycling ; Spodosol
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Bahiagrass (Paspalum notatum Flugge) pasture fertilization recommendations have traditionally been based upon clipping studies. Inclusion of P from manure, not originally considered when P recommendations were developed for pastures, may minimize the need for P fertilization without reducing bahiagrass production or P uptake. The objective of this research was to determine if manure contributes greatly to the P crop nutrient requirement. A 2-year field study utilized a factorial arrangement of 0 and 6.9 Mg air-dried manure ha-1 with 0, 17, 34, 51, and 68 kg inorganic P ha-1 from triple superphosphate to evaluate bahiagrass yield, root distribution, and P uptake response on a Myakka fine sand (sandy, siliceous, hyperthermic Aeric Alaquod). Because air-dried manure was used in the field study, a greenhouse study was employed to confirm that there were no differences in bahiagrass yield or P uptake from either air-dried or fresh cattle (Bos spp.) manure sources. There were no manure or manure by P interaction effects on yield or P uptake of bahiagrass indicating that manure source did not effect grass production in the greenhouse. In the field study, bahiagrass roots were distributed into the Bh horizon, and the Bh horizon had at least four times more Mehlich-1 extractable P than that of the Ap horizon. This horizon was most likely acting as a main source for P-uptake by the grass. This observation was further confirmed by no yield response to levels of inorganic P application in 1989. A linear-response-and-plateau (R2=0.196) relationship with a critical point of 15.4 kg P ha-1 was found in 1990. Bahiagrass yield and P uptake were not dependent on P fertilization, either from manure or inorganic P, due to the availability of P from the Bh horizon.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1009831011649
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  • 14
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    Isolation and PCR Amplification of Genomic DNA from Market Samples of Dry Tea (1999)
    Springer
    Plant molecular biology reporter 17 (1999), S. 171-178 
    Details
    ISSN: 1572-9818
    Keywords: Camellia sinensis ; DNA isolation ; PCR ; RAPD ; Tea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A simple procedure for DNA isolation from processed dried commercial samples of tea is described. The method involves a modified CTAB procedure employing extensive washing, use of 1% PVP to remove polyphenolics and a single phenol:chloroform extraction step. The average yield ranges from 164–494 μg/g tea sample for various market samples. The DNA obtained from 11 different brands of tea using this procedure were consistently amplifiable (using both RAPD primers as well as defined sequences as primers) and digestible with restriction endonucleases.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1007562802361
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  • 15
    Electronic Resource
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    Somaclonal variation in cryopreserved embryogenic clones of white spruce [Picea glauca (Moench) Voss.] (1999)
    Springer
    Plant cell reports 18 (1999), S. 948-953 
    Details
    ISSN: 1432-203X
    Keywords: Key words Somaclonal variation ; Picea glauca ; RAPD ; Somatic embryogenesis ; Cryopreservation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Trees were regenerated from six white spruce embryogenic clones after cryopreservation for 3 and 4 years, respectively. Genetic stability was evaluated using randomly amplified polymorphic DNA (RAPD) fingerprints. Somaclonal variation was detected in some in vitro embryogenic cultures 2 and 12 months after they were re-established following cryopreservation but not in the corresponding regenerated trees. These results suggest that trees regenerated from cryopreserved cultures in subsequent years are primarily genetically stable in the genomic regions tested and that variation observed due to the in vitro culture process infrequently affects trees regenerated from normally maturing and germinating somatic embryos. However, trees regenerated from somatic embryos that matured or germinated abnormally in in vitro culture exhibited altered RAPD fragment patterns.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002990050689
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  • 16
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    Detection of somaclonal variation in garlic (Allium sativum L.) using RAPD and cytological analysis (1999)
    Springer
    Plant cell reports 18 (1999), S. 473-477 
    Details
    ISSN: 1432-203X
    Keywords: Key wordsAllium sativum ; Garlic ; Genetic instability ; RAPD ; Somaclonal variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Plants were regenerated by somatic embryogenesis from long-term callus cultures derived from five garlic (Allium sativum L.) cultivars. Thirty-five of these plants were subjected to RAPD analysis. The frequency of variation was found to be cultivar dependent: approximately 1% in the two clones Solent White and California Late and around 0.35% in another three clones, Chinese, Long Keeper and Madena. Certain band changes were found in regenerants of different cultivars, suggesting the existence of a mutation-sensitive part of the garlic genome. The karyotypes of another 75 regenerants derived from the same callus cultures of three parental garlic clones were examined. Of these plants, 9.3% were found to be tetraploids, 4% aneuploid and 2.6% showed a change in the position of the secondary constriction. No association could be shown between the rate of variation for molecular and cytological characters either by comparing cultivars or examining individual regenerants.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002990050606
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  • 17
    Electronic Resource
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    Regulation of cardiomyocyte apoptosis in ischemic reperfused mouse heart by glutathione peroxidase (1999)
    Springer
    Molecular and cellular biochemistry 196 (1999), S. 13-21 
    Details
    ISSN: 1573-4919
    Keywords: apoptosis ; DNA fragmentation ; GSHPx-1 knockout mice ; GSHPx-1 transgenic mice ; ischemia/repurfusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Apoptosis, a genetically controlled programmed cell death, has been found to play a role in ischemic reperfusion injury in several animal species including rats and rabbits. To examine whether this is also true for other animals, an isolated perfused mouse heart was subjected to 30 min of ischemia followed by 2 h of reperfusion. Experiments were terminated before ischemia (baseline), after ischemia, and at 30, 60, 90 and 120 min of reperfusion. At the end of each experiment, hearts were processed for the evaluation of apoptosis and DNA laddering. The in situ end labeling (ISEL) technique was used to detect apoptotic cardiomyocyte nuclei while DNA laddering was evaluated by subjecting the DNA obtained from the cardiomyocytes to 1.8% agarose gel electrophoresis followed by photographing under UV illumination. The results of our study revealed that apoptotic cells appear only after 60 min of reperfusion as demonstrated by the intense fluorescence of the immunostained genomic DNA when observed under fluorescence microscopy. None of the ischemic hearts showed any evidence of apoptosis. These results were corroborated with the findings of DNA fragmentation showing increased ladders of DNA bands in the same reperfused hearts representing integer multiples of the internucleosomal DNA length (about 180 bp). Since our previous studies showed a role of glutathione peroxidase (GSHPx) in apoptotic cell death, we performed identical experiments using isolated hearts from GSHPx-l knockout mice and transgenic mice overexpressing GSHPx-l. GSHPx-l knockout mice showed evidence of apoptotic cell death even after 30 min of reperfusion. Significant number of apoptotic cells were found in the cardiomyocytes as compared to non-transgenic control animals. To the contrary, very few apoptotic cells were found in the hearts of the transgenic mice overexpressing GSHPx-l. Hearts of GSHPx-l knockout mice were more susceptible to ischemia/reperfusion injury while transgenic mice overexpressing GSHPx- 1 were less susceptible to ischemia reperfusion injury compared to non-transgenic control animals. The results of this study clearly demonstrate a role of GSHPx in ischemia/reperfusion-induced apoptosis in mouse heart.
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    URL: http://dx.doi.org/10.1023/A:1006905910140
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    Human DU145 prostate cancer cells overexpressing mitogen-activated protein kinase phosphatase-1 are resistant to Fas ligand-induced mitochondrial perturbations and cellular apoptosis (1999)
    Springer
    Molecular and cellular biochemistry 199 (1999), S. 169-178 
    Details
    ISSN: 1573-4919
    Keywords: MKP-1 ; Fas ligand ; Fas ; apoptosis ; prostate cancer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Recent studies have suggested that MAP kinase phosphatase 1 (MKP-1) is overexpressed in prostate cancer. To evaluate the role of MKP-1 in regulating cell death and tumor growth in prostate cancer, MKP-1 was conditionally overexpressed in the human prostate cancer cell line DU145. Overexpression of MKP-1 in DU145 cells blocked activation of stress-activated protein kinase (SAPK/JNK). MKP-1 overexpression in DU-145 cells was also found to inhibit Fas ligand (FasL)-induced apoptosis, as well as block the activation of caspases by Fas engagement. In addition, MKP-1 blocked the activation of apoptosis by transfected MEKK-1 and ASK-1, presumably through its inhibition of the SAPK/JNK family of enzymes. MKP-1 blocked the ability of FasL to induce loss of mitochondrial transmembrane potential (Δγm), suggesting that MKP-1 acts upstream of mitochondrial pro-apoptotic events induced by FasL and that the SAPK/JNK pathway may form the signaling link between Fas receptor and mitochondrial dysfunction. Thus, MKP-1 overexpression in prostate cancer may play a role in promoting prostate carcinogenesis by inhibiting FasL-induced cell death.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1006980326855
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  • 19
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    Poly(ADP-ribosylation) and apoptosis (1999)
    Springer
    Molecular and cellular biochemistry 199 (1999), S. 125-137 
    Details
    ISSN: 1573-4919
    Keywords: apoptosis ; ADP-ribosylation ; caspases ; PARP ; PARG
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Poly(ADP-ribosylation) is a post-translational modification playing a relevant role in DNA damage recovery, DNA replication and viral integration. Several reports also suggest a modulation of this process during cell death by apoptosis. The aim of this review is to discuss the possible involvement of poly(ADP-ribosylation) during apoptosis, by dealing with general considerations on apoptosis, and further examining the correlation between NAD consumption and cell death, the regulation of poly(ADP-ribose) metabolism in apoptotic cells, the effect of poly(ADP-ribose) polymerase inhibition on cell death occurrence and the use of enzyme cleavage as a marker of apoptosis. Finally, the future prospects of the research in this area will be addressed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1006962716377
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  • 20
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    A dual approach in the study of poly (ADP-ribose) polymerase: In vitro random mutagenesis and generation of deficient mice (1999)
    Springer
    Molecular and cellular biochemistry 193 (1999), S. 53-60 
    Details
    ISSN: 1573-4919
    Keywords: DNA binding protein ; NAD metabolism ; cellular response to DNA damage ; γ-rays ; alkylating agents ; genomic instability ; apoptosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract A dual approach to the study of poly (ADP-ribose)polymerase (PARP) in terms of its structure and function has been developed in our laboratory. Random mutagenesis of the DNA binding domain and catalytic domain of the human PARP, has allowed us to identify residues that are crucial for its enzymatic activity. In parallel PARP knock-out mice were generated by inactivation of both alleles by gene targeting. We showed that: (i) they are exquisitely sensitive to γ-irradiation, (ii) they died rapidly from acute radiation toxicity to the small intestine, (iii) they displayed a high genomic instability to γ-irradiation and MNU injection and, (iv) bone marrow cells rapidly underwent apoptosis following MNU treatment, demonstrating that PARP is a survival factor playing an essential and positive role during DNA damage recovery and survival.
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    URL: http://dx.doi.org/10.1023/A:1006947707713
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  • 21
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    Involvement of PARP and poly(ADP-ribosyl)ation in the early stages of apoptosis and DNA replication (1999)
    Springer
    Molecular and cellular biochemistry 193 (1999), S. 137-148 
    Details
    ISSN: 1573-4919
    Keywords: PARP ; poly(ADP-ribosyl)ation ; apoptosis ; DNA replication
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract We have focused on the roles of PARP and poly(ADP-ribosyl)ation early in apoptosis, as well as during the early stages of differentiation-linked DNA replication. In both nuclear processes, a transient burst of PAR synthesis and PARP expression occurs early, prior to internucleosomal DNA cleavage before commitment to apoptosis as well as at the round of DNA replication prior to the onset of terminal differentiation. In intact human osteosarcoma cells undergoing spontaneous apoptosis, both PARP and PAR decreased after this early peak, concomitant with the inactivation and cleavage of PARP by caspase-3 and the onset of substantial DNA and nuclear fragmentation. Whereas 3T3-L1, osteosarcoma cells, and immortalized PARP +/+ fibroblasts exhibited this early burst of PAR synthesis during Fas-mediated apoptosis, neither PARP-depleted 3T3-L1 PARP-antisense cells nor PARP -/- fibroblasts showed this response. Consequently, whereas control cells progressed into apoptosis, as indicated by induction of caspase-3-like PARP-cleavage activity, PARP-antisense cells and PARP -/- fibroblasts did not, indicating a requirement for PARP and poly(ADP-ribosyl)ation of nuclear proteins at an early reversible stage of apoptosis. In parallel experiments, a transient increase in PARP expression and activity were also noted in 3T3-L1 preadipocytes 24 h after induction of differentiation, a stage at which ~95% of the cells were in S-phase, but not in PARP-depleted antisense cells, which were consequently unable to complete the round of DNA replication required for differentiation. PARP, a component of the multiprotein DNA replication complex (MRC) that catalyzes viral DNA replication in vitro, poly(ADP-ribosyl)ates 15 of ~40 MRC proteins, including DNA pol α, DNA topo I, and PCNA. Depletion of endogenous PARP by antisense RNA expression in 3T3-L1 cells results in MRCs devoid of any DNA pol α and DNA pol δ activities. Surprisingly, there was no new expression of PCNA and DNA pol α, as well as the transcription factor E2F-1 in PARP-antisense cells during entry into S-phase, suggesting that PARP may play a role in the expression of these proteins, perhaps by interacting with a site in the promoters for these genes.
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    URL: http://dx.doi.org/10.1023/A:1006988832729
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    Bcl-2, survivin and variant CD44 v7-v10 are downregulated and p53 is upregulated in breast cancer cells by progesterone: Inhibition of cell growth and induction of apoptosis (1999)
    Springer
    Molecular and cellular biochemistry 202 (1999), S. 53-61 
    Details
    ISSN: 1573-4919
    Keywords: breast cancer cells ; anti-apoptotic genes ; apoptosis ; progesterone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Progesterone inhibits the proliferation of normal breast epithelial cells in vivo, as well as breast cancer cells in vitro. But the biologic mechanism of this inhibition remains to be determined. We explored the possibility that an antiproliferative activity of progesterone in breast cancer cell lines is due to its ability to induce apoptosis. Since p53, bcl-2 and survivin genetically control the apoptotic process, we investigated whether or not these genes could be involved in the progesterone-induced apoptosis. We found a maximal 90% inhibition of cell proliferation with T47-D breast cancer cells after exposure to 10 μM progesterone for 72 h. Control progesterone receptor negative MDA-231 cancer cells were unresponsive to 10 μM progesterone. The earliest sign of apoptosis is translocation of phosphatidylserine from the inner to the outer leaflet of the plasma membrane and can be monitored by the calcium-dependent binding of annexin V in conjunction with flow cytometry. After 24 h of exposure to 10 μM progesterone, cytofluorometric analysis of T47-D breast cancer cells indicated 43% were annexin V-positive and had undergone apoptosis and no cells showed signs of cellular necrosis (propidium iodide negative). After 72 h of exposure to 10 μM progesterone, 48% of the cells had undergone apoptosis and 40% were annexin V positive/propidium iodide positive indicating signs of necrosis. Control untreated cancer cells did not undergo apoptosis. Evidence proving apoptosis was also demonstrated by fragmentation of nuclear DNA into multiples of oligonucleosomal fragments. After 24 h of exposure of T47-D cells to either 1 or 10 μM progesterone, we observed a marked down-regulation of protooncogene bcl-2 protein and mRNA levels. mRNA levels of survivin and the metastatic variant CD44 v7-v10 were also downregulated. Progesterone increased p53 mRNA levels. These results demonstrate that progesterone at relative high physiological concentrations, but comparable to those seen in plasma during the third trimester of human pregnancy, exhibited a strong antiproliferative effect on breast cancer cells and induced apoptosis.
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    URL: http://dx.doi.org/10.1023/A:1007081021483
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    Role of nitric oxide in the induction of apoptosis by smokeless tobacco extract (1999)
    Springer
    Molecular and cellular biochemistry 200 (1999), S. 51-57 
    Details
    ISSN: 1573-4919
    Keywords: smokeless tobacco ; apoptosis ; nitric oxide
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Smokeless tobacco usage is, a growing public health concern in the United States. Lesions of the oral cavity have been clearly linked to smokeless tobacco use. The objective of this study was to determine the biochemical effects of smokeless tobacco extract (STE) exposure upon hamster cheek pouch cell (HCPC-1) cultures. HCPC-1 cells were exposed to a 5 -fold dose-range of STE (0.5, 1.0 and 2.5%) over a time-course of 24-96 h. Following each exposure we measured various biochemical parameters of cell proliferation and cell death. Cell viability, cell cycle progression and S-phase DNA synthesis were measured as markers of cell proliferation. We measured lactate dehydrogenase leakage as a marker of cell membrane damage and cell death due to necrosis. No significant alterations were observed in cell cycle progression and cell proliferation as a result of exposure to STE. LDH measured colorimetrically indicated no significant effect with the lower doses (0.5, 1.0 and 2.5% STE). Apoptosis measured as the A0 peak and by the TUNEL procedure revealed that STE caused significant rates of apoptosis. Maximal apoptosis was noted between 48-96 h. In order to probe the mechanism further we measured the levels of nitrites as an indicator of nitric oxide (NO) in the media. NO levels were significantly elevated at the doses that caused an induction of apoptosis. The results from this study indicate that STE causes a dose-dependent induction of apoptosis and that this is mediated by nitric oxide.
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    URL: http://dx.doi.org/10.1023/A:1006985700851
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    Clostridial toxins: Molecular probes of Rho-dependent signaling and apoptosis (1999)
    Springer
    Molecular and cellular biochemistry 193 (1999), S. 37-42 
    Details
    ISSN: 1573-4919
    Keywords: Rho ; GTPase ; toxins ; Clostridium ; signal transduction ; apoptosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The Rho family small GTPases are members of the Ras superfamily of small GTPases. Rho proteins were first determined to act as key regulators of many types of actin cytoskeletal-dependent cellular functions. Recent work by several investigators indicates that Rho GTPases are also critical modulators of several important intracellular and nuclear signal transduction pathways. Certain clostridial toxins and exoenzymes covalently modify, and thereby inactivate, specific types of Rho family GTPases. As such, these microbial enzymes have proven invaluable in helping to identify structural and functional attributes of Rho GTPases.
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    URL: http://dx.doi.org/10.1023/A:1006939505896
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    Functional analysis of poly(ADP-ribose) polymerase in Drosophila melanogaster (1999)
    Springer
    Molecular and cellular biochemistry 193 (1999), S. 103-108 
    Details
    ISSN: 1573-4919
    Keywords: Poly(ADP-ribose) polymerase ; Drosophila melanogaster ; alternative splicing ; apoptosis ; DNA repair ; development
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Poly(ADP-ribose) polymerase (PARP) is conserved in eukaryotes. To analyze the function of PARP, we isolated and characterized the gene for PARP in Drosophila melanogaster. The PARP gene consisted of six translatable exons and spanned more than 50 kb. The DNA binding domain is encoded by exons 1-4. Although the consensus cleavage site of CED-3 like protease during apoptosis is conserved from human to Xenopus laevis PARPs, it is neither conserved in the corresponding region of Drosophila nor Sarcophaga peregrina. There are two cDNAs species in Drosophila. One cDNA could encode the full length PARP protein (PARP I), while the other is a truncated cDNA which could encode a partial-length PARP protein (PARP II), which lacks the automodification domain and is possibly produced by alternative splicing. The expression of these two forms of PARP in E. coli demonstrated that while PARP II has the catalytic NAD-binding domain and DNA-binding domain it is enzymatically inactive. On the other hand PARP I is active. A deletion mutant of PARP gene could grow to the end of embryogenesis but did not grow to the adult fly. These results suggest that the PARP gene plays an important function during the development of Drosophila.
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    URL: http://dx.doi.org/10.1023/A:1006920429095
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    Newly discovered anti-inflammatory properties of the benzamides and nicotinamides (1999)
    Springer
    Molecular and cellular biochemistry 193 (1999), S. 119-125 
    Details
    ISSN: 1573-4919
    Keywords: benzamides ; nicotinamides ; apoptosis ; inflammation ; NF-kB ; DNA repair
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Our laboratory has concentrated on the possible regulation the benzamides and nicotinamides may have on the processes of DNA repair and apoptosis. Recent reports [14-16] have suggested that both apoptosis and inflammation are regulated by the transcription factor NF-kB. We have initiated studies regarding the hypothesis that the benzamides and nicotinamides could inhibit the production of tumor necrosis factor alpha (TNFalpha) and the inflammatory response as well as induce apoptosis via inhibition of NF-kB. Our data have shown that nicotinamide and two N-substituted benzamides, metoclopramide (MCA) and 3-chloroprocainamide (3-CPA), gave dose dependent inhibition of lipopolysacharide induced TNFalpha in the mouse within the dose range of 10-500 mg/kg. Moreover, lung edema was prevented in the rat by 3 ï 50 mg/kg doses of 3-CPA or MCA, and 100-200 μM doses of MCA could also inhibit NF-kB in Hela cells. Taken together these data strongly support the notion that benzamides and nicotinamides have potent anti-inflammatory and antitumor properties, because their primary mechanism of action is regulated by inhibition at the gene transcription level of NF-kB, which in turn inhibits TNFalpha and induces apoptosis.
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    URL: http://dx.doi.org/10.1023/A:1006932714982
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    Growth arrest and induction of apoptosis in breast cancer cells by antisense depletion of protein kinase A-RIα subunit: p53-independent mechanism of action (1999)
    Springer
    Molecular and cellular biochemistry 195 (1999), S. 25-36 
    Details
    ISSN: 1573-4919
    Keywords: antisense oligonucleotide ; apoptosis ; cAMP-dependent protein kinase ; cancer cells ; growth inhibition
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The enhanced expression of the RIα subunit of cyclic AMP-dependent protein kinase type 1 (PKA-I) has been correlated with cancer cell growth. We have investigated the effects of sequence-specific inhibition of RIα gene expression on the growth of MCF-7 human breast cancer cells. We report that RIα antisense treatment results in a reduction in RIα expression at both mRNA and protein levels and inhibition of cell growth. The growth inhibition was accompanied by changes in cell morphology, cleavage of poly(ADP-ribose) polymerase (PARP) and appearance of apoptotic nuclei. In addition, bcl-2 protein level was reduced and p53 expression increased in growth arrested cells. Interestingly, RIα antisense inhibited cell viability and induced apoptosis in the absence of p53, suggesting that these actions of RIα antisense are exerted independent of p53. In contrast, two- and four-base mismatched control oligonucleotides had no effect on either cell growth or morphology. These results demonstrate that the RIα antisense, which efficiently depletes the growth stimulatory molecule RIα, induces cell differentiation and apoptosis, providing a new approach to combat breast cancer cell growth.
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    URL: http://dx.doi.org/10.1023/A:1006990231186
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    TNF-α induced altered signaling mechanism in human neutrophil (1999)
    Springer
    Molecular and cellular biochemistry 197 (1999), S. 97-108 
    Details
    ISSN: 1573-4919
    Keywords: neutrophil ; PKC ; TNF-α ; apoptosis ; DNA fragmentation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract In the present study we investigated the TNF-α induced signal transduction mechanism in human neutrophil. Exogenously added TNF-α affects both PKC activity and its translocation from cytosol to the membrane. Endogenous protein phosphorylation pattern is inhibited in TNF-α induced neutrophil in Ca-dependent and Ca-independent manner, including a major 47 and 66 kDa cytosolic proteins, which may be implicated in superoxide anion generation. However TNF-α dose dependently enhances the expression of ζ-PKC isotype but not the β-PKC. Morphology and cell cytotoxicity are studied in TNF-α treated neutrophil to understand the TNF-α induced cell death or apoptosis and these experiment is further confirmed by DNA fragmentation analysis. These results clearly demonstrate that TNF-α induces cellular death of human neutrophil at least in part by enhanced expression of Ca-independent ζ-PKC. These observations provide an insight towards understanding the function of ζ-PKC in apoptotic pathway.
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    URL: http://dx.doi.org/10.1023/A:1006935114624
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    Diminished energy metabolism and enhanced apoptosis in livers of B6C3F1 mice treated with the antihepatocarcinogen rotenone (1999)
    Springer
    Molecular and cellular biochemistry 201 (1999), S. 25-32 
    Details
    ISSN: 1573-4919
    Keywords: rotenone ; apoptosis ; oncogenes ; liver cancer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Rotenone decreases the incidence of hepatocellular carcinoma and lowers rates of hepatocellular proliferation. In an effort to delineate mechanisms involved, the in vivo effect of rotenone on liver mitochondrial metabolism, apoptotic machinery as well as elements of the hepatic signal transduction pathways were investigated. Mitochondria from livers of male B6C3F1 mice fed a standard diet containing 600 ppm rotenone for 7 days were uncoupled or inhibited when succinate or glutamate plus malate were used as the substrate, respectively. These livers also showed a significant increase in apoptosis compared with control livers. Furthermore, rotenone increased the expression of c-myc mRNA to 5-fold of control values within 3 days, an effect which was still observed (3-fold) after 7 days. Levels of p53 mRNA were also increased 3-fold after 1 day, but declined to control levels by 7 days. Rotenone also caused a transient, yet marked increase in liver particulate glyceraldehyde phosphate dehydrogenase (GAPDH) protein expression, while it did not alter the expression of the cytosolic form of the enzyme. Conversely, mRNA of the proto-oncogene H-ras showed a decline of 35% after 3 days of rotenone treatment, and remained diminished for the duration of the experiment. These data suggest that rotenone may act as an anticancer agent by diminishing mitochondrial bioenergetics which prevents basal hepatocyte proliferation and lowers the threshold for liver cells with DNA damage to undergo apoptosis.
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    URL: http://dx.doi.org/10.1023/A:1007024905046
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    Preparative High-Resolution Two-Dimensional Electrophoresis Enables the Identification of RNA Polymerase B Transcription Factor 3 as an Apoptosis-Associated Protein in the Human BL60–2 Burkitt Lymphoma Cell Line (1999)
    Springer
    The protein journal 18 (1999), S. 225-231 
    Details
    ISSN: 1573-4943
    Keywords: Two-dimensional electrophoresis ; MALDI-MS ; apoptosis ; RNA polymerase B transcription factor 3
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Apoptosis or programmed cell death is essential in the process of controlling lymphocyte growth and selection. We identified RNA polymerase B transcription factor 3 (BTF3), which is associated with anti-IgM antibody-mediated apoptosis, using a subclone of the human Burkitt lymphoma cell line BL60. To identify the transcription factor BTF3, which is expressed only in minor amounts, we used preparative high-resolution two-dimensional gel electrophoresis (2DE) employing carrier ampholytes for isoelectric focusing. Comparison of the 2DE protein patterns from apoptotic and nonapoptotic cells showed BTF3 as a predominantly altered protein spot. The characterization of the differentially expressed transcription factor and 13 marker proteins described in this study were performed by internal Edman microsequencing and/or by peptide mass fingerprinting using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The proteome analysis was significantly improved by performing the newly developed preparative high-resolution two-dimensional gels employing high protein concentrations.
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    URL: http://dx.doi.org/10.1023/A:1020636308270
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    Genetics of resistance to ascochyta blight (Ascochyta lentis) of lentil and the identification of closely linked RAPD markers (1999)
    Springer
    Theoretical and applied genetics 98 (1999), S. 93-98 
    Details
    ISSN: 1432-2242
    Keywords: Key words Ascochyta lentis ; Lens culinaris ssp. culinaris ; Bulked segregant analysis ; Resistance genes ; RAPD ; QTL analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Foliar resistance to Ascochyta lentis is controlled at a single major locus by a dominant gene (AbR 1 ) in the lentil accession ILL5588 (cv ‘Northfield’). Flanking RAPD markers that are closely linked to the resistance locus in coupling phase were identified by bulked segregant analysis. Out of 261 decanucleotide primers screened 7 produced a polymorphic marker that segregated with the resistance locus, and all markers were found to exist within a single linkage group. Five of the seven RAPD markers were within 30 cM of the resistance locus. Log likelihood analysis for detecting QTL associated with the foliar resistance revealed that a single narrow peak accounted for almost 90% of the variance of resistance between the bulks. Preliminary mapping in an F3 population revealed that the closest flanking markers were approximately 6 and 14 centiMorgans (cM) away from the resistance locus. These markers should be useful for the discrimination of resistant germplasm through marker-assisted selection in future breeding programmes and represent the first essential step towards the map-based cloning of this resistance gene.
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    URL: http://dx.doi.org/10.1007/s001220051044
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    Linkage relationships among molecular markers and storage root traits of carrot (Daucus carota L. ssp. sativus) (1999)
    Springer
    Theoretical and applied genetics 99 (1999), S. 58-64 
    Details
    ISSN: 1432-2242
    Keywords: Key words Genetic map ; RFLP ; AFLP ; RAPD ; SAMPL ; Daucus carota L. ssp. sativus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  A 109-point linkage map consisting of three phenotypic loci (P 1, Y 2, and Rs), six restriction fragment length polymorphisms (RFLPs), two random amplified polymorphic DNAs (RAPDs), 96 amplified fragment length polymorphisms (AFLPs), and two selective amplification of microsatellite polymorphic loci (SAMPL) was constructed for carrot (Daucus carota L. ssp. sativus; 2n=2x=18). The incidence of polymorphism was 36% for RFLP probes, 20% for RAPD primers, and 42% for AFLP primers. The overall incidence of disturbed segregation was 18%. Linkage relationships at a LOD score of 4.0 and θ=0.25 indicated 11 linkage groups. The total map length was 534.4 cM and the map was clearly unsaturated with markers spaced at 4.9 cM. AFLP P6B15 was 1.7 cM from P 1, AFLP P1B34 was 2.2 cM from Y 2, and AFLP P3B30XA was 8.1 cM from Rs.
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    URL: http://dx.doi.org/10.1007/s001220051208
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    Random amplified polymorphic DNA (RAPD) analysis reveals genetic relationships among the annual Cicer species (1999)
    Springer
    Theoretical and applied genetics 98 (1999), S. 657-663 
    Details
    ISSN: 1432-2242
    Keywords: Key words Cicer ; Species relationships ; DNA fingerprinting ; RAPD ; Chickpea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Random amplified polymorphic DNA markers were used to distinguish between nine different Cicer taxa representing the cultivated chickpea and eight other related annual wild species. Of the 75 random10-mer primers tested, only 8 amplified genomic DNA across all the species. A total of 115 reproducibly scorable RAPD markers were generated, all except 1 polymorphic, and these were utilized to deduce genetic relationships among the annual Cicer species. Four distinct clusters were observed and represented C. arietinum, C. reticulatum and C. echinospermum in first cluster followed by C. chorassanicum and C. yamashitae in the second cluster, while C. pinnatifidum, C. judaicum and C. bijugum formed the third cluster. Cicer cuneatum did not cluster with any of the species and was most distantly placed from the cultivated species. Except for the placement of C. chorassanicum and C. yamashitae, deduced species’ relationships agreed with previous studies. In addition, species-diagnostic amplification products specific to all the nine species were identified. The results clearly demonstrate a methodology based on random-primed DNA amplification that can be used for studying Cicer phylogeny and chickpea improvement.
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    URL: http://dx.doi.org/10.1007/s001220051117
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    Development of a genetic composite map of Vicia faba using F2 populations derived from trisomic plants (1999)
    Springer
    Theoretical and applied genetics 98 (1999), S. 736-743 
    Details
    ISSN: 1432-2242
    Keywords: Key words Vicia faba ; Genetic map ; Trisomics ; RAPD ; Seed-protein genes ; QTLs
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Seven F2 families of faba bean descendent from plants trisomic for chromosomes 3, 4, 5 and 6 were analyzed for isozyme markers and two of these were also studied for morphological and RAPD markers and seed-protein genes. Linkage analysis revealed 14 linkage groups, 8 of which were unambiguously assigned to specific chromosomes. Several QTLs for seed weight were identified, the most important of which, located on chromosome 6, explained approximately 30% of the total phenotypic variation. Comparison of results from Vicia faba with the maps of the related species Pisum sativum L. and Cicer arietinum L. revealed one possible new case of linkage conservation. A composite linkage analysis based on 42 markers analyzed in this and previous studies, where line Vf 6 was also used as the female parental, allowed the new assignment of previously independent linkage groups and/or markers to specific chromosomes. Thus, the number of linkage groups was reduced to 13, each comprising an increased number of markers. No contradictory results were detected, indicating the suitability of the statistical procedure and methodology used so far in the development of the map of this species.
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    Variation among and within Capsicum species revealed by RAPD markers (1999)
    Springer
    Theoretical and applied genetics 99 (1999), S. 147-156 
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    ISSN: 1432-2242
    Keywords: Key words Capsicum ; Diagnostic markers ; Genetic diversity ; Germplasm ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Germplasm characterization is an important link between the conservation and utilization of plant genetic resources. A total of 134 accessions from six Capsicumspecies maintained at the Asian Vegetable Research and Development Center were characterized using 110 randomly amplified polymorphic DNA (RAPD) markers. Ten pairs of potentially duplicated accessions were identified. Multidimensional scaling analysis of the genetic distances among accessions resulted in clustering corresponding to a previous species assignment except for six accessions. Diagnostic RAPDs were identified which discriminate among the Capsicumspecies. The diagnostic markers were employed for improved taxonomic identification of accessions since many morphological traits used in the identification of Capsicumare difficult to score. Three Capsicumaccessions, misclassified based on morphological traits, were reassigned species status based on diagnostic RAPDs. Three accessions, not previously classified, were assigned to a species based on diagnostic RAPDs. Definitive conclusions about the species assignment of three other accessions were not possible. The level of diversity between Capsicum annuumaccessions from the genebank and the breeding program were compared and no differences were observed either for RAPD variation or diversity. The utilization of genetic resources as a source of variance for useful traits in the breeding program may be the reason for the similarity of these two groups.
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    URL: http://dx.doi.org/10.1007/s001220051219
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    Genetic relationships among Native American maize accessions of the Great Plains assessed by RAPDs (1999)
    Springer
    Theoretical and applied genetics 99 (1999), S. 1061-1067 
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    ISSN: 1432-2242
    Keywords: Key words Native American maize ; RAPD ; Genetic relationships ; Reproducibility ; Geography and evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Genetic variation among 15 accessions of Native American maize from the Great Plains was investigated using random amplified polymorphic DNA (RAPD). RAPDs revealed very high levels of polymorphism among accessions. Banding patterns ranged in percentage polymorphism from 46.7% to 86.2% with an overall mean of 70.7% for the primers analyzed. The construction of genetic relationships using cluster analysis and principal coordinates analysis revealed that RAPDs are successful in confirming hypothesized relationships and in identifying misclassified specimens. Furthermore, the phenogram fails to reveal a strong correspondence between genetic relationships and the geographical position of Native Americans prior to contact. This provides support for the hypothesis that multiple introductions of maize into the Great Plains via trade may have resulted in the great morphological variation found among accessions in the region. Based on these data, it is unlikely that a separate Great Plains race of maize can be distinguished. In general, we conclude that RAPDs are potentially very useful in organizing seed collections and understanding intraspecific genetic differentiation.
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    URL: http://dx.doi.org/10.1007/s001220051415
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    High-resolution genetic and physical mapping of the region containing the Bs2 resistance gene of pepper (1999)
    Springer
    Theoretical and applied genetics 99 (1999), S. 1201-1206 
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    ISSN: 1432-2242
    Keywords: Key words Bs2 resistance gene ; Pepper ; RAPD ; AFLP ; Positional cloning
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The Bs2 resistance gene of pepper confers resistance against the bacterial pathogen Xanthomonas campestris pv. vesicatoria. As a first step toward isolation of the Bs2 gene, molecular markers tightly linked to the gene were identified by randomly amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) analysis of near-isogenic lines. Markers flanking the locus were identified and a high-resolution linkage map of the region was developed. One AFLP marker, A2, was found to cosegregate with the locus, while two others, F1 and B3, flank the locus and are within 0.6 cM. Physical mapping of the A2 and F1 markers indicates that these markers may be within 150 kb of each other. Together, these results indicate that the Bs2 region may be cloned either by chromosome walker or landing. The linked markers were also used to characterize gamma-irradiation-induced mutants at the Bs2 locus.
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    URL: http://dx.doi.org/10.1007/s001220051325
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    Optimization of the choice of molecular markers for varietal identification in Vitis vinifera L. (1999)
    Springer
    Theoretical and applied genetics 98 (1999), S. 171-177 
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    ISSN: 1432-2242
    Keywords: Key words Varietal identification ; RAPD ; Microsatellite ; Vitis vinifera L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The aim of this study was to develop a cultivar identification tool based on molecular analysis and a statistical approach. From the PIC parameter we defined the D parameter, which evaluates the efficiency of a primer for the purpose of identification of varieties; i.e. the probability that two randomly chosen individuals have different patterns. D can be used to compare different types of markers even if only the allelic frequencies are known. We used this parameter to develop an algorithm for selecting the optimal combination of primers necessary to identify a set of varieties. The optimal combination of primers determined for a small elite group of varieties applied on a larger set induces a risk of confusion involving 1 of the elite varieties. We estimated the risk of confusion using the D value of each primer of the combination. We applied this methodology on a set of 224 varieties of Vitis vinifera screened with 21 RAPD primers and two microsatellite loci. The discriminating power of the primers did not only depend on the number of patterns it generates but also on the frequencies of the different patterns. A combination of 8 primers (6 RAPD and two microsatellite) was found to be optimum for the discrimination of these 224 varieties. A subset of 38 elite varieties was also investigated. The determined optimal combination of 4 primers (3 RAPD and one microsatellite) applied on the 224 varieties gave 9 risks of confusion involving 1 of the elite varieties. Confusion can happen between varieties with the same origin as well as between varieties of very diverse geographical origins.
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    URL: http://dx.doi.org/10.1007/s001220051054
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    Intertribal hexaploid somatic hybrid plants regeneration from electrofusion between diploids of Citrus sinensis and its sexually incompatible relative, Clausena lansium (1999)
    Springer
    Theoretical and applied genetics 98 (1999), S. 581-585 
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    ISSN: 1432-2242
    Keywords: Key words Somatic hybridization ; Hexaploid ; RAPD ; Chromosome number variation ; Genetic improvement ; Aurantioideae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Chinese wampee [Clausena lansium (Lour.) Skeels], a sexually incompatible relative of citrus, is commercially cultivated in South China. In this study, embryogenic protoplasts of ‘Bonanza’ navel orange [Citrus sinensis (L.) Osbeck] were electrically fused with leaf protoplasts isolated from ‘Chicken Heart’ Chinese wampee. After 8 months of culture, fusion products regenerated into shoots. More than 70% of the shoots unexpectedly rooted well. Chromosome counting of several shoot- and root-tips revealed that their chromosome numbers were not 2n=4x=36 as expected, but 2n=6x=54, suggesting that chromosome doubling occurred rather than chromosome elimination in this intertribal fusion combination. RAPD analysis of embryoids and the leaves of unrooted and rooted shoots verified their hybridity. This is the first report of hexaploid somatic hybrid plant regeneration from fusion between diploids in Aurantioideae.
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    Mean, genetic variance, and usefulness of selfing progenies from intra- and inter-pool crosses in faba beans (Vicia faba L.) and their prediction from parental parameters (1999)
    Springer
    Theoretical and applied genetics 98 (1999), S. 569-580 
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    ISSN: 1432-2242
    Keywords: Key words Vicia faba L. ; RAPD ; Mahalanobis genetic distance ; Usefulness ; Genetic variance ; Mid-parent heterosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Determining the genetic potential of a base population from the properties of their parental lines would improve the efficiency of a breeding program. In the present study, we investigated whether the means of the parents and the genetic distance determined from RAPD data (GD) or multivariate analysis (Mahalanobis D2), mid-parent heterosis (MPH), and the absolute difference between means of the parents (∣P1−P2∣) can be used for predicting the means and genetic variances (σ^2 g ) of F3:4 lines derived from different crosses in faba beans. The material comprised 18 intra- and 18 inter-pool crosses among lines from the Minor, Major, and Mediterranean germplasm pools. Fifty F3:4 lines from each cross were evaluated for days to anthesis, plant height, seeds per plant, and seed yield in German (GE) and Mediterranean (ME) environments. GD estimates between parent lines ranged from 0.38 to 0.58, while D2 ranged from 45.5 to 134.7. Correlations between means of the parents and F3:4 lines were highly significant for most traits. Estimates of σ2 g for all traits showed non-significant correlations with MPH, GD, D2. In one ME, ∣P1−P2∣ had significant associations with σ^2 g for seed yield and days to anthesis. The predicted usefulness of crosses, defined as the sum of the population mean and selection responses, was most closely associated with the means of F3:4 lines. We conclude from this study that the means of F3:4 lines can be predicted from the means of the parents, whereas the prediction of genetic variance is still an unsolved problem
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    Development and characterization of Hordeum chilense chromosome-specific STS markers suitable for wheat introgression and marker-assisted selection (1999)
    Springer
    Theoretical and applied genetics 98 (1999), S. 721-727 
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    ISSN: 1432-2242
    Keywords: Key words Addition lines ; Multiplexed PCR ; RAPD ; Sequence tagged site ; Tritordeum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  RAPD markers were developed for octoploid×Tritordeum (amphiploid Hordeum chilense×Triticum aestivum) and its parents. Addition lines were used to identify specific RAPD markers for the Hordeum chilense chromosomes detectable in a wheat background. Twelve RAPD fragments have been cloned, sequenced and converted into STS markers. Eleven of these STSs have maintained both the chromosome specificity and the possibility of detection in a wheat background. The use of these markers in multiplexed PCRs facilitates both the efficient and reliable screening of new addition lines as well as the monitoring of introgression of H. chilense in bread and durum wheat.
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    Effects of reforestation methods on genetic diversity of lodgepole pine: an assessment using microsatellite and randomly amplified polymorphic DNA markers (1999)
    Springer
    Theoretical and applied genetics 98 (1999), S. 793-801 
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    ISSN: 1432-2242
    Keywords: Key words Pinus contorta ; Silviculture ; Reforestation ; Gene conservation ; RAPD ; SSR ; DNA analyses
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  We examined the effects of different methods of forest regeneration on the genetic diversity of lodgepole pine (Pinus contorta var ‘latifolia’) using two different DNA-based molecular markers [randomly amplified polymorphic DNA (RAPDs) and microsatellites or simple sequence repeats (SSRs)]. Genetic diversity was estimated for 30 individuals in each of four populations for the following three stand types: (1) mature lodgepole pine (〉100 years); (2) 20- to 30-year-old harvested stands left for natural regeneration; (3) 20- to 30-year-old planted stands (4 stands of each type); and one group of 30 operationally produced seedlings. There was no significant effect of stand type on expected heterozygosity, although allelic richness and diversity were much higher for SSRs than for RAPDs. Expected heterozygosity ranged from 0.39 to 0.47 based on RAPDs and from 0.67 to 0.77 based on SSRs. The number of alleles per locus for SSRs ranged from 3 to 34 (mean 21.0), and there was a significant relationship between sequence repeat length and the number of alleles at a locus. Both marker types showed that over 94% of the variation was contained within the populations and that the naturally regenerated stands sampled had lower (not significant) expected heterozygosity than the planted or unharvested stands. The group of seedlings (assessed by RAPDs only) had expected heterozygosity and allele frequencies similar to those of the unharvested stands. Genetic distance measures were higher than obtained previously in the species using isozyme markers. There was no correlation between the two marker types for pair-wise genetic distances based on populations analyzed by both methods. Pair-wise genetic distance measures and an ordination of allele frequencies for both marker types showed little effect of geographic location or stand type on genetic similarity.
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    Development and polymorphism of microsatellite markers for Fagus crenata and the closely related species, F. japonica (1999)
    Springer
    Theoretical and applied genetics 99 (1999), S. 11-15 
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    ISSN: 1432-2242
    Keywords: Key words Fagus crenata ; Fagus japonica ; Microsatellite ; RAPD ; RAHM ; SSR
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  We have developed microsatellite markers (SSRs) applicable to Fagus crenata using the RAHM method and investigated their polymorphisms. We also applied the SSRs in an analysis of a closely related species, F. japonica. Here we describe the isolation and characterization of nine polymorphic microsatellite markers, of which eight are applicable to both species. Among 30 individuals of each of F. crenata and F. japonica we detected a total of 79 and 77 alleles, respectively, with an average of 9.9 and 8.6 alleles per locus. The mean expected heterozygosity (He) was 0.615 (range: 0.216–0.925) in F. crenata and 0.660 in F. japonica (range: 0.259–0.827). The He values were considerably higher than those previously found for isozymes. Paternity exclusion probabilities for multiple loci, calculated over all loci, were extremely high (0.999 and 0.998 in F. crenata and F. japonica, respectively): sufficiently high to study pollen flow in both species.
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    Genetic variation of petaloid male-sterile cytoplasm of carrots revealed by sequence-tagged sites (STSs) (1999)
    Springer
    Theoretical and applied genetics 99 (1999), S. 837-843 
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    ISSN: 1432-2242
    Keywords: Key words Daucus carota spp. sativus ; RAPD ; Cytoplasmic male sterility (CMS) ; Asymmetric cell fusion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The mitochondrial DNA of various carrot lines was characterized by random amplified polymorphic DNA (RAPD) analysis, and six sequence-tagged sites (STSs) led to identification of the petaloid type of cytoplasmic male sterility (CMS). Using six STS primer combinations, we were able to classify five CMS lines into two groups and eight fertile carrots into six groups. Both the STS1 and the STS4 primer combinations differentiated CMS cytoplasms from the fertile cytoplasms, and the STS2 primer combination revealed two different types of CMS cytoplasms – of Wisconsin Wild and Cornell origins. Cybrid carrot lines with petaloid flowers which had been obtained by asymmetric cell fusion could also be separated from fertile cybrids by the STS1 primer combination. The STS1 fragment contained a homologous sequence with the orfB gene. DNA gel blot analysis indicated that homologous regions to the STS1 fragment existed in fertile types as well as the CMS types, although the restriction fragment size patterns differed. These observations demonstrate that rearrangements involving this region occurred in the mitochondrial genome. The STS4 fragment had a more complicated gene structure, including retrotransposon-like sequences and small segments of chloroplast genome.
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    Identification of molecular markers associated with leptine production in a population of Solanum chacoense Bitter (1999)
    Springer
    Theoretical and applied genetics 98 (1999), S. 39-46 
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    ISSN: 1432-2242
    Keywords: Key words Glycoalkaloids ; Potato ; Metabolic pathways ; RAPD ; Leptine ; Insect resistance ; Solanum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract   Solanum chacoense Bitter, a wild relative of the cultivated potato, produces several glycoalkaloids, including solanine, chaconine, and the leptines. The foliar-specific leptine glycoalkaloids are believed to confer resistance to the Colorado Potato Beetle (CPB). Using two bulked DNA samples composed of high- and low-percent leptine individuals from a segregating F1 population of S. chacoense, we have identified two molecular markers that are closely linked to high percent solanine+chaconine and, conversely, to nil/low percent leptine. One of these, a 1,500-bp RAPD product (UBC370-1500), had a recombination value of 3% in the F1 progeny, indicating tight linkage. UBC370-1500 mapped to the end of the short arm of potato chromosome 1, in the region of a previously mapped major QTL for solanidine, from a S. tuberosum (solanidine)×S. berthaultii (solasodine) cross. Taken together, these results suggest that either (1) a major locus determining solanidine accumulation in Solanum spp. is on chromosome 1 in the region defined by the RFLP markers TG24, CT197, and CT233, or (2) this region of chromosome 1 may harbor two or more important genes which determine accumulation of steroidal aglycones. These findings are important for the genetics of leptine (as well as other glycoalkaloid) accumulation and for the development of CPB-resistant potato varieties.
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    Identification of DNA markers linked to the male sex in dioecious hemp (Cannabis sativa L.) (1999)
    Springer
    Theoretical and applied genetics 98 (1999), S. 86-92 
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    ISSN: 1432-2242
    Keywords: Key words Cannabis sativa ; Dioecy ; Sex ; RAPD ; SCAR
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  A 400-bp RAPD marker generated by a primer of random decamer sequence has been found associated with the male sex phenotype in 14 dioecious cultivars and accessions of hemp (Cannabis sativa L.). The primer OPA8 generates a set of bands, most of which polymorphic among all the individual plants tested, and 1 of which, named OPA8400, present in all male plants and absent in female plants. A screening of 167 plants belonging to different genotypes for the association of the OPA8400 marker with the sex phenotype revealed that only in 3 cases was the 400-bp band was present in plants phenotypically female; on the contrary, in male plants the band was never missing, while in monoecious plants it was never present. Despite this sex-specific association, the sequences corresponding to OPA8400 were present in both staminate and carpellate plants, as revealed by Southern blotting and hybridization with the cloned RAPD band. The RAPD marker was sequenced, and specific primers were constructed. These primers generated, on the same genotypes used for RAPD analysis, a SCAR marker 390 bp in length and male-specific. This SCAR is suitable for a precise, early and rapid identification of male plants during breeding programs of dioecious and monoecious hemp.
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    Morphological and RAPD markers show a highly skewed distribution in a pair of reciprocal crosses between hemisexual dogrose species, Rosa sect. Caninae (1999)
    Springer
    Theoretical and applied genetics 98 (1999), S. 557-563 
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    ISSN: 1432-2242
    Keywords: Key words Rosa sect. Caninae ; Biometrics ; Heterogamy ; RAPD ; Segregation distortion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The dogroses, Rosa sect. Caninae, are polyploid and characterized by their unique meiosis with an unequal number of chromosomes in the male and female gametes. The pollen cells have 7 chromosomes and the egg cells 21, 28 or 35 depending on the ploidy level of the species. The resulting matroclinal inheritance was studied with both morphological and molecular markers in a pair of reciprocal crosses between R. dumalis and R. rubiginosa (2n=35). A canonical discriminant analysis based on seven morphological characters showed only a minor overlapping between the two progeny groups. In addition, the R. dumalis×R. rubiginosa offspring were more heterogeneous than the offspring from the reciprocal cross in each of the characters analysed. Eleven RAPD markers specific for the R. dumalis parent and 10 RAPD markers specific for the R. rubiginosa parent were scored in the offspring. Each of the offspring exhibited either all, or all-but-one, of the seed parent markers. The average number of pollen donor markers found in the offspring was 3.2 (R. dumalis×R. rubiginosa) and 2.7 (R. rubiginosa×R. dumalis). About half of the pollen donor markers were never transmitted to the progeny. This is, to our knowledge, the first time the highly skewed chromosome distribution in Rosa sect. Caninae has been demonstrated with statistically evaluated morphological data and with molecular markers.
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    The use of RAPD analysis to classify Triticum accessions (1999)
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    Theoretical and applied genetics 98 (1999), S. 602-607 
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    ISSN: 1432-2242
    Keywords: Key words Triticum ; Germplasm ; RAPD ; Misclassification ; Duplication
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Crop germplasm collections contain a considerable percentage of misclassified accessions which may affect the use of germplasm for agricultural crop improvement. The objective of this study was to determine if random amplified polymorphic DNA (RAPD) analysis could be used to reclassify misclassified Triticum accessions. Twelve accessions suspected to be misclassified, based on morphological characters, as either macha or vavilovii wheat were studied using RAPD and cytological analyses. In the RAPD analysis, a dendrogram, based on Jaccard genetic similarity coefficients, grouped 5 dicoccum-like, 1 timopheevii-like, and 6 monococcum-like accessions with Triticum dicoccum, T. timopheevii, and T. monococcum accessions, respectively. These results were confirmed by the cytological analysis. A RAPD marker specific to the D genome was also detected. This study suggests that RAPD analysis can be used to classify germplasm and to distinguish some species in Triticum.
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    RAPD markers on seed bulks efficiently assess the genetic diversity of a Brassica oleracea L. collection (1999)
    Springer
    Theoretical and applied genetics 98 (1999), S. 1029-1035 
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    ISSN: 1432-2242
    Keywords: Key words Brassica oleracea L. ; RAPD ; Seed bulk ; Genetic resources ; Genetic variability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The concept of a core collection was elaborated to fit the necessity of optimizing the management, for both conservation and use, of genetic resources in sizeable collections. This approach requires an analysis of how the genetic variability is structured among the accessions. The large number of heterogeneous populations in our collection of Brassica oleracea makes genetic diversity studies based on plant-to-plant analysis impracticable. To overcome this limitation, the variability analysis by RAPD on seed bulks was investigated for its efficiency in assessing the structure of the genetic diversity of this collection. The optimal bulk size and the bulking or sampling variation were evaluated with bulks of different size and with replicated samples. A mixture of known genotypes was also used to characterise the band detection in bulks, and to compare the plant-to-plant and the bulk methods. Forty seeds were chosen to represent each population. In such a bulk, the detection of bands depended on the proportion of the genotype they were derived from in the mixture. Intense and frequent bands were detected in the bulk with a 15% detection limit. The observed bulking or sampling variation within populations was smaller than the variation between populations, leading to an efficient separation of populations with a clustering of all samples of the same population. The distances calculated from bulk data were highly correlated with the distances based on the plant-to-plant analysis. We demonstrated that RAPD on seed bulks can be used to describe the genetic diversity between populations.
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    RAPD variation within and among natural populations of outcrossing willow-leaved foxglove (Digitalis obscura L.) (1999)
    Springer
    Theoretical and applied genetics 98 (1999), S. 985-994 
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    ISSN: 1432-2242
    Keywords: Key words Digitalis obscura ; AMOVA ; HOMOVA ; Population genetics ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Random amplified polymorphic DNA (RAPD) markers were used to assess levels and patterns of genetic diversity in Digitalis obscura L. (Scrophulariaceae), an outcrossing cardenolide-producing medicinal plant species. A total of 50 plants from six natural populations on the Iberian Peninsula were analysed by six arbitrarily chosen decamer primers resulting in 96 highly reproducible RAPD bands. To avoid bias in parameter estimation, analyses of population genetic structure were restricted to bands (35 of 96) whose observed frequencies were less than 1–3/n in each population. The analysis of molecular variance (AMOVA) with distances among individuals corrected for the dominant nature of RAPDs (genotypic analysis) showed that most of the variation (84.8%) occurred among individuals within populations, which is expected for an outcrossing organism. Of the remaining variance, 9.7% was attributed to differences between regions, and 5.5% for differences among populations within regions. Estimates of the Wright, Weir and Cockerham and Lynch and Milligan FST from null-allele frequencies corroborated AMOVA partitioning and provided significant evidence for population differentiation in D. obscura. A non-parametric test for the homogeneity of molecular variance (HOMOVA) also showed significant differences in the amount of genetic variability present in the six populations. UPGMA cluster analyses, based on Apostol genetic distance, revealed grouping of some geographically proximate populations. Nevertheless, a Mantel test did not give a significant correlation between geographic and genetic distances. This is the first report of the partitioning of genetic variability within and between populations of D. obscura and provides important baseline data for optimising sampling strategies and for conserving the genetic resources of this medicinal species.
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    Development and utility of cleaved amplified polymorphic sequences (CAPS) and restriction fragment length polymorphisms (RFLPs) linked to the Fom-2 fusarium wilt resistance gene in melon (Cucumis melo L.) (1999)
    Springer
    Theoretical and applied genetics 99 (1999), S. 453-463 
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    ISSN: 1432-2242
    Keywords: Key words Cucumis melo ; Molecular markers ; RAPD ; CAPS ; RFLP ; Fusarium oxysporum ; Fusarium resistance ; Marker-assisted selection (MAS)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Fusarium wilt, caused by Fusarium oxysporum Schlecht f. sp. melonis Snyder & Hans, is a worldwide soil-borne disease of melon (Cucumis melo L.). Resistance to races 0 and 1 of Fusarium wilt is conditioned by the dominant gene Fom-2. To facilitate marker-assisted backcrossing with selection for Fusarium wilt resistance, we developed cleaved amplified polymorphic sequences (CAPS) and restriction fragment length polymorphisms (RFLP) markers by converting RAPD markers E07 (a 1.25-kb band) and G17 (a 1.05-kb band), respectively. The RAPD-PCR polymorphic fragments from the susceptible line ’Vedrantais’ were cloned and sequenced in order to construct primers that would amplify only the target fragment. The derived primers, E07SCAR-1/E07SCAR-2 from E07 and G17SCAR-1/G17SCAR-2 from G17, yielded a single 1.25-kb fragment (designated SCE07) and a 1.05-kb fragment (designated SCG17) (the same as RAPD markers E07 and G17), respectively, from both resistant and susceptible melon lines, thus demonstrating locus-specific associated primers. Potential CAPS markers were first revealed by comparing sequence data between fragments amplified from resistant (PI 161375) and susceptible (’Vedrantais’) lines and were then confirmed by electrophoresis of restriction endonuclease digestion products. Twelve restriction endonucleases were evaluated for their potential use as CAPS markers within the SCE07 fragment. Three (BclI, MspI, and BssSI) yielded ideal CAPS markers and were subsequently subjected to extensive testing using an additional 88 diverse melon cultigens, 93 and 119 F2 individuals from crosses of ’Vedrantais’ x PI 161375 and ’Ananas Yokneam’×MR-1 respectively, and 17 families from a backcross BC1S1 population derived from the breeding line ’MD8654’ as a resistance source. BclI- and MspI-CAPS are susceptible-linked markers, whereas the BssSI-CAPS is a resistant-linked marker. The CAPS markers that resulted from double digestion by BclI and BssSI are co-dominant. Results from BclI- and MspI-CAPS showed over 90% accuracy in the melon cultigens, and nearly 100% accuracy in the F2 individuals and BC1S1 families tested. This is the first report of PCR-based CAPS markers linked to resistance/susceptibility for Fusarium wilt in melon. The RFLP markers resulting from probing with a clone-derived 1.05-kb SCG17 PCR fragment showed 85% correct matches to the disease phenotype. Both the CAPS and RFLP markers were co-dominant, easier to score, and more accurate and consistent in predicting the melon phenotype than the RAPD markers from which they were derived.
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    URL: http://dx.doi.org/10.1007/s001220051257
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    RAPD variations among pure and hybrid populations ofPicea mariana, P. rubens, andP. glauca (Pinaceae), and cytogenetic stability ofPicea hybrids: Identification of species-specific RAPD markers (1999)
    Springer
    Plant systematics and evolution 215 (1999), S. 229-239 
    Details
    ISSN: 1615-6110
    Keywords: Pinaceae ; Picea mariana ; P. rubens ; P. glauca ; RAPD ; genetic relationship ; interspecific hybrids ; mitotic stability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Random amplified polymorphic DNA (RAPD) analysis was used to determine genetic relationships amongP. mariana (black spruce),P. rubens (red spruce), andP. glauca (white spruce) and to assess the degree of polymorphism within populations from different provenances and among spruce hybrids. Eleven arbitrary decamer primers were used to amplify genomic DNAs extracted from embryogenic cultures and seedlings. Species-specific RAPD markers were identified.Picea mariana andP. rubens showed similar RAPD profiles confirming their close genetic relationship. Species-specific RAPD markers were identified and were useful in distinguishing white spruce from black and red spruces. RAPD differentiation between populations within each species was small. The level of polymorphism was much higher in spruce hybrid populations than in the pure species. Cytological analysis ofP. mariana ×P. rubens hybrids showed normal mitotic behaviour at prophase, metaphase, anaphase, and telophase. All the hybrids analyzed from different cross combinations were euploids.
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    URL: http://dx.doi.org/10.1007/BF00984657
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    Identification of hybrids betweenQuercus petraea andQ. robur (Fagaceae): Results obtained with RAPD markers confirm allozyme studies based on theGot-2 locus (1999)
    Springer
    Plant systematics and evolution 217 (1999), S. 137-146 
    Details
    ISSN: 1615-6110
    Keywords: Fagaceae ; Quercus ; Hybridization ; RAPD ; allozymes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract RAPDs were employed as genetic markers to detect interspecific hybridization between the closely related oak speciesQuercus robur andQ. petraea. Fourteen primers were used in order to check the genetic status (“pure” or hybrid) of individuals classified morphologically. Among the 147 PCR fragments obtained 11 appear to be species-specific. In the phenotypically intermediate individuals different combinations of these species-specific bands were obtained. The patterns in these putative hybrids were not additive, which may be either the result of repeated backcrossing and introgression between the two species or of heterozygosity within the parental species. The results of the RAPD study are consistent with morphological analyses and allozyme data obtained for theGot-2 locus. Thus the RAPD markers used in this study may provide a powerful genetic tool for the identification of hybrids and the discrimination between the two “pure” species.
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    URL: http://dx.doi.org/10.1007/BF00984926
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    Retention of nutrients in river basins (1999)
    Springer
    Aquatic ecology 33 (1999), S. 29-40 
    Details
    ISSN: 1573-5125
    Keywords: denitrification ; nature restoration ; nitrogen ; phosphorus ; riparian areas ; sedimentation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In Denmark, as in many other European countries, the diffuse losses of nitrogen (N) and phosphorus (P) from the rural landscape are the major causes of surface water eutrophication and groundwater pollution. The export of total N and total P from the Gjern river basin amounted to 18.2 kg ha−1 and 0.63 kg P ha−1 during June 1994 to May 1995. Diffuse losses of N and P from agricultural areas were the main nutrient source in the river basin contributing 76% and 51%, respectively, of the total export. Investigations of nutrient cycling in the Gjern river basin have revealed the importance of permanent nutrient sinks (denitrification and overbank sedimentation) and temporary nutrient storage in watercourses. Temporary retention of N and P in the watercourses thus amounted to 7.2–16.1 g N m−2 yr−1 and 3.7–8.3 g P m−2 yr−1 during low-flow periods. Deposition of P on temporarily flooded riparian areas amounted from 0.16 to 6.50 g P m−2 during single irrigation and overbank flood events, whereas denitrification of nitrate amounted on average to 7.96 kg N yr−1 per running metre watercourse in a minerotrophic fen and 1.53 kg N yr−1 per linear metre watercourse in a wet meadow. On average, annual retention of N and P in 18 Danish shallow lakes amounted to 32.5 g N m−2 yr−1 and 0.30 g P m−2 yr−1, respectively, during the period 1989–1995. The results indicate that permanent nutrient sinks and temporary nutrient storage in river systems represent an important component of river basin nutrient budgets. Model estimates of the natural retention potential of the Gjern river basin revealed an increase from 38.8 to 81.4 tonnes yr−1 and that P-retention increased from −0.80 to 0.90 tonnes yr−1 following restoration of the water courses, riparian areas and a shallow lake. Catchment management measures such as nature restoration at the river basin scale can thus help to combat diffuse nutrient pollution.
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    URL: http://dx.doi.org/10.1023/A:1009947907811
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    Clonal structure of Rubus chamaemorus populations: comparison of different molecular methods (1999)
    Springer
    Plant ecology 143 (1999), S. 123-128 
    Details
    ISSN: 1573-5052
    Keywords: Clonal structure ; Cloudberry ; Genetic variation ; DNA fingerprinting ; RAPD ; Rubus chamaemorus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The clonal structure of Rubus chamaemorus populations was investigated using DNA fingerprinting. The PCR-based methods included the use of 10-base RAPD primers and 16-base simple sequence repeat primers. In the hybridization method variation was studied using hypervariable multilocus probes, one derived from the M13 bacteriophage and the other a synthetic (AC)/(TG) polynucleotide. Although R. chamaemorus expresses clear variation in morphology, the level of genetic differentiation appears to be fairly low. The observed numbers of clones in the three populations examined in Finland varied from 2 to 4. The total number of genotypes across populations was 5, of which one was unique. The results obtained using the two fingerprinting methods were comparable but lead to a slightly different grouping of clones.
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    URL: http://dx.doi.org/10.1023/A:1009898209220
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    Variation in nutrient availability and plant species diversity across forb and graminoid zones of a Northern New England high salt marsh (1999)
    Springer
    Plant ecology 143 (1999), S. 219-228 
    Details
    ISSN: 1573-5052
    Keywords: mineralization ; nitrogen ; phosphorus ; salinity ; stress
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Plant zonation patterns across New England salt marshes have been investigated for years, but how nutrient availability differs between zones has received little attention. We investigated how N availability, P availability, and plant N status varied across Juncus gerardii, Spartina patens, and mixed forb zones of a Northern New England high salt marsh. We also investigated relationships between several edaphic factors and community production and diversity across the high marsh. P availability, soil salinity, and soil moisture were higher in the mixed forb zone than in the two graminoid zones. NH+ 4-N availability was highest in the J. gerardii zone, but NO− 3-N availability and mid season net N mineralization rates did not vary among zones. Plant tissue N concentrations were highest in the mixed forb zone and lowest in the S. patens zone, reflecting plant physiologies more so than soil N availability. Community production was highest in the J. gerardii zone and was positively correlated with N availability and negatively correlated with soil moisture. Plant species diversity was highest in the mixed forb zone and was positively correlated with P availability and soil salinity. Thus, nutrient availability, plant N status, and plant species diversity varied across zones of this high marsh. Further investigation is needed to ascertain if soil nutrient availability influences or is a result of the production and diversity differences that exist between vegetation zones of New England high salt marshes.
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    URL: http://dx.doi.org/10.1023/A:1009887727465
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    Protein Kinase C Targeting in Antineoplastic Treatment Strategies (1999)
    Springer
    Investigational new drugs 17 (1999), S. 227-240 
    Details
    ISSN: 1573-0646
    Keywords: apoptosis ; protein kinase C ; sphingoid bases ; safingol ; diglyceride ; bryostatin 1 ; staurosporine ; 7-hydroxy staurosporine (UCN-01) ; 4′-N-benzoyl staurosporine (CGP-41251) ; calphostin C (UCN-1028c)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Abstract Neoplastic cell survival is governed by a balance between pro-apoptotic and anti-apoptotic signals. Noteworthy among several anti-apoptotic signaling elements is the protein kinase C (PKC) isoenzyme family, which mediates a central cytoprotective effect in the regulation of cell survival. Activation of PKC, and subsequent recruitment of numerous downstream elements such as the mitogen-activated protein kinase (MAPK) cascade, opposes initiation of the apoptotic cell death program by diverse cytotoxic stimuli. The understanding that the lethal actions of numerous antineoplastic agents are, in many instances, antagonized by cytoprotective signaling systems has been an important stimulus for the development of novel antineoplastic strategies. In this regard, inhibition of PKC, which has been shown to initiate apoptosis in a variety of malignant cell types, has recently been the focus of intense interest. Furthermore, there is accumulating evidence that selective targeting of PKC may prove useful in improving the therapeutic efficacy of established antineoplastic agents. Such chemosensitizing strategies can involve either (a) direct inhibition of PKC (e.g., following acute treatment with relatively specific inhibitors such as the synthetic sphingoid base analog safingol, or the novel staurosporine derivatives UCN-01 and CGP-41251) or (b) down-regulation (e.g., following chronic treatment with the non-tumor-promoting PKC activator bryostatin 1). In preclinical model systems, suppression of the cytoprotective function(s) of PKC potentiates the activity of cytotoxic agents (e.g., cytarabine) as well as ionizing radiation, and efforts to translate these findings into the clinical arena in humans are currently underway. Although the PKC-driven cytoprotective signaling systems affected by these treatments have not been definitively characterized, interference with PKC activity has been associated with loss of the mitogen-activated protein kinase (MAPK) response. Accordingly, recent pre-clinical studies have demonstrated that pharmacological disruption of the primary MEK-ERK module can mimic the chemopotentiating and radiopotentiating actions of PKC inhibition and/or down-regulation.
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    URL: http://dx.doi.org/10.1023/A:1006328303451
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    Activation-Induced Apoptosis of Peripheral Lymphocytes Treated with 7-Hydroxystaurosporine, UCN-01 (1999)
    Springer
    Investigational new drugs 17 (1999), S. 335-341 
    Details
    ISSN: 1573-0646
    Keywords: UCN-01 ; IL-2 receptor ; Fas ; Fas-ligand ; apoptosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
    Notes: Abstract 7-hydroxystaurosporine (UCN-01) is a new anticancer agentwhich exerts an inhibitory effect on cell cycle check points andis currently under phase I clinical trials in US and Japan.Preliminary clinical data indicated that UCN-01 remained inplasma at high concentrations for long periods of time. Thisunavoidable high plasma drug exposure is likely to lead tohematological toxicities in patients. In the present study,cultured human peripheral blood lymphocytes (PBLs) were used toevaluate the possible hematological toxicities of UCN-01treatment. UCN-01 induces apoptosis, and the induction ofapoptosis-related surface markers were also examined toinvestigate the involvement of these molecules in UCN-01-inducedapoptosis in PBLs. in vitroviability of PBLs wasdecreased by high dose of UCN-01 (25 μM, 3-day exposure). Thiseffect of UCN-01 was significantly suppressed by the presence ofhuman serum, suggesting that some specific inhibitory factor(s)in human serum may antagonize the lympholytic effect of UCN-01.The percentage of annexin V-positive PI-negative cells increasedwith exposure to UCN-01 in a time- and dose-dependent manner; byup to 30.3% after exposure to 25 μM UCN-01 for 3 days.At the same time, the expression of both interleukin-2 receptor(IL-2R, CD25) and Fas (CD95), analyzed by flow cytometry, wasinduced. Con A-stimulated PBLs were more sensitive toUCN-01-induced apoptosis than non-stimulated lymphocytes andUCN-01 increased the sFas-L released into culture medium from conA-stimulated PBLs. Therefore, lymphocyte depletion mediated byactivation-induced apoptosis is likely to occur in patientstreated with UCN-01 at high doses.
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    Influence of bcl-2 on antibody productivity in high cell density perfusion cultures of hybridoma (1999)
    Springer
    Cytotechnology 30 (1999), S. 95-106 
    Details
    ISSN: 1573-0778
    Keywords: apoptosis ; Bcl-2 ; fixed-bed ; hollow fibre ; hybridoma ; perfusion ; protein-free medium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Apoptosis is an active, genetically determined death mechanism which can be induced by a wide range of physiological factors and by mild stress. It is the predominant form of cell death during the production of antibodies from murine hybridoma cell lines. A number of studies have now demonstrated that the suppression of this death pathway, by means of over-expression of survival genes such as bcl-2, results in improved cellular robustness and antibody productivity during batch culture. In the present study, the influence of bcl-2 expression on hybridoma productivity in two high density perfusion bioreactor systems was investigated. In the first system, a fixed-bed reactor, the DNA content in the spent medium was 25% higher in the control (TB/C3-pEF) culture than that found in the bcl-2 transfected (TB/C3-bcl2) cultures at all perfusion rates. This is indicative of a higher level of cell death in the control cell line. The average antibody concentration for the TB/C3-pEF cell line was 14.9 mg L-1 at perfusion rates of 2.6 and 5.2 d-1. However, for the TB/C3-bcl2 cell line it was 33 mg L-1 at dilution rates of 2 and 4 d-1. A substantial increase in antibody concentration was also found in the Integra Tecnomouse hollow fibre reactor. The antibody titre in the TB/C3-bcl2 cassette was nearly 100% higher than that in the TB/C3-pEF cassette during the cultivation period which lasted 6 weeks. Clearly, these results demonstrate the positive impact of bcl-2 over-expression on production of antibody in hybridoma perfusion cultures.
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    URL: http://dx.doi.org/10.1023/A:1008055702079
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    Effect of meteorology and nutrient load on oxygen depletion in a Danish micro-tidal estuary (1999)
    Springer
    Aquatic ecology 33 (1999), S. 55-64 
    Details
    ISSN: 1573-5125
    Keywords: estuaries ; nitrogen ; oxygen depletion ; phosphorus ; regression model ; vertical mixing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In a detailed analysis of oxygen saturation in a shallow Danish estuary it was possible to separate the effect of meteorological forcings (i.e. wind and solar radiation) and nutrient loads on oxygen depletion in bottom water. Regression analysis showed that oxygen saturation tied to nitrogen load rather than to phosphorus load. During summer periods of stratification the oxygen saturation could be attributed to the time elapsed after the onset of stratification and the accumulated nitrogen loading 10 month prior to measurement. Using a 10-year meteorological database and an empirical model it was calculated that a 25% reduction in nitrogen loading would reduce the number of days with severe oxygen depletion (i.e. 〈15% of saturation) by more than 50%.
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    URL: http://dx.doi.org/10.1023/A:1009956210537
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    Co-expression of bcl-2 and bag-1, apoptosis suppressing genes, prolonged viable culture period of hybridoma and enhanced antibody production (1999)
    Springer
    Cytotechnology 31 (1999), S. 143-151 
    Details
    ISSN: 1573-0778
    Keywords: antibody productivity ; apoptosis ; BAG-1 ; Bcl-2 ; cell survival ; hybridoma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Human bcl-2 and bag-1 DNA were introduced into mouse hybridoma 2E3- O cells and expressed. The expression of bcl-2 in BCMGneo-bcl2 transfectants was confirmed by ELISA and that of bag-1 in pZeo-bag1 was confirmed by western blotting. In batch cultures, the over-expression of bcl-2 prolonged the culture period by 2 days and co-expression of bcl-2 and bag-1 prolonged the culture period by 3 days. The delayed increase in the dead cell number in culture of the bcl-2 and bag-1 cotransfectant indicated the additional antiapoptosis effect of bcl-2 and bag-1 cotransfection in comparison with the bcl-2 only transfection. The bcl-2 transfectants (2E3O-Bcl2) produced antibody twofold per batch culture in comparison with 2E3-O cells transfected with BCMGSneo (2E3O-Mock). Enhancement of this MoAb production was due to the improved survival of the cells and was not due to stimulation of antibody production rate per cell by Bcl-2 expression. And the bcl-2 and bag-1 co-transfectant (2E3O-Bcl2-BAG1) produced antibody approximately fourfold of 2E3O-Mock per batch culture. Enhancement of this MoAb production was due to the improved survival of the cells and was partly due to stimulation of MoAb production rate per cell in the non-growing phase by the cotransfection. The method to engineer hybridoma cells genetically with bcl-2 and bag-1 for increasing viability and productivity would be widely applied for improving antibody productivity of hybridoma cultures.
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    URL: http://dx.doi.org/10.1023/A:1008080407581
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    A novel 96-well scintillation proximity assay for the measurement of apoptosis (1999)
    Springer
    Cytotechnology 31 (1999), S. 271-282 
    Details
    ISSN: 1573-0778
    Keywords: annexin V ; Apo-2 ligand ; apoptosis ; Cytostar-T® scintillating microplates ; flow cytometry ; lymphotoxin (LT)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract The translocation of phospholipids across the plasma membrane has been widely documented as one of the earliest measurable biochemical events of apoptosis. Using fluorescently labelled annexin V, which preferentially binds phosphatidylserine (PS) in the presence of Ca2+, the externalization of PS can be measured and apoptosis quantified using flow cytometry. Conventional detection methods utilizing annexin V, while faster than in situ DNA end-labelling or DNA laddering, require extensive sample preparation which may compromise samples and makes rapid, high volume screening prohibitive. This paper describes a novel assay for the measurement of apoptosis based upon binding of radiolabelled annexin V to apoptotic cells attached to the growth surface of a 96-well scintillating microplate (Cytostar-T®). We compared measurements of apoptosis made by flow cytometry to those obtained with the scintillating microplate in three model systems, treatment of: mouse connective tissue (L-M) cells with lymphotoxin (LT), human lung carcinoma (H460) cells with Apo-2 ligand and human umbilical vein endothelial (HUVE) cells with staurosporine. In this assay, we compare both direct and indirect labelling methods by utilizing either iodinated annexin V or biotinylated annexin V/[35S] streptavidin to radiolabel apoptotic cells. The signal detected is a direct consequence of the binding of annexin V to externalized PS on apoptotic cells and the proximity of the label to the base of the plate. Using this method, separation of bound and unbound radiolabel signal occurs directly within the well resulting in a sensitive assay that requires minimal manipulation and can accomodate a large number of samples.
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    Comparison of PCR-based molecular marker analyses of Musa breeding populations (1999)
    Springer
    Molecular breeding 5 (1999), S. 233-244 
    Details
    ISSN: 1572-9788
    Keywords: Musa ; plantain ; RAPD ; VNTR ; AFLP ; breeding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Progress in the breeding of plantain and banana has been restricted by the complex genetic structure and behaviour of cultivated polyploid Musa. Genetic improvement has been hindered due to the large amount of space required for growth and maintenance of plant populations, in addition to the long growth cycle and the low levels of fertility and seed viability characteristic of cultivated genotypes. Molecular marker assisted breeding has the potential to dramatically enhance the pace and efficiency of genetic improvement in Musa. This study was conducted to compare different PCR-based marker systems (RAPD, VNTR and AFLP) for the analysis of breeding populations generated from two diverse Musa breeding schemes. All three assays detected a high level of polymorphism between parental genotypes and within progeny populations. As expected, AFLP assays had by far the highest multiplex ratio while VNTR analysis detected the highest levels of polymorphism. AFLP analysis of a full-sib tetraploid hybrid population confirmed previous reports based on VNTR analysis, of a high frequency of recombination during 2n (3x) gamete formation by a triploid plantain landrace. In addition, both VNTR and RAPD analyses of a full-sib triploid hybrid population suggested a high frequency of homoeologous recombination during n (2x) gamete formation by tetraploid hybrids. In general, there was a poor correlation between estimates of genetic similarity based on different types of marker. The implications of these findings for the molecular breeding of Musa crops are discussed.
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    Evaluation of AFLP for genetic mapping in Pinus radiata D. Don (1999)
    Springer
    Molecular breeding 5 (1999), S. 275-281 
    Details
    ISSN: 1572-9788
    Keywords: AFLP ; DNA markers ; genetic mapping ; marker-aided selection ; Pinus radiata ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Efficient construction of reasonable density genetic linkage maps is an essential component of QTL detection programmes. The AFLP technique has been used to produce genetic linkage maps in a range of species. We have developed protocols to generate reproducible AFLP profiles in Pinus radiata and have evaluated the inheritance and informativeness of AFLP markers in this important timber species. The large genome size of P. radiata necessitated increased levels of selection at both the pre-amplification and selective amplification steps of the AFLP protocol to generate reproducible AFLP profiles. Once optimised ca. 41.3 scorable AFLP bands were resolvable through denaturing gels, of which 48.4% were polymorphic in a screen of eight unrelated trees. This level of polymorphism is ca. three times higher than with RAPD markers. The total number of bands and the number of polymorphismic bands per PCR were ca. halved when AFLPs were electrophoresed on non-denaturing gels and stained with ethidium bromide. Using the protocols developed, AFLP is an efficient method for generating the DNA markers required for genetic linkage map construction in P. radiata.
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    URL: http://dx.doi.org/10.1023/A:1009612709760
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    Effects of Potato-cyst Nematodes (Globodera pallida) and Soil pH on Root Growth, Nutrient Uptake and Crop Growth of Potato (1999)
    Springer
    European journal of plant pathology 105 (1999), S. 61-76 
    Details
    ISSN: 1573-8469
    Keywords: core sampling ; foliar nutrient concentrations ; minirhizotrons ; nitrogen ; phosphorus ; potassium ; Rhizolab
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Potato-cyst nematodes (Globodera pallida) cause severe yield losses in potato. Plants infected with potato-cyst nematodes generally have reduced concentrations of nitrogen, phosphorus and potassium in the foliage. This study investigated whether reduced growth of nematode-infected potato is caused by nutrient limitation. Experiments in the field and in containers showed that phosphorus concentration correlated best with total crop biomass at early stages of growth. The role of phosphorus in nematode damage was further investigated in the field and in the Wageningen Rhizolab. The experimental field was infested with potato-cyst nematodes and two levels of nematode density were established by fumigation with a nematicide. Prior applications of calcium carbonate resulted in pHKCl levels of 4.8 and 6.1. Two levels of phosphorus fertiliser were applied: either 0 or 225 kg P ha−1. In the Wageningen Rhizolab, soil of both pH levels from the field was used after treatment with 1 MRad gamma irradiation to kill the nematodes. Subsequently, half of the soil was inoculated with cysts to give a nematode density of 30 viable juveniles per gram of soil. In the field, nine weeks after planting, the total crop biomass ranged from 107 g m−2 for the treatment with nematodes at pHKCl 6.1 without phosphorus fertiliser to 289 g m−2 for the fumigated treatment at pHKCl 4.8 with phosphorus fertiliser. The differences in total biomass for the various treatments were explained by differences in foliar phosphorus concentration. Nematodes induced or aggravated P deficiency and reduced total biomass. This was not the only damage mechanism as at high, non-limiting levels of foliar phosphorus concentration, nematodes still reduced total biomass. In the Wageningen Rhizolab, directly after planting, the number of roots visible against minirhizotrons was reduced by nematodes. However, the increase of root number in the nematode treatment continued longer than in the control, until root number was higher than that of the control. The compensary root growth of the nematode treatment was restricted to the top 30 cm and nematodes reduced rooting depth. High soil pH reduced growth, mainly by reducing the availability of phosphate. Both nematodes and high soil pH reduced nutrient uptake per unit root length. Our results lead us to suggest an interaction between nematodes and soil pH, with nematode damage being higher at pHKCl 6.1 than at pHKCl 4.8.
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    Verotoxin/Globotriaosyl Ceramide Recognition: Angiopathy, Angiogenesis and Antineoplasia (1999)
    Springer
    Bioscience reports 19 (1999), S. 345-354 
    Details
    ISSN: 1573-4935
    Keywords: Glycolipid ; apoptosis ; intracellular traffic ; multidrug resistance ; ovarian carcinoma ; astrocytoma ; post transplant lymphoproliferative disease ; bone marrow purging
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Verotoxin (VT) is involved in the etiology of both hemorrhagic colitis and the hemolytic uremic syndrome which are microvasculopathies of the colon and pediatric renal glomerulus respectively. Thus, VT can be considered a vasotoxin. Cell sensitivity in vitro varies according to the receptor glycolipid (globotriaosyl ceramide-Gb3) expression and also to intracellular trafficking of the receptor/toxin complex, such that in highly sensitive cells, the toxin is targeted to the endoplasmic reticulum and nuclear envelope. Such cells include tumor cells which have become drug resistant. Thus Gb3 is upregulated in certain tumors and when such tumor cells become drug resistant, their sensitivity to verotoxin increases. This may be due to a direct role of the MDR1 drug efflux pump in glycolipid biosynthesis. In addition to the tumor tissue, the toxin receptor may also be expressed in the tumor neovasculature suggesting that activated endothelial cells may be verotoxin sensitive. Thus VT may have both a direct and indirect antineoplastic potential. VT has proved highly effective in a xenograft cancer model and the possible therapeutic use of VT is discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1020299819637
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    Ceramide Glycanase Activities in Human Cancer Cells (1999)
    Springer
    Bioscience reports 19 (1999), S. 449-460 
    Details
    ISSN: 1573-4935
    Keywords: ceramide glycanase ; cancer cells ; glycosphingolipid ; sphingosine ; ceramide ; apoptosis ; PPMP ; PDMP
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Ceramide glycanase (CGase) activities have been detected in different human tumor cells (colon, carcinoma Colo-205; neuroblastoma, IMR-32; breast cancer lines, SKBr3 and MCF7). However, the level of enzymatic activity is lower in these cells compared to that present in other mammalian tissues reported before (Basu, M., Kelly, P., Girzadas, M. A., Li, Z., and Basu, S. Methods Enzymol. (in press)). The majority of CGase activity was found in the 100,000g soluble supernatant fraction isolated from all these cell lines and tissues. Using the soluble enzyme, the requirement for optimum CGase activity was found to be consistent with previous observations found for rat and rabbit tissues (Basu, M., Dastgheib, S., Girzadas, M. A., O'Donnell, P. H., Westervelt, C. W., Li, Z., Inokuchi, J. I., and Basu, S. (1998) Acta Pol. Biochim. 42:327). The CGase activities from both Colo-205 and IMR-32 cells are optimum at a protein to detergent ratio of one. All the mammalian CGases, including human cancer cells, show an optimum pH between 5.5 and 5.8 in sodium acetate buffer. The CGase activities from cancer cells are found to be cation-independent; however, mercury, zinc, and copper ions seem to inhibit the enzyme activity substantially in both tumor cells lines. The mercury ion inhibition of CGase activities from all different sources indicates a possible structural homology in the CGase proteins. Radiolabeled substrates, labeled at the sphingosine double bond or at the 3-position of sphingosine without modifying double bond of sphingosine were used in this investigation. Both were active substrates with all enzyme preparations isolated from different cancer cells (apparent Km, 500 μM for nLcOse5[3H-DT]Cer and 350 μM for GgOse4[sph-3-3H]Cer with Colo-205 enzyme). Structural analogues of ceramide and sphingosine (L-PPMP, L-PDMP, alkylamines, and Tamoxifen) inhibited cancer cell CGase activities in vitro.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1020220524180
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    Adenosine-induced cell death: evidence for receptor-mediated signalling (1999)
    Springer
    Apoptosis 4 (1999), S. 197-211 
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    ISSN: 1573-675X
    Keywords: Adenosine ; apoptosis ; necrosis ; physiopathological implications.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Adenosine modulates the proliferation, survival and apoptosis of many different cell types, ranging from epithelial, endothelial and smooth muscle cells, to cells of the immune and neural lineages. In this review, we critically discuss the available in vitro and in vivo data which support a role for adenosine in both development-associated apoptosis, and in diseases characterized by either pathologically increased cell death (e.g., ischemia, trauma and aging-associated neurodegeneration) or abnormally reduced spontaneous apoptosis (e.g., cancer). Particular emphasis is given to the possible role of extracellular adenosine receptors, since these may represent novel and attractive molecular targets for the pharmacological modulation of apoptosis. In some instances, adenosine-induced cell death has been demonstrated to require entry of the nucleoside inside cells; however, in many other cases, activation of specific adenosine extracellular receptors has been demonstrated. Of the four G protein-coupled adenosine receptors so far identified, the A2A and the A3 receptors have been specifically implicated in modulation of cell death. For the A3 receptor, results obtained by exposing both cardiomyocytes and brain astrocytes to graded concentrations of selective agonists suggest induction of both cell protection and cell death. Such opposite effects, which likely depend on the degree of receptor activation, may have important therapeutic implications in the pharmacological modulation of cardiac and brain ischemia. For the A2A receptor, recent intriguing data suggest a specific role in immune cell death and immunosuppression, which may be relevant to both adenosine-deaminase-immunodeficiency syndrome (a pathology characterized by accumulation of adenosine to toxic levels) and in tumors where induction of apoptosis via activation of specific extracellular receptors may be desirable. Finally, preliminary data suggest that, in a similar way to the adenosine-deaminase-immunodeficiency syndrome, the abnormal accumulation of adenosine in degenerative muscular diseases may contribute to muscle cell death. Although the role of adenosine receptors in this effect still remains to be determined, these data suggest that adenosine-induced apoptosis may also represent a novel pathogenic pathway in muscular dystrophies.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1009666707307
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    Apoptin®-induced apoptosis: a review (1999)
    Springer
    Apoptosis 4 (1999), S. 317-319 
    Details
    ISSN: 1573-675X
    Keywords: Anti-tumor therapy ; Apoptin® ; apoptosis ; Bcl-2 ; p53.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Apoptin, a protein encoded by an avian virus, induces apoptosis in various cultured human tumorigenic and/ or transformed cell lines, e.g. derived from breast and lung tumor, leukemia, lymphoma, osteosarcoma melanoma, cholangiocarcinoma, and hepatoma. In such cells, Apoptin induces p53-independent apoptosis, and the proto-oncogene Bcl-2 can accelerate this effect. The latter is surprising for, in general, Bcl-2 is known to inhibit e.g., p53-induced apoptosis. On the other hand, in normal non-transformed human cells, Apoptin is unable to induce apoptosis, even when Bcl-2 is over-expressed. In animal models Apoptin-induced apoptosis appears to be a safe and efficient anti-tumor agent. These data, in continuation with the observations that Apoptin is specifically stimulated by Bcl-2 in tumor cells, does not need p53, and is not inhibited by Bcr-Abl in these cells, imply that Apoptin is a potential anti-tumor therapy.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1009687019221
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    Apoptosis in ventricular myocytes: the role of tumor suppressor proteins (1999)
    Springer
    Apoptosis 4 (1999), S. 229-234 
    Details
    ISSN: 1573-675X
    Keywords: Adenovirus ; apoptosis ; Bcl-2 ; p53 ; Rb ; ventricular myocytes.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Apoptosis or programmed cell death is an important physiologic event crucial for the selective removal of damaged or unwanted cells from body tissues. In the cardiovascular system, apoptosis has been observed in the vasculature and myocardium. Untimely or inappropriate myocardial cell loss through an apoptotic process may contribute to ventricular remodeling and the ultimate demise of ventricular function following injury. Therapeutic interventions designed to modulate or prevent myocardial apoptotic cell loss may therefore prove beneficial in maintaining cardiac function. Incite into the molecular mechanisms that govern apoptosis in mammalian cells has led to the identification of several key factors that promote or prevent the apoptotic process. In this report, we discuss putative regulators of cardiac cell apoptosis with specific reference to the tumor suppressor proteins, p53 and Rb. The interplay between these factors, as well as the anti-apoptotic molecules related to the Bcl-2 the family are discussed in the context of the heart under normal and disease conditions.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1009640124029
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    Tau protein is involved in the apoptotic process induced by anti-microtubule agents on neuroblastoma cells (1999)
    Springer
    Apoptosis 4 (1999), S. 47-58 
    Details
    ISSN: 1573-675X
    Keywords: Anti-microtubule agents ; apoptosis ; doxorubicin ; neuroblastoma ; tau ; taxoid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Paclitaxel and docetaxel are potent anti-microtubule and antimitotic agents that induce apoptosis in bone marrow-derived cells and epithelial cells. This study examined apoptosis induced by anti-microtubule agents in the neuroblastoma SK-N-SH cell line with a special focus on tau protein which is one of the main Microtubule-Associated- Proteins (MAPs) in neuronal cells. In time, treatment with 1 μM paclitaxel successively induced formation of bundles, then pseudo-asters concomitantly with mitotic block and phosphorylation of bcl-2 (48 h), then phosphorylation of tau and externalization of phosphatidylserine at the early phase of apoptosis (72 h) and finally DNA fragmentation (96 h). Similar results were obtained with 0.5 μM vinorelbine. Paclitaxel induced a lower increase in tau phosphorylation in differentiated SK-N-SH/RA+ cells which are less sensitive to apoptosis. Moreover, doxorubicin whose mechanism of action is independent of microtubules also induced immunostaining of tau at 72 h treatment. In conclusion, our results on neuroblastoma cells show that overexpression of hyperphosphorylated tau is involved in the apoptotic process induced by anti-microtubule agents and may be extended to others cytostatic drugs. Thus, tau protein may play a role in the cellular events observed in neuroblastoma cells undergoing apoptosis.
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    URL: http://dx.doi.org/10.1023/A:1009682116158
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    IL-3 induces apoptosis in a ras-transformed myeloid cell line (1999)
    Springer
    Apoptosis 4 (1999), S. 71-80 
    Details
    ISSN: 1573-675X
    Keywords: abl ; apoptosis ; interleukin-3 ; oncogenes ; ras.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Growth factors promote cell survival and proliferation. Homeostasis is maintained by programmed cell death which occurs when the growth stimulus is withdrawn, in response to negative growth regulators such as interferons, TNF-α and CD95 ligand, or following differentiation. Although acutely-transforming oncogenes often overcome the need for growth factors, growth regulatory cytokines can influence proliferative responses of transformed cells. In this study we investigated the effects of IL-3 on the proliferative responses of parental bone marrow-derived 32D cells and cells transformed with ras and abl oncogenes. We show that treatment of ras-transformed 32D cells with IL-3 reduced proliferative responses and decreased colony-forming ability. These effects were exacerbated in the absence of serum and associated with inhibition of tyrosine kinase activity, down-regulation of RAS and MYC expression, and induction of apoptosis as indicated by DNA fragmentation. In contrast, treatment of parental 32D cells with IL-3, which is obligatory for cell survival and proliferation, increased tyrosine kinase activity, upregulated MYC and RAS expression and maintained DNA integrity. With abl-transformed cells, proliferation and colony-forming ability were also inhibited by IL-3. Tyrosine kinase activity and MYC expression were reduced, but early apoptosis was not evident. Calcium uptake however, was stimulated by IL-3 in both parental and oncogene-transformed cells. These results suggest that threshold levels of tyrosine kinase activity are necessary for cell survival and proliferation and that with ras-transformed cells, IL-3 treatment may result in this threshold being breached. We conclude that in some situations, growth-promoting cytokines can inhibit proliferation of transformed cells and induce cell death by apoptosis.
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    URL: http://dx.doi.org/10.1023/A:1009686220607
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    Ameloblast apoptosis and IGF-1 receptor expression in the continuously erupting rat incisor model (1999)
    Springer
    Apoptosis 4 (1999), S. 441-447 
    Details
    ISSN: 1573-675X
    Keywords: ameloblasts ; amelogenesis ; apoptosis ; insulin-like growth factor.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Enamel-producing cells (ameloblasts) pass through several phenotypic and functional stages during enamel formation. In the transition between secretory and maturation stages, about one quarter of the ameloblasts suddenly undergo apoptosis. We have studied this phenomenon using the continuously erupting rat incisor model. A special feature of this model is that all stages of ameloblast differentiation are presented within a single longitudinal section of the developing tooth. This permits investigation of the temporal sequence of gene and growth factor receptor expression during ameloblast differentiation and apoptosis. We describe the light and electron microscopic morphology of ameloblast apoptosis and the pattern of insulin-like growth factor-1 receptor expression by ameloblasts in the continuously erupting rat incisor model. In the developing rat incisor, ameloblast apoptosis is associated with downregulated expression of the insulin-like growth factor-1 receptor. These data are consistent with the hypothesis that ameloblasts are “hard wired” for apoptosis and that insulin-like growth factor-1 receptor expression is required to block the default apoptotic pathway. Possible mechanisms of insulin-like growth factor-1 inhibition of ameloblast apoptosis are presented. The rat incisor model may be useful in studies of physiological apoptosis as it presents apoptosis in a predictable pattern in adult tissues.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1009600409421
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    Apoptotic cell death and related gene expression in metastatic tumors of AKR lymphomas of varying malignancy (1999)
    Springer
    Apoptosis 4 (1999), S. 429-440 
    Details
    ISSN: 1573-675X
    Keywords: AKR lymphoma malignancy variants ; apoptosis ; apoptosis-related gene expression ; tumor progression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Resistance to apoptosis may be related to tumor progression, due to the implications it might have on both tumor mass and genetic instability. We compared the tendency to spontaneous apoptosis and the proliferative capacity of metastatic growths of several AKR lymphoma variants (TAU-45, TAU-47, TAU-44, TAU-33, TAU-42 and TAU-46, in the order of increasing metastatic potential). We further compared the expression of several apoptosis-related genes. Cell proliferative capacity did not appear to determine malignant behavior since, on the whole, a decrease in S + G2M fraction was observed with increasing malignancy. Sensitivity to apoptotic cell death decreased with increasing malignancy when comparing the TAU-45, TAU-47, TAU-44 and TAU-33 variants, suggesting a role of reduced apoptosis in this T-cell lymphoma. An increase in Bcl-2 content with increasing aggressiveness among these variants, implicates this protein in this tumor progression-related resistance to apoptosis. However, the two variants of highest malignancy, TAU-42 and TAU-46, did not follow the same trend, since they displayed a relatively high content in apoptotic cells and a low Bcl-2 content. Fas receptor expression did not correlate with tendency to apoptosis, indicating that malignant behavior in the AKR lymphoma does not depend on CD95/Fas/APO1 downregulation. Overexpression of p53 was observed only in one of the variants of lowest malignancy.
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    URL: http://dx.doi.org/10.1023/A:1009648325350
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    Signalling steps in apoptosis by ether lipids (1999)
    Springer
    Apoptosis 4 (1999), S. 419-427 
    Details
    ISSN: 1573-675X
    Keywords: Anti-oxidant defence ; apoptosis ; ether lipids ; glutathione ; ionising radiation ; stress.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We have investigated the mechanisms of induction of apoptosis by the antineoplastic ether lipid ET-18-OCH3 (ALP) in sensitive S49wt mouse lymphoma cells and ALP-resistant S49ar variants, both with wild-type p53, and in related L1210 cells with mutated p53. Ether lipid-resistant S49ar cells were cross-resistant to extracellular stress factors (cold shock, heat shock, H2O2, dimethylsulfoxide) and to radiation-induced apoptosis but not to physiological apoptotic signals (dexamethasone, growth factor deprivation, thapsigargin, C2-ceramide) and expressed similar levels of the apoptosis-regulating proteins Bcl-2, Bcl-X, Bax, Bad and Bak as did the parent S49wt cells. The uptake of [3H]-ALP was strongly reduced in the stress-resistant cells but this was not associated with significant differences in membrane cholesterol:phospholipid content nor in membrane microviscosity. In S49ar cells the activity of the antioxidant enzyme glutathione peroxidase (GSH-Px) was increased 4-fold and depletion of glutathione with the drug L-buthionine-S-R-sulfoximine (L-BSO) lowered the resistance of S49ar cells to ALP, stress factors and ionising radiation. The results indicate that ether lipids induce apoptosis by imposing a special form of physico-chemical stress, mediated by reactive oxygen species but independent of p53 status. The capacity of glutathione-dependent anti-oxidant defence appeared an important and shared determinant of the sensitivity to ether lipids, several types of extracellular stress and ionising radiation.
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    URL: http://dx.doi.org/10.1023/A:1009644208512
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    Sequential occurrence of mitochondrial and plasma membrane alterations, fluctuations in cellular Ca2+ and pH during initial and later phases of cell death (1999)
    Springer
    Apoptosis 4 (1999), S. 455-460 
    Details
    ISSN: 1573-675X
    Keywords: Annexin V ; apoptosis ; flow cytometry ; intracellular Ca2+ ; intracellular pH ; mitochondrial membrane potential.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The sequential occurrence of plasma and mitochondrial membrane alterations, intra-cellular pH shifts and changes in intracellular Ca2+ concentration after induction of cell death was monitored by flow cytometry in Jurkat and HSB2-cells. Cell death was induced by treatment with anti-Fas antibodies or by irradiation. Phosphatidylserine (PS) exposure and plasma membrane integrity were measured with FITC-Annexin V adhesion and by Propidium Iodide exclusion. Transition of the mitochondrial membrane potential was monitored by the occurrence of decay of DiOC6 fluorescence. Intracellular pH shifts were monitored by changes in the ratio of fluorescence at 575 nm and at 635 nm of SNARF-1-AM. Fluctuations in intracellular Ca2+ concentration were established by changes in Fura red quenching. The Jurkat cells were sensitive to anti-Fas treatment, while HSB-2 cells were not. HSB-2 cells appeared more sensitive to radiation damage than Jurkat cells. In all experiments the transition of mitochondrial membrane potential occurred first, almost immediately followed by PS exposure. Fluctuations in intracellular Ca2+ concentration occurred later and were less outspoken. A decrease in intracellular pH occurred not earlier than 24 hours after anti-Fas treatment. Chelation of intracellular Ca2+ concentration with BAPTA-AM had no effect on the time sequence of cell death related events.
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    URL: http://dx.doi.org/10.1023/A:1009604510329
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    The globalization of N deposition: ecosystem consequences in tropical environments (1999)
    Springer
    Biogeochemistry 46 (1999), S. 67-83 
    Details
    ISSN: 1573-515X
    Keywords: acidification ; anthropogenic nitrogen ; cations ; nitrate leaching ; nitric oxide ; nitrous oxide ; nutrient limitation ; phosphorus ; productivity ; tropical ecosystems
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract Human activities have more than doubled the inputs of nitrogen (N) into terrestrial systems globally. The sources and distribution of anthropogenic N, including N fertilization and N fixed during fossil fuel combustion, are rapidly shifting from the temperate zone to a more global distribution. The consequences of anthropogenic N deposition for ecosystem processes and N losses have been studied primarily in N-limited ecosystems in the temperate zone; there is reason to expect that tropical ecosystems, where plant growth is most often limited by some other resource, will respond differently to increasing deposition. In this paper, we assess the likely direct and indirect effects of increasing anthropogenic N inputs on tropical ecosytem processes. We conclude that anthropogenic inputs of N into tropical forests are unlikely to increase productivity and may even decrease it due to indirect effects on acidity and the availability of phosphorus and cations. We also suggest that the direct effects of anthropogenic N deposition on N cycling processes will lead to increased fluxes at the soilwater and soil-air interfaces, with little or no lag in response time. Finally, we discuss the uncertainties inherent in this analysis, and outline future research that is needed to address those uncertainties.
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    URL: http://dx.doi.org/10.1007/BF01007574
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    Ecosystem constraints to symbiotic nitrogen fixers: A simple model and its implications (1999)
    Springer
    Biogeochemistry 46 (1999), S. 179-202 
    Details
    ISSN: 1573-515X
    Keywords: carbon dioxide ; grazing ; nitrogen fixation ; nitrogen limitation ; phosphorus ; shade
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract The widespread occurrence ofN limitation to net primary production (NPP) and other ecosystem processes, despite the ubiquitous occurrence ofN-fixing symbioses, remains a significant puzzle in terrestrial ecology. We describe a simple simulation model for an ecosystem containing a generic nonfixer and a symbioticN fixer, based on: (1) a higher cost forN acquisition byN fixers than nonfixers; (2) growth of fixers and fixation ofN only when lowN availability limits the growth of nonfixers, and other resources are available; and (3) losses of fixedN from the system only when the quantity of availableN exceeds plant and microbial demands. Despite the disadvantages faced by theN fixer under these conditions,N fixation and loss adjustN availability close to the availability of other resources, and biomass and NPP in this simple model can be substantially but only transientlyN limited. We then modify the model by adding: (1) losses ofN in forms other than excess availableN (e.g., dissolved organicN, trace gases produced by nitrification); and (2) constraints to the growth and activity ofN fixers imposed by differential effects of shading,P limitation, and grazing. The combination of these processes is sufficient to describe an open system, with input from both precipitation andN fixation, that is nevertheless stronglyN-limited at equilibrium. This model is useful for exploring causes and consequences of constraints toN fixation, and hence ofN limitation, and we believe it will also be useful for evaluating howN fixation and limitation interact with elevatedCO 2 and other components of global enviromental change.
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    URL: http://dx.doi.org/10.1007/BF01007579
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    The impact of accelerating land-use change on the N-Cycle of tropical aquatic ecosystems: Current conditions and projected changes (1999)
    Springer
    Biogeochemistry 46 (1999), S. 109-148 
    Details
    ISSN: 1573-515X
    Keywords: estuaries ; lakes ; marine ; nitrogen ; phosphorus ; rivers ; streams ; temperate ; tropics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract Published data and analyses from temperate and tropical aquatic systems are used to summarize knowledge about the potential impact of land-use alteration on the nitrogen biogeochemistry of tropical aquatic ecosystems, identify important patterns and recommend key needs for research. The tropical N-cycle is traced from pre-disturbance conditions through the phases of disturbance, highlighting major differences between tropical and temperate systems that might influence development strategies in the tropics. Analyses suggest that tropical freshwaters are more frequently N-limited than temperate zones, while tropical marine systems may show more frequent P limitation. These analyses indicate that disturbances to pristine tropical lands will lead to greatly increased primary production in freshwaters and large changes in tropical freshwater communities. Increased freshwater nutrient flux will also lead to an expansion of the high production, N- and light-limited zones around river deltas, a switch from P- to N-limitation in calcareous marine systems, with large changes in the community composition of fragile mangrove and reef systems. Key information gaps are highlighted, including data on mechanisms of nutrient transport and atmospheric deposition in the tropics, nutrient and material retention capacities of tropical impoundments, and N/P coupling and stoichiometric impacts of nutrient supplies on tropical aquatic communities. The current base of biogeochemical data suggests that alterations in the N-cycle will have greater impacts on tropical aquatic ecosystems than those already observed in the temperate zone.
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    URL: http://dx.doi.org/10.1023/A:1006156213761
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    Detection of genetic relationships among fourArtemia species using randomly amplified polymorphic DNA (RAPD) (1999)
    Springer
    International journal of salt lake research 8 (1999), S. 139-147 
    Details
    ISSN: 1573-8590
    Keywords: Artemia ; genetic polymorphism ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geography
    Notes: Abstract We have applied the technique of random amplification of polymorphic DNA (RAPD) to the analysis of the relationships among four species of brine shrimp:Artemia franciscana, A. urmiana, A. sinica, andA. parthenogenetica. Seventy ten-base synthetic oligonucleotides were used to amplify a total of 458 distinct fragments. DNA polymorphisms were found in all the species examined; the highest percentage of polymorphic bands was found inA. parthenogenetica, with 28.8 per cent. Each species was scored for the presence or absence of every amplification product and the data entered into a binary data matrix. Cluster analysis was then performed to create a dendrogram using UPGMA by the NTSYS program. There are significant differences between bisexual species and parthenogenetic populations.A. parthenogenetica provided 94 specific molecular markers, while bisexual species gave 27 specific molecular markers.A. sinica is a species distinct from the other Old World bisexual species.
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    URL: http://dx.doi.org/10.1007/BF02442127
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    Detection of genetic relationships among four Artemia species using randomly amplified polymorphic DNA (RAPD) (1999)
    Springer
    International journal of salt lake research 8 (1999), S. 139-147 
    Details
    ISSN: 1573-8590
    Keywords: Artemia ; genetic polymorphism ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geography
    Notes: Abstract We have applied the technique of random amplification of polymorphic DNA (RAPD) to the analysis of the relationships among four species of brine shrimp: Artemia franciscana, A. urmiana, A. sinica, and A. parthenogenetica. Seventy ten-base synthetic oligonucleotides were used to amplify a total of 458 distinct fragments. DNA polymorphisms were found in all the species examined; the highest percentage of polymorphic bands was found in A. parthenogenetica, with 28.8 per cent. Each species was scored for the presence or absence of every amplification product and the data entered into a binary data matrix. Cluster analysis was then performed to create a dendrogram using UPGMA by the NTSYS program. There are significant differences between bisexual species and parthenogenetic populations. A. parthenogenetica provided 94 specific molecular markers, while bisexual species gave 27 specific molecular markers. A. sinica is a species distinct from the other Old World bisexual species.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1009039815660
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    Gene and Genotypic Diversity of Phytophthora cinnamomi in South Africa and Australia Revealed by DNA Polymorphisms (1999)
    Springer
    European journal of plant pathology 105 (1999), S. 667-680 
    Details
    ISSN: 1573-8469
    Keywords: asexual reproduction ; mating types ; oomycetes ; origin ; RAPD ; RFLP ; population genetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Phytophthora cinnamomi isolates from South Africa and Australia were compared to assess genetic differentiation between the two populations. These two populations were analysed for levels of phenotypic diversity using random amplified polymorphic DNAs (RAPDs) and gene and genotypic diversity using restriction fragment length polymorphisms (RFLPs). Sixteen RAPD markers from four decanucleotide Operon primers and 34 RFLP alleles from 15 putative loci were used. A few isolates from Papua New Guinea known to posses alleles different from Australian isolates were also included for comparative purposes. South African and Australian P. cinnamomi populations were almost identical with an extremely low level of genetic distance between them (Dm=0.003). Common features for the two populations include shared alleles, low levels of phenotypic/genotypic diversity, high clonality, and low observed and expected levels of heterozygosity. Furthermore, relatively high levels of genetic differentiation between mating type populations (Dm South Africa=0.020 and Dm Australia=0.025 respectively), negative fixation indices, and significant deviations from Hardy–Weinberg equilibrium, all provided evidence for the lack of frequent sexual reproduction in both populations. The data strongly suggest that both the South African and Australian P. cinnamomi populations are introduced.
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    URL: http://dx.doi.org/10.1023/A:1008755532135
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    Pharmacodynamics and Toxicodynamics of Drug Action: Signaling in Cell Survival and Cell Death (1999)
    Springer
    Pharmaceutical research 16 (1999), S. 790-798 
    Details
    ISSN: 1573-904X
    Keywords: MAPK ; caspases ; chemopreventive agents ; phase II drug metabolizing enzymes ; apoptosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract In therapeutic response to drugs, the plasma concentration range leads to the establishment of a safe and effective dosage regimen. Our hypothesis is that by studying drug concentration-dependent effect on signal transduction mechanisms, a better understanding of the beneficial pharmacodynamic and adverse toxicodynamic responses elicited by the drug may be achieved. Using two classes of chemopreventive compounds (phenolic antioxidants and isothiocyanates), we illustrate the potential utility of two signal transduction pathways elicited by these agents to predict the pharmacodynamic effect (induction of Phase II drug metabolizing enzymes) and the potential toxicodynamic response (stimulation of caspase activity and cytotoxic cell death). At lower concentration, phenolic antioxidants and isothiocyanates activate mitogen-activated protein kinase (MAPK; extracellular signal-regulated protein kinase 2, ERK2; and c-Jun N-terminal kinase 1, JNK1) in a concentration-and time-dependent manner. The activation of MAPK by these compounds may lead to the induction of cell survival/protection genes such as c-jun, c-fos, or Phase II drug metabolizing enzymes. However, at higher concentrations, these agents activate another signaling molecule, ICE/Ced3 cysteine protease enzymes (caspases) leading to apoptotic cell death. The activation of these pathways may dictate the fate of the cells/tissues upon exposure to drugs or chemicals. At lower concentrations, these compounds activate MAPK leading to the induction of Phase II genes, which may protect the cells/tissues against toxic insults and therefore may enhance cell survival. On the other hand, at higher concentrations, these agents may activate the caspases, which may lead to apoptotic cell death, and have toxicity. Understanding the activation of these and other signal transduction events elicited by various drugs and chemicals may yield insights into the regulation of gene expression of drug metabolizing enzymes and cytotoxicity. Thus, the study of signaling events in cell survival (hemeostasis) and cell death (cytotoxicity) may have practical application during pharmaceutical drug development.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1011953431486
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    The limits of phosphorus removal in wetlands (1999)
    Springer
    Wetlands ecology and management 7 (1999), S. 165-175 
    Details
    ISSN: 1572-9834
    Keywords: flow ; loading ; models ; phosphorus ; removal limits ; wetlands
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The phosphorus concentrations exported from wetlands are explored via data and a first order model. The graph of outlet concentration versus areal phosphorus loading is used to display these data and the model. For a given wetland, data and models show that P concentrations show an ‘S’ curve response to increasing P loadings. The lower plateau is the background concentration and the upper plateau is the inlet concentration. The position of the ascending limb between the two plateaus is positioned differently for different wetlands. Phosphorus (P) removal in wetlands is often typified by a stable decreasing gradient of P concentrations from inlet to outlet, and an accompanying stable decreasing gradient in P assimilation. Limits to removal are inherent in the physical, chemical and biological processes. A lower outlet concentration limit exists because of the P cycle in the un-impacted wetland. The loading at which the outlet concentration departs from background, the lower knee in the loading curve, varies from wetland to wetland. An upper outlet concentration limit is imposed by the source concentration for extremely high inflows. The first order mass balance equation interpolates between these limits. The model gives further insights about performance within an overall envelope. The water carrying capacity of the wetland precludes flows in excess of the hydraulic conveyance capacity, thus limiting the possible P loadings to the system. Conversely, extremely low hydraulic loadings cause the wetland to be dominated by atmospheric additions and losses. The central tendency of inter-system data in the North American Database is shown to be inadequate to draw generalized conclusions about ecosystem processes in an individual wetland. The proposed ‘one gram rule’ of Richardson, et al. (1997) is shown to be an over-simplification.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1008415401082
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  • 85
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    Intraspecific classification of melons (Cucumis melo L.) in view of their phenotypic and molecular variation (1999)
    Springer
    Plant systematics and evolution 217 (1999), S. 313-332 
    Details
    ISSN: 1615-6110
    Keywords: Cucumis melo ; melon ; intra-specific classification ; RAPD ; Inter-SSR ; DNA fingerprinting
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cucumis melo L. (melon) genotypes differ widely in morphological and biochemical traits. Intraspecific classification of such variability has been difficult, and most taxonomists still rely on the work of Naudin (1859). A collection of 54 accessions representing diverse genotypes from 23 countries was surveyed. Morphological traits related to the vegetative and flowering stages and mature fruit morphology and quality parameters, e.g., taste, aroma, sugar composition and pH, were scored. These were used to construct a “botanical-morphological” dendrogram that generally reflected the classification ofCucumis melo into several horticultural varieties. DNA polymorphism among the accessions was assessed using the Inter-SSR-PCR and RAPD techniques that detected abundant DNA polymorphism among melon genotypes. Cluster analysis indicated that the largest divergence was between North American and Europeancantalupensis andinodorus cultivars as one group, and the more “exotic” varieties:conomon, chito, dudaim, agrestis andmomordica, as a second group. The molecular phylogeny agreed, broadly, with the classification of melon into two subspecies, and did not contradict the division into “horticultural varieties”. It was apparent, however, that the infra-specific division is rather loose, molecular variation being distributed continuously between and within cultivar groups. We suggest that despite the morphological diversity, separation between varietal-groups may be based on a too small number of genes to enable unambiguous infra-specific classification based on DNA diversity.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00984373
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  • 86
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    Effects of phosphorus nutrition on tiller emergence in wheat (1999)
    Springer
    Plant and soil 209 (1999), S. 283-295 
    Details
    ISSN: 1573-5036
    Keywords: leaf emergence ; phosphorus ; photosynthesis ; tillering ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Phosphorus (P) deficiency limits the yield of wheat, particularly by reducing the number of ears per unit of area because of a poor tiller emergence. The objectives of this work were to (i) determine whether tiller emergence under low phosphorus availability is a function of the availability of assimilates for growth or a direct result of low P availability, (ii) attempt to establish a quantitative relation between an index of the availability of P in the plant and the effects of P deficiency on tiller emergence, and (iii) to provide a better understanding of the mechanisms involved in tiller emergence in field-grown wheat. Wheat (Triticum aestivum L., cv. INTA Oasis), was grown in the field under drip irrigation on a typic Argiudol, low in P (5.5 μg P g-1 soil Bray & Kurtz I) in Balcarce, Argentina. Treatments consisted of the combination of three levels of P fertilization 0, 60 and 200 kg P2O5 ha-1, and two levels of assimilate availability, a control (non-shaded) and 65% of reduction in incident irradiance from seedling emergence until the end of tillering (shaded). Phosphorus treatments significantly modified the pattern of growth and development of the plants. Shading reduced the growth and concentration of water-soluble carbohydrates in leaves and stems. Leaf photosynthetic rate at saturating irradiance was reduced by P deficiency, but was not affected by shading. At shoot P concentrations less than 4.2 g P kg-1 the heterogeneity in the plant population increased with respect to the number of plants bearing a certain tiller. At a shoot P concentration of 1.7 g P kg-1 tillering ceased completely. Phosphorus deficiency directly altered the normal pattern of tiller emergence by slowing the emergence of leaves on the main stem (i.e. increasing the phyllochron), and by reducing the maximum rate of tiller emergence for each tiller.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1004690404870
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    The effect of regeneration burns on the growth, nutrient acquisition and mycorrhizae of Eucalyptus regnans F. Muell. (mountain ash) seedlings (1999)
    Springer
    Plant and soil 210 (1999), S. 273-283 
    Details
    ISSN: 1573-5036
    Keywords: ectomycorrhizae ; Eucalyptus regnans ; forest burns ; nitrogen ; phosphorus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract This study was conducted to compare the effects on the growth of Eucalyptus regnans seedlings of unheated soil and soil heated to different extents (as indicated by soil colour–bright red or black) in burnt logging coupes, and to separate the effects of heating of the soil on direct nutrient availability and on morphotypes and effectiveness of ectomycorrhizae. Burnt soils were collected from three logging coupes burnt 2, 14 and 25 months previously and unbumt soil from adjacent regrowth forests. Compared to unburnt soil, the early seedling growth was stimulated in black burnt soil from all coupes (burnt 2, 14 and 25 months previously). Seedling growth was generally poor in red burnt soil, especially in soil collected 2 months after burning. However, the concentration of extractable P was extremely high in red burnt soil, especially in soil collected 2 months after burning. In black burnt soil, extractable P was increased in soil 2 months after burning, but not in the soils collected 14 or 25 months after burning. However, both total P content and concentration in seedlings were increased in all collections of black burnt soil. Frequency of ectomycorrhizae was high in seedlings grown in all black burnt soils, but the mycorrhizal mantles were poorly developed in seedlings in black burnt soil collected 2 months after burning. Seedlings were also ectomycorrhizal in red burnt soil, except in soil collected 2 months after burning. Fine root inocula from seedlings grown in black burnt soils collected 14 and 25 months after burning significantly stimulated both seedling growth and P uptake compared with the uninoculated control, whereas the fine root inocula from the seedlings grown in all the other soils did not. These results suggest that, in black burnt soil, both direct nutritional changes and changes in the ectomycorrhizae may contribute to seedling growth promotion after regeneration burns. The generally poor seedling growth in red burnt soils is likely to have been due to N deficiency as the seedlings in these soils were yellow-green and the tissue concentrations of N were significantly lower than in other treatments.
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    URL: http://dx.doi.org/10.1023/A:1004609912315
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    Detection of molecular markers associated with linolenic and erucic acid levels in spring rapeseed (Brassica napur L.) (1999)
    Springer
    Euphytica 105 (1999), S. 173-181 
    Details
    ISSN: 1573-5060
    Keywords: Brassica napur ; doubled haploids ; RAPD ; linolenic acid ; erucic acid ; marker assisted selection ; rapeseed breeding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Undesirable characteristic of rapeseed oil is a relatively high level of linolenic acid (18:3), which is easily oxidized leading to rancidity and a shortened shelf life of the oil. Previous attempts to reduce linolenic acid levels in rapeseed oil through breeding have been impaired by complex genetics and strong environmental sensitivity of this trait. Therefore, our objective was to develop molecular markers for low linolenic acid that could facilitate the breeding of low linolenic rapeseed. Bulked segregant analysis was employed to identify two RAPD markers associated with 18:3 in a doubled haploid population segregating for linolenic and erucic acid levels. Based on analysis of individual DH lines, the markers RM350 and RM574, representing two independent loci, accounted for a total of 39% of the genetic variability in this population. This marker RM350 alone accounted for 25% genetic variation for this trait with no evidence of recombination. Significant interlocus interaction found between the markers RM350 and RM574 suggested that epistasis was involved in the genetic control of 18:3 level in this population. Another marker designated as RM322, which was independent of the other two, was found significantly associated with the erucic acid level and oil content. RAPD markers identified in this study should be a useful tool for the early detection of low linolenic, or low or high erucic acid genotypes in rapeseed breeding programs based on doubled haploids.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1003494217074
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    Identification of RAPD markers linked to fusarium crown and root rot resistance (Frl) in tomato (1999)
    Springer
    Euphytica 105 (1999), S. 205-210 
    Details
    ISSN: 1573-5060
    Keywords: crown rot ; fusarium crown and root rot ; genetic linkage ; Lycopersicon peruvianum ; molecular markers ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract In tomato ( Lycopersicon esculentum Mill.) a single dominant gene ( Frl) on chromosome 9 confers resistance to fusarium crown and root rot (crown rot) incited by Fusarium oxysporum f. sp. radicis-lycopersici. To identify randomly amplified polymorphic DNA (RAPD) markers linked to Frl, crown rot susceptible and resistant tomato lines were screened for polymorphisms using 1000 random 10-mer primers and three reliable RAPD markers were found linked to Frl (UBC #'s 116, 194, and 655). A codominant polymorphic PCR marker of TG101, a restriction fragment length polymorphic (RFLP) marker linked to Frl, was developed to facilitate the linkage studies. Using TG101 and the four RAPD markers, on a Frl segregating backcross population of 950 plants indicated that all belong to the same linkage group. The polymorphic allele order was found to be TG101 – 655 – 116 – 194 – Frl. UBC 194 was found to be 5.1 cM from Frl in this population. Furthermore, it was the only marker found in the resistant genotypes ‘Mocis’ and Fla 7226, whereas resistant genotypes ‘Momor’, Ohio 89-1, and Fla 7464 all had UBC 194 and UBC #'s 116, 194, and 655.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1003497719705
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    Identification of a RAPD marker linked to a brown planthopper resistance gene in rice (1999)
    Springer
    Euphytica 107 (1999), S. 23-28 
    Details
    ISSN: 1573-5060
    Keywords: Bph-1 ; linkage analysis ; mapping ; RAPD ; RFLP ; rice
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract We report the tagging of a brown planthopper (BPH) resistance gene (Bph–1) in rice using RAPD and RFLP markers. The Korean rice variety ‘Gayabyeo’ has dominant duplicate genes including Bph–1 conferring resistance to biotype 1 of BPH. Bulked segregant RAPD analysis was employed for rapid identification of DNA markers linked to resistance genes. For tagging these two genes, an F2F3 population from a ‘Gayabyeo’ × ‘Nagdongbyeo’ cross was developed and evaluated for BPH resistance. Three bulked DNAs from two groups of homozygous BPH resistant (each for Bph–1 and the other unknown gene) and homozygous susceptible F2 plants were analyzed by RAPD using 140 random oligomers. One primer, OPD–7 yielded a 700-bp fragment that was present in Gayabyeo and resistant F2 plants (homozygous for Bph-1 locus) but absent in Nagdongbyeo and susceptible F2 plants. Cosegregation of this marker with Bph-1 was verified using an F2 population segregating for Bph-1. Chromosomal regions surrounding the Bph-1 were examined with additional RFLP and microsatellite markers on chromosome 12 to define the location of the RAPD marker and Bph-1. Use of this RAPD marker could facilitate early selection of resistant lines for BPH.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1003506830735
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  • 91
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    Persistence of tree related patterns in soil nutrients following slash-and-burn disturbance in the tropics (1999)
    Springer
    Plant and soil 209 (1999), S. 137-157 
    Details
    ISSN: 1573-5036
    Keywords: fire ; nitrogen ; phosphorus ; soil nutrient heterogeneity ; tree effects ; tropical dry forest
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Individual trees are known to influence soil chemical properties, creating spatial patterns that vary with distance from the stem. The influence of trees on soil chemical properties is commonly viewed as the agronomic basis for low-input agroforestry and shifting cultivation practices, and as an important source of spatial heterogeneity in forest soils. Few studies, however, have examined the persistence of the effects of trees on soil after the pathways responsible for the effects are removed. Here, we present evidence from a Mexican dry forest indicating that stem-related patterns of soil nutrients do persist following slash-and-burn removal of trees and two years of cropping. Pre-disturbance concentrations of resin extractable phosphorus (P), bicarbonate extractable P, NaOH extractable P, total P, total nitrogen (N) and carbon (C), KCl extractable nitrate (NO3 -), and net N mineralization and nitrification rates were higher in stem than dripline soils under two canopy dominant species of large-stemmed trees with contrasting morphologies and phenologies (Caesalpinia eriostachys Benth. and Forchhammeria pallida Liebm.). These stem effects persisted through slash burning and a first growing season for labile inorganic and organic P, NaOH inorganic P, and plant-available P, and through a second growing season for labile organic P, NaOH organic P, and plant-available P. While stem effects for extractable NO3 -, net nitrification rates, total N and C disappeared after felling and slash burning, these stem effects returned after the first growing season. These results support the view that tree-influenced patterns of soil nutrients do persist after tree death, and that trees contribute to the long-term spatial heterogeneity of forest soils.
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    URL: http://dx.doi.org/10.1023/A:1004503023973
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    Nitrogen and phosphorus requirement of the microbiota in soils of the Bornhöved Lake district (1999)
    Springer
    Plant and soil 212 (1999), S. 173-181 
    Details
    ISSN: 1573-5036
    Keywords: nitrogen ; phosphatase activity ; phosphorus ; protease activity ; soil microbial biomass ; substrate-induced respiration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Estimating in situ N and P status of the soil microbiota is complicated because microbiological features reflect potentials rather than field conditions. Complementary microbiological assays were, therefore, combined to evaluate the N and P requirement of the microbiota in seven agricultural, grassland and forest topsoils of the Bornhöved Lake district as follows: (i) the sensitivity of the substrate-induced respiration (SIR) to supplemental addition of N and P was monitored during microbial growth and (ii) soil protease and phosphatase activities were analysed and related to soil mass and microbial biomass content. Nitrogen addition increased the maximal SIR rate in all except one soil indicating that the growth of organisms is restricted by this element when easily degradable C source is present. Supplemental N (and in some cases also P) retarded the respiratory response within the first 24 h which suggests microbial sensitivity and/or greater anabolic efficiency. With additional N the maximal SIR rate was most strongly enhanced in topsoils of the beech forest and the dystric alder forest. Thus, the microbial growth in these soils that were below litter horizons seems to be mostly restricted by N. Supplemental P positively affected respiratory response of soils under monoculture, wet grassland and dystric alder forest. In the dystric alder forest soil, high rates of alkaline and unbuffered phosphatase activity were observed when activity was related to either soil mass or microbial biomass content. The data of proteolytic and phospholytic enzymes are discussed with reference to nutrient deficiency and microbial strategy for N and P adsorption.
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    URL: http://dx.doi.org/10.1023/A:1004677427165
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    Root-induced modifications of the exchange of phosphate ion between soil solution and soil solid phase (1999)
    Springer
    Plant and soil 211 (1999), S. 103-110 
    Details
    ISSN: 1573-5036
    Keywords: bioavailability ; isotopic evaluation ; phosphate ion exchange ; phosphorus ; rhizosphere ; soil solution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The uptake of phosphorus (P) by roots results in a depletion of phosphate ions (PO4) in the rhizosphere. The corresponding decrease in PO4 concentration in the soil solution (CP) gives rise to a replenishment of P from the solid phase which is time- and CP-dependent. This PO4 exchange which reflects the buffer power of the soil for PO4 also varies with the composition and the physico-chemical conditions of the soil. As root activity can modify these physico-chemical conditions in the rhizosphere, the question arises whether these modifications affect the ability of PO4 bound to the soil solid phase to exchange with PO4 in soil solution. The aim of the present work was to measure and compare the parameters which describe the amount of PO4 bound to soil solid phase that is capable to replenish solution P for both rhizosphere and bulk soils. The soil sample was a P-enriched, calcareous topsoil collected from a long-term fertiliser trial. Rhizosphere soil samples were obtained by growing dense mats of roots at the surface of 3 mm thick soil layer for one week. Three plant species were compared: oilseed rape (Brassica napus L., cv Goeland) pea (Pisum sativum L., cv. Solara) and maize ( Zea mays L., cv. Volga). The time- and CP-dependence of the PO4 exchange from soil to solution were described using an isotopic dilution method. The measured CP values were 0.165 mg P L−1 for bulk soil and 0.111, 0.101 and 0.081 mg P L−1 for rhizosphere soils of maize, pea and rape, respectively. The kinetics of the PO4 exchange between liquid and solid phases of soil were significantly different between rhizosphere and bulk soils. However, when changes in CP were accounted for, the parameters describing the PO4 exchange with time and CP between soil solution and soil solid phase were found to be very close for bulk and rhizosphere soils. For this calcareous and P-enriched soil, plant species differed in their ability to deplete PO4 in solution. The resulting changes in the ability of the soil solid phase to replenish solution PO4 were almost fully explained by the depletion of soil solution P.
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    URL: http://dx.doi.org/10.1023/A:1004485432261
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    Analyses of genetic diversity in Cuban rice varieties using isozyme, RAPD and AFLP markers (1999)
    Springer
    Euphytica 109 (1999), S. 107-115 
    Details
    ISSN: 1573-5060
    Keywords: AFLP ; DNA fingerprinting ; isozymes ; RAPD ; rice
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A survey of the genetic diversity among the major cuban rice cultivars was conducted using isozyme, RAPD and AFLP markers. Polymorphisms were detected for esterases, peroxidases, alcohol dehydrogenases and polyphenoloxidases systems; 21 RAPD primers and four AFLP primer combinations. Heterozygosity arithmetic mean value (Hav(p)), the effective multiplex ratio (EMR) and the marker index (MI), were calculated for isozyme, RAPD and AFLP markers. The mean value of genetic similarity among the different varieties was 0.92 for isozyme, 0.73 for RAPD and 0.58 for AFLP analyses. Thus, AFLP were able to detect polymorphisms with higher efficiency than RAPD (+15%) and isozyme (+34%). Data from the isozyme, RAPD and AFLP analyses were used to compute matrices of genetic similarities. The efficiency of the UPGMA for the estimation of genetic relatedness among varieties was supported by cophenetic correlation coefficients. The resulting values indicated that the distortion level for the estimated similarities was minimal. The correlation coefficients obtained by the Mantel matrix correspondence test, which was used to compare the cophenetic matrices for the different markers, showed that estimated values of genetic relationship given for isozyme and RAPD markers (r = 0.89), as well as for AFLP and RAPD markers (r = 0.82) were properly related. However, AFLP and isozyme data showed only moderate correlation (r = 0.63). Although the genetic variability found among the different cultivars was low, both RAPD and AFLP markers proved to be efficient tools in assessing the genetic diversity of rice genotypes.
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    URL: http://dx.doi.org/10.1023/A:1003672729948
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    Genetic diversity of plums characterized by random amplified polymorphic DNA (RAPD) analysis (1999)
    Springer
    Euphytica 109 (1999), S. 143-147 
    Details
    ISSN: 1573-5060
    Keywords: classification ; DNA ; plum varieties ; Prunus ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract We investigated the genetic diversity of 42 plum varieties by RAPD analysis. Twenty primers discriminated all plum varieties excepting two synonymous pairs: 'Botankyou and ‘Kelsey’, and ‘Chairn’ and ‘Tragedy’. Two North American plums, ‘Beach Plum’ and ‘Glow’, were genetically distinct from the other plums by cluster analysis. Overlaps observed between the ‘European plum group’ and the ‘Japanese plum group’, were perhaps due to intercrossing. We could also discriminate ‘Sordum’ from 'Late Sordum and ‘Bansei Sordum’, although ‘Late Sordum’ and ‘Bansei Sordum’ are thought to be derived from bud mutants of ‘Sordum’.
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    URL: http://dx.doi.org/10.1023/A:1003728201100
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    Effect of ethrel on apoptosis in carrot protoplasts (1999)
    Springer
    Plant growth regulation 27 (1999), S. 119-123 
    Details
    ISSN: 1573-5087
    Keywords: apoptosis ; carrot protoplast ; DNA ladder ; ethrel ; ethylene ; nucleus condensation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract In recent years, apoptosis has been reported to exist in plants during normal development and in response to stress. However, little is known about the relation of hormones to this form of programmed cell death. Here, we report examination of characteristics of apoptosis in carrot protoplasts induced by ethylene evolved from ethrel (2-chloroethylphosphonic acid). Nucleus condensation and DNA ladders were observed, and neutral comet assay, which detects DNA cleavage, also provided evidence that ethrel treatment resulted in nuclear DNA fragmentation. Strikingly, a close correlation between the incidence of DNA comets and the percentage of apoptotic protoplasts was shown in ethrel-treated carrot protoplasts. In conclusion, this study demonstrated that ethylene is an active inducer of apoptosis in carrot protoplasts, and that ethylene-induced plant cell death showed characteristics similar to those of apoptosis in animals.
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    URL: http://dx.doi.org/10.1023/A:1006105101813
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    The genetic structure and conservation of aus, aman and boro rices from Bangladesh (1999)
    Springer
    Genetic resources and crop evolution 46 (1999), S. 587-598 
    Details
    ISSN: 1573-5109
    Keywords: diversity ; genetic resources ; GIS ; Oryza sativa ; RAPD ; rice
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A diverse set of 115 rice varieties from Bangladesh was surveyed using 35 polymorphic RAPD (randomly amplified polymorphic DNA) markers and the genetic structure of this germplasm, encompassing the principal rice ecotypes of Bangladesh (aus, aman and boro), was determined using multivariate analysis. The level of genetic diversity was evaluated and compared with the levels of diversity found within other rice growing areas of the world. Geographical information systems analysis using Atlas-GIS was employed to analyse and present the geographic distribution of genetic diversity across Bangladesh, and cluster analysis was used to test the efficiency of selection of material for a core collection.
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    URL: http://dx.doi.org/10.1023/A:1008749532171
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    Marker-assisted sampling of the cultivated Andean potato Solanum phureja collection using RAPD markers (1999)
    Springer
    Genetic resources and crop evolution 46 (1999), S. 547-555 
    Details
    ISSN: 1573-5109
    Keywords: core collection ; germplasm ; molecular marker ; potato ; RAPD ; Solanum phureja
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The potato crop originated in the Andean highlands where numerous farmer's varieties and non-cultivated wild species exist. An Andean potato collection is held in trust at the International Potato Center (CIP) to preserve the biodiversity of this crop and ensure the supply of germplasm for potato improvement worldwide. A core collection representing the biodiversity of the Andean potato germplasm is under construction using morphological, molecular, and geographic data. One of the eight cultivated potato species, Solanum phureja, has been genotyped using the RAPD technique. A protocol suitable for large germplasm collection genotyping has been developed to process numerous samples at reasonable costs. From 106 RAPD primers evaluated, we have selected 12 primers yielding 102 polymorphic markers, which unambiguously discriminated all 128 accessions but 2 that are possible duplicates. The S. phureja germplasm collected throughout the Andean countries appears to have a homogeneous genetic constitution. There was no clear geographic pattern as indicated by cluster analysis of the RAPD data. A sub-group of 20 accessions has been identified on the basis of the marker data and selected to maximize molecular (RAPD) variance and polymorphism. The probability of capturing equal amounts of marker polymorphism in this sub-group of 20 accessions by random sampling is less than 40%. This set accessions represents our first group of accessions that may constitute a core of the S. phureja collection. This tentative core will be challenged for diversity content by alternate markers and agronomic traits. Hence, the methodology for sampling less than 10% of the base collection, proposed for core collections by Brown (1989), can be based on molecular marker data provided cost-efficient fingerprints are developed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1008724007888
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  • 99
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    Qualitative and quantitative investigations of phytoplankton in Lake Ringsjön, Scania, Sweden (1999)
    Springer
    Hydrobiologia 404 (1999), S. 27-40 
    Details
    ISSN: 1573-5117
    Keywords: phytoplankton ; fish reduction ; phosphorus ; biomanipulation ; blue-green algae ; cyanobacteria ; Microcystis ; Anabaena ; Aphanizomenon ; Aulacoseira ; cryptomonads
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The algal record from Lake Ringsjön covers a period of more than 100 years. Towards the end of the 19th century, the lake had a well-functioning commercial fishery, when a brownish mess began to appear in the water, clogging-up the fishing nets and making fishing difficult. This was the first record of algal problems in the lake. Following complaints from the fishermen, the algal flora was investigated and the mess was stated to be a mass development of diatoms belonging to the genus Melosira ( Aulacoseira). Diatom maxima then appeared regularly in spring and autumn, whereas blue-green algae only occurred occasionally and in low abundances. Between 1900 and 1950, nutrient concentrations slowly increased in the lake. Algal blooms of Anabaena lemmermannii and Gloeotrichia echinulata began to appear in summer. Between 1960 and 1980, the lake developed into a hypertrophic status with extensive blooms of blue-green algae from May to October, including high biomasses of mainly Microcystis spp. This severe pollution started with increased usage of the lake, increased tourism and recreation, intensified farming with the introduction of artificial fertilization, and also the diversion of sewage water from a sewage treatment plant. In 1968, cattle death was reported and in 1981, algal toxicity was verified by mouse bioassay. After a period of extremely poor water quality, the lake began to show signs of improvement as a result of the nutrient and fish reduction programmes. The very heavy blooms of blue-green algae decreased and the duration of blooms became shorter. The Microcystis species were to a certain extent replaced by Anabaena and Aphanizomenon species, and biodiversity of algae increased. With respect to the algal community, the lake has improved considerably, but is still eutrophic and suffers summer blooms of toxic blue-green algae.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1003757504155
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  • 100
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    Synthesis of theoretical and empirical experiences from nutrient and cyprinid reductions in Lake Ringsjön (1999)
    Springer
    Hydrobiologia 404 (1999), S. 145-156 
    Details
    ISSN: 1573-5117
    Keywords: biomanipulation ; nutrient reduction ; zooplankton ; phytoplankton ; bottom-up ; top-down ; phosphorus ; submerged vegetation ; benthic macrofauna
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The reduction in external phosphorus load to Lake Ringsjön during the 1980s, did not result in improved water transparency during the following ten-year period. Furthermore, a fish-kill in the Eastern Basin of the lake, in addition to a cyprinid reduction programme (biomanipulation; 1988–1992), in contrast to theory, did not lead to any increase in zooplankton biomass or size. This absence of response in the pelagic food chain may have been attributed to the increase in abundance of YOY (0+) fish, following the fish reduction programme. Despite the lack of effect on zooplankton, there was a decrease in phytoplankton biomass, a change in species composition and an increase in water transparency following biomanipulation. In 1989, one year after the fish-kill in Eastern Basin, the Secchi depth (summer mean) increased from 60 cm to 110 cm. In the following years, water transparency increased further, despite an increase in phosphorus loading. An unexpected effect of the biomanipulation was an increase in benthic invertebrate and staging waterfowl abundances, which occurred 2–4 years after fish reduction. Hence, the response in the benthic community following biomanipulation was considerably stronger than in the pelagic community. A likely explanation is that reduction in abundance of the benthic feeding fish species bream (Abramis brama), strongly affected the benthic invertebrate fauna. In this paper, we present what we believe happened in Lake Ringsjön, and which processes are likely to have been important at various stages of the restoration process.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1003788900521
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