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  • Articles  (485)
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  • 1
    Electronic Resource
    Electronic Resource
    Nitrogen limitation of production in a Californianannual grassland: The contribution of arbuscularmycorrhizae (2000)
    Springer
    Biogeochemistry 49 (2000), S. 37-51 
    Details
    ISSN: 1573-515X
    Keywords: Arbuscular mycorrhizae/Vesicular-arbuscular mycorrhizae (AM/VAM) ; annual grasslands ; benomyl ; nitrogen limitation ; phosphorus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract Nutrient availability limits plant production acrossa wide range of terrestrial ecosystems. In this studyof a Californian annual grassland community, theinfluence of arbuscular mycorrhizal (AM) associationson plant nutrient acquisition was investigated usingfactorial combinations of nitrogen (N), phosphorus (P)and benomyl fungicide. N additions resulted in asignificant increase in shoot biomass demonstratingthat plant productivity in these soils was N-limited. The effect of P additions consistently depended onfungicide treatment. In the absence of benomyl, shootP accumulation was unaltered by P addition. In thepresence of benomyl, P addition significantlyincreased shoot P accumulation and was associated witha consistent trend towards enhanced shoot biomass.The induction of P-deficiency with benomylapplication suggests that effective P acquisition byAM may contribute to the strong N-limitation ofproduction observed in many terrestrial ecosystems.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1006282803693
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  • 2
    Electronic Resource
    Electronic Resource
    Nutrient losses in runoff from grasslandand shrubland habitats in southern New Mexico: II. Field plots (2000)
    Springer
    Biogeochemistry 49 (2000), S. 69-86 
    Details
    ISSN: 1573-515X
    Keywords: Bouteloua eriopoda ; Chihuahuan Desert ; desertification ; hydrology ; Larrea tridentata ; nitrogen ; nutrient budgets ; phosphorus ; runoff
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract Losses of dissolved nutrients (N, P, K, Ca, Mg, Na,Cl, and SO4) in runoff were measured on grasslandand shrubland plots in the Chihuahuan desert ofsouthern New Mexico. Runoff began at a lowerthreshold of rainfall in shrublands than ingrasslands, and the runoff coefficient averaged 18.6%in shrubland plots over a 7-year period. In contrast,grassland plots lost 5.0 to 6.3% of incidentprecipitation in runoff during a 5.5-year period. Nutrient losses from shrubland plots were greater thanfrom grassland plots, with nitrogen losses averaging0.33 kg ha−1 yr−1 vs0.15 kg ha−1 yr−1, respectively, during a 3-year period. Thegreater nutrient losses in shrublands were due tohigher runoff, rather than higher nutrientconcentrations in runoff. In spite of these nutrientlosses in runoff, all plots showed net accumulationsof most elements due to inputs from atmosphericdeposition. Therefore, loss of soil nutrients byhillslope runoff cannot, by itself, account for thedepletion of soil fertility associated withdesertification in the Chihuahuan desert.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1006246126915
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  • 3
    Electronic Resource
    Electronic Resource
    Mercury speciation in natural waters: Measurement of dissolved gaseous mercury with a field analyzer (2000)
    Springer
    Biogeochemistry 48 (2000), S. 237-259 
    Details
    ISSN: 1573-515X
    Keywords: gaseous ; kinetics ; mercury ; methods ; speciation ; waters
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract Mercury evasion from water is commonly modeled using measurements of dissolved gaseous mercury (DGM). We developed a method using a recently available automated field-ready mercury vapor analyzer to rapidly measure the concentrations of DGM in surface waters. We summarize here results of laboratory tests of the method, field intercomparisons with a manual method, and selected data from recent sampling campaigns in Florida and Michigan. The method uses the 1.5 lpm flow of a Tekran® Model 2537A mercury analyzer to purge and analyze discrete water samples, generating near real time (5-min) data on DGM in samples and blanks. Application of the Tekran allowed for detailed analysis of DGM removal kinetics and short-term diel studies characterizing the influence of sunlight and precipitation on DGM production in surface waters. Gas removal kinetics for dozens of samples indicates a first-order rate constant, and supports a 20-min. purge time for surface water samples from Florida (40-min for Michigan samples). Blanks are measured during a second such purge. Our results indicate that DGMs determined by both automated and manual methods are generally comparable, and that DGM in Florida samples is unstable during storage (loss rate constant ∼0.1--0.2 h-1), probably due to oxidation. This suggests that rapid in-field analysis is preferred to storage with delayed analysis. Our data indicate that DGM at the Florida site is influenced by inputs of reactive Hg in rainwater, and by production of surface DGM during photoreduction of oxidized Hg in the water column.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1006228612872
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  • 4
    Electronic Resource
    Electronic Resource
    The Relationship Between Rate of Administration of an Intubating Dose of Rocuronium and Time to 50% and 90% Block at the Adductor Pollicis Muscle (2000)
    Springer
    Journal of clinical monitoring and computing 16 (2000), S. 219-222 
    Details
    ISSN: 1573-2614
    Keywords: Neuromuscular relaxants ; rocuronium ; kinetics ; distribution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Computer Science , Medicine
    Notes: Abstract Objective.To determine the relationship between the rate ofrocuronium injection and the onset time of neuromuscular block. Methods.After intravenous induction, 60 female patients (ASA I–II) wereassigned randomly into 3 groups for rocuronium administration within1–15, 15–30 or 30–60 seconds. Acceleromyography of the thumbwas performed using train-of-four (TOF) stimulation. Times to 50% and 90%twitch depression of the first twitch of the TOF stimulation (T1) wererecorded. Results.Injection time significantly influences time to 50%relaxation, but not time to 90% relaxation. Body mass index is negativelycorrelated with time to 50% and 90% relaxation. Conclusions.Weconclude that rate of injection influences only the initial phase ofdevelopment of the block and that slower injection times do not significantlyaffect time to 90% relaxation at the adductor pollicis muscle.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1009964305762
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  • 5
    Electronic Resource
    Electronic Resource
    Non-Steady State Monitoring by Respiratory Gas Exchange (2000)
    Springer
    Journal of clinical monitoring and computing 16 (2000), S. 351-360 
    Details
    ISSN: 1573-2614
    Keywords: carbon dioxide ; oxygen ; kinetics ; non-steady state ; cardiac output ; PEEP ; pulmonary embolism ; pulmonary gas exchange monitoring
    Source: Springer Online Journal Archives 1860-2000
    Topics: Computer Science , Medicine
    Notes: Abstract Traditionally, the study of CO2 and O2 kinetics in the body has been mostly confined to equilibrium conditions. However, the peri-anesthesia period and the critical care arena often involve conditions of non-steady state. The detection and explanation of CO2 kinetics during non-steady state pathophysiology have required the development of new methodologies, including the CO2 expirogram, average alveolar expired PCO2, and CO2 volume exhaled per breath. Several clinically relevant examples of non-steady state CO2 kinetics perturbations are examined, including abrupt decrease in cardiac output, application of positive end-expiratory pressure during mechanical ventilation, and occurrence of pulmonary embolism. The lesser known area of non-steady state O2 kinetics is introduced, including the measurement of pulmonary O2 uptake per breath. Future directions include the study of the respiratory quotient per breath, where the anaerobic threshold during anesthesia is identified by increasing respiratory quotient.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1011447500984
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  • 6
    Electronic Resource
    Electronic Resource
    Effects of the emergent macrophyte Juncus effusus L. on the chemical composition of interstitial water and bacterial productivity (2000)
    Springer
    Biogeochemistry 48 (2000), S. 307-322 
    Details
    ISSN: 1573-515X
    Keywords: aquatic plants ; dissolved organic carbon ; nutrient cycling ; phosphorus ; sediments ; wetlands
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract Release of oxygen from the roots ofaquatic macrophytes into anaerobic sediments canaffect the quantity of interstitial dissolved organicmatter and nutrients that are available to bacteria. Nutrient and dissolved organic carbon (DOC)concentrations were compared between subsurface(interstitial) waters of unvegetated sediments andsediments among stands of the emergent herbaceousmacrophyte Juncus effusus L. in a lotic wetlandecosystem. Concentrations of inorganic nitrogen(NH4 +, NO3 -, and NO2 -)were greater from sediments of the unvegetatedcompared to the vegetated zone. DOC concentrations ofinterstitial waters were greater in sediments of theunvegetated zone both in the winter and springcompared to those from the vegetated zone. AlthoughDOC concentrations in hydrosoils collected from bothzones increased from winter to spring, bacterialproductivity per mg DOC in spring decreased comparedto winter. Greater initial bacterial productivityoccurred on DOM collected from the vegetated comparedto the unvegetated zone in winter samples (days 1 and4), with increased bacterial productivity on samplescollected from the unvegetated zone at the end of thestudy (day 20). Bacterial productivity wassignificantly greater on all sampling days on DOM fromvegetated samples compared to unvegetated samples. In nutrient enrichment experiments, bacterialproductivity was significantly increased (p 〈 0.05)with phosphorus but not nitrogen only amendments.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1006208213821
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  • 7
    Electronic Resource
    Electronic Resource
    Carbon, nitrogen and phosphorus cycling in rivermarginal wetlands; a model examination of landscapegeochemical flows (2000)
    Springer
    Biogeochemistry 50 (2000), S. 45-71 
    Details
    ISSN: 1573-515X
    Keywords: carbon ; landscape geochemical flows ; model ; nitrogen ; phosphorus ; wetland
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract The importance of landscape geochemical flows wasinvestigated using a dynamic model simulating carbon,nitrogen and phosphorus cycling in riverine wetlands,which has been described in a previous paper. Thehydro-geomorphic unit (HGMU) concept was incorporatedin the model by defining a separate, completeunit-model for each unit (HGMU) within the wetland.These unit-models were connected by defining the flowsof nitrogen and phosphorus between them. These flows,also called landscape geochemical flows, usuallyconsist of flows of water containing N and P. The model was applied to a site at Kismeldon Meadows,in south-western England. This site consists of twounits, a slope and a floodplain, separated by a ditch,which catches most of the run off and shallowgroundwater flows from the slope. Only an estimated1% of the N and P that leaves the slope unit in thewater outflow reaches the floodplain unit; the rest iscaught in the system of ditches, which prevent thegeochemical flows taking their natural course. Toexamine the influence of this system of ditches, themodel was run for the same site, but without theditches. This is comparable to a situation of arestored site, where run off and shallow groundwaterflows containing nutrients, can freely get from theslope to the floodplain. The computer simulation experiment reconnecting theslope and floodplain showed that this (1) increasedthe nutrient input into the floodplain, causing ahigher biomass production, and (2) increased thewetness of the floodplain, causing slowerdecomposition, which together (3) led to a faster soilorganic matter accumulation in the floodplain.Nutrient inflows became relatively more importantcompared to atmospheric deposition, especially forphosphorus. By connecting the slope and the floodplainmore nitrogen and less phosphorus flowed into theriver.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1006360315792
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  • 8
    Electronic Resource
    Electronic Resource
    Internal eutrophication in peat soils through competition between chloride and sulphate with phosphate for binding sites (2000)
    Springer
    Biogeochemistry 50 (2000), S. 183-194 
    Details
    ISSN: 1573-515X
    Keywords: chloride ; internal eutrophication ; nitrogen ; peat soil ; phosphorus ; phytometer ; sulphate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract Inputs of surface waters high in chloride and sulphateincreased the availability of nutrients in fenpeatlands. This `internal eutrophication' wasdemonstrated with test plants (`phytometers') andthrough water and soil analysis. Three experiments arepresented in which the level of chloride and/orsulphate was increased to 3 mmolc l−1. Inexperiment 1 chloride levels were increased from 0.5to 3 mmolc l−1 as CaCl2 or NaCl. Inexperiment 2 and 3 similar increased levels forchloride and sulphate (3 mmolc l−1; as NaCland Na2SO4) were used. The following resultswere found: (i) No differences in soil total-N and total-P werefound before and after the treatments in any of thethree experiments. (ii) Experiment 1 showed a significant increase inBio-Available P (BAP) in pots planted with Anthoxanthum odoratum as well as in bare pots for theNaCl and CaCl2 treatments. The plants in thesetreatments had taken up much more P. (iii) Experiment 2 showed an increase in soil BAPafter treatment with chloride and sulphate in potsplanted with Anthoxanthum odoratum. The chloridetreatment had no effect on plant biomass, whereas thesulphate treatment resulted in a reduction in rootbiomass and root N and P content. The shoots showedan increase in P content in the sulphate and chloridetreatments, while N content remained the same. (iv) In experiment 3, treatments with chloride andsulphate led to significantly increased biomass and Puptake of Anthoxanthum odoratum. Again, noeffects on N uptake were found. These experiments provide evidence for distinctlyincreased availability of phosphate in peat soils whenthese come into contact with water with evenmoderately increased sulphate or chloride levels.Surface water originating from the Rhine river, whichis enriched in chloride and sulphate, is oftensupplied to fen reserves in The Netherlands, tocompensate for water losses due to agriculturaldrainage in the region. The results of this study showthat phosphate availability to the vegetation may risedrastically, with detrimental effects on the speciesdensity and the occurrence of rare species in thevegetation. Hence, supply of this water should beavoided.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1006374018558
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  • 9
    Electronic Resource
    Electronic Resource
    Al and Fe Biogeochemistry in a floodplain forest: Implications for P retention (2000)
    Springer
    Biogeochemistry 51 (2000), S. 1-32 
    Details
    ISSN: 1573-515X
    Keywords: aluminum ; floodplain forests ; iron ; organic matter ; P sorption capacity ; phosphorus ; soil ; wetlands
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract We examined spatial and temporal variationsin soil chemistry in a floodplain forest landscape todetermine the effects of flooding on aluminum (Al) andiron (Fe) oxide biogeochemistry and inorganicphosphorus (Pi) sorption capacity. Whenpreviously sorbed Pi was considered, the sorptioncapacities of floodplain and adjacent upland soilswere comparable, suggesting that floodplain soilsrepresent a second line of defense protectingdownstream aquatic ecosystems from agriculturalrun-off. Pi sorption capacity was highlycorrelated with oxalate-extractable Al (Alo)(rs = 0.78); Alo and percent organic matter(OM) were also highly correlated (rs = 0.72),suggesting the importance of OM-Al complexes in thesesoils. The correlation of oxalate-extractable Fe(Feo) with OM (rs = 0.64) was improved(rs = 0.80) by removing lower elevation (swale)soils, suggesting that flooding inhibits theassociation of Feo with OM. Fe oxidecrystallinity decreased during seasonal flooding, buttotal extractable Fe did not change significantly. Fesolubilized during flooding was either replaced bysediment deposition (252 ± 3 mmol kg−1yr−1), and/or reprecipitated locally. Al oxidecrystallinity also decreased during flooding due to asignificant decline in NaOH-extractable Al (AlN). AlN concentrations subsequently returned topre-flooding levels, but sediment Al inputs (57 ±3 mmol kg−1 yr−1), were insufficient to account for this recovery. Observed Fetransformations suggest the importance offlooding-induced declines in soil redox potential toFe biogeochemistry; observed Al transformationssuggest the importance of complexation reactions withsoil OM to Al biogeochemistry in this floodplainforest.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1006302600347
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  • 10
    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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  • 11
    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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  • 12
    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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  • 13
    Electronic Resource
    Electronic Resource
    Effects of phosphatidylinositol 4,5-bisphosphate and neomycin on phospholipase D: Kinetic studies (1999)
    Springer
    Molecular and cellular biochemistry 197 (1999), S. 195-201 
    Details
    ISSN: 1573-4919
    Keywords: phospholipase D ; phosphatidylinositol 4,5-bisphosphate ; neomycin ; kinetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The kinetics of phosphatidylcholine-specific phospholipase D activated by phosphatidylinositol 4,5-bisphosphate (PIP2) and inhibition by neomycin were studied in an enzyme preparation partially purified from human hepatocarcinoma cell line. It was found that phospholipase D was marginally activated by phosphatidyl-4-phosphate (PIP) and phosphatidylethanolamine (PE). In contrast, it was considerably activated by PIP2 in different concentration of phosphatidylcholine (PC). Sphingomyelin (SM), lysophosphatidylcholine (LPC) and phosphatidylserine (PS) were neither substrates nor inhibitors of the phospholipase D. PIP2 induced an allosteric effect on phospholipase D and a negative cooperative effect with respect to phosphatidylcholine as indicated in the Lineweaver-Burk plot. In the absence of PIP2, a straight line was obtained, whereas a downward concave curve was observed in the presence of 25 μM of PIP2. The Hill coefficient and the apparent Km of phosphatidylcholine in the presence of 25 μM PIP2 were calculated to be 0.631 and 10.79 mM, respectively. PIP2 also increased the maximal velocity (Vmax) of the phospholipase D reaction, suggesting that the affinity of substrate to enzyme was decreased, and the turnover number of the enzyme (kcat) was increased by PIP2. The activation of phospholipase D by PIP2 was dose dependent up to 50 μM of PIP2. The Ka of PIP2 was 15.8 mM. Neomycin, a polycationic glycoside, was shown to be an uncompetitive inhibitor of phospholipase D, and revealed the formation of a neomycin-PIP2 complex. The Ki of neomycin was estimated to be 8.7 mM.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1006930706311
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  • 14
    Electronic Resource
    Electronic Resource
    Regulatory mechanisms of poly(ADP-ribose) polymerase (1999)
    Springer
    Molecular and cellular biochemistry 193 (1999), S. 19-22 
    Details
    ISSN: 1573-4919
    Keywords: poly(ADP-ribose)polymerase ; kinetics ; allosterism ; regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract Here, we describe the latest developments on the mechanistic characterization of poly(ADP-ribose) polymerase (PARP) [EC 2.4.2.30], a DNA-dependent enzyme that catalyzes the synthesis of protein-bound ADP-ribose polymers in eucaryotic chromatin. A detailed kinetic analysis of the automodification reaction of PARP in the presence of nicked dsDNA indicates that protein-poly(ADP-ribosyl)ation probably occurs via a sequential mechanism since enzyme-bound ADP-ribose chains are not reaction intermediates. The multiple enzymatic activities catalyzed by PARP (initiation, elongation, branching and self-modification) are the subject of a very complex regulatory mechanism that may involve allosterism. For instance, while the NAD+ concentration determines the average ADP-ribose polymer size (polymerization reaction), the frequency of DNA strand breaks determines the total number of ADP-ribose chains synthesized (initiation reaction). A general discussion of some of the mechanisms that regulate these multiple catalytic activities of PARP is presented below.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1006979220009
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  • 15
    Electronic Resource
    Electronic Resource
    The surface-connected canalicular system in the sinus endothelial cells of rat spleen (1999)
    Springer
    Cell & tissue research 296 (1999), S. 439-442 
    Details
    ISSN: 1432-0878
    Keywords: Key words Spleen ; Sinus endothelial cells ; Surface-connected canalicular system ; Lanthanum ; Three-dimensional reconstruction ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The existence of a surface-connected canalicular system in the splenic sinus endothelial cells of the rat has been demonstrated by transmission electron microscopy with lanthanum nitrate acting as a tracer for the extracellular space. In addition, the three-dimensional arrangement of the canaliculi has been revealed by computer-aided reconstruction. The surface-connected canalicular system of the sinus endothelial cells consists of slender canaliculi that are branched, anastomosed, and that show continuity with the plasma membrane. They twist in and out among the organelles and are often found in close apposition to the spherical invaginations of the plasma membrane and run alongside them. Canaliculi which are not infiltrated by lanthanum nitrate take the form of electron-lucent tubules and are accompanied by numerous spherical invaginations of the plasma membrane. From a computer-aided reconstruction, the canaliculi, which invaginate from various sites of the plasma membrane, have been found to be continuous with each other and to penetrate to the surface of the sinus endothelial cell; they also branch and anastomose to form a complex network in the cytoplasm. Although the surface-connected canalicular system in blood platelets and thrombocytes is believed to function as the main route for the discharge of granules and the uptake of foreign materials and also to take part in the storage and transport of calcium, it is unclear at present whether the network of the surface-connected canalicular system in splenic sinus endothelial cells has any physiological significance.
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    Elongated pericyte-like cells connect discrete capillaries in the cochlear stria vascularis of gerbils and rats (1999)
    Springer
    Cell & tissue research 296 (1999), S. 673-676 
    Details
    ISSN: 1432-0878
    Keywords: Key words Basal lamina ; Immunohistochemistry ; Confocal laser microscopy ; Cochlea ; Mongolian gerbil ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Bridging structures between discrete capillaries in the stria vascularis of the cochlea were studied morphologically in gerbils and rats. Serial thin sections for transmission electron microscopy revealed (1) that elongated cells surrounded by the basal lamina provided the structural basis for the bridging structure, (2) that the basal lamina surrounding the elongated cell extended to the basal lamina around the capillary endothelial cell, (3) that the electron density of the cytoplasm was similar to that of the pericytes around the capillaries, and (4) that the cell was attached to the capillaries at both ends only. Visualization of the basal lamina by immunofluorescent methods revealed (1) that capillaries were often bent at the site of attachment of the bridging cell, (2) that the bridging cell bifurcated occasionally, and (3) that the density of the bridging cell was much higher in the stria vascularis than in the underlying spiral ligament. Filamentous actin visualized by fluorescent phalloidin was not apparent in the bridging cell. We propose that the bridging cell provides mechanical strength to the tortuous capillary network in the stria vascularis and participates in the specific function of the stria vascularis in cooperation with other types of cells.
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    URL: http://dx.doi.org/10.1007/s004410051327
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    Immunohistochemical evidence for the presence of tryptophan hydroxylase and serotonin in the rodent Harderian gland (1999)
    Springer
    Cell & tissue research 296 (1999), S. 517-523 
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    ISSN: 1432-0878
    Keywords: Key words Harderian gland ; Tryptophan hydroxylase ; Serotonin ; Immunohistochemistry ; Rat (Wistar) ; Syrian hamster ; Mesocricetus auratus ; Djungarian hamster ; Phodopus sungorus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The Harderian gland is considered as being an extrapineal source of melatonin. In most rodents, the Harderian gland contains two epithelial cell types (I and II). The aim of this study has been to define which cell type is involved in indoleamine synthesis. The presence and localization of serotonin (melatonin precursor) and tryptophan hydroxylase (the rate-limiting enzyme for serotonin synthesis) have been investigated by immunohistochemistry in male Wistar rats, Syrian hamsters and Djungarian hamsters. The results of the present study show that immunoreactivity for tryptophan hydroxylase and serotonin is confined to the type I cell, suggesting that this cell type is involved in indoleamine synthesis in the rodent Harderian gland.
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    Expression of neurotrophin receptors trkB and trkC and their ligands in rat adrenal gland and the intermediolateral column of the spinal cord (1999)
    Springer
    Cell & tissue research 296 (1999), S. 271-279 
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    ISSN: 1432-0878
    Keywords: Key words Brain-derived neurotrophic factor ; Neurotrophin-3 ; Sympathetic preganglionic neurons ; Chromaffin cells ; Adrenal cortex ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Neurotrophins and their trk receptors constitute major classes of signaling molecules with important actions in the developing and adult nervous system. With regard to the sympathoadrenal cell lineage, which gives rise to sympathetic neurons and chromaffin cells, neurotrophin-3 (NT-3) and nerve growth factor (NGF) are thought to influence developing sympathetic neurons. Neurotrophin requirements of chromaffin cells of the adrenal medulla are less well understood than those for NGF. In order to provide the bases for understanding of putative functions of neurotrophins for the development and maintenance of chromaffin cells and their preganglionic innervation, in situ hybridization has been used to study the expression of brain-derived neurotrophic factor (BDNF) and NT-3, together with their cognate receptors trkB and trkC, in the adrenal gland and in the intermediolateral column (IML) of the spinal cord. BDNF is highly expressed in the embryonic adrenal cortex and later in cells of the cortical reticularis zone. Adrenal medullary chromaffin cells fail to express detectable levels of mRNAs for BDNF, NT-3, and their cognate receptors trkB and trkC. Neurons in the IML express BDNF and trkB, and low levels of NT-3 and trkC. Our data make it unlikely that BDNF and NT-3 serve as retrograde trophic factors for IML neurons but suggest roles of BDNF and NT-3 locally within the spinal cord and possibly for sensory nerves of the adrenal cortex.
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    URL: http://dx.doi.org/10.1007/s004410051288
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    Innervation of the rat pineal gland by pituitary adenylate cyclase-activating polypeptide (PACAP)-immunoreactive nerve fibres (1999)
    Springer
    Cell & tissue research 296 (1999), S. 247-257 
    Details
    ISSN: 1432-0878
    Keywords: Key words Calcitonin gene-related peptide ; Vasoactive intestinal peptide ; Neuropeptide Y ; Colocalization ; Trigeminal ganglion ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Pituitary adenylate cyclase-activating polypeptide (PACAP)-immunoreactive nerve fibres were demonstrated in the rat pineal gland. These fibres entered the pineal gland through the conarian nerve at the distal tip of the gland. A high density of the fibres was observed in the capsule of the gland, from where the immunoreactive elements penetrated into the pineal perivascular spaces and parenchyma. The majority of PACAP-immunoreactive nerve fibres also contained calcitonin gene-related peptide (CGRP). Some PACAP-immunoreactive nerve fibres contained neuropeptide Y (NPY), but only occasionally was PACAP colocalized with vasoactive intestinal peptide (VIP). After removal of both superior cervical ganglia, a high number of PACAP-containing nerve fibres were still present in the gland. In the nervous system PACAP is present in two isoforms, PACAP-38 and PACAP-27. The concentration of PACAP-38 in the superficial pineal gland was determined by radioimmunoassay to be 20.4 pmol/g tissue at midday and 18.9 pmol/g tissue at midnight. The concentration of PACAP-27 was only about 3% of the concentration of PACAP-38. In summary, this study is the first demonstration of a PACAP-containing innervation of the rat pineal gland. The PACAP concentration in the pineal gland does not exhibit a day-night difference. The colocalization of PACAP with calcitonin gene-related peptide in the pinealopetal nerve fibres indicates that the majority of PACAP-immunoreactive nerve fibres might originate from the trigeminal ganglion.
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    URL: http://dx.doi.org/10.1007/s004410051286
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    Changes in fibre populations of the rat hairy skin following selective chemodenervation by capsaicin (1999)
    Springer
    Cell & tissue research 296 (1999), S. 471-477 
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    ISSN: 1432-0878
    Keywords: Key words Skin ; Sensory innervation ; Capsaicin ; Protein gene product 9.5 ; Neurogenic inflammation ; Sensory neuropeptides ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Perineural application of capsaicin results in a selective and permanent reduction in the sensitivity to noxious chemical and heat stimuli and elimination of the neurogenic inflammatory response. The present quantitative immunohistochemical study has been undertaken to reveal the populations of cutaneous afferent nerves that are affected by perineural capsaicin treatment. Areas of intact and chemodenervated skin were determined with the aid of the vascular labelling technique. In sections taken from intact skin areas, staining with antibodies against protein gene product 9.5 revealed a rich epidermal innervation. Fibres immunoreactive for growth-associated protein 43 were also abundant; nerve fibres immunoreactive for substance P and calcitonin gene-related peptide were less numerous. Somatostatin- and RT97-immunoreactive fibres were seen only in the subepidermal layer. In sections taken from skin areas supplied by the sciatic nerve treated with capsaicin 3 days previously, the number of epidermal nerve fibres immunoreactive to protein gene product 9.5, growth-associated protein 43, substance P and calcitonin gene-related peptide was reduced by 90%, 95%, 97% and 66%, respectively. These changes persisted for at least 42 days. The findings reveal that the majority of epidermal axons are capsaicin-sensitive and comprise a chemically heterogeneous population. Reductions in cutaneous fibre populations following perineural capsaicin treatment may result from both the degeneration of sensory axons and the depletion of neuron-specific macromolecules. In addition, most cutaneous nociceptive axons may not use the major sensory neuropeptides substance P and calcitonin gene-related peptide as afferent neurotransmitters.
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    Articular chondrocytes and synoviocytes in a co-culture system: influence on reactive oxygen species-induced cytotoxicity and lipid peroxidation (1999)
    Springer
    Cell & tissue research 296 (1999), S. 555-563 
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    ISSN: 1432-0878
    Keywords: Key words Chondrocyte ; Synoviocyte ; Co-culture ; Proliferation ; Lipid peroxidation ; Cytotoxicity ; Ultrastructure ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Objective: A new co-culture system of rat articular chondrocytes and synoviocytes (HIG-82; cell line) was incubated with phorbol myristate acetate (PMA), H2O2 or a combination of Fe2+ and ascorbic acid to simulate inflammation-like radical attacks in articular joints. Methods: Chondrocytes were characterized by immunocytochemistry against collagen type II, transmission electron (TEM) and light microscopy. Lipid peroxidation was investigated by measuring thiobarbituric-acid-reactive material in the supernatants, cytotoxicity by determining release of lactate dehydrogenase and proliferation by measuring [3H]thymidine incorporation, culture protein and DNA. Results: PMA or Fe2+ and ascorbic acid induced lipid peroxidation in chondrocytes and synoviocytes that was decreased significantly in co-cultures. PMA and H2O2 dose dependently induced release of lactate dehydrogenase in chondrocytes, which was lowered in co-cultures or in previously co-cultured chondrocytes to a nearly basal level. In contrast, conditioned media of synoviocyte cultures showed no lowering effect on the radical-induced toxicity. Protection against H2O2-induced damage of cellular membranes by co-culturing was also shown by TEM. Synoviocytes released chondrocyte-stimulating growth factors spontaneously without previous interaction. Conclusion: Chondrocytes establish protective mechanisms against reactive oxygen species via an interaction with synoviocytes. Our co-culture model presents a possible way to study mechanisms of inflammation in articular joints under defined conditions.
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    Changing patterns of spatial buffering of glutamate in developing rat retinae are mediated by the Müller cell glutamate transporter GLAST (1999)
    Springer
    Cell & tissue research 297 (1999), S. 57-66 
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    ISSN: 1432-0878
    Keywords: Key words D-aspartate ; Development ; Glutamate ; Retina ; Glutamate transporter (GLAST) ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The patterns of expression of the glutamate transporter GLAST were compared with the patterns of uptake of exogenous D-aspartate, which is a substrate for all glutamate transporters. At postnatal day 0, fine radial processes and end feet of presumptive Müller cells were weakly immunoreactive for GLAST. At postnatal day 3, intense labelling was associated with astrocytes enveloping newly formed blood vessels on the vitread surface of the retina. Between postnatal days 7 and 10, there was a rapid increase in the intensity of labelling in the Müller cells but clear stratification of GLAST-immunoreactive processes in the inner plexiform layer was not observed until postnatal day 14. By comparison, D-aspartate uptake was initially associated with a wide variety of cellular elements including most neuroblasts, presumptive Müller cells, and astrocytes associated with blood vessels but was absent from the somata of many neurons in the ganglion cell layer and amacrine cell layer. There was a gradual contraction in the numbers of cells that were able to take up D-aspartate, such that, by adulthood, uptake was restricted mainly to Müller cells and astrocytes. We conclude that, during early retinal development, the low levels of GLAST expression by Müller cells permit D-aspartate, and by inference, glutamate, to permeate the retina freely, thus allowing uptake by other glutamate transporters on other cell types. As the retina matures, increased expression of GLAST by Müller cells restricts the access of D-aspartate to other cellular compartments in the retina. This changing pattern of spatial buffering of glutamate by GLAST probably has significant implications regarding our understanding of the role of glutamate during processes such as retinal synaptogenesis.
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    Expression of mRNAs for type-I collagen, bone sialoprotein, osteocalcin, and osteopontin at different stages of osteoblastic differentiation and their regulation by 1,25 dihydroxyvitamin D3 (1999)
    Springer
    Cell & tissue research 297 (1999), S. 249-259 
    Details
    ISSN: 1432-0878
    Keywords: Key words Bone sialoprotein ; Osteopontin ; Osteocalcin ; Osteoblasts ; In situ hybridization ; Dexamethasone ; 1 ; 25 dihydroxyvitamin D3 ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  We have used in situ hybridization to evaluate the effects of 1,25 dihydroxyvitamin D3 (1,25 (OH)2 D3) on the expression of mRNA for bone-matrix proteins and to determine whether mature osteoblasts respond differently to 1,25 (OH)2 D3 than younger, newly differentiated osteoblasts. Rat calvaria cells were cultured for 7, 12, 15, and 19 days to obtain a range of nodules from very young to very mature. At each time point, some cultures were treated with 10 nM 1,25 (OH)2 D3 for 24 h prior to fixation. In control cultures, type-I collagen mRNA was detectable in osteoblastic cells in very young nodules and increased with increasing maturity of the nodules and the osteoblasts lining them. The bone sialoprotein mRNA signal was weak in young osteoblasts, increased in older osteoblasts, and decreased in mature osteoblasts. Weak osteocalcin and osteopontin signals were seen only in osteoblasts of intermediate and mature nodules. 1,25 (OH)2 D3 treatment markedly upregulated osteocalcin and osteopontin mRNAs and downregulated mRNA levels of bone sialoprotein and, to a lesser extent, type-I collagen in both young and mature osteoblasts. However, a marked diversity of signal levels for bone sialoprotein, osteocalcin, and osteopontin existed between neighboring mature osteoblasts, particularly after 1,25 (OH)2 D3 treatment, which may therefore selectively affect mature osteoblasts, depending on their differentiation status or functional stage of activity.
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    Correlation between gender and spontaneous C-cell tumors in the thyroid gland of the Wistar rat (1999)
    Springer
    Cell & tissue research 297 (1999), S. 451-457 
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    ISSN: 1432-0878
    Keywords: Key words Thyroid gland ; Calcitonin ; C-cell hyperplasia ; C-cell carcinoma ; Medullary thyroid carcinoma ; Sexual dimorphism ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  In many rat strains, C-cell hyperplasia occurs in an age-dependent manner and is often associated with multifocal C-cell carcinoma. The purpose of this study was to investigate the spectrum of spontaneous, proliferative C-cell disorders by gender in Wistar rats throughout their lifespan. The incidence of C-cell hyperplasia shows a significant increase with age (P〈0.001) and is much higher in female rats than in male rats (P〈0.05). From 3 to 24 months of life, 27.5% of female rats showed a normal C-cell pattern, 55.0% showed C-cell hyperplasia, and 17.5% showed C-cell tumors; while 57.5% of male rats showed a normal C-cell pattern, 32.5% showed C-cell hyperplasia, and 10% showed C-cell tumors. Although the overall frequency of C-cell neoplasms in females was nearly double that in males, these data are not statistically significant. However, the number of C-cell tumors showed a significant increase with age (P〈0.05). Therefore, we can conclude that there were significant differences in the incidence of the total spectrum of C-cell proliferative abnormalities in the thyroid gland of Wistar rats that were both age-dependent and gender-dependent.
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    Motor innervation by enteric nerve fibers containing both nitric oxide synthase and galanin immunoreactivities in the striated muscle of the rat esophagus (1999)
    Springer
    Cell & tissue research 295 (1999), S. 241-245 
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    ISSN: 1432-0878
    Keywords: Key words Enteric innervation ; Immunohistochemistry ; Nitric oxide synthase ; Galanin ; Striated muscle ; Esophagus ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The relationship between nitric oxide synthase (NOS)- and galanin-immunoreactive nerve terminals and the origin of NOS-immunoreactive nerve terminals on the motor endplates in the striated muscles of the rat esophagus was investigated. Double immunohistochemical staining revealed a dual innervation of motor endplates by calcitonin gene-related peptide (CGRP)-immunoreactive axons and by axons that were immunoreactive for both NOS and galanin. On average, 91% of NOS terminals were galanin immunoreactive. NOS-immunoreactive fibers were revealed at 67% of endplates, identified by the presence of CGRP terminals. The left vagus and superior laryngeal nerve were cut and 15 days allowed for terminals to degenerate. This caused a significant loss of CGRP fibers, but did not affect the density of innervation of the striated muscle by NOS-immunoreactive fibers. Thus the NOS/galanin fibers are deduced to originate from ganglia in the esophageal wall. This is supported by our observation of numerous NOS-immunoreactive nerve cell bodies in the myenteric ganglia of the esophagus, 74% of which were galanin immunoreactive. There were no CGRP-immunoreactive nerve cell bodies in the wall of the esophagus.
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    Oxidative stress response induced in rat primary hepatocyte monolayers by mechanical removal of adherent cells (1999)
    Springer
    Cell & tissue research 295 (1999), S. 363-367 
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    ISSN: 1432-0878
    Keywords: Key words Primary hepatocytes ; Oxidative stress ; Haem oxygenase ; Manganese superoxide dismutase ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract We show that mechanical separation of adherent rat primary hepatocytes after the monolayer-forming stage causes the induction of the oxidative stress genes HO-1 (haem oxygenase) and MnSOD (manganese superoxide dismutase). The procedures for enzymatically breaking up liver tissue structure and isolating hepatocytes do not cause HO-1 and MnSOD activation. Only after a 3-h incubation, during which hepatocytes form a monolayer on culture dishes, does the hydrodynamic shearing away of necrotic cells sticking to the monolayer surface activate these two genes. Analysis of this injury-response pathway shows that oxidative stress and mitochondrial dysfunction play a role, as activation can be repressed by antioxidants and by respiratory inhibitors. Recovery of the cells takes a further 24-h incubation during which HO-1 and MnSOD expression returns to basal levels.
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    Simultaneous apocrine and merocrine secretion in the rat coagulating gland (1999)
    Springer
    Cell & tissue research 295 (1999), S. 495-504 
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    ISSN: 1432-0878
    Keywords: Key words Coagulating gland ; Apocrine secretion ; Merocrine secretion ; Immunocytochemistry ; Immunoelectron microscopy ; Scanning electron microscopy ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The coagulating gland of the rat synthesizes two prevalent secretory proteins (transglutaminase and 115 K) that are discharched in a different manner, one being secreted in an apocrine fashion (transglutaminase) and the other one in a merocrine way (115 K). Differences in the intra- cellular pathway and the release of either protein were studied using immunofluorescence on semithin sections, immunoelectron microscopy of preembedding-processed chopper sections and postembedding-processed ultrathin sections of rat coagulating gland. Immunohistochemical staining using an anti-transglutaminase antibody resulted in dense labeling of the cytoplasm of secretory cells and their apical blebs, whereas the cisternae of the rough endoplasmic reticulum and the Golgi apparatus were completely unlabeled. When, on the contrary, the anti-115 K antiserum was used, dense labeling of the cisternae of the rough endoplasmic reticulum, the Golgi apparatus, and the secretory granules was seen. Intraluminal secretion was also labeled, but the secretory blebs remained unlabeled. Our findings show that, in the coagulating gland of the male rat, the two secretory proteins studied are processed in parallel, but at completely different intracellular pathways. They are released via different extrusion mechanisms. Transglutaminase is synthesized outside the endoplasmic reticulum, reaches the apical cell pole by free flow in the cytoplasm, and is released via apocrine blebs, the membranes of which appear to be derived from the apical plasma membrane. The protein 115 K, on the other hand, follows the classic route, being synthesized within the cisternae of rough endoplasmic reticulum, subsequently glycosylated in the Golgi apparatus, and released in a merocrine fashion. The mutual exclusion of the two secretory pathways and the regulation of the alternative release mechanism are still unresolved issues.
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    Immunocytochemical evidence of plasticity in the nervous structures of the rat lower lip (1999)
    Springer
    Cell & tissue research 297 (1999), S. 203-211 
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    ISSN: 1432-0878
    Keywords: Key words Growth-associated protein 43 (GAP-43) ; B50 ; Trigeminal sensory system ; Neuroplasticity ; Skin ; Mucosa ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  In this immunocytochemical study we investigated the distribution of nervous structures in the lower lip of adult rats. The region is characterized by a rich cutaneous and mucosal sensory innervation originating from terminal branches of the trigeminal system. Lower lip innervation was investigated by detection of the general neuronal marker protein gene product 9.5 (PGP 9.5) and the growth-associated protein 43 (GAP-43), a neurochemical marker of neuronal plasticity. The entire neural network of both cutaneous and mucosal aspects was stained by the antibody to PGP 9.5. In particular, nerve fibers were observed in the submucosal and the subepithelial plexuses. Thin immunoreactive fibers were observed within the epithelial layers ending as free fibers or as fibers associated with immunopositive Merkel cells. Well-identified anatomical structures receiving sensory or autonomic innervation were also surrounded by PGP 9.5-ir nerve fibers, in particular, hair follicles, vibrissae, glands, and blood vessels. GAP-43-immunostained nerve fibers were observed in all these structures; however, they were generally less numerous than the PGP 9.5-immunoreactive elements. An equal amount of PGP 9.5 and GAP-43 immunoreactivity occurred, in contrast, in the subepidermal and the submucosal plexuses, or in the epidermis and the mucosal epithelium. The present results show that GAP-43 is normally expressed in the mature trigeminal sensory system of the rat. Skin and oral mucosa are characterized by continuous remodeling that may also involve the sensory nervous apparatus. Continuous neural remodeling, regeneration and sprouting may be the reason for the observed expression of GAP-43.
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    Immunohistochemical localization of a water channel, aquaporin 7 (AQP7), in the rat testis (1999)
    Springer
    Cell & tissue research 295 (1999), S. 279-285 
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    ISSN: 1432-0878
    Keywords: Key words Spermatids ; Spermatozoa ; Spermatogenesis ; Cell volume reduction ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Cell volume reduction is one of the most distinct morphological changes during spermiogenesis and may be largely attributable to water efflux from the cell. A strong candidate for a water efflux route, aquaporin 7 (AQP7), which is a water channel, was studied immunohistochemically in the rat testis. Immunoreactivity was restricted within the elongated spermatids, testicular spermatozoa, and residual bodies remaining in the seminiferous epithelium. Weak but distinct immunoreactivity was first observed in the cytoplasmic mass of the spermatid at step 8 of spermiogenesis. The Golgi-like apparatus became steadily immunoreactive at step 10. The plasma membrane covering the cytoplasmic mass showed strong immunoreactivity after step 16. At this step, the middle piece of the tail also showed immunoreactivity at the portion protruding into the lumen. The whole head and distal tail, where the elongated spermatid had only a limited amount of cytoplasm, showed no immunoreactivity throughout spermiogenesis. After spermiation, the immunoreactivity of AQP7 remained at the middle piece and in the cytoplasmic droplet in the testicular spermatozoon. The present observations suggest that AQP7 contributes to the volume reduction of spermatids, since this water channel protein is localized on the plasma membrane covering the condensing cytoplasmic mass of the elongated spermatid, and since the seminiferous tubule fluid is hypertonic.
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    Aquaporin-5 (AQP5), a water channel protein, in the rat salivary and lacrimal glands: immunolocalization and effect of secretory stimulation (1999)
    Springer
    Cell & tissue research 295 (1999), S. 513-521 
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    ISSN: 1432-0878
    Keywords: Key words Water channel protein ; Aquaporin ; AQP5 ; Rat ; Salivary glands ; Immunolocalization ; Secretory stimulation ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Aquaporin-5 (AQP5) is a water channel protein and is considered to play an important role in water movement across the plasma membrane. We raised anti-AQP5 antibody and examined the localization of AQP5 protein in rat salivary and lacrimal glands by immunofluorescence microscopy. AQP5 was found in secretory acinar cells of submandibular, parotid, and sublingual glands, where it was restricted to apical membranes including intercellular secretory canaliculi. In the submandibular gland, abundant AQP5 was also found additionally at the apical membrane of intercalated duct cells. Upon stimulation by isoproterenol, apical staining for AQP5 in parotid acinar cells tended to appear as clusters of dots. These results suggest that AQP5 is one of the candidate molecules responsible for the water movement in the salivary glands.
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    Expression of a novel member of the TGF-β superfamily, growth/differentiation factor-15/macrophage-inhibiting cytokine-1 (GDF-15/MIC-1) in adult rat tissues (1999)
    Springer
    Cell & tissue research 297 (1999), S. 103-110 
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    ISSN: 1432-0878
    Keywords: Key words TGF-β ; CNS ; Epithelial tissues ; Macrophages ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  We have cloned a novel member of the transforming growth factor-β (TGF-β) superfamily from a human placental cDNA library. The sequence is identical to five very recently published sequences, of which only one (macrophage inhibitory cytokine-1, MIC-1) has been characterized in terms of function. In light of the present data demonstrating the wide distribution of the mRNA and putative multifunctionality, we propose to name this molecule growth/differentiation factor-15/MIC-1 (GDF-15/MIC-1). The deduced amino acid sequence reveals typical features of a secreted molecule. The epithelium of the choroid plexus is the only site in the adult brain expressing detectable levels of GDF-15/MIC-1 mRNA. Many epithelia of non-neural tissues including those of the prostate and intestinal mucosa, bronchi and bronchioli, secretory tubuli of the submandibular gland, and lactating mammary gland are prominent sites of GDF-15/MIC-1 synthesis. GDF-15/MIC-1 is also strongly expressed by macrophages in the adrenal gland. Thus, GDF-15/MIC-1, like many other members of the TGF-β superfamily, is widely distributed in adult tissues, being most strongly expressed in epithelial cells and macrophages.
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  • 32
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    Electronic Resource
    Immunocytochemical localization of substance P neurokinin-1 receptors in rat gingival tissue (1999)
    Springer
    Cell & tissue research 297 (1999), S. 213-222 
    Details
    ISSN: 1432-0878
    Keywords: Key words Neurokinin-1 receptor ; Substance P ; Junctional epithelium ; Neutrophil ; Endosomes ; Immunoelectron microscopy ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The distributions of substance P (SP) and the neurokinin-1 receptor (NK1-R), the receptor preferentially activated by SP, were examined in rat gingiva by immunocytochemical methods with light and electron microscopy. SP-immunoreactive nerve fibers were located preferentially in the junctional epithelium (JE) but few in the other oral and oral sulcular epithelia. NK1-R immunoreactivity was found in the endothelial cells (capillaries and postcapillary venules underlying the JE). NK1-R-labeled and -unlabeled unmyelinated nerve fibers were located close to the blood vessels and partially or completely covered by a Schwann cell sheath. In the JE, labeled naked axons without Schwann cell sheaths were observed. Neutrophils and macrophages in the connective tissue underlying the JE and in the JE were also labeled with NK1-R. Furthermore, NK1-R was found in the JE cells. Basically, immunoreaction products for NK1-R were found throughout various cells (endothelial cells, neutrophils, and JE cells) at invaginations of the plasma membrane and in vesicular and granular structures that are probably endosomes and are found close to both the plasma membrane and the nucleus. This is a first report, demonstrating the presence of NK1-R in the gingival tissue in the normal nonstimulated condition. Furthermore, it is thought that SP may modulate the permeability of blood vessels beneath the JE, the production of antimicrobial agents in neutrophils, and the proliferation and endocytotic ability of JE cells through NK1-R.
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  • 33
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    Cellular localization and distribution of glucocorticoid receptor immunoreactivity and the expression of glucocorticoid receptor messenger RNA in rat pituitary gland (1999)
    Springer
    Cell & tissue research 295 (1999), S. 207-214 
    Details
    ISSN: 1432-0878
    Keywords: Key words Glucocorticoid receptor (GR) ; Immunohistochemistry ; In situ hybridization ; Pituitary ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract By means of double immunohistochemical techniques and a nonradioisotopic in situ hybridization method, we determined the colocalization pattern of glucocorticoid receptor (GR) and pituitary hormones and the GR messenger RNA (mRNA) expression in the pituitaries of Wistar adult male rats. Immunoreactivity for GR was detected in the nuclei of cells in the anterior and posterior pituitary. Double immunohistochemistry revealed that the colocaliza- tion of GR and anterior pituitary hormones occurred in almost 99% of the growth hormone (GH)-producing cells and adrenocorticotropic hormone (ACTH)-producing cells, and in 67% of the thyroid stimulating hormone (TSH)-producing cells. Almost all of the folliculostellate cells (93%), marginal layer cells (94%) in the anterior pituitary, and pituicytes (96%) in the posterior pituitary immunostained for S100 protein antibody were also immunostained with GR. GR mRNA was abundant in the cytoplasm of anterior and intermediate pituitary cells but scattered sparsely in that of the posterior pituitary. These results suggest that glucocorticoids directly influence certain pituitary cells in order to regulate cell function, including the synthesis and/or secretion of hormones.
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  • 34
    Electronic Resource
    Electronic Resource
    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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  • 35
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    Electronic Resource
    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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  • 36
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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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  • 37
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    A simulation model of organic matter and nutrient accumulation in mangrove wetland soils (1999)
    Springer
    Biogeochemistry 44 (1999), S. 93-118 
    Details
    ISSN: 1573-515X
    Keywords: Everglades National Park ; mangrove soils ; organic matter ; nitrogen ; phosphorus ; sedimentation ; simulation model
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract The distribution and accumulation of organic matter, nitrogen (N) and phosphorus (P) in mangrove soils at four sites along the Shark River estuary of south Florida were investigated with empirical measures and a process-based model. The mangrove nutrient model (NUMAN) was developed from the SEMIDEC marsh organic matter model and parameterized with data from mangrove wetlands. The soil characteristics in the four mangrove sites varied greatly in both concentrations and profiles of soil carbon, N and P. Organic matter decreased from 82% in the upstream locations to 30% in the marine sites. Comparisons of simulated and observed results demonstrated that landscape gradients of soil characteristics along the estuary can be adequately modeled by accounting for plant production, litter decomposition and export, and allochthonous input of mineral sediments. Model sensitivity analyses suggest that root production has a more significant effect on soil composition than litter fall. Model simulations showed that the greatest change in organic matter, N, and P occurred from the soil surface to 5 cm depth. The rapid decomposition of labile organic matter was responsible for this decrease in organic matter. Simulated N mineralization rates decreased quickly with depth, which corresponded with the decrease of labile organic matter. The increase in organic matter content and decrease in soil bulk density from mangrove sites at downstream locations compared to those at upstream locations was controlled mainly by variation in allochthonous inputs of mineral matter at the mouth of the estuary, along with gradients in mangrove root production. Research on allochthonouns sediment input and in situ root production of mangroves is limited compared to their significance to understanding nutrient biogeochemistry of these wetlands. More accurate simulations of temporal patterns of nutrient characteristics with depth will depend on including the effects of disturbance such as hurricanes on sediment redistribution and biomass production.
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    URL: http://dx.doi.org/10.1007/BF00993000
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  • 38
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    Electronic Resource
    Nutrient budgets and biogeochemistry in an experimental agricultural watershed in Southeastern China (1999)
    Springer
    Biogeochemistry 45 (1999), S. 1-19 
    Details
    ISSN: 1573-515X
    Keywords: Chaohu Lake ; chemical fertilizer ; cycling ; denitrification ; multipond system ; nitrogen ; nutrient budget ; phosphorus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract During a two-year field study, an annual nutrient budget and cycles were developed for a small agricultural watershed. The study emphasized the integrated unit of the watershed in understanding the biogeochemistry. It was found that the total nutrient input was 39.1× 104 kg nitrogen and 3.91×104 kg phosphorus in the year 1995, of which the greatest input of nutrients to the watershed was chemical fertilizer application, reaching 34.7×104 kg (676 kg/ha) nitrogen and 3.88×104 kg (76 kg/ha) phosphorus. The total nutrient output from the watershed was 13.55×104 kg nitrogen and 0.40×104 kg phosphorus, while the largest output of nitrogen was denitrification, accounting for 44.1% of N output; the largest output of phosphorus was sale of crops, accounting for 99.4% of P output. The results show that the nutrient input is larger than output, demonstrating that there is nutrient surplus within the watershed, a surplus which may become a potential source of nonpoint pollution to area waters. The research showed that both denitrification and volatilization of nitrogen are key ways of nitrogen loss from the watershed. This suggests that careful management of fertilizer application will be important for the sustainable development of agriculture. The research demonstrated that a multipond system within the watershed had high retention rate for both water and nutrients, benefiting the water, nutrient and sediment recycling in the terrestrial ecosystem and helping to reduce agricultural nonpoint pollution at its source. Therefore, this unique watershed system should be recommended due to its great potential relevance for sustainable agricultural development.
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    URL: http://dx.doi.org/10.1007/BF00992870
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  • 39
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    Electronic Resource
    Nutrient losses in runoff from grassland and shrubland habitats in Southern New Mexico: I. rainfall simulation experiments (1999)
    Springer
    Biogeochemistry 45 (1999), S. 21-34 
    Details
    ISSN: 1573-515X
    Keywords: Chihuahuan desert ; desert ; desertification ; grassland ; nitrogen ; nutrient budgets ; phosphorus ; runoff
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract Rainfall simulation experiments were performed in areas of semiarid grassland (Bouteloua eriopoda) and arid shrubland (Larrea tridentata) in the Chihuahuan desert of New Mexico. The objective was to compare the runoff of nitrogen (N) and phosphorus (P) from these habitats to assess whether losses of soil nutrients are associated with the invasion of grasslands by shrubs. Runoff losses from grass- and shrub-dominated plots were similar, and much less than from bare plots located in the shrubland. Weighted average concentrations of total dissolved N compounds in runoff were greatest in the grassland (1.72 mg/1) and lowest in bare plots in the shrubland (0.55 mg/1). More than half of the N transported in runoff was carried in dissolved organic compounds. In grassland and shrub plots, the total N loss was highly correlated to the total volume of discharge. We estimate that the total annual loss of N in runoff is 0.25 kg/ha/yr in grasslands and 0.43 kg/ha/yr in shrublands — consistent with the depletion of soil N during desertification of these habitats. Losses of P from both habitats were very small.
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    URL: http://dx.doi.org/10.1007/BF00992871
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  • 40
    Electronic Resource
    Electronic Resource
    A simulation model of organic matter and nutrient accumulation in mangrove wetland soils (1999)
    Springer
    Biogeochemistry 44 (1999), S. 93-118 
    Details
    ISSN: 1573-515X
    Keywords: Everglades National Park ; mangrove soils ; organic matter ; nitrogen ; phosphorus ; sedimentation ; simulation model
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract The distribution and accumulation of organic matter, nitrogen (N) and phosphorus (P) in mangrove soils at four sites along the Shark River estuary of south Florida were investigated with empirical measures and a process-based model. The mangrove nutrient model (NUMAN) was developed from the SEMIDEC marsh organic matter model and parameterized with data from mangrove wetlands. The soil characteristics in the four mangrove sites varied greatly in both concentrations and profiles of soil carbon, N and P. Organic matter decreased from 82% in the upstream locations to 30% in the marine sites. Comparisons of simulated and observed results demonstrated that landscape gradients of soil characteristics along the estuary can be adequately modeled by accounting for plant production, litter decomposition and export, and allochthonous input of mineral sediments. Model sensitivity analyses suggest that root production has a more significant effect on soil composition than litter fall. Model simulations showed that the greatest change in organic matter, N, and P occurred from the soil surface to 5 cm depth. The rapid decomposition of labile organic matter was responsible for this decrease in organic matter. Simulated N mineralization rates decreased quickly with depth, which corresponded with the decrease of labile organic matter. The increase in organic matter content and decrease in soil bulk density from mangrove sites at downstream locations compared to those at upstream locations was controlled mainly by variation in allochthonous inputs of mineral matter at the mouth of the estuary, along with gradients in mangrove root production. Research on allochthonouns sediment input and in situ root production of mangroves is limited compared to their significance to understanding nutrient biogeochemistry of these wetlands. More accurate simulations of temporal patterns of nutrient characteristics with depth will depend on including the effects of disturbance such as hurricanes on sediment redistribution and biomass production.
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    URL: http://dx.doi.org/10.1023/A:1006076405557
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  • 41
    Electronic Resource
    Electronic Resource
    Nutrient budgets and biogeochemistry in an experimental agricultural watershed in Southeastern China (1999)
    Springer
    Biogeochemistry 45 (1999), S. 1-19 
    Details
    ISSN: 1573-515X
    Keywords: Chaohu Lake ; chemical fertilizer ; cycling ; denitrification ; multipond system ; nitrogen ; nutrient budget ; phosphorus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract During a two-year field study, an annual nutrient budget and cycles were developed for a small agricultural watershed. The study emphasized the integrated unit of the watershed in understanding the biogeochemistry. It was found that the total nutrient input was 39.1 × 104 kg nitrogen and 3.91 × 104 kg phosphorus in the year 1995, of which the greatest input of nutrients to the watershed was chemical fertilizer application, reaching 34.7 × 104 kg (676 kg/ha) nitrogen and 3.88 × 104 kg (76 kg/ha) phosphorus. The total nutrient output from the watershed was 13.55 × 104 kg nitrogen and 0.40 × 104 kg phosphorus, while the largest output of nitrogen was denitrification, accounting for 44.1% of N output; the largest output of phosphorus was sale of crops, accounting for 99.4% of P output. The results show that the nutrient input is larger than output, demonstrating that there is nutrient surplus within the watershed, a surplus which may become a potential source of nonpoint pollution to area waters. The research showed that both denitrification and volatilization of nitrogen are key ways of nitrogen loss from the watershed. This suggests that careful management of fertilizer application will be important for the sustainable development of agriculture. The research demonstrated that a multipond system within the watershed had high retention rate for both water and nutrients, benefiting the water, nutrient and sediment recycling in the terrestrial ecosystem and helping to reduce agricultural nonpoint pollution at its source. Therefore, this unique watershed system should be recommended due to its great potential relevance for sustainable agricultural development.
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    URL: http://dx.doi.org/10.1023/A:1006073408623
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  • 42
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    Electronic Resource
    The fate of NH4NO3 added toSphagnum magettanicum carpets at five European mire sites (1999)
    Springer
    Biogeochemistry 45 (1999), S. 73-93 
    Details
    ISSN: 1573-515X
    Keywords: atmospheric deposition ; moss ; bog ; nitrogen ; phosphorus ; water table
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract Nitrogen additions as NH4NO3 corresponding to 0 (N0), 1 (N1), 3 (N3) and 10 (N10) g N m−2 yr−1 were made toSphagnum magellanicurn cores at two-week intervalsin situ at four sites across Europe, i.e. Lakkasuo (Finland). Männikjärve (Estonia), Moidach More (UK) and Côte de Braveix (France). The same treatments were applied in a glasshouse experiment in Neuchâtel (Switzerland) in which the water table depth was artificially maintained at 7, 17 and 37 cm below the moss surface. In the field, N assimilation in excess of values in wet deposition occurred in the absence of growth, but varied widely between sites, being absent in Lakkasuo (moss N∶P ratio 68) and greatest in Moidach More (N∶P 21). In the glasshouse, growth was reduced by lowering the water table without any apparent effect on N assimilation. Total N content of the moss in field sites increased as the mean depth of water table increased indicating growth limitation leading to increased N concentrations which could reduce the capacity for N retention. Greater contents of NH4 + in the underlying peat at 30 cm depth, both in response to NH4NO3 addition and in the unamended cores confirmed poor retention of inorganic N by the moss at Lakkasuo. Nitrate contents in the profiles at Lakkasuo, Moidach More, and Côte de Braveix were extremely low, even in the N10 treatment, but in Männikjärve, where the mean depth of water table was greatest and retention absent, appreciable amounts of NO3 − were detected in all cores. It is concluded that peatland drainage would reduce the capture of inorganic N in atmospheric deposition bySphagnum mosses.
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  • 43
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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/10.1007/BF01007576
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  • 44
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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 soil-water 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/10.1023/A:1006152112852
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  • 45
    Electronic Resource
    Electronic Resource
    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 of N limitation to net primary production (NPP) and other ecosystem processes, despite the ubiquitous occurrence of N-fixing symbioses, remains a significant puzzle in terrestrial ecology. We describe a simple simulation model for an ecosystem containing a generic nonfixer and a symbiotic N fixer, based on: (1) a higher cost for N acquisition by N fixers than nonfixers; (2) growth of fixers and fixation of N only when low N availability limits the growth of nonfixers, and other resources are available; and (3) losses of fixed N from the system only when the quantity of available N exceeds plant and microbial demands. Despite the disadvantages faced by the N fixer under these conditions, N fixation and loss adjust N availability close to the availability of other resources, and biomass and NPP in this simple model can be substantially but only transiently N limited. We then modify the model by adding: (1) losses of N in forms other than excess available N (e.g., dissolved organic N, trace gases produced by nitrification); and (2) constraints to the growth and activity of N 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 and N fixation, that is nevertheless strongly N-limited at equilibrium. This model is useful for exploring causes and consequences of constraints to N fixation, and hence of N limitation, and we believe it will also be useful for evaluating how N fixation and limitation interact with elevated CO2 and other components of global enviromental change.
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    URL: http://dx.doi.org/10.1023/A:1006185020121
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  • 46
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    A comparison of fractionation methods for forms of phosphorus in soils (1999)
    Springer
    Biogeochemistry 47 (1999), S. 25-38 
    Details
    ISSN: 1573-515X
    Keywords: Hedley fractionation ; phosphorus ; Ruttenberg fractionation ; soil phosphorus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract We used l6 soils to compare the Hedley method for soil phosphorus fractionation to an alternative method recently developed by Ruttenberg to differentiate among P fractions in marine sediments. For forms of labile and Fe-bound P in soils, these methods were poorly correlated, with the Hedley fractionation showing a greater ability to discriminate among variations in plant-available P. For Ca-bound P, total organic P, and total P, the methods were well correlated (r2 = 0.93, 0.48, 0.74, respectively), although the sum of P measured in the Ruttenberg extractions is only 45% of the total P recovered by the Hedley fractionation. The Hedley fractionation seems superior when an index of plant-available phosphorus and a separation of organic and inorganic forms is needed, whereas the Ruttenberg method allows a separation of CaCO3-bound P from apatite-P, which is potentially useful in calcareous soils.
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    URL: http://dx.doi.org/10.1023/A:1006105420235
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  • 47
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    A comparison of fractionation methods for forms of phosphorus in soils (1999)
    Springer
    Biogeochemistry 47 (1999), S. 25-38 
    Details
    ISSN: 1573-515X
    Keywords: Hedley fractionation ; phosphorus ; Ruttenberg fractionation ; soil phosphorus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract We used 16 soils to compare the Hedley method for soil phosphorus fractionation to an alternative method recently developed by Ruttenberg to differentiate among P fractions in marine sediments. For forms of labile and Fe-bound P in soils, these methods were poorly correlated, with the Hedley fractionation showing a greater ability to discriminate among variations in plant-available P. For Ca-bound P, total organic P, and total P, the methods were well correlated (r2=0.93, 0.48, 0.74, respectively), although the sum of P measured in the Ruttenberg extractions is only 45% of the total P recovered by the Hedley fractionation. The Hedley fractionation seems superior when an index of plant-available phosphorus and a separation of organic and inorganic forms is needed, whereas the Ruttenberg method allows a separation of CaCO3-bound P from apatite-P, which is potentially useful in calcareous soils.
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    Absolute Rate Coefficients for the Gas-Phase Reactions of NO3 Radical with a Series of Monoterpenes at T = 298 to 433 K (1999)
    Springer
    Journal of atmospheric chemistry 33 (1999), S. 265-282 
    Details
    ISSN: 1573-0662
    Keywords: atmosphere ; kinetics ; nitrate radical ; monoterpenes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract The aim of this work is to study the reactivity of some naturally emitted terpenes, 2-carene, sabinene, myrcene, α-phellandrene, d-limonene, terpinolene and γ-terpinene, towards NO3 radical to evaluate the importance of these reactions in the atmosphere and their atmospheric impact. The experiments with these monoterpenes have been carried out under second-order kinetic conditions over the range of temperature 298–433 K, using a discharge flow system and monitoring the NO3 radical by Laser Induced Fluorescence (LIF). This work is the first temperature dependence study for the reactions of the nitrate radical with the above-mentioned monoterpenes. The measured rate constants at 298 K for the reaction of NO3 with such terpenes are as follows: 2-carene, 16.6 ± 1.8, sabinene 10.7 ± 1.6, myrcene 12.8 ± 1.1, α-phellandrene 42 ± 10, d-limonene 9.4 ± 0.9, terpinolene 52 ± 9 and γ-terpinene 24 ± 7, in units of 10-12 cm3 molecule-1 s-1. The proposed Arrhenius expressions, for the reactions of NO3 with 2-carene, sabinene, myrcene and α-phellandrene are, respectively k1 = (1.4 ± 0.7) × 10-12 exp[(741 ± 190/T)] (cm3 molecule-1 s-1), k2=(2.3 ± 1.3) × 10-10 exp[−(940 ± 200/T)] (cm3 molecule-1 s-1), k3 = (2.2 ± 0.2) × 10-12 exp[(523 ± 35/T)] (cm3 molecule1 s-1) and k4 = (1.9 ± 1.3) × 10-9 exp[−(1158 ± 270/T)] (cm3 molecule-1 s-1). A decrease in the rate constants when raising the temperature has also been found for the reaction of d-limonene with NO3 while an increase in the rate constant with temperature has been observed for the reactions of terpinolene and γ-terpinene with NO3. Tropospheric half-lives for these terpenes have been calculated at night and during the day for typical NO3 and OH concentrations showing that both radicals provide an effective tropospheric sink for these compounds and that the night-time reaction with NO3 radical can be an important, if not dominant, loss process for these naturally emitted organics and for NO3 radicals.
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    Laboratory Kinetic Studies of the Reactions of Cl Atoms with Species of Biogenic Origin: δ3-Carene, Isoprene, Methacrolein and Methyl Vinyl Ketone (1999)
    Springer
    Journal of atmospheric chemistry 34 (1999), S. 163-170 
    Details
    ISSN: 1573-0662
    Keywords: δ3-carene ; chlorine atoms ; isoprene ; kinetics ; methacrolein and methyl vinyl ketone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract The rate coefficients for the reaction between atomic chlorine and a number of naturally occurring species have been measured at ambient temperature and atmospheric pressure using the relative rate technique. The values obtained were (4.0 ± 0.8) × 10-10, (2.1 ± 0.5) × 10-10, (3.2 ± 0.5) × 10-10, and (4.9 ± 0.5) × 10-10 cm3 molecule-1 s-1, for reactions with isoprene, methyl vinyl ketone, methacrolein and δ3-carene, respectively. The value obtained for isoprene compares favourably with previously reported values. No values have been reported to date for the rate constants of the other reactions.
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    The fate of NH4NO3 added to Sphagnum magellanicum carpets at five European mire sites (1999)
    Springer
    Biogeochemistry 45 (1999), S. 73-93 
    Details
    ISSN: 1573-515X
    Keywords: atmospheric deposition ; moss ; bog ; nitrogen ; phosphorus ; water table
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract Nitrogen additions as NH4NO3 corresponding to 0 (N0), 1 (N1), 3 (N3) and 10 (N10) g N m-2 yr-1 were made to Sphagnum magellanicum cores at two-week intervals in situ at four sites across Europe, i.e. Lakkasuo (Finland), Männikjärve (Estonia), Moidach More (UK) and Côte de Braveix (France). The same treatments were applied in a glasshouse experiment in Neuchâtel (Switzerland) in which the water table depth was artificially maintained at 7, 17 and 37 cm below the moss surface. In the field, N assimilation in excess of values in wet deposition occurred in the absence of growth, but varied widely between sites, being absent in Lakkasuo (moss N:P ratio 68) and greatest in Moidach More (N:P 21). In the glasshouse, growth was reduced by lowering the water table without any apparent effect on N assimilation. Total N content of the moss in field sites increased as the mean depth of water table increased indicating growth limitation leading to increased N concentrations which could reduce the capacity for N retention. Greater contents of NH4+ in the underlying peat at 30 cm depth, both in response to NH4NO3 addition and in the unamended cores confirmed poor retention of inorganic N by the moss at Lakkasuo. Nitrate contents in the profiles at Lakkasuo, Moidach More, and Côte de Braveix were extremely low, even in the N10 treatment, but in Männikjärve, where the mean depth of water table was greatest and retention absent, appreciable amounts of NO3- were detected in all cores. It is concluded that peatland drainage would reduce the capture of inorganic N in atmospheric deposition by Sphagnum mosses.
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    Distribution and speciation of phosphorus along a salinity gradient in intertidal marsh sediments (1999)
    Springer
    Biogeochemistry 45 (1999), S. 197-221 
    Details
    ISSN: 1573-515X
    Keywords: intertidal marshes ; phosphorus ; sediments
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract We examined forms of solid phosphorus fractions in intertidal marsh sediments along a salinity (0–22‰) gradient in a river-dominated estuary and in a marine-dominated salt marsh with insignificant freshwater input. Freshwater marsh sediments had the highest ratio of organic N:P of between 28:1 and 47:1 mol:mol, compared to 21:1 to 31:1 mol:mol in the saltmarshes, which is consistent with a trend toward P-limitation of primary production in freshwater and N-limitation in salt marshes. However, total P concentration, 24.7 ± 11.1 µmol P g dw-1 (±1 SD) averaged over the upper meter of sediment, was greatest in the freshwater marsh where bioavailablity of P is apparently limited. In the freshwater marsh the greatest fraction of total P (24–51%) was associated with humic acids, while the importance of humic-P decreased with increasing salinity to 1–23% in the salt marshes. Inorganic P contributed considerably less to total sediment P in the freshwater marsh (15–40%) than in the salt marshes (33–85%). In reduced sediments at all sites, phosphate bound to aluminum oxides and clays was an important inorganic P pool irrespective of salinity. Inorganic P associated with ferric iron [Fe(III)] phases was most abundant in surface sediments of freshwater and brackish marshes, while Ca-bound P dominated inorganic P pools in the salt marshes. Thus, our results showed that particle-bound P in marsh sediments exhibited changes in chemical association along the salinity gradient of an estuarine system, which is a likely consequence of changes in ionic strength and the availability of iron and calcium.
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    Distribution and speciation of phosphorus along a salinity gradient in intertidal marsh sediments (1999)
    Springer
    Biogeochemistry 45 (1999), S. 197-221 
    Details
    ISSN: 1573-515X
    Keywords: intertidal marshes ; phosphorus ; sediments
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract We examined forms of solid phosphorus fractions in intertidal marsh sediments along a salinity (0–22%.) gradient in a river-dominated estuary and in a marine-dominated salt marsh with insignificant freshwater input. Freshwater marsh sediments had the highest ratio of organic N:P of between 28:1 and 47:1 mol:mol, compared to 21∶1 to 31∶1 mol∶mol in the saltmarshes, which is consistent with a trend toward P-limitation of primary production in freshwater and N-limitation in salt marshes. However, total P concentration, 24.7±11.1μmol P g dw−1 (±1 SD) averaged over the upper meter of sediment, was greatest in the freshwater marsh where bioavailablity of P is apparently limited. In the freshwater marsh the greatest fraction of total P (24–51%.) was associated with humic acids, while the importance of humic-P decreased with increasing salinity to 1–23%. in the salt marshes. Inorganic P contributed considerably less to total sediment P in the freshwater marsh (15–40%.) than in the salt marshes (33–85%.). In reduced sediments at all sites, phosphate bound to aluminum oxides and clays was an important inorganic P pool irrespective of salinity. Inorganic P associated with ferric iron [Fe(III)] phases was most abundant in surface sediments of freshwater and brackish marshes, while Ca-bound P dominated inorganic P pools in the salt marshes. Thus, our results showed that particle-bound P in marsh sediments exhibited changes in chemical association along the salinity gradient of an estuarine system, which is a likely consequence of changes in ionic strength and the availability of iron and calcium.
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    Nutrient losses in runoff from grassland and shrubland habitats in Southern New Mexico: I. rainfall simulation experiments (1999)
    Springer
    Biogeochemistry 45 (1999), S. 21-34 
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    ISSN: 1573-515X
    Keywords: Chihuahuan desert ; desert ; desertification ; grassland ; nitrogen ; nutrient budgets ; phosphorus ; runoff
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract Rainfall simulation experiments were performed in areas of semiarid grassland (Bouteloua eriopoda) and arid shrubland (Larrea tridentata) in the Chihuahuan desert of New Mexico. The objective was to compare the runoff of nitrogen (N) and phosphorus (P) from these habitats to assess whether losses of soil nutrients are associated with the invasion of grasslands by shrubs. Runoff losses from grass- and shrub-dominated plots were similar, and much less than from bare plots located in the shrubland. Weighted average concentrations of total dissolved N compounds in runoff were greatest in the grassland (1.72 mg/l) and lowest in bare plots in the shrubland (0.55 mg/l). More than half of the N transported in runoff was carried in dissolved organic compounds. In grassland and shrub plots, the total N loss was highly correlated to the total volume of discharge. We estimate that the total annual loss of N in runoff is 0.25 kg/ha/yr in grasslands and 0.43 kg/ha/yr in shrublands – consistent with the depletion of soil N during desertification of these habitats. Losses of P from both habitats were very small.
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    Molecular structure of trans-[Co(NH3)4(NH2CH3)Cl](ClO4)2 (1998)
    Springer
    Journal of chemical crystallography 28 (1998), S. 69-72 
    Details
    ISSN: 1572-8854
    Keywords: Co(III) complex ; crystal structure ; kinetics ; steric effects
    Source: Springer Online Journal Archives 1860-2000
    Topics: Geosciences , Physics
    Notes: Abstract The title compound crystallizes in the orthorhombic space group Pnma, with a = 7.9209(5), b = 9.818(1), c = 16.867(2) Å, and Z = 4. The structure was solved employing 1864 independent x-ray reflections with I〉2σ(I) by Patterson and difference Fourier techniques and refined by full-matrix least-squares to R = 0.036. The trans-[CO(NH3)4(NH2CH3)Cl](ClO4)2 molecule is on a crystallographic mirror plane. The cobalt ion is in an elongated octahedral coordination with four equatorial ammonia ligands [average Co–N distance equal to 1.966(2) Å], an axial methylamine [Co–N=1.965(3)Å], and an axial chlorine ion [Co–Cl=2.2771(9)Å]. Kinetic steric effects of the complex are interpreted in terms of structural results.
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    Variability of diatom-inferred phosphorus profiles in a small lake basin and its implications for histories of lake eutrophication (1998)
    Springer
    Journal of paleolimnology 20 (1998), S. 47-55 
    Details
    ISSN: 1573-0417
    Keywords: diatoms ; spatial variability ; canonical correspondence analysis ; lake eutrophication ; transfer functions ; phosphorus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences
    Notes: Abstract Diatom analyses were undertaken of sediment cores covering a range of water depths in a small eutrophic lake (Lough Augher, Co. Tyrone, N. Ireland). The significance of between-core variability in diatom relative frequency stratigraphy was assessed by Canonical Correspondence Analysis (CCA) where the ordination axes were constrained to external environmental variables (sediment depth, core location coordinates, water depth, effective fetch, distance-from-shore and distance-from-inflow). After the removal of the effect of sediment age by partialling it out, the resultant first two axes from the partial-CCA were significantly correlated with water depth and distance-from-shore, indicating non-uniform diatom stratigraphies across the lake. Despite this variability, all cores show the same succession of species and, therefore, record the eutrophication of the lake. Diatom-inferred total phosphorus (DI-TP) was inferred for six cores using weighted averaging regression and calibration. Apart from considerable differences of DI-TP in surficial sediment samples, there was good between-core repeatability of DI-TP profiles. These data support the use of DI-TP for establishing background nutrient concentrations for lakes, and associated implications for lake restoration schemes using single cores. Comparisons of DI-TP profiles and total diatom accumulation rate data for the individual cores indicate that diatom production peaked prior to the maximum TP concentrations in the lake.
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    Modern and historic accumulation rates of phosphorus in Lake Okeechobee, Florida (1998)
    Springer
    Journal of paleolimnology 20 (1998), S. 31-46 
    Details
    ISSN: 1573-0417
    Keywords: phosphorus ; Lake Okeechobee ; lead-210 dating ; eutrophication ; phosphorus loading
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences
    Notes: Abstract Phosphorus accumulation rates in depositional zone sediments of Lake Okeechobee were determined in 11 mud-zone cores and two peat-zone cores dated by 210Pb. Although difficulties were encountered in interpreting 210Pb data from some sites, reliable dating of sediments from the mud zone of this shallow lake is possible. Sediment accumulation rates in this zone have increased during the present century by an average of about twofold, and accumulation of organic sediments in the lake during pre-settlement times apparently was much slower than during the past century. Concentrations of all forms of sedimentary P but especially nonapatite inorganic-P and organic-P also have increased since pre-settlement times and especially since about 1940. Annual P accumulation rates in the lake's sediments have increased about fourfold during the 1900s, with most of the increase occurring in the past 40–50 years. The recent accumulation rate of sedimentary P (past ~ 10 years) agrees within a factor of 1.5 with the net retention of P in the lake calculated from published input-output mass balances.
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    Evidence for O-dealkylation of 7-pentoxyresorufin by cytochrome P450 2B1/2B2 isoenzymes in brain (1998)
    Springer
    Molecular and cellular biochemistry 189 (1998), S. 201-205 
    Details
    ISSN: 1573-4919
    Keywords: brain ; P450 ; PB ; PROD ; induction ; inhibition ; kinetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract O-dealkylation of 7-pentoxyresorufin (PR) was studied in rat brain to characterise the functional activity specific for cytochrome P450 2B1/2B2 isoenzymes in brain microsomes. Brain microsomes catalyzed the O-dealkylation of PR in the presence of NADPH. Pretreatment with phenobarbital (PB; 80 mg/kg body wt, i.p.× 5 days) resulted in 3-4 fold induction of pentoxyresorufin-O-dealkylase (PROD) activity while 3-methylcholanthrene (MC; 30 mg/kg body wt, i.p. × 5 days) did not produce any significant increase in enzyme activity. Kinetic studies revealed that the rate of velocity (Vmax) for the O-dealkylation of PR was significantly increased to 2.9 times higher in brain microsomes isolated from PB pretreated rats. In vitro studies using metyrapone, an inhibitor of P450 2B1/2B2 catalyzed reactions and antibody for hepatic PB inducible P450s (P450 2B1/2B2) significantly inhibited the activity of PROD in cerebral microsomes prepared from PB pretreated animals. These studies suggest that PB inducible isoenzymes of P450, i.e. P450 2B1/2B2 specifically catalyze the O-dealkylation of PR in brain microsomes.
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    Distinct populations of macrophages in the adult rat adrenal gland: a subpopulation with neurotrophin-4-like immunoreactivity (1998)
    Springer
    Cell & tissue research 291 (1998), S. 365-373 
    Details
    ISSN: 1432-0878
    Keywords: Key words Adrenal medulla ; Chromaffin cells ; Neurotrophins ; Neurotrophin receptors ; c-fos ; Neuro-endocrine-immune interactions ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Macrophages are widely distributed in lymphohaemopoietic and many other mammalian tissues, where they are mainly involved in host defence mechanisms, phagocytosis, wound repair, and secretion of growth factors. Increasing evidence suggests that secretory products of macrophages can influence adrenal gland functions. In the present study, we have used specific antibodies to ED1 (cytoplasmic antigen), ED2 (membrane antigen), ED8 (membrane antigen), and OX-6 (MHC class II/membrane antigen) as markers for macrophages to examine their distribution within the adult rat adrenal gland. ED2 and OX-6 recognize distinct subpopulations of adrenal gland macrophages, whereas macrophages immunoreactive (-ir) for ED1 and ED8 could not be detected. OX-6-ir macrophages were most numerous in the cortical reticularis and glomerulosa zones, while only few cells were found in the zona fasciculata and in the adrenal medulla. Macrophages immunoreactive for ED2 were restricted to the adrenal medulla. The majority of these macrophages were associated with vascular sinuses or chromaffin cells. By double-immunolabelling we found that most of ED2-ir medullary macrophages contain neurotrophin-4 (NT-4)-like ir. Attempts to clarify whether macrophages take up NT-4 from NT-4-ir chromaffin cells indicated that medullary macrophages are immunonegative for chromogranin A and neuropeptide Y, two major secretory products of chromaffin cells. In situ hybridizations and immunofluorescence showed expression of the neurotrophin receptor TrkA, but not TrkB in the adrenal medulla. In vitro studies indicated that NT-4, similar to nerve growth factor, can induce c-fos-ir in chromaffin cells. We conclude that chromaffin cells are putative targets for adrenal medullary NT-4, whose functions remain to be clarified.
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    Distribution of hydroxyindole-O-methyltransferase mRNA in the rat brain: an in situ hybridisation study (1998)
    Springer
    Cell & tissue research 291 (1998), S. 415-421 
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    ISSN: 1432-0878
    Keywords: Key words Hydroxyindole-O-methyltransferase ; Pineal gland ; In situ hybridisation ; Melatonin ; 5-Methoxyindoles ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Hydroxyindole-O-methyltransferase (HIOMT) is the enzyme involved in the last step of the melatonin synthesis pathway. Recently, a cDNA encoding HIOMT has been isolated from a rat pineal gland library. Using this cDNA, we developed a highly sensitive in situ hybridisation protocol to investigate the distribution of HIOMT mRNA in both the rat brain and dissociated pinealocytes maintained in primary cell culture. In the rat brain, HIOMT mRNA was only detected in the three parts of the pineal complex: the superficial pineal, the stalk and the deep pineal. No extra-pineal hybridisation labelling was observed. These results strongly suggest that melatonin synthesis also occurs in the deep part and the stalk of the pineal gland. HIOMT mRNA was markedly expressed in cultured pinealocytes. No particular subcellular area was observed to express HIOMT mRNA specifically, as the labelling was homogeneously distributed in the cytosol and in the axon-like processes. In conclusion, the use of in situ and in vitro hybridisation with a pineal riboprobe has detected notable HIOMT expression restricted to pinealocytes.
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    Differential myogenicity of satellite cells isolated from extensor digitorum longus (EDL) and soleus rat muscles revealed in vitro (1998)
    Springer
    Cell & tissue research 291 (1998), S. 455-468 
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    ISSN: 1432-0878
    Keywords: Key words EDL ; Soleus ; Satellite cells ; Proliferation ; Differentiation ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Following muscle damage, fast- and slow-contracting fibers regenerate, owing to the activation of their satellite cells. In rats, crush-induced regeneration of extensor digitorum longus (EDL, a fast muscle) and soleus (a slow muscle) present different characteristics, suggesting that intrinsic differences exist among their satellite cells. An in vitro comparative study of the proliferation and differentiation capacities of satellite cells isolated from these muscles is presented there. We observed several differences between soleus and EDL satellite cell cultures plated at high density on gelatin-coated dishes. Soleus satellite cells proliferated more actively and fused into myotubes less efficiently than EDL cells. The rate of muscular creatine kinase enzyme appeared slightly lower in soleus than in EDL cultures at day 11 after plating, when many myotubes were formed, although the levels of muscular creatine kinase mRNA were similar in both cultures. In addition, soleus cultures expressed higher levels of MyoD and myogenin mRNA and of MyoD protein than EDL satellite cell cultures at day 12. A clonal analysis was also carried out on both cell populations in order to determine if distinct lineage features could be detected among satellite cells derived from EDL and soleus muscles. When plated on gelatin at clonal density, cells from both muscles yielded clones within 2 weeks, which stemmed from 3–15 mitotic cycles and were classified into three classes according to their sizes. Myotubes resulting from spontaneous fusion of cells from the progeny of one single cell were seen regardless of the clone size in the standard culture medium we used. The proportion of clones showing myotubes in each class depended on the muscle origin of the cells and was greater in EDL- than in soleus-cell cultures. In addition, soleus cells were shown to improve their differentiation capacity upon changes in the culture condition. Indeed, the proportions of clones showing myotubes, or of cells fusing into myotubes in clones, were increased by treatments with a myotube-conditioned medium, with phorbol ester, and by growth on extra-cellular matrix components (Matrigel). These results, showing differences among satellite cells from fast and slow muscles, might be of importance to muscle repair after trauma and in pathological situations.
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    Effects of glutamate-induced excitotoxicity on calretinin-expressing neuron populations in the area postrema of the rat (1998)
    Springer
    Cell & tissue research 293 (1998), S. 227-233 
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    ISSN: 1432-0878
    Keywords: Key words Calretinin ; Calcium-binding proteins ; Monosodium glutamate ; Excitotoxicity ; Area postrema ; Circumventricular organs ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  We mapped the distribution of calretinin-immunoreactive neuron populations in a circumventricular organ of the rat, the area postrema, and investigated their sensitivity to excitotoxic stimuli mediated by subcutaneously administered monosodium glutamate. We were specifically interested to ascertain whether the presence of calretinin can, per se, confer an in vivo intrinsic resistance for area postrema neurons to glutamate excitotoxicity. We found that dense populations of calretinin-positive neurons displayed a subregional compartmentation in coronal sections of the area postrema along its rostrocaudal axis. We demonstrated that calretinin-positive neurons differ in their sensitivities to monosodium glutamate depending on their position within the area postrema. Neurons in the caudal area postrema were the most sensitive ones, while those in the rostral area postrema were spared of degeneration. We conclude that calretinin-positive neurons in the area postrema are not uniformly protected against glutamate excitotoxicity. It is possible that differences in the local concentrations of monosodium glutamate due to regional heterogeneities in density and permeability of the capillary bed rather than neuronal expression of calretinin account for the observed effects.
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    Neurocalcin-like immunoreactivity in the rat esophageal nervous system (1998)
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    Cell & tissue research 294 (1998), S. 57-68 
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    ISSN: 1432-0878
    Keywords: Key words Calcium-binding protein ; Nerve terminal ; Motor endplate ; Vagus nerve ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Neurocalcin is a newly identified neuronal calcium-binding protein. We tried here to investigate the immunohistochemical distribution of neurocalcin in the rat esophagus. Nerve cell bodies having neurocalcin immunoreactivity were found throughout the myenteric plexus. In the myenteric ganglia, two types of nerve terminals showed neurocalcin immunoreactivity. One was varicose terminals containing numerous small clear vesicles and forming a synapse with nerve cells. The other terminals were characterized by laminar or pleomorphic structure and many mitochondria. These laminar terminals were supposed to be sensory receptors of the esophageal wall. In the motor endplates of the striated muscles, nerve terminals containing many small clear vesicles and mitochondria also had neurocalcin immunoreactivity. After left vagus nerve cutting under the nodose ganglia, the number of immunopositive thick nerve fibers, laminar endings and nerve terminals on the striated muscles decreased markedly. Retrograde tracing experiments using Fast Blue showed extrinsic innervation of esophagus from ambiguus nucleus, dorsal motor nucleus of vagus, superior cervical ganglia, celiac ganglia, nodose ganglia and dorsal root ganglia. In the celiac ganglia, nodose ganglia and dorsal root ganglia, retrogradely labeled nerve cells were neurocalcin-immunoreactive. Neurons in the celiac ganglia may project varicose terminals, while nodose and dorsal root neurons project laminar terminals. Although cell bodies of motoneurons in the ambiguus nucleus lacked neurocalcin immunoreactivity, these neurons may contain neurocalcin only in the nerve terminals in the motor endplates. Neurocalcin immunoreactivity is distributed in many extrinsic and intrinsic neurons in the esophagus and this protein may play important roles in regulating calcium signaling in the neurons.
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    Osteogenic inhibition by rat periodontal ligament cells: modulation of bone morphogenic protein-7 activity in vivo (1998)
    Springer
    Cell & tissue research 294 (1998), S. 475-483 
    Details
    ISSN: 1432-0878
    Keywords: Key words Periodontal ligament ; α-Smooth muscle actin ; Osteopontin ; Bone sialoprotein ; Bone morphogenic protein ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Periodontal ligament width is precisely maintained throughout the lifetime of adult mammals but the biological mechanisms that inhibit ingrowth of bone into this soft connective tissue are unknown. As bone morphogenic proteins strongly stimulate osteogenesis and can induce ectopic bone formation in vivo, we tested the hypothesis that topical application of this powerful osteogenic agent will overwhelm the osteogenic inhibitory mechanisms of periodontal ligament cells and induce ankylosis. Wounds through the alveolar bone and periodontal ligament were created in 45 male Wistar rats. Defects were filled with either a collagen implant or collagen plus bone morphogenic protein (BMP-7), or were left unfilled (controls). Three animals per time period were killed on days 2, 5, 10, 21 and 60 after surgery for each wound type. Cellular proliferation and clonal growth in periodontal tissues were assessed by 3H-thymidine labeling 1 h before death, followed by radioautography. Cellular differentiation of soft and mineralizing connective tissue cell populations was determined by immunohistochemical staining of α-smooth muscle actin, osteopontin and bone sialoprotein. In regenerating periodontium, BMP-7 induced abundant bone formation by 21 days (2.5-fold greater than controls or collagen implant only; P〈0.001), but by day 60 the volume of the newly formed bone had returned to baseline levels and was similar for all groups. Independent of the type of treatment, periodontal ligament width was unchanged throughout the experimental period (P〉0.05). Animals treated with BMP-7 implants showed greatly increased cellular proliferation in the periodontal ligament adjacent to the wound site and in the regenerating alveolar bone at days 5 and 10 after wounding compared to the other treatment groups (P〈0.005). Animals in the BMP-7 group exhibited similar spatial and temporal staining patterns for α-smooth muscle actin, osteopontin and bone sialoprotein as controls. Collectively, these data show that BMP-7 promoted the proliferation of precursor cells in the periodontal ligament but did not induce osteogenic differentiation in this compartment. Consequently a powerful osteogenic stimulus like BMP-7 cannot significantly perturb the mechanisms that regulate periodontal ligament width and maintain periodontal homeostasis.
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    The role of voltage-gated Ca2+ channels in neurite growth of cultured chromaffin cells induced by extremely low frequency (ELF) magnetic field stimulation (1998)
    Springer
    Cell & tissue research 291 (1998), S. 217-230 
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    ISSN: 1432-0878
    Keywords: Key words: Amperometry  ; Calcium currents ; Catecholamines ; Chromaffin cells ; Differentiation ; Magnetic field ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The ion Ca2+ has been shown to play an important role in a wide variety of cellular functions, one of them being related to cell differentiation in which nerve growth factor (NGF) is involved. Chromaffin cells obtained from adrenals of 2- to 3-day-old rats were cultured for 7 days. During this time, these cells were subjected to the application of either NGF or extremely low frequency magnetic fields (ELF MF). Since this induced cell differentiation toward neuronal-like cells, the mechanism by which this occurred was studied. When the L-Ca2+ channel blocker nifedipine was applied simultaneously with ELF MF, this differentiation did not take place, but it did when an N-Ca2+ channel blocker was used. In contrast, none of the Ca2+ channel blockers prevented differentiation in the presence of NGF. In addition, Bay K-8644, an L-Ca2+ channel agonist, increased both the percentage of differentiated cells and neurite length in the presence of ELF MF. This effect was much weaker in the presence of NGF. [3H]-noradrenaline release was reduced by nifedipine, suggesting an important role for L-Ca2+ channels in neurotransmitter release. Total high voltage Ca2+ currents were significantly increased in ELF MF-treated cells with NGF, but these currents in ELF MF-treated cells were more sensitive to nifedipine. Amperometric analysis of catecholamine release revealed that the KCl-induced activity of cells stimulated to differentiate by ELF MF is highly sensitive to L-type Ca2+ channel blockers. A possible mechanism to explain the way in which the application of magnetic fields can induce differentation of chromaffin cells into neuronal-like cells is proposed.
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    Diurnal pattern of rat pancreatic acinar cell replication (1998)
    Springer
    Cell & tissue research 291 (1998), S. 277-283 
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    ISSN: 1432-0878
    Keywords: Key words Diurnal pattern ; Pancreas ; Acinar cell replication ; Cholecystokinin (CCK) ; Proliferating cell nuclear antigen (PCNA) ; 5-Bromodeoxyuridine (BrdU) ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Fully differentiated pancreatic acinar cells can enter the cell cycle under appropriate conditions in the rat. The aim of this study was to analyse the diurnal pattern of acinar cell proliferation as a function of food intake and the release of cholecystokinin (CCK), because the peptide hormone CCK is a major physiological regulator of rat pancreatic acinar cell replication. Pancreatic acinar cell replication was quantitated using an antibody against the S-phase marker proliferating cell nuclear antigen (PCNA). In addition, acinar cells in S-phase were detected after injecting bromodeoxyuridine (BrdU) and subsequent immunohistochemical staining of BrdU-positive nuclei. Rat pancreata were analysed during the day under standard diet conditions, as well as after various schedules of fasting and refeeding and after the application of the CCK receptor antagonist L-364,718. Between 6 a.m. and 6 p.m., the PCNA labeling index was 4.4±0.9%, while between 9 p.m. and 3 a.m. the PCNA labeling index was elevated and reached peak values of 11.4% (mean value: 7.8±2.5%) around midnight. BrdU-positive cells also doubled around midnight, compared to the 9:00 a.m. value. In fasted rats, acinar cell proliferation was completely suppressed and this suppression could be overcome by injection of the CCK analog cerulein. In addition, the CCK antagonist L-364,718 led to the same results as fasting. Here we show for the first time that there is a diurnal pattern of pancreatic acinar cell proliferation in rats, which is dependent on food intake and is mediated by CCK.
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    Intergranular bridges in the anterior pituitary cell and their possible involvement in Ca2+-induced granule-granule fusion (1998)
    Springer
    Cell & tissue research 292 (1998), S. 513-519 
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    ISSN: 1432-0878
    Keywords: Key words Intergranular bridges ; Granule-granule fusion ; Annexin II ; Anterior pituitary cells ; Multi granular exocytosis ; Clostridium perfringens enterotoxin ; Quick-freeze deep-etch microscopy ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Quick-freeze deep-etch electron microscopy showed the presence of bridge-like structures between adjacent secretory granules in rat anterior pituitary secretory cells. These intergranular bridges were variable in length and thickness. The finest bridges were 7–8 nm in length, while the longest ones were as long as 80 nm. Annexin II, one of the Ca2+-dependent phospholipid-binding proteins, is known to interlink between two membranes and induce aggregation of liposomes and chromaffin granules under the presence of Ca2+. In anterior pituitary cells, annexin II was detected by immunoelectron microscopy at the contact sites of secretory granules with other granules. The anterior pituitary cells treated under the presence of extracellular Ca2+ with Clostridium perfringens enterotoxin which induces Ca2+ influx showed multigranular exocytosis, i.e., multiple fusions of secretory granules with each other and with the plasma membrane. The granule-granule fusion in progress could be captured by the quick-freeze deep-etch technique. The membranes of adjacent secretory granules were partially fused at their contact sites where intergranular strands were no longer seen, while there existed intergranular strands between unfused portions of the granule membranes. From these results, we consider that the intergranular bridges, some of which may be composed of annexin II, are involved in Ca2+-induced granule-granule fusion in anterior pituitary cells.
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    Quantitative changes in rat renin secretory granules after acute and chronic stimulation of the renin system (1998)
    Springer
    Cell & tissue research 292 (1998), S. 563-571 
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    ISSN: 1432-0878
    Keywords: Key words Juxtaglomerular cells ; Stereology ; Disector ; Salt diets ; Enalapril ; Renal perfusion pressure ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  In order to study the cellular mode of renin secretion, stereological methods were used to estimate number and volume of rat renin secretory granules during stimulation of the renin system. An acute decrease in renal perfusion pressure to 40 mmHg for 5 min increased plasma renin concentration (PRC) twofold, but did not significantly change the number of renin granules per arteriole or the renin-containing volume of the arteriole. Chronic stimulation was achieved by a combination of low-salt diet and inhibition of angiotensin-converting enzyme (ACE) for 14 days, and resulted in a 36-fold increase in PRC, a 20-fold increase in the number of granules per arteriole, and a 17-fold increase in the arteriolar volume that contained renin. An acute decrease in renal perfusion pressure to 40 mmHg for 5 min in the chronically stimulated rats increased PRC further (1.6-fold), and significantly reduced the number of granules per arteriole by 4000 (45% reduction), but did not change the renin-containing arteriolar volume significantly. The average renin granule size was 0.35 μm3 with no significant differences among the groups. We conclude that recruited granular cells contribute significantly to renin release, and that all granular cells along the arteriole participate in secretory responses. The reduced number of renin granules after acute stimulation is compatible with exocytosis as the dominating mechanism of renin release.
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    Immunocytochemistry and in situ hybridization studies of pepsinogen C-producing cells in developing rat fundic glands (1998)
    Springer
    Cell & tissue research 293 (1998), S. 121-131 
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    ISSN: 1432-0878
    Keywords: Key words Pepsinogen C ; Ontogeny ; Mucous neck cell ; Chief cell ; Intermediate mucopeptic cell ; Immunocytochemistry ; In situ hybridization ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The ontogeny of pepsinogen C-producing cells in rat fundic glands was studied by means of light and electron microscopy using an antiserum raised against a synthetic peptide based on rat pepsinogen C. To confirm the immunocytochemistry results, the expression of rat pepsinogen C messenger RNA (mRNA) in the fundic gland was also examined by in situ hybridization using a digoxigenin-labeled RNA probe. In adult rats, pepsinogen C was produced by chief cells, mucous neck cells, and intermediate mucopeptic cells. Pepsinogen C-producing cells appeared in embryos as early as 18.5 days’ gestation. The development of these cells could be classified into four stages: (1) 18.5 days’ gestation to 0.5 days after birth; (2) 0.5 days to 2 weeks after birth; (3) 3–4 weeks after birth; (4) 4–8 weeks after birth. In embryos and young animals, pepsinogen C-producing cells were mucopeptic cells. By 4 weeks after birth, mucous neck cells could be distinguished morphologically. The maturation stages of the chief cells could be traced by electron microscopy along the longitudinal axis of the rat fundic gland by double-staining with anti-pepsinogen C antibody and periodic acid-thiocarbohydrazide-silver proteinate. Positive reactions for pepsinogen C and pepsinogen C mRNA expression were detected in mucous neck cells. Therefore, we conclude that mucous neck cells are precursor cells of chief cells. Mucous neck cells, intermediate cells, and chief cells are in the same differentiating cell lineage.
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    Ultrastructural and lanthanum tracer examination of rapidly resorbing rat alveolar bone suggests that osteoclasts internalize dying bone cells (1998)
    Springer
    Cell & tissue research 293 (1998), S. 173-176 
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    ISSN: 1432-0878
    Keywords: Key words Osteoclast ; Bone ; Bone resorption ; Alveolar bone ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Glutaraldehyde-formaldehyde fixed undecalcified alveolar bone from 7-day-old rats was prepared for light and electron microscopy. Colloidal lanthanum was used as an ultrastructural tracer, and both random and semi-serial sections were examined. Lanthanum penetrated the infoldings of the ruffled border and some nearby vacuoles and vesicles. The majority of vacuoles and vesicles were lanthanum-free. Some osteoclast profiles contained a large vacuole with a cell enclosed in its interior. The enclosed cell exhibited an irregular nucleus containing condensed peripheral chromatin, intact cytoplasmic organelles, conspicuous rough endoplasmic reticulum and large blebs on the cell surface. These features are characteristic of osteoblasts or bone-lining cells or immature osteocytes which may be undergoing apoptosis or necrosis. The observation of remnants of cellular structures within internalized osteoclast vacuoles, together with the above results, suggests that osteoclasts engulf and probably degrade dying osteoblasts/bone-lining cells or immature osteocytes.
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    Electron-immunocytochemical localization of P2X1 receptors in the rat cerebellum (1998)
    Springer
    Cell & tissue research 294 (1998), S. 253-260 
    Details
    ISSN: 1432-0878
    Keywords: Key words P2X1 receptor ; Ultrastructure ; Cerebellum ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The distribution of the P2X1 subtype of purinoceptors associated with the extracellular activities of ATP was studied in the rat cerebellum at the electron-microscope level. Receptors were labelled with peroxidase-antiperoxidase and the avidin-biotin-peroxidase complex for immunocytochemistry. Immunoreactivity to P2X1 receptors was localized in subpopulations of synapses between varicosities of parallel fibres of granule cells and dendritic spines of Purkinje cells. Unlabelled varicosities of parallel fibres formed asymmetric synapses with labelled dendritic spines, whereas labelled varicosities of parallel fibres formed asymmetric synapses with unlabelled dendritic spines. P2X1 immunoreactivity was also localized in some astrocyte processes. The functional significance of these findings is discussed.
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    Expression of transforming growth factor-β isoforms in the rat male accessory sex organs and epididymis (1998)
    Springer
    Cell & tissue research 294 (1998), S. 271-277 
    Details
    ISSN: 1432-0878
    Keywords: Key words Coagulating gland ; Epididymis ; Gene expression ; Prostate ; Seminal vesicle ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  We studied the expression and distribution of transforming growth factor-β (TGF-β) isoforms in the rat male accessory sex glands and the epididymis. Our data demonstrate the expression of both TGF-β1 and -β3 isoforms in ventral prostate (VP), seminal vesicle (SV), coagulating gland (CG), and epididymis (E) by Northern blot analysis. In addition, there was differential expression of TGF-β3 in the three regions of epididymis, the corpus region being the highest. Immunostaining data showed intense staining for latent TGF-β1 in all the male accessory glands. In contrast, no staining using antibodies specific for active TGF-β1 was observed. No expression of TGF-β2 was evident either by immunohistochemistry or Northern blot analysis. The presence of mature TGF-β3 protein was observed in the secretory epithelium of VP, CG, and corpus E. There was no detectable staining of TGF-β3 in the seminal vesicle and caput and cauda regions of epididymis. These data suggest possible differential regulation of TGF-β isoform expression in the male reproductive system and predict unique roles for individual TGF-β isoforms in sperm maturation and maintenance.
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    Expression of the gap-junction connexins 26 and 30 in the rat cochlea (1998)
    Springer
    Cell & tissue research 294 (1998), S. 415-420 
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    ISSN: 1432-0878
    Keywords: Key words Inner ear ; Gap junctions ; Connexins ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Gap junction channels which are responsible for direct intercellular communication are composed of connexin proteins. Different connexins are distributed in a tissue-specific manner. Up to now only connexin26 has been identified to be widely expressed in the inner ear. In order to investigate the role of additional gap junction proteins, the expression of connexin30 and 43 was investigated in the rat cochlea. Connexin26 and connexin30 were both expressed in the spiral limbus, the spiral ligament, the stria vascularis and between supporting cells of the organ of Corti. Double-labeling experiments suggest that both connexins are partly colocalized between cells. Weak staining of connexin43 could only be detected in the stria vascularis, the spiral ligament and between organ of Corti supporting cells. The corresponding transcripts for connexin26, 30 and 43 could be detected by Northern blot analysis. The expression of different gap junction channels in the cochlea suggests functional diversity. Gap junctions in the inner ear may control ion concentrations of cochlear fluids or act as conduits through which glucose and other metabolites diffuse.
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    Decline and recovery of olfactory receptor expression following unilateral bulbectomy (1998)
    Springer
    Cell & tissue research 294 (1998), S. 421-430 
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    ISSN: 1432-0878
    Keywords: Key words Odorant receptors ; Olfactory epithelium ; Regeneration ; Bulbectomy ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The effects of unilateral olfactory bulb ablation upon the odorant receptor expression were studied during the degeneration/regeneration process in the olfactory epithelium of adult rats. Using the in situ hybridization approach, we compared the time course of decay and recovery of expression for three different receptor subtypes (OR14, OR5, OR124). The number of neurons expressing receptor subtypes dramatically decreased in the olfactory epithelium on the lesioned side and reached a minimum at day 5 postsurgery. A progressive recovery was then observed from day 5 to day 15 postlesion, when a plateau was reached. Noticeable differences in the recovery level of receptor expression were observed according to the zonal patterning: the recovery level for neurons located in the lateral zone reached 70% of the control side value while the recovery levels in the dorsal and medial zones represented 35% and 53% of this value, respectively. Axotomy experiments suggest that zone-specific differences in receptor reexpression reported after bulbectomy might be related to the trophic influence of the olfactory bulb.
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    The effect of retinoic acid on the re-establishment of differentiated hepatocyte phenotype in primary culture (1998)
    Springer
    Cell & tissue research 293 (1998), S. 337-347 
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    ISSN: 1432-0878
    Keywords: Key words Hepatocytes ; Cell polarity ; Cytoskeleton ; Cytokeratin ; Bile canaliculus ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The usefulness of cultured hepatocytes is limited by the gradual loss of their typical physiological functions that occurs in vitro, mainly due to the absence of microenviromental conditions found in vivo. In this study we describe the effect of retinoic acid on the re-establishment of morphological characteristics and on the reorganization of the cytoskeletal network in cultured rat hepatocytes. Results obtained demonstrate that retinoic acid can influence hepatocyte differentiation, as regards the recovery of cell polarity, polyhedric shape and reformation of bile canaliculi and junctional complexes. The main target of this action appears to be the cytoarchitecture of cytoskeletal components, particularly cytokeratin filaments, which regain the configuration present in intact liver. The reorganization of the intermediate filaments does not seem to be dependent on the induction of higher levels of cytokeratin proteins, but rather appears to be due to post-translational regulation. The effect of retinoic acid on the cytoskeletal organization could determine the stabilization of intercellular contacts by means of junctions, leading to the appearance of morpho-functional characteristics typical of well-differentiated hepatocytes.
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    Thyroid hormone-induced chondrocyte terminal differentiation in rat femur organ culture (1998)
    Springer
    Cell & tissue research 293 (1998), S. 357-364 
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    ISSN: 1432-0878
    Keywords: Key words Thyroxine ; Cartilage ; Differentiation ; Type X collagen ; Organ culture ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Thyroid hormone plays a role in skeletal maturation. The present study was undertaken to investigate the effect of thyroid hormone on growth and maturation of the epiphyseal cartilage in the rat femur organ culture system. The femora increased in both weight and size over time in culture in the absence of serum. The growth rate was suppressed by thyroxine treatment. Thyroxine induced chondrocyte hypertrophy in the area adjacent to the articular surface in the femoral condyle. In this area, the expression of type X collagen, a marker of chondrocyte terminal differentiation, was detected after 21 days in culture by immunohistochemistry and reverse transcription/polymerase chain reaction. These data suggest that thyroxine suppresses cartilage growth by stimulating chondrocyte terminal differentiation.
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    Pulmonary intraepithelial vagal nodose afferent nerve terminals are confined to neuroepithelial bodies: an anterograde tracing and confocal microscopy study in adult rats (1998)
    Springer
    Cell & tissue research 293 (1998), S. 395-405 
    Details
    ISSN: 1432-0878
    Keywords: Key words Lung ; Neuroendocrine system ; Neuroepithelial bodies ; Innervation ; Neuronal tracing ; DiI ; Calcitonin gene-related peptide ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Our present understanding of the morphology of neuroepithelial bodies (NEBs) in mammalian lungs is comprehensive. Several hypotheses have been put forward regarding their function but none has been proven conclusively. Microscopic data on the innervation that appears to affect the reaction of NEBs to stimuli have given rise to conflicting interpretations. The aim of this study has been to check the validity of the hypothesis that pulmonary NEBs receive an extensive vagal sensory innervation. The fluorescent neuronal tracer DiI was injected into the vagal sensory nodose ganglion and NEBs were visualized in toto by using immunocytochemistry and confocal microscopy on 100-µm-thick frozen sections of the lungs of adult rats. The most striking finding was the extensive intraepithelial terminal arborizations of DiI-labelled vagal afferents in intrapulmonary airways, apparently always co-appearing with calcitonin gene-related peptide (CGRP)-immunoreactive NEBs. Not all NEBs received a traced nerve fibre. Intrapulmonary CGRP-containing nerve fibres, including those innervating NEBs, always appeared to belong to a nerve fibre population different from the DiI-traced fibres and hence did not arise from the nodose ganglion. Therefore, at least some of the pulmonary NEBs in adult rats are supplied with sensory nerve fibres that originate from the vagal nodose ganglion and form beaded ramifications between the NEB cells, thus providing support for the hypothesis of a receptor function for NEBs.
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    Uptake of lithium carmine by sinusoidal endothelial and Kupffer cells of the rat liver: new insights into the classical vital staining and the reticulo-endothelial system (1998)
    Springer
    Cell & tissue research 292 (1998), S. 395-410 
    Details
    ISSN: 1432-0878
    Keywords: Key words Sinusoidal endothelial cells ; Kupffer cells ; Vital staining ; Lithium carmine ; Endocytosis ; Reticulo-endothelial system ; Mononuclear phagocyte system ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Sinusoidal cells in the rat liver were studied in vivo and in vitro using the original vital staining with lithium carmine, which has contributed much to the development of the concept of the reticulo-endothelial system. Immunohistochemical and electron-microscopic studies revealed that the dye-incorporating cells were sinusoidal endothelial cells, Kupffer cells, and monocytes. The endothelial cells took up much more dye than did the Kupffer cells and bulged largely into the sinusoidal lumen. Electron microscopy revealed that small particles of lithium carmine were associated with coated vesicles of endothelial cells and ruffled membranes of Kupffer cells. In the endothelial cells, these particles were present in various concentrations within vacuolated structures and condensed in the lysosomes forming large aggregates of lithium carmine lumps. These lumps showed crystalline structures, within which the size of the individual particle was up to 30 nm in width and 50 nm in length. A few endothelial cells containing abundant dye underwent degeneration, and some were taken up by Kupffer cells. Liver endothelial cells isolated from lithium carmine-administered rats endocytosed fluorescence-labeled collagen. Isolated endothelial cells from normal rat liver, when cultured with lithium carmine, did not take up any dye, and their endocytosis of formaldehyde-treated albumin was inhibited dose-dependently. We conclude that in the liver, endothelial cells, but not Kupffer cells, predominantly take up lithium carmine. Furthermore, we propose the existence of a generalized cell system based on its vital staining capacity.
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    Nonvagal origin of galanin-containing nerve terminals innervating striated muscle fibers of the rat esophagus (1998)
    Springer
    Cell & tissue research 292 (1998), S. 453-461 
    Details
    ISSN: 1432-0878
    Keywords: Key words Nucleus ambiguus ; Enteric nervous system ; Motor endplate ; Vasoactive intestinal peptide (VIP) ; Calcitonin gene-related peptide (CGRP) ; Coinnervation ; Anterograde tracing ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  We investigated the origin of galanin-positive nerve fibers on motor endplates in rat esophagus using anterograde 1,1′-dioleyl-3,3,3′,3′-tetramethylindocarbocyanine methane sulfonate (DiI) tracing from the nucleus ambiguus combined with galanin immunocytochemistry and calcitonin gene-related peptide immunocytochemistry. To demonstrate spatial relationships of galanin-positive nerve fibers to vagal and enteric nerve fibers on motor endplates, we combined galanin immunocytochemistry with calcitonin gene-related peptide immunostaining for labeling of vagal terminals, and vasoactive intestinal peptide immunoreactivity and NADPH-diaphorase histochemistry for demonstration of enteric nerve fibers. Within fine varicose nerve fibers, galanin was colocalized with vasoactive intestinal peptide and NADPH-diaphorase to a high degree and turned out to be completely separated from calcitonin gene-related peptide-positive or anterogradely DiI-labeled vagal motor terminals. These results indicate that the enteric nervous system is the most important and possibly the only source of galanin-positive nerve terminals on motor endplates in rat esophagus. Galanin may be, in addition to nitric oxide and vasoactive intestinal peptide, a mediator of the enteric coinnervation of striated muscle in this organ.
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    URL: http://dx.doi.org/10.1007/s004410051074
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    Maturation of rat dendritic cells during intrahepatic translocation evaluated using monoclonal antibodies and electron microscopy (1998)
    Springer
    Cell & tissue research 294 (1998), S. 503-514 
    Details
    ISSN: 1432-0878
    Keywords: Key words Dendritic cells ; Maturation ; Intrahepatic translocation ; Immunohistochemistry ; Electron microscopy ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Specific populations of hepatic sinusoidal cells were stained with monoclonal antibodies that recognize monocytes/macrophages (ED1), tissue macrophages (Kupffer cells) (ED2), MHC class II (Ia) antigen (MRC OX6), and dendritic cells/γ,δ T-cells (MRC OX62) and analyzed by light and electron microscopy. The majority of ED1+ and/or ED2+ cells were localized to the hepatic parenchyma, whereas OX6+ and/or OX62+ cells were more densely distributed within Glisson’s sheath than in the hepatic parenchyma. Double-immunoperoxidase staining of normal liver for ED1, ED2, and OX6 identified dendritic cells (DC) of two different phenotypes, ED1+ED2–OX6+ and ED1–ED2–OX6+. DC can be classified into three different types based on ultrastructural characteristics. The first type (type I) is characterized by one or more long cytoplasmic processes and a well-developed lysosomal system. The second type (type II) has an inconspicuous lysosomal system, abundant hyaloplasm, and characteristic short cytoplasmic processes. The third type (type I–II) has cytologic features intermediate between those of type I and type II DC. At the electron-microscopic level, these three cell types are found in the sinusoidal lumen, whereas the majority of type II DC are located in the space of Disse and Glisson’s sheath. Furthermore, some OX6-labeled elongated DC appeared to traverse the lumen of sinusoids through endothelial pores to enter the space of Disse. One hour after intravenous injection of latex particles (0.81 μm in diameter), numerous latex-laden dendritic cells (ED1+OX6+, type I and type I–II) were detected in the lumen of hepatic sinusoids, but not in the space of Disse or Glisson’s sheath. These findings suggest that normal rat liver contains resident dendritic cells which downregulate phagocytic activity and mature into potent accessory cells during migration from the portal vein toward the central vein. These DC then traverse the sinusoidal lumen to the hepatic lymph system via the space of Disse.
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    URL: http://dx.doi.org/10.1007/s004410051201
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    Plant-soil Interactions in Deserts (1998)
    Springer
    Biogeochemistry 42 (1998), S. 169-187 
    Details
    ISSN: 1573-515X
    Keywords: aridisols ; desertification ; erosion ; geostatistics ; Larrea tridentata ; nitrogen ; phosphorus ; Prosopis glandulosa ; soil heterogeneity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract Geostatistical analyses show that the distribution of soil N, P and K is strongly associated with the presence of shrubs in desert habitats. Shrubs concentrate the biogeochemical cycle of these elements in ‘islands of fertility’ that are localized beneath their canopies, while adjacent barren, intershrub spaces are comparatively devoid of biotic activity. Both physical and biological processes are involved in the formation of shrub islands. Losses of semiarid grassland in favor of invading shrubs initiate these changes in the distribution of soil nutrients, which may promote the further invasion and persistence of shrubs and cause potential feedbacks between desertification and the Earth's climate system.
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    URL: http://dx.doi.org/10.1023/A:1005939924434
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    Peroxy Radical Kinetics Resulting from the OH-Initiated Oxidation of 1,3-Butadiene, 2,3-Dimethyl-1,3-Butadiene and Isoprene (1998)
    Springer
    Journal of atmospheric chemistry 29 (1998), S. 267-298 
    Details
    ISSN: 1573-0662
    Keywords: peroxy radicals ; kinetics ; conjugateddienes ; biogenic VOC ; degradation mechanisms ; tropospheric ozone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract The laser flash photolysis/UV absorption spectrometry technique has been used to investigate the kinetics of the peroxy radical permutation reactions (i.e. self and cross reactions) arising from the OH-initiated oxidation of isoprene (2-methyl-1,3-butadiene), and of the simpler, but related conjugated dienes, 1,3-butadiene and 2,3-dimethyl-1,3-butadiene. The results of the two simpler systems are analysed to provide values of the rate coefficients for the 6 peroxy radical permutation reactions of the three types of isomeric peroxy radical produced in each system (T = 298 K, P = 760 Torr). The rate coefficients are all significantly larger than values estimated previously by extrapolation of structure-reactivity relationships based on the kinetics of a limited dataset of simpler radicals containing similar structural features. The results are discussed in terms of trends in self and cross reaction reactivity of primary, secondary and tertiary peroxy radicals containing combinations of allyl, β-hydroxy and δ-hydroxy functionalities. Since the peroxy radicals formed in these systems are structurally very similar to those formed in the isoprene system, the kinetic parameters derived from the results of the simpler systems are used to assist the assignment of kinetic parameters to the 21 permutation reactions of the six types of isomeric peroxy radical generated in the isoprene system. Kinetic models describing the OH-initiated degradation of all three conjugated dienes to first generation products in the absence of NOx are recommended, which are also consistent with available end product studies. The model for isoprene is considered to be a further improvement on that suggested previously for its OH-initiated oxidation in the absence of NOx. The mechanism is further extended to include chemistry applicable to ‘NOx-present’ conditions, and calculated product yields are compared with those reported in the literature.
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    URL: http://dx.doi.org/10.1023/A:1005940332441
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    Kinetics of the Gas-Phase Reactions of OH and NO3 Radicals and O3 with the Monoterpene Reaction Products Pinonaldehyde, Caronaldehyde, and Sabinaketone (1998)
    Springer
    Journal of atmospheric chemistry 31 (1998), S. 281-297 
    Details
    ISSN: 1573-0662
    Keywords: hydroxyl radical ; nitrate radical ; ozone ; pinonaldehyde ; caronaldehyde ; sabinaketone ; kinetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Geosciences
    Notes: Abstract Using a relative rate method, rate constants have been measured for the gas-phase reactions of OH and NO3 radicals with pinonaldehyde, caronaldehyde and sabinaketone at 296 ± 2 K. The OH radical reaction rate constants obtained are (in units of 10−12 cm3 molecule−1 s−1): pinonaldehyde, 48 ± 8; caronaldehyde, 48 ± 8; and sabinaketone, 5.1 ± 1.4, and the NO3 radical reaction rate constants are (in units of 10−14 cm3 molecule−1 s−1): pinonaldehyde, 2.0 ± 0.9; caronaldehyde, 2.5 ± 1.1; and sabinaketone, 0.036 ± 0.023, where the error limits include the estimated overall uncertainties in the rate constants for the reference compounds. Upper limits to the O3 reaction rate constants were also obtained, of 〈2 × 10−20 cm3 molecule−1 s−1 for pinonaldehyde and caronaldehyde, and 〈5 × 10−20 cm3 molecule−1 s−1 for sabinaketone. These reaction rate constants are combined with estimated ambient tropospheric concentrations of OH radicals, NO3 radicals and O3 to calculate tropospheric lifetimes and dominant transformation process(es) of these and other monoterpene reaction products.
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    On the inhibition of camel retina acetylcholinesterase activity by cycloheximide in vitro (1998)
    Springer
    Cell biology and toxicology 14 (1998), S. 167-174 
    Details
    ISSN: 1573-6822
    Keywords: acetylcholinesterase ; cycloheximide ; inhibition ; kinetics ; retina
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The kinetic parameters of inhibition of camel retinal acetylcholinesterase (AChE) activity by cycloheximide (CH) were investigated. For the control system, the Michaelis–Menten constant (K m)for the hydrolysis of acetylthiocholine iodide was found to be 0.076 mmol/L and the V max was 0.547 μmol/min per mg protein. In contrast, these parameters were decreased in the CH-treated systems. Dixon and Lineweaver–Burk plots, and their secondary replots, indicated that the inhibition was of the linear mixed type, which seems to be a combination of partial competitive and pure noncompetitive inhibition. The values of K′i(slope) and K I(intercept) were estimated to be 3.50 and 5.68 mmol/L, respectively. K i was greater than K′i, indicating that CH has a greater binding affinity for the peripheral site than the active site.
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    Distribution and sources of phosphorus in tidal river sediments in the Washington, DC, Area (1997)
    Springer
    Environmental geology 30 (1997), S. 224-230 
    Details
    ISSN: 1432-0495
    Keywords: Key words Sediment ; Washington ; DC ; Pollution ; phosphorus ; nutrients
    Source: Springer Online Journal Archives 1860-2000
    Topics: Geosciences
    Notes: Abstract  Sediments in the rivers and basins around Washington, DC, have high concentrations of phosphorus, which, based on geographic distributions, is largely derived from urban runoff and municipal sewage. Dissolved-particulate phosphate exchange reactions and biological uptake of dissolved phosphorus from the water column may be an added source of phosphorus to the sediments. Concentrations of total sedimentary phosphorus ranged from 24 to 56 μm P/g-dw, and were highest in areas near combined sewer outfalls. As a part of this study, sedimentary phosphorus was fractionated into Fe-P, Ca-P, Al-P, and organic phases using a selective-sequential leaching procedure. The distribution of the phases in all sediments analyzed follow the order , Fe-P〉Ca-P〉Al-P. Spatial variations in the amounts of phosphorus in the different phases is related to the sources of phosphorus to the area. The proportions of occluded Al-P and organic P are 10–20% of the total P, respectively. This suggests that phosphorus from natural sources is small compared to anthropogenic inputs in this area. The high leachable Fe-P and Ca-P in these sediments might contribute a substantial amount of P to the water column under conditions of remobilization.
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    Bioavailability of a new effervescent tablet of ibuprofen in healthy volunteers (1997)
    Springer
    European journal of clinical pharmacology 52 (1997), S. 505-506 
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    ISSN: 1432-1041
    Keywords: Key words Ibuprofen; effervescent tablets ; kinetics ; bioavailability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Medicine
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    URL: http://dx.doi.org/10.1007/s002280050327
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    A 54-kDa protein related to ras-guanine nucleotide release factor expressed in the rat exocrine pancreas (1997)
    Springer
    Cell & tissue research 289 (1997), S. 505-515 
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    ISSN: 1432-0878
    Keywords: Key words: Acinus ; Guanine nucleotide release factor ; Pancreas ; exocrine ; ras ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The polymerase chain reaction was used to amplify a cDNA encoding the catalytic region of ras-guanine nucleotide release factor (ras-GRF1) from mouse embryonic stem cell mRNA. Antibodies directed against this protein were prepared and affinity purified. Western immunoblotting of rat tissue lysates revealed a 140-kDa protein in brain as expected but, in addition, a strongly immunoreactive 54-kDa protein, p54, was identified in pancreas. Expression of ras-GRF1 in pancreas was confirmed at the RNA level by reverse-transcriptase-coupled polymerase chain reaction analysis; p54 may therefore correspond to a form of ras-GRF1 or a closely related protein. The cellular and subcellular localization of p54 was investigated by enzyme-linked immunocytochemistry and immunogold electron microscopy. In the pancreas, p54 was expressed primarily in acinar cells, where it was localized along the basolateral and apicolateral plasma membranes. Indirect immunofluorescence microscopy of cultured acini further indicated that the plasma membrane localization of p54 was dependent on the maintenance of the acinar histotype and organized acinar structure. When primary acinar cells were permitted to dissipate into monolayer cultures devoid of zymogen granules, ras-GRF1 staining became cytosolic. Our results suggest that ras-GRF1 is involved in the structure and function of the pancreas.
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    Immunoreactivity for the Fas ligand in the mammalian enteric nervous system (1997)
    Springer
    Cell & tissue research 290 (1997), S. 21-29 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Intestines ; Neuropeptide Y ; Nitric oxide synthase ; Vasoactive intestinal polypeptide ; Guinea-pig ; Rat (Wistar) ; Mouse (C3H/HeJ ; gld) ; Ferret ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The Fas ligand induces apoptosis in activated immunocytes that express the Fas receptor. Fas-ligand transcripts have been found previously in murine intestine but the intestinal tissues that express Fas ligand have not been identified. We used immunohistochemistry to examine the expression of the Fas ligand in the enteric nervous system of rats, mice, guinea-pigs, ferrets and humans. Fas-ligand immunoreactivity was detectable in enteric nerve fibres and neurons in all species tested, representing 25%–50% of the neurons in rats, mice and guinea-pigs. An antigen of approximately 48 kDa was detected by Western blot analysis with Fas-ligand antiserum in the dissected enteric plexuses of duodenum from a C3H/HeJ mouse. In gld mice that harbour a Fas-ligand mutation, Fas-ligand immunoreactivity was slightly more intense in neurons and fibres and was also apparent in submucosal lymphocytes. In the myenteric plexuses of guinea-pig ileum and human colon, Fas-ligand immunoreactivity was not contained in neurons exhibiting nicotinamide-adenine dinucleotide phosphate-diaphorase activity. In the submucosal plexus of guinea-pig ileum, labelled neurons included some neuropeptide-Y-containing neurons but none with vasoactive intestinal polypeptide. We conclude that the Fas ligand is expressed by a large subset of enteric neurons and may provide the basis for cytotoxic neuroimmune interactions in the intestines.
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    Remodeling of pineal epithelium in the fetal rat as delineated by immunohistochemistry of laminin and cadherin (1997)
    Springer
    Cell & tissue research 287 (1997), S. 263-274 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Pineal gland ; Laminin ; Cadherin ; Synaptophysin ; BrdU ; Immunohistochemistry ; Embryology ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Epithelial remodeling in the rat pineal during fetal development was immunohistochemically analyzed by using antibodies for laminin and cadherin as molecular markers of basal lamina and intercellular junctions, respectively. The proliferation and differentiation of pinealocytes were also investigated in relation to the advance of epithelial remodeling. The pineal anlage of embryonic day 16 is completely covered by basal lamina immunolabeled for laminin. After embryonic day 17, local dissolution of the basal lamina occurs on the epithelial folds, which develop predominantly in the rostral pineal wall. Some pineal cells migrate through these interruptions and form cellular aggregations outside the basal lamina. Cadherin immunostaining reveals focal dissolution of intercellular junctions in epithelial regions protruding into the pineal lumen. Dissolution of the basal lamina and intercellular junctions accompanied by cellular migration into the stromal tissue or into the pineal lumen continues until birth. The distribution of mitotic cells immunolabeled for BrdU is homogeneous throughout the organ during the fetal period, whereas that of differentiating pinealocytes immunoreactive for synaptophysin shows striking regional heterogeneity in close correlation with the remodeling of the pineal epithelium. The migrating cell populations located either outside the basal lamina or inside the pineal lumen are more liable to become synaptophysin-positive than the rest of the epithelium. These results suggest that epithelial remodeling in the fetal pineal is induced, at least in part, by epithelial infolding and that this remodeling promotes the differentiation of pinealocytes.
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    Immunocytochemical analysis of the transport of arginine analogues into nitrergic neurons and other cells in the retina and pituitary (1997)
    Springer
    Cell & tissue research 290 (1997), S. 501-514 
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    ISSN: 1432-0878
    Keywords: Key words: Arginine ; Analogues ; Immunocytochemistry ; Nitric oxide ; Pituitary ; Retina ; Transport ; Rat (Wistar) ; Rabbit
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Nitric oxide is formed by the action of nitric oxide synthase upon l-arginine. The efficacy of some exogenously applied arginine analogues in inhibiting nitric oxide synthase and thus nitrergic transmission indicates that neurons producing nitric oxide may possess an arginine transport system. To investigate whether arginine analogues are preferentially transported into nitric oxide-utilising cells or into cells making other neurochemicals, we have raised highly specific antisera against a number of arginine analogues including NG-methyl arginine, d-arginine, NGnitro-l-arginine, NG-nitro-l-arginine methyl ester and canavanine. Retinae were incubated in physiological media containing these analogues and rats were given intraperitoneal injections of the analogues to study the pituitary. Immunocytochemistry and NADPH-diaphorase histochemistry revealed that many of these analogues could be transported preferentially, but not exclusively, into nitric oxide-generating cells. However, some nitric oxide-producing cells apparently lacked the ability to take up some arginine analogues. We conclude that nitric oxide-generating cells in the retina and pituitary possess one or more arginine transporters. Other subsets of neurons that use GABA or glutamate as a neurotransmitter may also accumulate arginine analogues, possibly as a substrate for formation of these neurochemicals.
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    Differential distribution of the mitochondrial heat-shock protein 60 in rat gastrointestinal tract (1997)
    Springer
    Cell & tissue research 287 (1997), S. 343-350 
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    ISSN: 1432-0878
    Keywords: Key words: Heat-shock protein 60 (hsp60) ; Chaperonin ; Mitochondria ; Gastrointestinal tract ; Epithelium ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Immunohistochemical studies on various parts of the rat gastrointestinal tract by means of an antibody against the mitochondrial chaperonin, heat-shock protein 60 (hsp60), has revealed a cell-specific distribution pattern. The active form of hsp60 is a heptameric complex that is involved in the import and refolding of nuclear-encoded proteins destined for the mitochondrial matrix. This chaperonin is detectable in highly replicating cells, e.g., keratinizing cells of the esophagus and short-living epithelial cells of the intestine. In the stomach, some of the oxyntic cells contain hsp60-positive mitochondria lying near intracellular canaliculi. Neuronal cells of the enteric nervous system present intense positive staining for hsp60 in some areas. All other non-epithelial cells of the digestive tract show weak or no hsp60 immunoreactivity. The presence of hsp60 in mitochondria seems to reflect two different forms of mitochondrial renewal: (1) total reformation of mitochondria and their content after mitotic division and (2) regeneration of these organelles following high activity, e.g., ATP synthesis.
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    Ramification of microglia, monocytes and macrophages in vitro: influences of various epithelial and mesenchymal cells and their conditioned media* (1997)
    Springer
    Cell & tissue research 287 (1997), S. 447-458 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Microglia ; Monocytes ; Macrophages ; Astroglia ; Epithelial cells ; Fibroblasts ; Ramification ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Microglial cells are able to switch between an ”active” amoeboid and a ramified ”resting” morphology during development and after experiencing lesions. We have previously shown that in vitro microglial morphology is controlled by their cellular environment, i.e. cells become ramified in astrocyte coculture but amoeboid on monolayers of fibroblasts. In the present study we have extended the analysis of the control of macrophage morphology by maintaining macrophages of different origins in coculture with different epithelial or mesenchymal cells and their conditioned media. Microglia, monocytes and spleen macrophages seeded onto monolayers of astrocytes, kidney epithelia or hepatoma cells developed the ramified morphology but remained amoeboid in fibroblast coculture. Ramification was also induced by media conditioned by these cells as well as by phorbolic esters, i.e. activators of protein kinase C. In double coculture assays, even small numbers of fibroblasts were able to override the ”epithelial” influence. Likewise, microglia remained amoeboid, when incubated on several constituents of the extracellular matrix. These results indicate that macrophage ramification is an active process initiated by diffusible factors secreted by various epithelial cells, possibly acting upon a protein-kinase-C-related receptor. We interprete the modification of macrophage morphology as a functional adaptation to the surrounding type of tissue that is enforced by its constituent cells. Thus, the specific morphologies of microglia, hepatic von Kupffer’s cells or peritubular kidney macrophages could be explained by similar epithelium–macrophage interaction.
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    Fibroblast growth factor-2 (FGF-2) and FGF-receptor (FGFR-1) immunoreactivity in embryonic spinal autonomic neurons (1997)
    Springer
    Cell & tissue research 287 (1997), S. 471-480 
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    ISSN: 1432-0878
    Keywords: Key words: FGF-2 ; FGF receptor-1 ; Autonomic nervous system ; Sympathetic preganglionic neurons ; Spinal cord ; Development ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The development of the nervous system appears to be under the control of multiple growth factors, neurotrophins and cytokines, which may be expressed either continuously or transiently throughout defined stages of cellular generation, proliferation or differentiation. Fibroblast growth factor (FGF) cytokines and their receptors are abundantly expressed in the embryonic nervous system but their localization at autonomic levels in the fetal spinal cord has not yet been detailed. Immunoreactivity to FGF-2, probably the best characterized member of the FGF family (FGF-1 to FGF-10) and of one of its high affinity receptors, FGFR-1, was found in autonomic neurons at embryonic day E14, the peak day of generation and proliferation in the common ventral motoneuron pool. It was also continuously present throughout the investigated subsequent stages (E15 to postnatal day P30). Immunogold electron microscopy revealed the cytoplasmic localization of FGF-2 and FGFR-1 in intermediolateral neurons, the major group of sympathetic preganglionic neurons in the spinal cord. In these neurons, immunocytochemistry from E14 onwards showed the co-distribution of both markers at the period of axonal outgrowth to peripheral targets, e.g. the adrenal medulla. Our findings suggest autocrine and/or paracrine actions of FGF-2 for sympathetic preganglionic development but do not support its role as a target-derived neurotrophic factor for autonomic neuron development.
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    2-Acetylaminofluorene-produced selective cytotoxic damage of a ductal compartment and its repair in the submandibular gland of rats (1997)
    Springer
    Cell & tissue research 288 (1997), S. 371-379 
    Details
    ISSN: 1432-0878
    Keywords: Key words: 2-Acetylaminofluorene ; Selective ductal cytotoxicity ; Submandibular gland ; Regenerative repair ; Immunohistochemistry ; Systemic effects ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Salivary glands of rodents are rarely affected with spontaneous and induced malignancies. This may be linked with low physiologic cell renewal and the infrequency of cytolethal actions by xenobiotics. The genotoxic 2-acetylaminofluorene, carcinogenic for other organs, causes acute damage in salivary ducts. In the submandibular glands the damage is limited to the granulated convoluted tubules. They produce and release regulatory peptides including epidermal growth factor and nerve growth factor. The partial chemical sialoadenectomy is repaired by sequential cell proliferation in the basal cell layer of interlobular ducts and in dilated intralobular ducts (day 4 and 6), in intermediate duct-like structures (day 6 and 8), and lastly in acini (day 8 and 12). This is associated with a transient loss of structural characteristics of striated ducts and acini (up to day 6) and of the immunoreactivity for S-100 protein (up to day 4). Actin immunoreactivity at the acinar base is increased from day 6 to 20. Analogous to the late postnatal differentiation of the granulated convoluted tubules, their structural characteristics and immunoreactivity for epidermal growth factor do not recover within 20 days. The acute lesion of the endocrine ductal segment is suggested to be causally involved with other systemic effects following treatment with 2-acetylaminofluorene. First, hypophagia with loss of body, liver and thymus weight may result from disturbed saliva production. Second, previous studies have shown a mitotic burst of the biliary epithelium and bloodborne lymphocyte-stimulating activities. Either effect could be brought about by regulatory peptides (see above), probably after elevated circulatory release from necrotic granulated convoluted tubules.
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    Expression of basic fibroblast growth factor-like immunoreactivity in the nuclei of regenerating hepatocytes (1997)
    Springer
    Cell & tissue research 288 (1997), S. 517-527 
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    ISSN: 1432-0878
    Keywords: Key words: Liver ; Subcellular fractions ; Immunohistochemistry ; Electron microscopy ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. This study, utilizing rats subjected to two-thirds partial hepatectomy or sham operation, was designed (1) to investigate the content of basic fibroblast growth factor (bFGF) in the subcellular fractions of regenerating and sham-operated rat livers by immunoblot experiments and enzyme-linked immunosorbent assay (ELISA), (2) to show that bFGF immunoreactivity and proliferating cell nuclear antigen (PCNA) immunoreactivity are markers for hepatocellular mitosis before and after partial hepatectomy, and (3) to observe the location and fine structure of the bFGF immunoreaction within the regenerating liver with special attention to bFGF immunoreactivity in the nuclei of regenerating hepatocytes. Immunoblot experiments and ELISA showed a transient increase in high-molecular-weight forms of bFGF in the nuclear subcellular fraction of regenerating liver 48 h after partial hepatectomy. By light microscopy, bFGF and PCNA immunoreactivities were detected in the nuclei of regenerating hepatocytes. Electron microscopy demonstrated bFGF-like immunoreactivity mainly in the nuclear euchromatin and rarely in the heterochromatin or nucleoli of regenerating hepatocytes. The transient increase in high-molecular-weight forms of bFGF in the nuclear euchromatin of regenerating hepatocytes, together with the concomitant expression of PCNA in the regenerating liver, suggests an important role of the high-molecular-weight forms of bFGF in hepatocyte proliferation and/or mitosis, although authentic bFGF with a molecular form of 18 kDa is not considered to be involved in hepatic regeneration.
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    Simultaneous restitution of matrix and cells in gastric ulcer: use of a combined in-situ hybridization and immunohistochemistry technique applicable to paraffin-embedded tissue (1997)
    Springer
    Cell & tissue research 287 (1997), S. 601-610 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Collagen ; Combined non-radioactive in-situ hybridization and immunohistochemistry ; Gastric ulcer ; Immunohistochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The restoration of gastric tissue after ulceration involves cellular and matrix components. Our aim was to investigate the kinetics of collagen expression and cellular proliferation in an animal model of gastric ulcer. To demonstrate the expression of type I and IV collagen mRNAs by proliferating cells, a method combining in-situ hybridization and immunohistochemistry was devised. In order to avoid the disadvantages of radioisotopes, digoxigenin-labeled RNA-riboprobes were utilized and combined with single-step immunohistochemistry. This method proved sensitive enough to detect type I and IV procollagen mRNA transcripts in the submucosal area beneath the ulcer crater or adjacent to the ulcer rim. In addition, a subset of cells transcribing either procollagen type I or IV RNA was concomitantly positive for proliferating cell nuclear antigen by immunohistochemistry. Focal proliferation of cells simultaneously expressing extracellular matrix components may therefore occur in the gastric submucosa after ulceration, starting as soon as 3 days after the insult and continuing for several weeks. The devised method of combined in-situ hybridization and immunohistochemistry can be used with standard paraffin-embedded tissues, yields results within 2 days, and avoids radioisotopes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004410050783
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  • 96
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    Electronic Resource
    Filamentous contacts: the ultrastructure and three-dimensional organization of specialized non-synaptic interneuronal appositions in thalamic relay nuclei (1997)
    Springer
    Cell & tissue research 288 (1997), S. 43-57 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Thalamus ; Intercellular junctions ; Synapse ; Synaptic glomeruli ; Agranular endoplasmic reticulum ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Filamentous contacts are non-synaptic interneuronal junctions characteristic of thalamic relay nuclei. Symmetrical filamentous contacts occur between two dendrites, two somata or a dendrite and a soma; asymmetrical filamentous contacts occur between axon terminals and dendrites, or occasionally somata, chiefly between the large specific afferent axon terminals of the synaptic glomeruli and the shafts of relay cell dendrites. Both are arranged as extensive net-like (reticular) specializations. The strands of the network enclose fenestrae of variable shape and size and, in perpendicular thin sections, appear as stretches of slightly widened intercellular space containing an electron-dense material and bounded by plasma membranes, the cytoplasmic surfaces of which are coated by electron-dense material into which microfilaments appear to insert. The lamina of cytoplasmic material in dendrites and somata is thicker than that in axon terminals and contains distinct electron-dense sub-units. Regular synaptic junctions may be situated like islands within the territory of an asymmetrical filamentous contact, and small spot-like close membrane appositions resembling gap junctions are occasionally seen in the fenestrae adjacent to the strands of both varieties of contact. Bundles of neurofilaments running in different directions, but in a plane parallel to the plasma membrane, are prominent on either side of the symmetrical filamentous contact and on the dendritic side of the asymmetrical variety. The agranular reticulum also exhibits differences between the contact types. Because of their highly specialized ultrastructure and specific distribution, filamentous contacts probably do not serve a purely adhesive function. Their possible role in the establishment and maintenance of orderly connections between cells is discussed but not favoured. Filamentous contacts probably mediate some form of intercellular communication, possibly involving gap junctions.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004410050791
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  • 97
    Electronic Resource
    Electronic Resource
    Neurochemical differentiation of rat enteric neurons during pre- and postnatal life (1997)
    Springer
    Cell & tissue research 288 (1997), S. 11-23 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Nitric oxide synthase (NOS) ; Vasoactive intestinal peptide (VIP) ; Pituitary-adenylyl-cyclase-activating peptide (PACAP) ; α-Internexin ; Peripherin ; Immunohistochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The presence of nitric oxide synthase (the enzyme which synthesizes nitric oxide), vasoactive intestinal peptide (VIP) and pituitary adenylyl-cyclase-activating peptide (PACAP) in the rat enteric nervous system between term (E18) and 90 days postpartum (P90) was investigated by immunohistochemistry. Neuronal maturity was ascertained by emanining for the presence of two novel neuron intermediate filament proteins: α-internexin, which is transiently expressed in developing neurons, and peripherin, which is expressed in differentiating as well as in mature neurons. The alimentary canal of the foetus at E18, unfed newborn, suckling, weaning and adult rats was studied. Throughout the rat gut the myenteric neurons were recognized by the time the foetal stage was reached and the submucous neurons by the suckling period. α-Internexin was present at E18; its levels increased during ageing and were markedly reduced at P30. Peripherin was first detected at birth; its levels increased with ageing and pratically almost all of the neurons and nerve fibres were labelled from P30. Nitric oxide synthase was present at E18 and the number of labelled neurons gradually increased with ageing, while both VIP and PACAP could be detected by the end of the suckling period in an equal number of neurons to that in adulthood. These data indicate that in the rat gut the onset of the mature neuronal phenotype is not yet achieved at birth and that the neurochemical differentiation is accomplished during the first month of postnatal life.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004410050788
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  • 98
    Electronic Resource
    Electronic Resource
    Developmental patterns of proteoglycan-containing extracellular matrix in perineuronal nets and neuropil of the postnatal rat brain (1997)
    Springer
    Cell & tissue research 288 (1997), S. 33-41 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Ontogenesis ; Perineuronal nets ; Extracellular matrix ; Proteoglycans ; Brain ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The extracellular matrix is involved in various morphogenetic processes which are accompanied by changes in its physicochemical properties and spatial organization. In the adult brain it contributes to cellular communication and the regulation of neuronal activity. The present study deals with the postnatal appearance and transformation into adult distribution patterns of extracellular matrix components related to chondroitin-sulphate proteoglycans (CSPGs) in the rat brain. The differential accumulation of these components in neuropil and in perineuronal nets (PNs) enriched in certain regions was examined in 0-, 7-, 14-, 21- and 35-day-old rats and adult animals using the N-acetylgalactosamine-binding Wisteria floribunda agglutinin (WFA) and immunocytochemical detection of CSPGs. The lectin stained the olfactory-bulb glomerular layer and layer Ia of piriform and entorhinal cortex already in newborn animals. On postnatal day 7 diffuse neuropil staining was additionally found in certain subcortical nuclei and in deep neocortical layers. The first sharply contoured PNs were detected at this age in the brain stem, indicating the more advanced maturation of matrix components in subcortical regions. CSPG immunoreactivity yielded staining patterns largely identical to WFA-binding patterns but appeared only between postnatal day 14 and 21. The adult-like stage was revealed with both methods between 21 and 35 days after birth. The results provide further evidence that the accumulation of certain CSPGs in the extracellular space is spatiotemporally related to distinct patterns of neuronal activity at the regional and cellular level.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004410050790
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  • 99
    Electronic Resource
    Electronic Resource
    Correlation of function and structure in developing rat distal colon (1997)
    Springer
    Cell & tissue research 288 (1997), S. 95-99 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Distal colon ; Aldosterone adaptation ; Sodium absorption ; Intestinal development ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The distal colon of suckling (14-day-old) and weanling (27-day-old) rats absorbs Na+ via channel-mediated, electrogenic amiloride-sensitive Na+ transport which disappears after weaning. This transport pathway is induced by aldosterone in superficial cells of colonic epithelium. The purpose of the present study was to correlate the changes in distal colon function with changes in the apical and basolateral cell membrane surface areas of superficial enterocytes. The basolateral but not apical membrane surface density (i.e. surface areas of the basolateral and apical membranes of superficial enterocytes per unit volume of superficial enterocytes) was found to increase between postnatal day 14 and 27 (i.e. during the weaning period). The plasma concentrations of aldosterone were very high during this period and electrogenic amiloride-sensitive Na+ transport and Na, K-ATPase activity were increased significantly. High dietary salt intake during the weaning period prevented the developmental increase of basolateral membrane surface density, inhibited electrogenic amiloride-sensitive Na+ transport and significantly depressed plasma aldosterone levels and Na, K-ATPase activity. Apical cell membrane surface density did not change significantly after a sodium load. Thus, high plasma concentrations of aldosterone and/or high colonic Na+ transport during weaning exert an important and selective effect on the basolateral membrane and are responsible for the developmental changes of this cell membrane surface during weaning. Concomitant changes in the morphology of superficial colonic enterocytes and in colonic Na+ transport suggest that the structure of the immature epithelium is related to its function.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004410050796
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  • 100
    Electronic Resource
    Electronic Resource
    Transient expression of GAP-43 within the hippocampus after global brain ischemia in rat (1997)
    Springer
    Cell & tissue research 288 (1997), S. 225-238 
    Details
    ISSN: 1432-0878
    Keywords: Key words: Cerebral ischemia ; Plasticity ; GAP-43 ; Hippocampus ; mRNA ; Immunohistochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Neuroanatomical methods have been used to study selective vulnerability after global brain ischemia. A consistent pattern of ischemic neuronal damage is found in the rodent hippocampus with loss of CA1 neurons and of some cells in the hilus of the dentate gyrus. Very little is known about plastic changes that would be expected in ischemia-resistant areas such as CA3 neurons and granule cells. Neuronal plasticity after lesions may be indicated by changes in labeling with antibodies to the growth-associated protein 43 (GAP-43). Expression of GAP-43 as a marker for neuronal plasticity was studied here in the hippocampus after global brain ischemia. Halothane-anesthetized rats were subjected to 20 min of transient forebrain ischemia using four-vessel occlusion. In situ hybridization was used to study GAP-43 mRNA at 1, 3, 6, and 12 h and at 1, 3, and 7 days after ischemia. Immunostaining was carried out with two different antibodies to GAP-43 in brains which were perfusion-fixed after 1, 2, 4, and 7/8 days. In the control hippocampus, GAP-43 mRNA was localized to CA1–CA3 and the hilus. Moderate increases in cellular signals were seen in hilar cells and granule cells early after ischemia, and some changes occurred in CA3 at late stages. Hybridization was lost in CA1 due to cell death. With immunostaining, GAP-43 was not seen in the cytoplasm of neurons, whereas dense labeling occurred in a differentiated pattern in the axonal and dendritic layers. At 1 day after ischemia, neurons in the hilus of the dentate gyrus and in the stratum pyramidale and lucidum of CA3 showed strong cytoplasmic labeling for GAP-43. Few cells were labeled in these regions at 2 days, and none at later stages. Pyramidal cells in CA1 and CA3 areas and granule cells were never labeled. These studies demonstrate a transient expression of GAP-43 mRNA and protein in a subset of vulnerable neurons after transient brain ischemia. The cytoplasmic localization in hilar neurons could be due to increased synthesis of GAP-43 or to changes in axoplasmic transport. It is suggested that axonal damage occurs in hilar cells which stimulates GAP-43 expression. The increased production of trophic factors after ischemia in granule cells could also cause plastic changes in hilar cells. Since hilar neurons are in a strategic position to control the excitability of the dentate area, increased expression of GAP-43 may indicate an important pathophysiological process. In seizure experiments, strong expression of GAP-43 mRNA in granule cells was associated with abnormal mossy fiber sprouting and development of chronic epilepsy. The relevance of the minor GAP-43 mRNA upregulation after ischemia must be considered. The changes in CA3 neurons at several days after ischemia might represent a plastic response to a loss of CA1 neurons.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004410050808
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