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  • phosphorus  (250)
  • Immunocytochemistry
  • Springer  (416)
  • National Academy of Sciences
  • 1995-1999  (416)
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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Journal of applied electrochemistry 29 (1999), S. 1171-1176 
    ISSN: 1572-8838
    Keywords: alloys ; cyclic voltammetry ; electrodeposition ; electroless deposition ; nickel ; phosphorus ; zinc
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Electrical Engineering, Measurement and Control Technology
    Notes: Abstract Electroless Ni–Zn–P alloy deposition from a sulphate bath, containing sodium hypophosphite as reducer, was investigated. To increase the plating rate, the deposition parameters were optimized. The effect of process parameters (T, pH and [Zn2+]) on the plating rate and deposit composition was examined and it was found that the presence of zinc in the bath has an inhibitory effect on the alloy deposition. As a consequence, the percentage of zinc in the electroless Ni–Zn–P alloys never reaches high values. Using cyclic voltammetry the electrodeposition mechanism of Ni–Zn–P alloys was investigated. It was observed that the zinc deposition inhibits the nickel discharge and, as a consequence, its catalytic activity on hypophosphite oxidation. It was also found that increase in temperature or pH leads to the deposition of nickel rich alloys.
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  • 2
    ISSN: 1572-8854
    Keywords: Octahedral ; phosphorus ; chloride
    Source: Springer Online Journal Archives 1860-2000
    Topics: Geosciences , Physics
    Notes: Abstract The title compound [P(tpp)Cl2]+Cl− crystallizes in the space group P21/n witha=10.701(2),b=24.860(2),c=14.799(2), β=94.24(2)°,Z=4. The phosphorus atom has an octahedral coordination geometry formed by the four nitrogen atoms (Np) of the porphyrinato group and the two chloride ions. The average phosphorus-chloride distance is 2.150(1) Å, with phosphorus situated 0.006 Å below the porphyrin ring.
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  • 3
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    Journal of applied electrochemistry 27 (1997), S. 1198-1206 
    ISSN: 1572-8838
    Keywords: alloy ; amorphous ; anomalous ; hydrogen ; iron ; nickel ; phosphorus ; plating
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Electrical Engineering, Measurement and Control Technology
    Notes: Abstract In this study we have investigated the electrodeposition of amorphous iron–nickel–phosphorus alloys from a sulfate electrolyte. Fe-Ni alloys are known to exhibit an ’anomalous‘ type of plating behaviour in which deposition of the less noble metal is favoured. We have found that the codeposition of phosphorus from hypophosphite in the electrolyte led to a reversal to a ’normal‘ behaviour. This reversal was due both to the suppression of iron and enhancement of nickel partial currents. The overall deposition process is dominated by the hydrogen evolution reaction. This is exacerbated by the low pH needed to codeposit sufficient phosphorus to achieve an amorphous structure.
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  • 4
    ISSN: 1572-8838
    Keywords: alloys ; electroless ; microstructure ; morphology ; nickel ; phosphorus ; zinc
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Electrical Engineering, Measurement and Control Technology
    Notes: Abstract Electroless Ni–Zn–P alloy coatings were obtained on an iron substrate from a sulfate bath at various pH values. The effects of changes in bath pH on alloy composition, morphology, microstructure and corrosion resistance were studied. Scanning electron microscopy was performed to observe the morphological change of the deposits with bath pH. Coating crystallinity was investigated by grazing incidence asymmetric Bragg X-ray diffraction and transmission electron microscopy. A transition from an amorphous to polycrystalline structure was observed on increasing the bath alkalinity, and thus decreasing the phosphorus content of the alloys. A single crystalline phase corresponding to face-centred-cubic nickel was identified in the alloys obtained from a strong alkaline solution. An increase in zinc percentage up to 23% in the deposits does not change the f.c.c. nickel crystalline structure. Corrosion potential and polarization resistance measurements indicated that the corrosion resistance of electroless Ni–Zn–P alloys depends strongly on the microstructure and chemical composition. The deposits obtained at pH 9.0–9.5 and with 11.4–12.5% zinc and 11.8–11.2% phosphorous exhibited the best corrosion resistance.
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  • 5
    ISSN: 1572-879X
    Keywords: selective catalytic reduction ; nitric oxide reduction ; phosphorus ; acid property
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract To examine the influence of phosphorus on the commercial V2O5(WO3)/TiO2 SCR catalyst, measurements were carried out by means of infrared and Raman spectroscopy, XPS, and NO reduction measurement as a function of phosphorus loading. Phosphorus added to the catalyst was found to disperse well over the catalyst without a significant agglomeration up to 5 wt% P2O5 addition. The number of the hydroxyl groups bonded to the vanadium and titanium species decreased readily with increasing amount of phosphorus. Correspondingly, the hydroxyl groups bonded to the phosphorus species were formed. NH3 adsorbed on both hydroxyl groups bonded to vanadium and phosphorus as ammonium ions, implying that the P–OH groups formed are also responsible for the Brønsted acidity. The NO reduction activity was found to be decreased with increasing amount of phosphorus; however, the influence of phosphorus was relatively small irrespective of the large amount of phosphorus addition. The deactivation might be caused by the change in the nature of the surface hydroxyl groups as Brønsted acid sites. Phosphorus species might partially wrap the surface V=O and W=O groups, which might also contribute to the deactivation.
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  • 6
    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.
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  • 7
    ISSN: 1436-5073
    Keywords: aluminium oxide ; phosphorus ; XRF ; RBS ; FTIR
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Phosphorus-doped aluminium oxide thin films were deposited in a flow-type ALE reactor from AlCl3, H2O and from either P2O5 or trimethyl-phosphate. Structural information of the films was obtained from Fourier transform infrared (FTIR) spectra. Rutherford backscattering spectroscopy (RBS) was used to quantitatively determine the composition of the films. The P/Al intensity ratios calculated from X-ray fluorescence (XRF) results were in a linear relation with the P/Al concentration ratios calculated from RBS results. For comparison, the intensity ratios of the phosphorus peak (P=O) at about 1250 cm−1 and the aluminium peak (Al-O) at about 950 cm−1 were determined from the IR absorption spectra. The calibration of FTIR peak intensities was done by plotting the intensity ratios of phosphorus and aluminium peaks against the P/Al concentration ratios measured by RBS. FTIR gave also a linear calibration curve with RBS but the method is less suitable for routine analysis of P/Al ratio than XRF.
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  • 8
    ISSN: 1573-0417
    Keywords: diatoms ; eutrophication ; lake management ; paleolimnology ; British Columbia ; lakes ; phosphorus ; training sets
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Geosciences
    Notes: Abstract Eighteen lakes were added to a published training set of 46 British Columbia (BC) lakes in order to expand the original range of total phosphorus (TP) concentrations. Canonical correspondence analysis (CCA) was used to analyze the relationship between diatom assemblages and environmental variables. Specific conductivity and [TP] each explained significant (P≤0.05) directions of variance in the distribution of the diatoms. The relationship between diatom assemblages and [TP] was sufficiently strong to warrant the development of a weighted-averaging (WA) regression and calibration model that can be used to infer past trophic status from fossil diatom assemblages. The relationship between observed and inferred [TP] was not improved by the addition of more eutrophic lakes, however the [TP] range and the number of taxa used in the transfer function are now superior to the original model. Diatom species assemblages changed very little in lakes with TP concentrations greater than 85 µg 1−1, so we document the development of a model containing lakes with TP≤85 µg 1−1. The updated model uses 59 training lakes and covers a range of species optima from 6 to 41.9 µg 1−1 TP, and a total of 150 diatom taxa. The updated inference model provided a more realistic reconstruction of the anthropogenic history of a highly eutrophic BC lake. The model can now be used to infer past nutrient conditions in other BC lakes in order to assess changes in trophic status.
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  • 9
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    Journal of paleolimnology 20 (1998), S. 47-55 
    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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  • 10
    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.
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  • 11
    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.
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  • 12
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    Journal of paleolimnology 20 (1998), S. 31-46 
    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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  • 13
    ISSN: 1573-0832
    Keywords: ELISA ; Endodermis ; H. annosum ; Immunocytochemistry ; Root rot ; Vascular tissues
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Vascular disintegration mainly of medulla rays of spruce roots is of major significance in root rot disease of spruce caused byH. annosum. Using seedling roots as an experimental model, the possible routes and initial host reactions preceding invasion of vascular tissues was investigated. Transmission electron microscopy showed that penetration through the endodermis was an obvious route but not without host resistance. Using antibodies againstH. annosum hyphal materials, some labelling of vascular tissues remote from sites of fungal colonization suggest the release of fungal secretory products partly active in tissue disintegration. Similarly, intense labelling was also observed in severely colonized host tissues at late stages of infection. Strong labelling recorded at 3 d p.i. mainly on fungal hyphae and scant gold particles on invaded host tissues could imply that induction of host antifungal metabolites may have been a late event. A correlation was found between total antigenic material in root homogenates measured by ELISA, density of tissue labelling by immunocytochemistry and severity of disease symptoms. The importance of this in relation to diagnosis of biotic root rot diseases in the field is discussed.
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  • 14
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    Nutrient cycling in agroecosystems 41 (1995), S. 167-178 
    ISSN: 1573-0867
    Keywords: phosphorus ; workshop ; environment ; review
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A workshop was held in 1990 in Muscle Shoals, Alabama to discuss current and future research on phosphorus in agriculture. Twenty four presentations were given in areas ranging from basic to applied research. For five of the research areas presented at the workshop, this paper presents a literature review, a review of presentations at the workshop, and a discussion of future research ideas.
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  • 15
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    Nutrient cycling in agroecosystems 43 (1995), S. 109-115 
    ISSN: 1573-0867
    Keywords: phosphorus ; European network ; maintenance fertilization ; fixation capacity ; comparison of methods
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract After three years of a research network project on mineral phosphorus fertilization including five experimental fields located in Europe the first results are discussed. Crop response was very significant to TSP application in the alluvial calcareous polder soil of Netherlands, and in the brown silty acid soil of Scotland, both having a low level of P availability and a high fixation capacity. In the alluvial sandy loam on chalk in England, a response was observed to the first fertilization level equal to the previous crop export of phosphorus. In the brown sandy-silty soil on sand in Germany the highest rate of TSP led to a response in the third year. No effect on the final yields was observed in the brown silt loam of Belgium characterised by a textural B horizon with a high P fixation capacity. The critical values for phosphorus fertilization are discussed as the amount of P needed to maintain a target value of soil phosphorus. Concerning the supply of the different soils, no balance was reached in the Dutch and Scottish soils, a steady state was reached in the English soil with the return of the previous crop removal and the critical value for P was lower than the return of the previous crop export in the German and Belgian soils. According to the eight methods of P determination compared in the network, the P contents in the plow layer were raised in the soils of Netherlands, England and Scotland. They remained at the same level or fluctuated depending on the soil testing methods in Germany and in Belgium. High correlations exist between the different methods used in routine analysis, except for the calcium cloride and calcium acetate lactate method. Annual fluctuations in the soil P were detected at different depths depending on analytical methods and need further research.
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  • 16
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    Nutrient cycling in agroecosystems 43 (1995), S. 131-136 
    ISSN: 1573-0867
    Keywords: phosphorus ; titanium ; fertilizer efficiency ; plant nutrition
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract To study the titanium effect on P nutrition, a greenhouse experiment withCapsicum annuum L., cv. Bunejo plants growing under differential P fertilization was conducted. All the plants were grown under identical conditions and they only differred in the P fertilization and in Ti supply. Plant biomass production of the Ti-untreated plants was affected by the diminution of the P-feed, but the plants growing under the lowest P supply did not showed any deficiency symptom during the crop cycle. All the Ti-treated plots showed a significative increase of the plant biomass against their corresponding untreated references. The biomass enhancement was mainly caused by the increase of the fruit yield with an absolute enhancement of 62% in the plants growing under the lowest P feed, and of 45% in the plants with a complete P support.
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  • 17
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    Nutrient cycling in agroecosystems 43 (1995), S. 209-215 
    ISSN: 1573-0867
    Keywords: phosphorus ; saturation ; inventory ; leaching ; eutrophication
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The last three decades, pig breeding has evolved towards a specialised, large scaled, land independent bio-industry in the province of West-Flanders. Subsequently, in certain regions, very high amounts of liquid pig manure are produced each year. This pig slurry is used as a fertilizer at a rate which very often exceeds normal agricultural practices. Because of the nonequilibrium between the phosphorus crop requirements and the P-inputs, phosphates accumulate in the soil. However, the phosphate sorption capacity of a soil is limited. Once the sorption capacity is exceeded, phosphates will start leaching through the soil profile. Since, during winter, in these areas, the groundwater table is situated at a depth of less than 1.0 m, phosphate breakthrough might take place. In the sandy loam soil region (± 1000 km2) of the province, an inventory of the P status of the soil was made. The region was sampled according to a regular grid with 2 km intervals. At random, some sample points were only 500 m apart. This resulted in a total of 296 samplings. In view of fertilizer recommendations, lactate extractable P of the plough layer (0-30 cm) was determined. A maximum value of 101 mg P 100 g−1 of air dry soil, a minimum value of 6 mg P 100 g−1 and a median value of 31 mg P 100 g−1 were found, indicating that for half of the spots monitored, the P status of the soil is high to very high. An oxalate extraction was done to investigate the phosphate saturation of the soil profile (0-90 cm). Based on a critical phosphate saturation degree of 30%, more than half of the soil profiles are phosphate saturated. Phosphate leaching at a rate higher than 0.1 mg ortho-P 1−1 at a depth of 90 cm can be expected. Therefore, a restriction of the P fertilization should be highly recommended. The geostatistical processing of the data using block kriging resulted in a spatial continuous estimate of the phosphate saturation degree. A good agreement was found between the pig density and the phosphate saturation degree of the soil profile.
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  • 18
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    Nutrient cycling in agroecosystems 45 (1995), S. 221-233 
    ISSN: 1573-0867
    Keywords: fertilizer recovery ; modelling ; nitrogen ; nutrient efficiency ; nutrient surplus ; phosphorus ; Poland
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Research on nutrient losses from agricultural systems should try to relate these losses to farm characteristics. This was done for private farms in two districts in Poland. Using data from a farm survey, nutrient surpluses and Nutrient Use Efficiency (NUE, defined as the ratio of outgoing and incoming nutrients) were calculated for nitrogen and phosphorus. Both nutrient surplus and NUE are relatively high. A model was developed to estimate surpluses and NUE from farm characteristics like location, farm size, fertilizer application level, animal density, grass production and sugar beet or potato area. The results of the model are satisfying for nutrient surplus (R2=0.9) and nitrogen NUE (R2=0.4). Estimation of phosphorus NUE was not satisfactory. High surpluses are associated with high fertilizer applications, high animal density and high grass production while an increasing share of sugar beets leads to lower surpluses. A high nitrogen NUE is associated with low fertilizer applications, low animal density and little grass production, and with a high sugar beet area share. Results suggest that, with exception of sugar beet, fertilizer recovery in Poland is very low. Sugar beet, however, combines high fertilizer applications with low surpluses and high NUE. The outcome of the model can be used in the design of environmental policies. The paper ends with some remarks on the type of measures that can be taken, and the effects these will have on private farms in Poland.
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  • 19
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    Nutrient cycling in agroecosystems 46 (1996), S. 81-90 
    ISSN: 1573-0867
    Keywords: elemental sulfur ; granule size ; nitrogen ; phosphorus ; potassium ; S oxidation ; sulfur fertilizers
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Experiments were undertaken to determine the effect of granule size and nutrients in granulated compound fertilizers fortified with finely divided elemental sulfur (So) on the rate of So oxidation. In one experiment, So was banded together with or apart from triple superphosphate (TSP) while in two others, So was granulated with nutrient and inert carriers. A fourth experiment examined response to S in an So-fortified TSP from a range of granule sizes. Response and, in some cases, So recovery (using 35S labels) by test crops (maize, wheat, upland rice) was measured. In all experiments, P mixed with So increased plant growth and S recovery above treatments in which P and So were physically separated. There was however, no effect of distance of separation on S recovery. In one experiment, N as urea and N and P as diammonium phosphate (DAP) were also found to enhance response to So although to a lesser degree than P alone. These observations were attributed to a nutritional requirement of So-oxidizing microorganisms for P and N. Granulation of So with carriers also influenced oxidation rate, as inferred from the fertilizer S recovery. For a given So concentration, the effect was inversely proportional to the mean diameter of granules. It is shown that this relationship can be explained if one assumes that So particles in granules collapse into a fixed number of aggregates per granule irrespective of granule size when the soluble nutrient carrier dissolves and diffuses away from the point of application.
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  • 20
    ISSN: 1573-0867
    Keywords: long-term experiments ; phosphorus ; rice ; nutrient balance ; phosphorus uptake ; fertilizer P response ; soil testing ; ion-exchange resin ; phosphorus supplying capacity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Data from long-term experiments at 11 sites in Asia with a wide range of nutrient input treatments and yield levels were used to quantify crop P requirements of rice (Oryza sativa L.) and the P balance in intensive, irrigated rice systems. Uptake of 1.8–4.2 kg P was required to produce one ton of grain yield. Physiological P use efficiency varied between 220 to 900 kg grain kg P-1. Without added P, there was a net loss of 7 to 8 kg P ha-1 per crop; with added P there was a net gain of 4 to 5 kg P ha-1 per crop. Phosphorus adsorption kinetics on mixed-bed ion-exchange resin capsules provided an integrative measure of soil P status, P diffusion, and acid-induced P solubilization. The resin capsule was a sensitive tool to characterize buildup or depletion of soil P as a result of different P balances. Both Olsen-P and the resin capsule were suitable methods to predict P uptake of tropical lowland rice. It is hypothesized that both methods measure a similar soil P pool which is soluble under alkaline, aerobic conditions but transformed into acid-soluble P froms as a result of submergence and reduction. Present recommendations for P fertilizer use on rice of 20–25 kg P ha-1 are adequate to maintain yields of 5–6 t ha-1, but sustaining higher yields of 7–8 t ha-1 will require farm-specific management strategies based on knowledge of the long-term P balance and soil P-supplying capacity.
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  • 21
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    Nutrient cycling in agroecosystems 46 (1996), S. 71-79 
    ISSN: 1573-0867
    Keywords: ammonium poly-phosphate ; diammonium orthophosphate ; fertilizer reaction ; gram ; Indian soils ; phosphorus ; P uptake ; single superphosphate ; triple superphosphate ; yield
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Laboratory studies on the characterization of soil-fertilizer P reaction products were carried out by reacting three-soils occurring in a toposequence in the plateau region of Bihar (India) with saturated solutions of diammonium orthophosphate (DAP), triple superphosphate (TSP) and ammonium polyphosphate (APP) for 1 hour and 24 hours. The reaction products (precipitates) formed in the solutions after 120 days of incubation were isolated and identified through X-ray diffraction technique. Results indicate the formation of Brushite [CaHPO4 · 2H2O, Strengite (FePO4 · 2H2O), Variscite (AIPO4 · 2H2O) and Fe4(P2O7)3 as major soil-fertilizer P reaction products in these soils with ortho-and polyphosphates as source of phosphorus. Pot cultures were used to evaluate the relative efficiency of reaction products (Struvite, Brushite, Variscite and Strengite), orthophosphates (DAP and SSP) and polyphosphate (APP) as sources of P for gram (Cicer arietinum L.) in a typical acid soil. Results indicate significant response of gram to different sources and level of added P. The dry weight and P uptake at 0, 6 and 12 mg P kg-1 soil were 0.406, 0.519 and 0.609 (g pot-1); and 0.289, 0.428 and 0.575 (mg P pot-1), respectively. Among the sources , struvite proved to be superior or equally effective as APP, DAP or SSP as sources of P for gram. Uptake of P also varied significantly with different P sources and levels of P application. Strengite was least effective in enhancing yield and P uptake by the crop.
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    Nutrient cycling in agroecosystems 44 (1995), S. 1-8 
    ISSN: 1573-0867
    Keywords: aerobic incubation ; cation-anion-exchange resin ; phosphorus ; resin beads ; resin membranes ; suspension incubation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Six Portuguese soils of varying P sorption capacity were incubated aerobically at 30° C without and with added P in order to give 0.1.mg P L−1 in the soil solution. Two methods of measuring extractable P were compared: (i) mixed-bed cation-anion-resin beads in bags and (ii) a simpler method with anion-resin membrane only. The bag method extracted about twice and 1.5 times as much as the strip method, respectively, without and with added P. The relationships were much closer after one extraction for 2 hours (r = 0.982, p 〈 0.01) instead of the cumulative extraction of 24 hours (r = 0.635,p 〉 0.05.). P recovery after incubation was inversely related to some soil properties as organic matter, buffer capacity, selective dissolution Al forms (Alox and Ald) and P sorption. It is suggested that the simpler resin membrane method is more adequate to assess P for many studies of P reaction with soil. A simpler incubation method was tried, consisting of incubation as a soil suspension in water at a high temperature (50° C). The results suggested that this method gave similar results to aerobic incubation, with the advantage that there was no need to measure the required and final water contents of incubated soil.
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    Nutrient cycling in agroecosystems 46 (1996), S. 179-187 
    ISSN: 1573-0867
    Keywords: electrical conductivity ; leaching ; nitrogen ; pH ; phosphorus ; potassium ; release pattern ; slow-release fertilizers ; temperature
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract We studied the effect of temperature on the release of N, P, and K from slow-release fertilizers (SRF). The study was conducted in micro-lysimeters filled with moist peat medium. Increasing the temperature from 4 to 12°C slightly increased N release from three different slow-release N (SRN) carriers with different particle sizes and coating thicknesses. At 21°C the rate of release was significantly different than the other two temperatures. Urea formaldehyde (UF), sulphur coated urea (SCU) and coated calcium nitrate (CCN), incubated in sphagnum moss peat, released between 3 and 20% of the applied N in six weeks. For eight synthetic and organic NPK carriers, the release pattern was similar to UF and SCU. However, the leaching losses of N from the NPK fertilizers were up to twenty times more than for the SRN products. Except for Osmocote® and Duna, which released 30–40% of the applied N as mineral-N within six weeks, all other slow-release and slowly mineralized NPK carriers acted like readily water-soluble compound NPK. Temperature did not affect the nutrient release from NPK fertilizers.
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    Nutrient cycling in agroecosystems 50 (1998), S. 321-324 
    ISSN: 1573-0867
    Keywords: nutrient modelling ; leaching ; nitrogen ; phosphorus ; schematization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract In context of preparing the Fourth National Policy Document on Water Management in the Netherlands effects of different scenarios of fertilizer management on nitrogen (N) and phosphorus (P) leaching from rural areas into Dutch surface waters were analyzed. The manuscript offers insight into the model instrument that is used to simulate the different scenarios. Main parts of the modelinstrument are: a procedure to schematize the Netherlands in horizontal areal units, field scale mechanistic models for water and nutrient behaviour in the soil and an empirical model for fertilizer additions.
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  • 25
    ISSN: 1573-0867
    Keywords: agriculture ; catchment ; fertilizer ; historical ; manure ; nitrate ; phosphorus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A suggested increase in the growth of macrophytic algae within the Ythan estuary (N.E. Scotland) over recent years has been linked to the increased amounts of nitrogen in the form of NO3–N entering the estuary from the river. The increased NO3 concentration in the river has been associated with recent changes in farming practices in this predominantly agricultural catchment. Terrestrially derived phosphorus is also considered to contribute increasingly to eutrophication of fresh waters. Historical agricultural census data together with appropriate surveys of fertilizer practice were used to calculate the total quantities of fertilizer and manure derived N and P applied annually over the wholeYthan catchment during the period 1960 – 1990. While the total agricultural land area has remained similar, significant changes in cropping practice have occurred. In particular, a greater proportion of land is given to autumn sown crops while the area of grassland has declined. These changes in farming practice are associated with differences in both the total amounts and timing of fertilizer applied. The use of inorganic N in the catchment has trebled since 1960 and is currently approximately 6400 tonnes (104 kg N/ha). The use of P has decreased by more than a quarter to 1274 tonnes (21 kg P/ha) over a similar time period. There has been no obvious change in total quantity of N and P derived from animal manures, estimated to be 44 and 11 kg per ha, respectively, when averaged over the area of agricultural land. Cattle and sheep numbers have remained relatively constant and together account for approximately 80% of the manure N and 70% of the manure P produced annually. However, poultry have declined by 70% since 1960 while pig numbers have increased six-fold. The average annual application rate of manure derived N over the whole catchment (44 kg/ha) is considerably below that proposed at the farm scale in the EC Nitrate Directive (210–170 kg/ha). However, on a local scale difficulties may arise for large manure producing concerns such as dairy or pig units.
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    Nutrient cycling in agroecosystems 55 (1999), S. 7-14 
    ISSN: 1573-0867
    Keywords: fertiliser formulation ; nutrients ; phosphorus ; relative humidity ; soil moisture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Phosphorus lost in runoff from agricultural land leads to the enrichment of surface waters and contributes to algal blooms. Fertilisers are one source of this P. To compare the water available P of different fertiliser formulations in the laboratory it is necessary to control environmental conditions, temperature, relative humidity and soil water content, prior to simulating rainfall. Two chambers were designed in which relative humidity and soil water content were controlled using salt solutions. An initial design comprising a sealed chamber with three layers of soil samples over a salt bath was found to be inferior to a single layer design. The changes in water content of soil samples were used to test the single layer chamber in a constant temperature environment (15 °C) using a saturated KCl solution (90% relative humidity). Based on the final soil water content of the samples, the spatial variation within the chamber was within tolerable limits. The single layer chamber was used for a simulation experiment comparing the water available P of two commercial fertilisers. Using a saturated resorcinol solution (95% relative humidity) soil samples were equilibrated at 15 °C for 21 days, fertiliser added, and the water available P measured up to 600 h after fertiliser application. The results indicate that the amount of water available P was related to the fertiliser compound and exponentially related to the time since fertiliser application. It was concluded that the single layer chamber is suitable for controlling relative humidity and soil water content in trials such as these where the water available P of fertilisers are being compared.
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  • 27
    ISSN: 1573-0867
    Keywords: dairy systems ; feeds ; fertilizers ; phosphorus ; P surplus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Inputs of phosphorus (P) above requirements for production on dairy farms lead to surplus P with increased risk of P transfer in land run-off to surface waters causing eutrophication. The impact of reducing surplus P inputs in purchased feeds and fertilizers on milk and forage production was investigated in a comparison of three dairy farm systems on chalkland soils in southern England over a 3-year period. In accordance with current commercial practice, no attempt was made to regulate P inputs in system 1, which accumulated an average annual surplus of 23 kg P ha-1. Progressive reductions in purchased feed and/or fertilizer inputs into systems 2 and 3 decreased surplus P to 17 and 3 kg ha-1, respectively, without apparently limiting either milk or herbage dry matter production. The estimated reduction in faecal P output from system 3 cows fed a low P diet compared to system 1 cows fed a high P diet was 26%. Milk P concentrations significantly (P 〈0.001) increased in systems 2 and 3 which included maize in the diet. Output of P in milk and meat products, as a proportion of the total dietary P inputs, increased from 28% in system 1 to 36% in system 3. Surplus P was greatest in continuous maize fields receiving both dairy manure and starter P fertilizer. Withholding P fertilizer in system 3 did not reduce P offtake in cut herbage on soils of moderate P fertility. Total annual losses of P in storm run-off and leaching were no greater than annual inputs of P from the atmosphere (0.5 kg ha-1). The results indicate there is scope to reduce surplus P on commercial dairy farms without sacrificing production targets at least in the short term. Purchased feeds are the largest of the P inputs on intensive dairy farms, yet these are rarely quantified on commercial holdings.
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  • 28
    ISSN: 1573-0867
    Keywords: leaching ; phosphorus ; poultry litter ; soil
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract To determine P loadings, added through poultry litter, sufficient to cause downward movement of P from the cultivated layer of a sandy soil, six rates of poultry litter were applied annually for four years to a site in central England. (total loading 0 – 1119 kg P ha-1). A single extra plot also received an extra 1000 kg ha-1 as triple superphosphate (TSP; total loading 2119 kg P ha-1) and three other treatments received 200 – 800 kg ha-1 P as TSP only. Annual soil sampling in 30-cm increments to 1.5-m depth provided information on P build-up in the topsoil and P movement to depth. There were strong linear trends between P balance (P applied – P removed in crops) and total P, Olsen bicarbonate extractable P and water-soluble P in the topsoil. Phosphorus from TSP and poultry litter fell on the same regression lines, suggesting that both would be equally effective as fertilizer sources. We calculated that 100 kg ha-1 surplus total P would increase the Olsen extractable P content by c. 6 mg kg-1 and the water-soluble P by c. 5 mg kg-1. Thus, relatively large amounts of P would need to be applied to raise soil P status. We found some evidence of P movement into the soil layers immediately below cultivation depth. However, neither soil sampling nor soil solution extracted through Teflon water samplers showed evidence of movement into the deep subsoil (1 m) despite large P loadings.
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    Nutrient cycling in agroecosystems 54 (1999), S. 259-266 
    ISSN: 1573-0867
    Keywords: bahiagrass ; manure ; pasture fertilization ; phosphorus ; phosphorus cycling ; Spodosol
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Bahiagrass (Paspalum notatum Flugge) pasture fertilization recommendations have traditionally been based upon clipping studies. Inclusion of P from manure, not originally considered when P recommendations were developed for pastures, may minimize the need for P fertilization without reducing bahiagrass production or P uptake. The objective of this research was to determine if manure contributes greatly to the P crop nutrient requirement. A 2-year field study utilized a factorial arrangement of 0 and 6.9 Mg air-dried manure ha-1 with 0, 17, 34, 51, and 68 kg inorganic P ha-1 from triple superphosphate to evaluate bahiagrass yield, root distribution, and P uptake response on a Myakka fine sand (sandy, siliceous, hyperthermic Aeric Alaquod). Because air-dried manure was used in the field study, a greenhouse study was employed to confirm that there were no differences in bahiagrass yield or P uptake from either air-dried or fresh cattle (Bos spp.) manure sources. There were no manure or manure by P interaction effects on yield or P uptake of bahiagrass indicating that manure source did not effect grass production in the greenhouse. In the field study, bahiagrass roots were distributed into the Bh horizon, and the Bh horizon had at least four times more Mehlich-1 extractable P than that of the Ap horizon. This horizon was most likely acting as a main source for P-uptake by the grass. This observation was further confirmed by no yield response to levels of inorganic P application in 1989. A linear-response-and-plateau (R2=0.196) relationship with a critical point of 15.4 kg P ha-1 was found in 1990. Bahiagrass yield and P uptake were not dependent on P fertilization, either from manure or inorganic P, due to the availability of P from the Bh horizon.
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    Nutrient cycling in agroecosystems 47 (1996), S. 243-250 
    ISSN: 1573-0867
    Keywords: fertilizer value ; nitrogen ; phosphorus ; poultry manure ; urea ; wetland rice
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Poultry manure applied alone or in combination with urea at different N levels was evaluated as a N source for wetland rice grown in a Fatehpur loamy sand soil. Residual effects were studied on wheat which followed rice every year during the three cropping cycles. In the first year, poultry manure did not perform better than urea but by the third year, when applied in quantities sufficient to supply 120 and 180 kg N ha−1, it produced significantly more rice grain yield than the same rates of N as urea. Poultry manure sustained the grain yield of rice during the three years while the yield decreased with urea. Apparent N recovery by rice decreased from 45 to 28% during 1987 to 1989 in the case of urea, but it remained almost the same (35, 33 and 37%) for poultry manure. Thus, urea N values of poultry manure calculated from yield or N uptake data following two different approaches averaged 80, 112 and 127% in 1987, 1988 and 1989, respectively. Poultry manure and urea applied in 1:1 ratio on N basis produced yields in between the yields from the two sources applied alone. After three cycles of rice-wheat rotation, the organic matter in the soil increased with the amount of manure applied to a plot. Olsen available P increased in soils amended with poultry manure. A residual effect of poultry manure applied to rice to supply 120 or 180 kg N ha−1 was observed in the wheat which followed rice and it was equivalent to 40 kg N ha−1 plus some P applied directly to wheat.
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  • 31
    ISSN: 1573-0867
    Keywords: pasture fertilization ; phosphorus ; potassium ; nutrient budget ; nutrient efficiency
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Dairy farming is the main agricultural activity of the Basque Country. A dairy farm is characterized as a system with soils and crops, forage, cattle and manure as main components, and in such a system, nutrient cycling is very important to maintain soil fertility and optimize forage production. To quantify nutrient transfers in the cycle, a simple system was developed and has been applied to seventeen farms to examine its ability to achieve a balanced P and K fertilization. These farms have provided data on inputs (fertilizer, feeds, concentrates), pasture and manure management, and outputs (milk production), and soil samples have been taken from farm pastures. Phosphorus and K in excreta and uneaten pasture is used with a relatively high efficiency as suggested by the relatively high efficiency of P and K utilization by the pasture that usually ranges from 70 to 90%. Concentrate feeding (3000 kg cow−1 yr−1) represents one of the main P and K inputs in Basque Country dairy farms, averaging 26 and 66 kg ha−1, respectively. Besides, release of K in the soil through slow liberation from non-exchangeable sites was estimated as 30 kg ha−1. Thus, a high efficiency in excreta recycling would diminish substantially P and K mineral fertilizer needs. Farm nutrient budgets appear to be a convenient tool for determining nutrient shortages and surpluses at farm level, and thus they are considered as a first step to support a better management of maintenance fertilization of permanent pastures.
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    Nutrient cycling in agroecosystems 45 (1995), S. 193-197 
    ISSN: 1573-0867
    Keywords: cation activity ; phosphorus ; potassium chloride ; soil solution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The electrolyte concentration of the soil solution affects the availability of some nutrients in the soil, especially of P, but it is not know at what salt concentration the reactions start to be significantly affected and their magnitude. This study was carried out to evaluate the effect of rates of potassium chloride (KCl) on some soil parameters that determine supplying of P, K, Ca, Mg, and Al in an unlimed acid soil. Increasing rates of KCl (from zero up to 2000 mg K kg−1) were applied to soil samples fertilized with 360 mg P kg−1. Solution (Cli) and exchangeable (Csi) forms of P, Ca, Mg, K, and Al were determined in the treated soil samples after 30-days of incubation; cation activity in solution and their selectivity coefficients were then calculated. Addition of KCl at rates equal to or above 500 mg K kg−1 caused a large relative increase on P in the soil solution (Pli) but a small and insignificant increase on the absolute value of Pli. All forms of soil K increased with increases on K applied, and buffer power for K varied according to the range of soil K. At all KCl rates, K displaced Ca, Mg, and Al from the solid phase to the soil solution, but had no effect on the extractable values. The relative preference of cations for the adsorption sites increased with increase on cation valency, and only those selectivity coefficients involving K were affected by K applied.
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    Mangroves and salt marshes 2 (1998), S. 37-42 
    ISSN: 1572-977X
    Keywords: mangrove ; phosphorus ; distribution ; dynamics ; exportation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The distribution and dynamics of phosphorus have been studied in the mangroves of Sepetiba Bay, Brazil. Leaf fall contributes 3.0 kg P ha=1yr=1to the sediment. The total above ground biomass of the R. mangle stand was about 65.3 t ha=1, the P accumulation was 3.9 kg P ha=1where 63% of the total P-biomass was accumulated in the leaves. The biomass of below ground roots was about 8.2 tha=1 and accumulated 16% of total P-biomass. Sediment contained 452 kg P ha=1 where P combined with calcium (P-Ca) was the main fraction (260 kg ha=1). The annual flux of P as litter fall was small (〈 1%) compared to total P in the sediment reservoir. The annual export of P by macrodetritus corresponds to 0.05% of the total sediment reservoir.
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    Environmental geology 30 (1997), S. 224-230 
    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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    Trees 13 (1999), S. 138-151 
    ISSN: 0931-1890
    Keywords: Key words Cytoskeleton ; Immunocytochemistry ; Model systems ; Populus ; Secondary vascular system
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract  Trees represent a, probably the, major component of the biosphere and have a unique place in the history of Mankind. One of their most fascinating features is the process of secondary growth which is effected principally by the secondary vascular system, the developmental continuum of secondary phloem, vascular cambium, and secondary xylem. However, for too long assumptions about the developmental biology of trees have had to be based upon studies of primary growth systems within annual, herbaceous species because study of the secondary vascular system had been largely ignored. Even when attempts are made to understand some of the most fundamental features of the secondary vascular system, such as xylogenesis, the current model system, isolated Zinnia mesophyll cells, is not entirely appropriate to the situation in the intact tree. Some deficiencies of the Zinnia system are discussed, and the advantages of the genus Populus as a model for study of the hardwood secondary vascular system are considered. Some of the new approaches which are poised to lead to significant advances in our knowledge of the cell bio-logy of the secondary vascular system of trees – spe-cifically of the cell wall, the plasmalemma, and the cytoskeleton – are discussed. The value of one of these new techniques – immunocytochemistry – is demonstrated by a consideration of recent work on the role of the cytoskeleton in the hardwood secondary vascular system.
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    Histochemistry and cell biology 106 (1996), S. 9-17 
    ISSN: 1432-119X
    Keywords: Silver enhancement ; Immunogold-silver staining ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The immunogold silver staining method (IGSS) is widely used as a sensitive and specific immunohistochemical visualisation technique. IGSS involves the specific deposition of metallic silver at the site of immunogold labelling and provides a means of visualisation at low magnification by light or electron microscopy. Silver developers for IGSS rapidly deposit metallic silver only at the site of heavy metals, including gold and silver, because of their catalytic activity. The developing solution contains the silver ions and reducing agent necessary for this reaction. Using different silver salts as ion donors and by selecting an appropriate temperature and pH, visible amounts of silver can be deposited in a few minutes at the site of colloidal gold labelling while little non-specific background deposition occurs. Inclusion of protective colloids in the solution can also be used to control the reaction. Although studies of the chemical basis of silver deposition around unlabelled colloidal gold date back to 1939, immunogold enhancement by silver was established in 1983. The IGSS method evolved from the combination of disparate photographic, histochemical and immunogold techniques which have been effectively combined and optimised over the last 10 years to provide a visualisation system which is well suited to many immunohistochemical studies.
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    Histochemistry and cell biology 106 (1996), S. 9-17 
    ISSN: 1432-119X
    Keywords: Key words Silver enhancement ; Immunogold-silver staining ; Immunocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The immunogold silver staining method (IGSS) is widely used as a sensitive and specific immunohistochemical visualisation technique. IGSS involves the specific deposition of metallic silver at the site of immunogold labelling and provides a means of visualisation at low magnification by light or electron microscopy. Silver developers for IGSS rapidly deposit metallic silver only at the site of heavy metals, including gold and silver, because of their catalytic activity. The developing solution contains the silver ions and reducing agent necessary for this reaction. Using different silver salts as ion donors and by selecting an appropriate temperature and pH, visible amounts of silver can be deposited in a few minutes at the site of colloidal gold labelling while little non-specific background deposition occurs. Inclusion of protective colloids in the solution can also be used to control the reaction. Although studies of the chemical basis of silver deposition around unlabelled colloidal gold date back to 1939, immunogold enhancement by silver was established in 1983. The IGSS method evolved from the combination of disparate photographic, histochemical and immunogold techniques which have been effectively combined and optimised over the last 10 years to provide a visualisation system which is well suited to many immunohistochemical studies.
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    Environmental and resource economics 10 (1997), S. 341-362 
    ISSN: 1573-1502
    Keywords: Baltic Sea ; eutrophication ; nitrogen ; phosphorus ; cost effective
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering , Economics
    Notes: Abstract Due to eutrophication caused by heavy loads of nitrogen and phosphorus, the biological conditions of the Baltic Sea have been disturbed: large sea bottom areas without any biological life, low stocks of cods, and toxic blue green algaes. It is recognized that the nitrogen and phosphorus loads to the Baltic Sea must be reduced by 50% in order to restore the sea. The main purpose of this paper is to calculate cost effective nitrogen and phosphorus reductions to the Baltic Sea from the nine countries surrounding the Baltic Sea. The results show a significant difference in minimum costs of decreasing nitrogen and phosphorus loads to the Sea: approximately 12 000 millions of SEK per year and 3 000 millions of SEK respectively for reductions by 50%. It is also shown that a change from a policy of cost-effective nutrient reductions to a policy where each country reduces the nutrient loads by 50% increase total costs for both nitrogen and phosphorus reductions by about 300%. The results are, however, sensitive to several of the underlying assumptions and should therefore be interpreted with much caution.
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  • 39
    ISSN: 1573-2932
    Keywords: phosphorus ; P flux ; microbial activity ; redox ; simulation ; Lake Kinneret ; sediment ; accumulative P release
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract Different factors which interactively control the flux of soluble reactive phosphorus (SRP) at the sediment-water interface (SWI) of Lake Kinneret were studied seasonally. The influence of pH, Eh and microbial activity on SRP flux at the SWI was investigated by manipulating the conditions in the overlying water of intact sediment cores. The calculated diffusive SRP flux out of the sediment was lower in cores sampled during winter and spring than during the period of amixis. Potential SRP release, as measured in the absence of microbial activity, was strongly enhanced upon the transition from oxic to anoxic conditions indicating P release from iron(III)-bound phosphorus. In spring and summer cores, an enhanced SRP flux from sediments at pH 7 in comparison to pH 8 indicated P release from carbonate-bound P which sedimented previously as result of high pH values during the algal spring bloom. Microbial uptake at the SWI was the most important sink for SRP and no net-flux occured under oxic conditions. The higher net-flux of P under anoxic conditions was linked to carbon limitation of the bacteria at the SWI.
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    Water, air & soil pollution 99 (1997), S. 457-464 
    ISSN: 1573-2932
    Keywords: sediment ; phosphorus ; fractionation ; release ; humic lake
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract Lake Flosek (north-eastern part of Poland) is a small shallow and without outflow lake which has been limed in 1970. The concentration of Ca was increased from 3-4 mg L-1 to 17 mg L-1 in the water and from 0.2-0.3% dry weight to 0.9-1.7% dry weight in sediments (5 cm upper layer) due to CaCO3 addition to the lake. In the spring-summer seasons of 1992 and 1993, an experimental study was conducted in Lake Flosek to assess the capacity of bottom sediments to uptake and release mineral phosphorus. The rate of phosphorus exchange between sediments and near-bottom water was experimentally measured under conditions of high (100%), and of reduced (10%) oxygen saturation in near-bottom water. To determine the component of sediments responsible for the uptake of most phosphorus, the proportions of phosphorus forms in sediments were analysed. Sediments of Lake Flosek showed a slight tendency to release phosphates. The rate of this process was similar under high (100%) and low (10%) oxygen saturations ranging from - 0.161 to + 0.200 mg P m-2 d-1. This is much lower (by 1-2 orders of magnitude) than reported from other harmonic, non-humic lakes. In the total phosphorus pool, the highest content of phosphorus was found in the organic and residual phosphorus fractions (over 70% of the total phosphorus in sediments). The largest part of the readily extractable phosphorus was found in the fraction bound to Al and humic substances (41%). Both these fractions determine a weak exchange of phosphorus between sediments and water. No difference in P-release related to P-fraction compound was found in the cores taken from three sites in the lake.
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  • 41
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    Water, air & soil pollution 99 (1997), S. 477-486 
    ISSN: 1573-2932
    Keywords: sediment ; nitrogen ; phosphorus ; organic matter ; cluster analysis ; Gulf of Finland ; estuaries
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract Dry weight (DW), ignition loss (IL) and concentrations of total nitrogen (TN) and total phosphorus (TP) of the sediment surface layer (0 to 10 cm, 1 cm slices) were analyzed from 20 sites in the eastern Gulf of Finland. The distance of the sampling sites from the mouth of the River Neva explained the nutrient concentrations of the sediments well, while the effect of water depth was negligible. The increase of TN and the decrease of TP along the transect from the river mouth towards the open Gulf were caused by the diminishing share of allochthonous material supplied from the River Neva. The mean TN concentration of the different accumulation areas was about 40 % higher in the sediment surface than in the deeper layer (9 to 10 cm). The corresponding difference for TP varied from 53 to 56 %. The results suggest considerable netflux of nutrients from sediment to water. The net sediment accumulation of nutrients were estimated as 6.0 g m-2 a-1 of N and 1.7 g m-2 a-1 of P corresponding 22 000 t a-1 of N and 6 100 t a-1 of P for the whole eastern Gulf.
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  • 42
    ISSN: 1573-2932
    Keywords: Adriatic Sea ; nutrients ; benthic fluxes ; carbon ; nitrogen ; silicon ; phosphorus ; budgets
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract Benthic fluxes of dissolved inorganic N, Si and P nutrients, alkalinity, dissolved inorganic C (DIC), and O2 from sediments in the Gulf of Trieste (northern Adriatic, Italy) were measured monthly in the period September 1995 – August 1996 using in situ incubated light benthic chambers. The highest efluxes of DIC, NH4 +, PO4 3−, Si(OH)4, and NO3 − influxes encountered in late summer — early autumn were the consequence of degradation of benthic microalgae, and in autumn mostly of sedimented phytoplankton. High NO3 − efflux was observed in spring. Only NH4 + and Si(OH)4 fluxes were significantly correlated with temperature. This correlation suggests that the rate of downward input and the quality and quantity of sedimentary organic matter (autochthonous and allochthonous) were superimposed on the temperature fluctuations. High DIC, NH4 + and Si(OH)4 effluxes observed in July 1996 were due to the late spring — early summer degradation of sedimentary organic matter produced by benthic microalgae, while the autumn phytoplankton bloom was quickly reflected in enhanced benthic fluxes due to higher temperature. Significant correlations between NH4 +, PO4 3− and Si(OH)4 fluxes suggested their parallel regeneration and utilization at the sediment-water interface. The nutrient fluxes were linked to O2 consumption, suggesting that aerobic oxidation processes were important at the sediment-water interface in the Gulf. The N, P and Si nutrients released from sediment pore waters are probably utilized in benthic microalgal and bottom-water primary production. This indicates that pelagic and benthic communities in the central part of the Gulf of Trieste function relatively independently of each other.
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  • 43
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    Water, air & soil pollution 99 (1997), S. 633-640 
    ISSN: 1573-2932
    Keywords: zebra mussel ; Dreissena polyrnorpha ; phosphorus ; trophic state ; recovery ; take Como
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract A large scale study on the western basin of Lake Como (N. Italy) was started in 1995 to examine the effects of the zebra mussel colonization which began in early '90. Our results have been related to '91–92 data (pre-Dreissena period), before the maximum colonization of zebra mussel. In spring and summer of the post-Dreissena period total phosphorus, P-PO4, nitrate and chlorophyll values decreased, while ammonium and tranparency increased at every sampling station. Zebra mussel does not modify the trophic state of this sub-basin but it plays an important role in nutrient cycling. The entire population can filter epilimnetic craters 2.1 times per year and can produce 2.9 × 104 t y of pseudofaeces which are transferred to sediments.
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  • 44
    ISSN: 1432-2048
    Keywords: Key words:β-1 ; 3-Glucanase ; Immunocytochemistry ; Leaf rust pathogen ; Resistance ; Triticum (pathogen resistance)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. An antiserum raised against the purified 33-kDa β-1,3-glucanase of wheat (Triticum aestivum L.) was employed to investigate the ultrastructural localization of the enzyme in wheat leaves infected with Puccinia recondita Rob. ex Desm. f.sp. tritici Eriks. and Henn. using a post-embedding immunogold labelling technique. In both compatible and incompatible interactions, β-1,3-glucanase was detected in the host plasmalemma and in the domain of the host cell wall near the plasmalemma of the mesophyll cells, but higher concentrations of the enzyme were detected in infected resistant wheat leaves than in infected susceptible ones. β-1,3-Glucanase was also found in the secondary thickening of xylem vessels and in the walls of guard cells, epidermal cells and phloem elements, while no labelling was observed in host organelles, viz. vacuoles, mitochondria, endoplasmic reticulum, Golgi bodies, nuclei and chloroplasts. A low concentration of the enzyme was detected on the intercellular hyphal wall and in the hyphal cytoplasm. In the compatible interaction, β-1,3-glucanase was demonstrated to accumulate predominantly in the haustorial wall and extrahaustorial matrix. In the incompatible interaction, strong labelling for β-1,3-glucanase was found in host cell wall appositions, in the extracellular matrix in the intercellular space, and in electron-dense structures of host origin which occurred in the incompatible interaction only.
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  • 45
    ISSN: 1432-1424
    Keywords: Water channels ; Vasopressin ; Rat kidney ; Immunocytochemistry ; Microtubules ; Cell polarity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Aquaporin 2 is a collecting duct water channel that is located in apical vesicles and in the apical plasma membrane of collecting duct principal cells. It shares 42% identity with the proximal tubule/thin descending limb water channel, CHIP28. The present study was aimed at addressing three questions concerning the location and behavior of the AQP2 protein under different conditions. First, does the AQP2 channel relocate to the apical membrane after vasopressin treatment? Our results show that AQP2 is diffusely distributed in cytoplasmic vesicles in collecting duct principal cells of homozygous Brattleboro rats that lack vasopressin. In rats injected with exogenous vasopressin, however, AQP2 became concentrated in the apical plasma membrane of principal cells, as determined by immunofluorescence and immunogold electron microscopy. This behavior is consistent with the idea that AQP2 is the vasopressin-sensitive water channel. Second, is the cellular location of AQP2 modified by microtubule disruption? In normal rats, AQP2 has a mainly apical and subapical location in principal cells, but in colchicine-treated rats, it is distributed on vesicles that are scattered throughout the entire cytoplasm. This is consistent with the dependence on microtubules of apical protein targeting in many cell types, and explains the inhibitory effect of microtubule disruption on the hydroosmotic response to vasopressin in sensitive epithelia, including the collecting duct. Third, is AQP2 present in neonatal rat kidneys? We show that AQP2 is abundant in principal cells from neonatal rats at all days after birth. The detection of AQP2 in early neonatal kidneys indicates that a lack of this protein is not responsible for the relatively weak urinary concentrating response to vasopressin seen in neonatal rats.
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  • 46
    ISSN: 1432-0878
    Keywords: Oxytocin ; Neurophysin ; Vasotocin ; Mesotocin ; Suprachiasmatic nucleus ; Medial nucleus of the infundibular recess ; Immunocytochemistry ; Natrix maura (Serpentes) ; Mauremys caspica (Chelonia)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The probable presence of oxytocin in the hypothalamo-hypophysial system of two reptilian species, the snake Natrix maura and the turtle Mauremys caspica, was re-investigated. A high-pressure liquid chromatographic analysis of the turtle neural lobe revealed the existence of vasotocin, mesotocin, and a third compound co-eluting with oxytocin. Brains from both species were fixed by vascular perfusion with Bouin's fluid. Adjacent paraffin sections were immunostained using antisera against the following substances: (1) bovine oxytocin-neurophysin; (2) a mixture of bovine oxytocin-neurophysin and vasopressin-neurophysin; (3) dogfish neurophysins; (4) oxytocin; (5) arginine-vasotocin; (6) mesotocin; (7) somatostatin. Immunoreactivity against oxytocin was found in parvocellular neurons of the snake suprachiasmatic nucleus and cerebrospinal-fluid contacting neurons of the medial nucleus of the infundibular recess of both species, the latter immunoreactivity being much more conspicuous in the turtle. Numerous fibers containing immunoreactive oxytocin extended between the medial nucleus of the infundibular recess, and the internal region of the medium eminence and the neural lobe. The oxytocin-immunoreactivity in all locations was completely abolished by preabsorption of the anti-oxytocin serum with three different oxytocin preparations. None of the neurons of the suprachiasmatic and medial nucleus of the infundibular recess, including the oxytocin-immunoreactive elements, reacted with either the antineurophysin sera used, or the anti-vasotocin or anti-mesotocin antibodies. The possible existence of a reptilian oxytocin-neurophysin is discussed. The alternative that, in the reptilian hypothalamus, neurons synthesize a compound closely related to, but different from oxytocin is also considered.
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  • 47
    ISSN: 1432-0878
    Keywords: Pituitary gland ; Gonadotropin ; Subunits ; Gonadotropes ; Immunocytochemistry ; Immunoblotting ; Oncorhynchus mykiss (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Salmon gonadotropin (GTH II) is a heterodimeric glycoprotein hormone (α and IIβ subunits), serving as a maturational GTH, and is produced in a specific gonadotropic cell-type (GTH II-cells) containing small granules and large globules. In trout GTH II-cells, double immunolabeling for the α- and IIβ-subunits shows that colocalization of the α- and IIβ-immunolabeling is confined to the small granules, indicating storage of functional GTH II. On the other hand, α-immunolabeling is absent in the large globules, even though IIβ labeling is abundant throughout the period of seasonal gametogenesis. The α-specific antiserum recognizes the intact α-subunit as well as the reduced and deglycosylated α-subunits by immunoblotting. These results indicate that an accumulation of the IIβ-subunit is specifically generated in the large globules of these cells. In fact, with sexual maturity, the quantity of IIβ-subunits becomes elevated in the trout pituitary due to a marked increase in GTH II-cells containing many large globules. However, the derivation and function of the large globules and the fate of their contained IIβ-subunits remains unknown.
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  • 48
    ISSN: 1432-0878
    Keywords: Trans-differentiation ; Proliferation ; Bromodeoxyuridine ; Immunocytochemistry ; Regeneration ; Ciona intestinalis (Tunicata)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract In this study, we use three monoclonal antibodies that recognise antigens present in the central nervous system of the ascidian Ciona intestinalis to study regeneration and post-metamorphic development of the neural ganglion. We have also used bromodeoxyuridine labelling to study generation of the neuronal precursor cells. The first antibody, CiN 1, recognises all neurones in the ganglion, whereas the second, CiN 2, recognises only a subpopulation of the large cortical neurones. Western blotting studies show that CiN 2 recognises two membrane-bound glycoproteins of apparent Mr 129 and 100 kDa. CiN 1 is not reactive on Western blots. Immunocytochemical studies with these antibodies show that CiN 1-immunoreactive neurone-like cells are present at the site of regeneration as early as 5–7 days post-ablation, a sub-population of CiN 2-immunoreactive cells being detected by 9–12 days post-ablation. The third antibody, ECM 1, stains extracellular matrix components and recognises two diffuse bands on Western blots of whole-body and ganglion homogenates. The temporal and spatial pattern of appearance of CiN 1 and CiN 2 immunoreactivity both during post-metamorphic development and in regeneration occurs in the same sequence in both processes. Studies with bromodeoxyuridine show labelled nuclei in some neurones in the regenerating ganglion. Plausibly these originate from the dorsal strand, an epithelial tube that reforms by cell proliferation during the initial phases of regeneration. A second population of cells, the large cortical neurones, do not incorporate bromodeoxyuridine and thus must have been born prior to the onset of regeneration. This latter finding indicates a mechanism involving trans-differentiation of other cell types or differentiation of long-lived totipotent stem cells.
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  • 49
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    Cell & tissue research 280 (1995), S. 541-548 
    ISSN: 1432-0878
    Keywords: Key words: Musle ; striated ; skeletal ; Regeneration ; Myosin ; Immunocytochemistry ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Indirect immunofluorescence was used to localize embryonic myosin heavy chains in soleus, adductor longus, tibialis anterior, plantaris, and extensor digitorum longus muscles of 6-month-old rats. A monoclonal antibody (2B6), specifically recognizing rat embryonic myosin, was applied to unfixed, transverse, frozen sections. The number of embryonic myosin-positive (EMP) extrafusal fibers was expressed as a percentage of the total number of fibers. EMP extrafusal fibers were only seen in the soleus and adductor longus muscles, both postural muscles. Approximately 1% of the soleus muscle fibers appeared positively stained for embryonic myosin. The majority of such fibers had a small diameter (〈500 μ2), appeared intensely fluorescent, and typically contained central nuclei. Re-expression of embryonic myosin due to spontaneous fiber denervation is not a likely factor in this study, since alpha-bungarotoxin and N-CAM localization were restricted to the motor end-plate region of EMP fibers. Since embryonic myosin was shown to disappear in all normal-sized myofibers by 2 to 3 months of age, the results suggest that the EMP extrafusal fibers seen in postural muscles of 6 to 12-month-old animals are regenerating myofibers. We speculate that a small number of muscle fibers may be regenerating in normal, adult postural muscles, in response to fiber damage possibly caused by excessive recruitment or overloading.
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  • 50
    ISSN: 1432-0878
    Keywords: Key words: AKH/RPCH peptide family ; Insect brain ; Corpora cardiaca ; Immunocytochemistry ; Enzyme immunoassay ; Periplaneta americana ; Leucophaea maderae (Insecta) ; Tegenaria atrica (Chelicerata)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. An antiserum against the octapeptide Pea-CAH-I, a member of the adipokinetic hormone/red pigment-concentrating hormone family, has been produced for immunocytochemical staining in insects and various other invertebrate species. The anti-Pea-CAH-I serum stains the glandular corpora cardiaca cells of those insect species that synthesize identical or structurally similar peptides. In the corpora cardiaca of species producing peptides with a different C-terminus, these cells remain unstained. Pea-CAH-I-like immunoreactivity has also been found in neurons of the central nervous system of all invertebrate orders studied. The antiserum recognizes the C-terminal sequence Pro-Asn-Trp-NH2 of the Pea-CAH-I molecule as established by enzyme immunoassay. The widespread Pea-CAH-I-like immunoreactivity in all nervous systems of the studied animals probably does not reflect the presence of Pea-CAH-I but the occurrence of peptides carrying similar epitopes.
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  • 51
    ISSN: 1432-0878
    Keywords: Key words: Melanin-concentrating hormone ; Immunocytochemistry ; Development ; ontogenetic ; Sparus auratus (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The development of the hypothalamic melanin-concentrating hormone (MCH) system of the teleost Sparus auratus has been studied by immunocytochemistry using an anti-salmon MCH serum. Immunoreactive perikarya and fibers are found in embryos, larvae, and juvenile specimens. In juveniles, most labeled neurons are present in the nucleus lateralis tuberis; some are dispersed in the nucleus recessus lateralis and nucleus periventricularis posterior. From the nucleus lateralis tuberis, MCH neurons project a conspicuous tract of fibers to the ventral hypothalamus; this penetrates the pituitary stalk and reaches the neurohypophysis. Most fibers end close to the cells of the pars intermedia, and some reach the adenohypophysial rostral pars distalis. Immunoreactive fibers can also be seen in extrahypophysial localizations, such as the preoptic region and the nucleus sacci vasculosi. In embryos, MCH-immunoreactive neurons first appear at 36 h post-fertilization in the ventrolateral margin of the developing hypothalamus. In larvae, at 4 days post-hatching, perikarya can be observed in the ventrolateral border of the hypothalamus and in the mid-hypothalamus, near the ventricle. At 26 days post-hatching, MCH perikarya are restricted to the nucleus lateralis tuberis. The neurohypophysis possesses MCH-immunoreactive fibers from the second day post-hatching. The results indicate that MCH plays a role in larval development with respect to skin melanophores and cells that secrete melanocyte-stimulating hormone.
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  • 52
    ISSN: 1432-0878
    Keywords: Key words: GABA (gamma-aminobutyric acid) ; Nervous system ; central ; Nervous system ; peripheral ; Immunocytochemistry ; Lumbricus terrestris (Annelida)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The distribution of neurons immunoreactive for γ-aminobutyric acid was studied in the nervous system of Lumbricus terrestris (Oligochaeta). In the cerebral ganglion, the 86 cells immunoreactive for γ-aminobutyric acid represented 4.0% of the nerve cells in the brain, had a diameter of 12–50 μm, and were arranged in seven groups. Small-sized (18–30 μm) immunoreactive neurons occurred in the circumpharyngeal connectives. The axons of most immunoreactive neurons of the cerebral ganglion richly arborized in the ventral part of the neuropil and some could also be traced in the circumpharyngeal connectives. The subesophageal ganglion contained 94 immunoreactive cells (6.7% of the cells of this ganglion), also divided into seven groups, and with a diameter of 8–55 μm. The axons of the labeled neurons ran to the central neuropil giving both contra- and ipsilateral processes. Altogether 108 neurons in each ganglion (8.0% of their cells) of the ventral cord were immunopositive. Four labeled cell groups were present in the rostral and caudal part of each ganglion. Axons of these immunoreactive cells arborized in the central neuropil and projected to the segmental nerves. The stomatogastric ganglia and the enteric plexus also contained immunoreactive neurons. Many small elongated immunoreactive cells occurred in the gut epithelium. Postembedding immunogold electron microscopy revealed that immunoreactive varicosities mainly contained small pleomorphic (24 nm) agranular synaptic vesicles and some small granular (50 nm) vesicles.
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  • 53
    ISSN: 1432-0878
    Keywords: Key words: Neuropeptide Y ; Gastroenteropancreatic (GEP) endocrine system ; Development ; ontogenetic ; Vitellointestinal duct ; Pancreas ; exocrine ; Pancreas ; endocrine ; Immunocytochemistry ; Scyliorhinus torazame (Elasmobranchii)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. This immunocytochemical study was carried out to elucidate the ontogenetic development of neuropeptide Y-like-immunoreactive cells in the gastroenteropancreatic endocrine system of the cloudy dogfish, Scyliorhinus torazame. Immunostained cells first appeared in the pancreas of the embryo at the 15-mm stage, and were also detected in the vitellointestinal duct of the yolk stalk at the 20-mm stage. These cells were polymorphic, with occasional processes that were sometimes directed toward the vascular wall or into the cavity of the vitellointestinal duct. At the 34-mm stage, immunostained cells could also be found in the proximal part of the spiral intestine and, by the 74-mm stage, immunopositive cells were present in the gastric mucosa. In the gut and pancreas, the cells gradually increased in number with development, whereas in the vitellointestinal duct and internal yolk sac, they decreased and seemed to disappear following hatching. Thus, in juveniles, the distribution of the neuropeptide Y-like-immunoreactive cells in the gastroenteropancreatic endocrine system had attained that of adults. Electron-microscopic immunocytochemistry demonstrated that, in the labeled cells of the vitellointestinal duct, the neuropeptide Y-like antigen was located in cytoplasmic granules, as in the cells of the gut and pancreas.
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  • 54
    ISSN: 1432-0878
    Keywords: Key words: Esophagus ; Epithelial cells ; Intestinal lectin ; L-36 ; RI-H fragment ; Immunocytochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Using an affinity purified antibody raised against the RI-H fragment of rat intestinal lectin L-36, the latter protein has been identified within the esophageal epithelium by means of ultracryotomy followed by immunogold labeling. The epithelium consists of 4 morphologically distinct cell-types, namely, the basal, spiny, granular and squamous cells, and each of these exhibits a different immunolabeling pattern. The basal cells form a layer on the basal lamina, and in these a diffuse cytoplasmic staining is observed. This basal cell layer is overlaid by spiny cells that extend many cell processes into wide intercellular spaces. In these cells, immunogold particles are found only on small granular inclusions consisting of an electron-lucent homogeneous substance. The granular cells form a third layer over the spiny cells, and are characterized by a number of large granular inclusions with an electron-dense core rimmed by a less electron-dense substance. Immunogold labeling is found on these granules, both on the core and peripheral region. Squamous cell-types constitute the most superficial layer of the epithelium. They are without granular inclusions, and immunogold labeling is confined to the cytoplasmic surface of the thickened plasma membrane. These findings suggest that L-36 is produced in the basal cells as free cytosolic protein, then becomes progressively aggregated into the granular inclusions of the spiny and granular cells, and is eventually transferred onto the cytoplasmic surface of the squamous cell plasma membrane where it may interact with complementary glycoconjugate(s) located at this site. The membrane lining substance thus formed may play a role in stabilizing the squamous cell membranes, thereby maintaining the structural integrity of the epithelium against mechanical stress coming from the esophageal lumen.
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  • 55
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    Cell & tissue research 283 (1996), S. 461-468 
    ISSN: 1432-0878
    Keywords: Key words: Gills ; Na+ ; K+-ATPase ; Immunocytochemistry ; DASPMI ; Cell culture ; Oncorhynchus mykiss (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The Na+,K+-ATPase (the sodium pump) plays a crucial role in ion transport in the fish gill. An immunocytochemical method has been optimized, using the mouse monoclonal antibody IgG α5, raised against the αsubunit of the avian sodium pump, to localize Na+,K+-ATPase in fish gill cells. The method appears to be successful for the immunolocalization of Na+,K+-ATPase in both paraffin-embedded gill tissue sections and primary cultures of fish gill epithelial cells. The immunostaining has demonstrated that Na+,K+-ATPase-positive cells are mainly localized on the primary lamellae, in the interlamellar region, which is in agreement with the distribution of ion-transporting cells, also called chloride cells, as shown by electron microscopy. Na+,K+-ATPase-positive cells have been demonstrated for the first time in primary cultures of gill epithelial cells. Comparative labeling studies of primary cultures have shown that sites of Na+,K+-ATPase-positive cells correspond to sites of cells labeled with dimethylaminostyrylmethyl-pyridiniumiodine, a fluorescent mitochondrial probe for ion-transporting cells. The immunocytochemical detection method for Na+,K+-ATPase in cells is proposed as an easy and specific Na+-transport-related method to characterize and localize ion-transporting cells in primary cultures and in tissue sections of fish gill epithelium.
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  • 56
    ISSN: 1432-0878
    Keywords: Key words: Met-enkephalin ; Opioids ; Immunocytochemistry ; Lymnaea stagnalis (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The distribution of an opioid peptide related to YGGFMRF was determined in the CNS and other organs of the pond snail, Lymnaea stagnalis, by RIA and immunocytochemistry. RIA revealed the highest levels in the CNS (1 pmol/organ) and penis (400 fmol/organ). There were also significant levels in the haemolymph, most of which was not associated with haemocytes (580 fmol/ml). Both serial section and whole-mount immunocytochemistry of the CNS revealed immunoreactive cells in every ganglion with the majority in the cerebral and pedal ganglia. In the pedal ganglia some of the immunoreactive cells were close to the cells of the A-cluster, which are known to respond to opioids, and could innervate them. In the cerebral ganglia the immunoreactive cells included a group of neurosecretory cells, the caudo dorsal cells (CDCs) and the terminals of these cells in the cerebral commissure were also stained. The CDCs secrete peptides into the haemolymph and so could be the source of the YGGFMRF immunoreactivity. Immunoreactivity (including the CDCs) was observed in locations that correspond to those reported for other fragments of proenkephalin, such as Met- and Leu-enkephalin, suggesting that they may share a common precursor, a Lymnaea proenkephalin. A map of the 358 YGGFMRF-immunoreactive cells in the CNS is presented, many of which have not been previously identified.
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  • 57
    ISSN: 1432-0878
    Keywords: Key words: Chromaffin cells ; Neurotrophic factor ; RT-PCR ; Immunocytochemistry ; Western blotting ; Rat (Hannover-Wistar) ; Bovine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Glial cell line-derived neurotrophic factor (GDNF) is a widely distributed member of the transforming growth factor-β superfamily and a potent neurotrophic molecule for several neuron populations in the peripheral and central nervous system. We show here that adrenal medullary chromaffin cells synthesize GDNF mRNA and contain immunoreactive GDNF protein. GDNF immunoreactivity can be found as early as embryonic day 16 in chromaffin progenitor cells of the rat adrenal gland and becomes more prominent with age. Most of the chromaffin cells within the adult rat adrenal medulla are GDNF immunoreactive, including both the noradrenergic and adrenergic subpopulations. The functions of adrenal medullary GDNF are still enigmatic but may include both auto/paracrine roles and retrograde trophic support of preganglionic neurons in the spinal cord or of sensory neurons that innervate chromaffin cells.
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  • 58
    ISSN: 1432-0878
    Keywords: Key words: Calsensin ; Calcium-binding protein ; Immunocytochemistry ; Neurons ; Leeches ; Hirudo medicinalis ; Haemopis marmorata ; Macrobdella decora (Annelida)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. By immunocytochemistry the distribution and developmental expression of the small EF-hand calcium-binding protein calsensin in the peripheral (PNS) and central nervous system (CNS) of the three hirudinid leech species Haemopis, Hirudo, and Macrobdella was compared. Labeling with calsensin-specific antibodies demonstrated that there was a pronounced difference in the distribution of calsensin immunoreactivity in the CNS of these leeches. In Haemopis more than 70 neurons were labeled, whereas the number in Hirudo was 51 and in Macrobdella only 8. Furthermore, the expression of calsensin in identified cells common to all three leech species also differed. Immunoblot analysis indicated that this variability was not likely to be due to multiple proteins or isoforms being recognized by the calsensin antibody. Labeling of embryos in various stages of development shows that the ontogeny of calsensin expression in the CNS is a gradual process with some neurons expressing calsensin immediately after completion of neurogenesis, about one-third of the way through embryogenesis, and others expressing calsensin only postembryonically. In contrast to the variability in the pattern and temporal expression by CNS neurons, the early embryonic calsensin expression in a small subgroup of sensillar PNS neurons was a shared feature by all three leech species. These findings suggest that calsensin may have different functional properties in CNS and PNS neurons.
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  • 59
    ISSN: 1432-0878
    Keywords: Key words Estrogen receptor ; Progesterone receptor ; Gonadotropin-releasing hormone ; Preoptic area ; Hypothalamus ; Immunocytochemistry ; Mink (Mustela vison)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The distribution of gonadal steroid (estrogen, progesterone) receptors in the brain of the adult female mink was mapped by immunocytochemistry. Using a monoclonal rat antibody raised against human estrogen receptor (ER), the most dense collections of ER-immunoreactive (IR) cells were found in the preoptic/anterior hypothalamic area, the mediobasal hypothalamus (arcuate and ventromedial nuclei), and the limbic nuclei (amygdala, bed nucleus of the stria terminalis, lateral septum). Immunoreactivity was mainly observed in the cell nucleus and a marked heterogeneity of staining appeared from one region to another. A monoclonal mouse antibody raised against rabbit uterine progesterone receptor (PR) was used to identify the PR-IR cells in the preoptic/anterior hypothalamic area and the mediobasal hypothalamus (arcuate and ventromedial nuclei). This study also focused on the relationship between cells containing sex-steroid receptors and gonadotropin-releasing hormone (GnRH) neurons on the same sections of the mink brain using a sequential double-staining immunocytochemistry procedure. Although preoptic and hypothalamic GnRH neurons were frequently in close proximity to perikarya containing ER or PR, they did not themselves possess receptor immunoreactivity. The present study provides neuroanatomical evidence that GnRH cells are not the major direct targets for gonadal steroids and confirms for the first time in mustelids the results previously obtained in other mammalian species.
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  • 60
    ISSN: 1432-0878
    Keywords: Key words Immunochemistry ; Chromatography ; Immunocytochemistry ; PACAP ; VIP ; CGRP ; NOS ; GRP ; Gastointestinal tract ; Capsaicin ; Denervation ; Rat (Sprague Dawley ; Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The expression of pituitary adenylate cyclase activating polypeptide (PACAP) was studied in the gastrointestinal tract (GI-tract) of normal rats using radioimmunoassay, chromatography, immunocytochemistry, and in situ hybridization. PACAP-38, PACAP-27, and PACAP-related peptide were demonstrated in all parts of the GI-tract, PACAP-38 being the predominant form confirmed by chromatography. PACAP-immunoreactive nerve fibers and nerve cell bodies were found in the myenteric ganglia throughout the GI-tract. PACAP-containing nerve cell bodies were also demonstrated in the submucous ganglia of the small and large intestine. The synthesis of PACAP in intrinsic neurons was confirmed by in situ hybridization. Double immunostaining showed that PACAP is present in calcitonin gene-related peptide-containing sensory nerve fibers as well as in vasoactive intestinal polypeptide (VIP)- or VIP/gastrin-releasing peptide (GRP)-containing (intramural) nerve fibers in the upper GI-tract and in anally projecting, intrinsic VIP-and VIP/nitric oxide syntase-containing nerve cell bodies and nerve fibers in the small and large intestine. Neonatal treatment with capsaicin significantly reduced the concentration of PACAP-38 in the esophagus, stomach, and colon. Extrinsic denervation decreased the PACAP-38 concentration in the stomach, while no change was observed in the small intestine. These results indicate that PACAP- immunoreactive nerve fibers in the GI-tract originate from both intrinsic (enteric) and extrinsic (presumably sensory) sources suggesting that PACAP may have diverse gastrointestinal functions.
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  • 61
    ISSN: 1432-0878
    Keywords: Key words: Neurofilaments ; Phosphorylation ; Differentiation ; Immunocytochemistry ; Brain storage ; Fixation ; Microwave ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Neurofilaments, which are exclusively found in nerve cells, are one of the earliest recognizable features of the maturing nervous system. The differential distribution of neurofilament proteins in varying degrees of phosphorylation within a neuron provides the possibility of selectively demonstrating either somata and dendrites or axons. Non-phosphorylated neurofilaments typical of somata and dendrites can be visualized with the aid of monoclonal antibody SMI 311, whereas antibody SMI 312 is directed against highly phosphorylated axonal epitopes of neurofilaments. The maturation of neuronal types, the development of area-specific axonal networks, and the gradients of maturation can thus be demonstrated. Optimal immunostaining with SMI 311 and SMI 312 is achieved when specimens are fixed in a mixture of paraformaldehyde and picric acid for up to 3 days and sections are incubated free-floating. Neurons, with their dendritic domains immunostained by SMI 311 in a Golgi-like manner, can be completely visualized in relatively thick sections. The limitations of Golgi-preparations, such as glia-labeling, artifacts, and the staining of only a small non-representative percentage of existing neurons, are not apparent in SMI preparations, which additionally provide the possibility of selectively staining axonal networks. The results achieved in normal fetal brain provide the basis for studies of developmental disturbances.
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  • 62
    ISSN: 1432-0878
    Keywords: Key words Annexin 5 ; Immunocytochemistry ; Pituitary ; Ovary ; Testis ; Adrenal gland ; Thyroid gland ; Rat (wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Annexin 5, a unique calcium- and phospholipid-binding protein, has been investigated for its specific distribution in rat endocrine organs by immunocytochemistry with a specific antiserum to recombinant rat annexin 5. Follicular epithelial cells and parafollicular cells of the thyroid gland, adrenocortical cells of the zona fasciculata and zona reticularis, luteal cells, testicular interstitial cells, and Sertoli cells were shown to contain annexin 5. To examine whether the synthesis of annexin 5 would be affected by a change in humoral signal, the distribution of annexin 5 in the anterior pituitary was examined three weeks after ovariectomy. The withdrawal of ovarian hormones induced huge castration cells in the anterior pituitary gland, which contained abundant annexin 5. Annexin 5 was not detected in the pineal gland, the parathyroid gland, the islet of Langerhans, the adrenal medulla, zona glomerulosa cells, and granulosa cells. Since annexin 5 was shown to exist in many of the endocrine tissues examined, to be localized in specific cell types, and to be abundant in castration cells, it is suggested that annexin 5 contributes to secretory cell functions, which may be common to endocrine cells secreting chemically different hormones.
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  • 63
    ISSN: 1432-0878
    Keywords: Key words Co-expression of mRNAs ; Gonadotropin-releasing hormone ; Immunocytochemistry ; In situ hybridization ; Oncorhynchus nerka (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The localization of two salmon-type gonadotropin-releasing hormone (sGnRH) precursors, pro-sGnRH-I (short type) and pro-sGnRH-II (long type), was investigated by using in situ hybridization techniques in the brain of the landlocked sockeye salmon, Oncorhynchus nerka. We used 30-mer oligonucleotide probes complementary to pro-sGnRH-I and pro-sGnRH-II cDNA. No significant differences were observed in the localization of sGnRH neurons expressing pro-sGnRH-I and pro-sGnRH-II mRNAs; both were expressed in the olfactory nerve, the olfactory bulbs, the regions between the olfactory bulb and telencephalon, the ventral telencephalon, the preoptic area, and the hypothalamus. Almost all sGnRH neurons examined co-expressed both precursors. The expression of two sGnRH precursors in the same neuron and the wide distribution of such neurons in the brain suggest that there are no functional differences between the two precursors.
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  • 64
    ISSN: 1432-0878
    Keywords: Key words Pancreas ; exocrine ; Hibernation ; Amylase ; Immunocytochemistry ; Muscardinus avellanarius (Rodentia)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Pancreatic acinar cells of euthermic, hibernating and arousing individuals of the hazel dormouse Muscardinus avellanarius (Gliridae) have been observed at the electron-microscopic level and analysed by means of ultrastructural morphometry and immunocytochemistry in order to investigate possible fine structural changes of cellular components during periods of strikingly different degrees of metabolic activity. During hibernation, the cisternae of the rough endoplasmic reticulum (RER) flatten assuming a parallel pattern, the Golgi apparatus is extremely reduced and the mitochondria contain many electron-dense particles. The cell nuclei appear irregularly shaped, with deep indentations containing small zymogen granules. They also contain abundant coiled bodies and unusual constituents, such as amorphous bodies and dense granular bodies. Large numbers of zymogen granules occur in all animals. However, the acinar lumina are open and filled with zymogen only in euthermic animals, whereas, in hibernating and arousing individuals, they appear to be closed. Morphometrical analyses indicate that, in pancreatic acinar cells, nuclei and zymogen granules significantly decrease in size from euthermia to hibernation, probably reflecting a drastic decrease of metabolic activities, mainly protein synthesis and processing. In all the studied animals, immunocytochemistry with specific antibodies has revealed an increasing gradient in α-amylase content along the RER-Golgi-zymogen granule pathway, reflecting the protein concentration along the secretory pathway. Moreover, during deep hibernation, significantly larger amounts of α-amylase accumulate in RER and zymogen granules in comparison to the other seasonal phases analysed. Upon arousal, all cytoplasmic and nuclear constituents restore their euthermic aspect and all morphometrical and immunocytochemical parameters exhibit the euthermic values, thereby indicating a rapid resumption of metabolic activities.
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  • 65
    ISSN: 1432-0878
    Keywords: Key words Adrenomedullin ; Pancreas ; Development ; Immunocytochemistry ; Colocalizations ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Adrenomedullin is an α-amidated 52-amino acid peptide involved in many physiological actions, among others the regulation of insulin secretion. Using immunohistochemical methods, we found that adrenomedullin immunoreactivity first appears at day 11.5 of embryonic development in the rat, coinciding with the appearance of pancreatic glucagon. The early appearance of adrenomedullin in the developing pancreas may indicate an active involvement in either the morphogenesis of the organ or its endocrine/paracrine/autocrine hormone regulation during intrauterine life. We also investigated the pattern of colocalizations of adrenomedullin with the other pancreatic hormones. At some point during development all the cell types express adrenomedullin, progressively evolving towards the adult pattern where only the pancreatic polypeptide cells contain a strong immunoreactivity for adrenomedullin. At this point the remaining cells of the islet are, in general, weakly stained. This sequential and time-dependent expression of adrenomedullin suggests a tight regulation similar to that observed for other modulatory substances responsible for embryonic morphogenesis.
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  • 66
    ISSN: 1432-0878
    Keywords: Key words Fish ; Zebrafish ; Immunocytochemistry ; Keratin ; Cytoskeleton ; Danio rerio (Teleostei)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  We have identified a number of type I and type II keratins in the zebrafish Danio rerio by two-dimensional polyacrylamide gel electrophoresis, complementary keratin blot-binding assay and immunoblotting. These keratins range from 56 kDa to 46 kDa in molecular mass and from pH 6.6 to pH 5.2 in isoelectric point. Type II zebrafish keratins exhibit significantly higher molecular masses (56–52 kDa) compared with the type I keratins (50–48 kDa), but the isoelectric points show no significant difference between the two keratin subclasses (type II: pH 6.0–5.5; type I: pH 6.1–5.2). According to their occurrence in various zebrafish tissues, the identified keratins can be classified into “E” (epidermal) and “S” (simple epithelial) proteins. A panel of monoclonal anti-keratin antibodies has been used for immunoblotting of zebrafish cytoskeletal preparations and immunofluorescence microscopy of frozen tissue sections. These antibodies have revealed differential cytoplasmic expression of keratins; this not only includes epithelia, but also a variety of mesenchymally derived cells and tissues. Thus, previously detected fundamental differences in keratin expression patterns between higher vertebrates and a salmonid, the rainbow trout Oncorhynchus mykiss, also apply between vertebrates and the zebrafish, a cyprinid. However, in spite of notable similarities, trout and zebrafish keratins differ from each other in many details. The present data provide a firm basis from which the application of keratins as cell differentiation markers in the well-established genetic model organism, the zebrafish, can be developed.
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  • 67
    ISSN: 1432-0878
    Keywords: Key words: Leu-callatostatins ; Callatostatins ; Allatostatins ; Neuropeptides ; Peptide purification ; Immunocytochemistry ; Myoinhibition ; Codling moth ; Cydia pomonella (Tortricidae: Lepidoptera) (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. A neuroendocrine peptide of the Leu-callatostatin family, LPVYNFGL-NH2, has been isolated from tissue extracts of 5th instar larvae of the codling moth, Cydia pomonella (Lepidoptera). It is identical to a peptide previously isolated from the blowfly, Calliphora vomitoria (Diptera). The distribution of this peptide within the tissues of C. pomonella has been mapped by immunocytochemistry using antisera raised against LPVYNFGL-NH2. Midgut endocrine cells contain Leu-callatostatin immunoreactivity, as do several paired Leu-callatostatin neurones in the brain and ventral nerve cord. Within the visceral nervous system, the frontal ganglion contains four Leu-callatostatin neurones. Axons from these cells combine with others originating from neurones in the brain and project within the nervi cardiostomatogastrici to innervate the tissues of the foregut. In particular, the oesophageal valve has a prominent ring of Leu-callatostatin-immunoreactive fibres. The synthetic peptide, LPVYNFGL-NH2, has a potent reversible inhibitory effect in vitro on all visible forms of spontaneous contractile activity of the foregut, including closure of the oesophageal valve. Complete myoinhibition is observed at peptide concentrations from 10−10 to 10−16 M. These results, in conjunction with the results of similar studies on cockroaches, crickets and flies, suggest that the Leu-callatostatins are a ubiquitous family of insect neuroendocrine peptides with an important role in the control of gut motility.
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  • 68
    ISSN: 1432-0878
    Keywords: Key words: Pineal organ ; Pinealocytes ; Pineal photoreceptors ; Sensory structures ; Photopigment ; Pinopsin ; Immunocytochemistry ; Chicken
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Pinopsin is a photoreceptive molecule cloned from the chicken pineal organ. An antibody highly specific for pinopsin was applied in light- and electron-microscopic immunocytochemical studies of the pineal organ of 1 to 2-month-old chickens. Intense immunoreactivity was found in the follicular lumen at the light-microscopic level. In addition, small immunoreactive spherical or fibrous structures were diffusely distributed at the parafollicular aspect of the pineal organ. To identify immunoreactive elements precisely, we used pre-embedding immunoelectron microscopy. These studies revealed immunoreactive outer segments of pinealocytes arranged closely side by side in the follicular lumina. The thin initial portion of the outer segment arose from a basal body located in the inner segment. Immunoreactive pear-shaped outer segments occupied small lumina. Follicular lumina displayed immunonegative arrays of whorl-like lamellar membranes. Occasionally, these immunonegative structures were surrounded by immunoreactive concentric lamellar complexes. In the parafollicular pineal parenchyma, long slender cilium-like structures or enlarged cilia and concentric lamellar arrays showed intense immunoreactivity. All immunoreactive structures observed in this study were considered to represent outer segments of pinealocytes of the chicken pineal organ.
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  • 69
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    Cell & tissue research 289 (1997), S. 265-274 
    ISSN: 1432-0878
    Keywords: Key words: Retina ; Amacrine cells ; Neurotransmitters ; Immunocytochemistry ; Tyrosine hydroxylase ; Dopamine ; Ichthyophis kohtaoensis (Gymnophiona)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Ichthyophis kohtaoensis, a member of the limbless Gymnophiona, has a specialized subterranean burrowing mode of life and a predominantly olfactory-guided orientation. The only visually guided behavior seems to be negative phototaxis. As these animals possess extremely small eyes (only 540 μm in diameter in adults), functional investigations of single retinal cells by electrophysiological methods have so far failed. Therefore, the content and distribution of retinal transmitters have been investigated as indications for a functioning sense organ in an animal that is supposed to be blind. In this study, the organization and development of the dopaminergic system have been examined in the retinae of embryonic, larval, and adult I. kohtaoensis, by using an antiserum against tyrosine hydroxylase, the rate-limiting enzyme in the catecholamine synthetic pathway, and an antiserum against dopamine itself. Labeled somata are situated in the inner nuclear layer and in the ganglion cell layer. Dopamine-positive fibers form a dense diffuse plexus, that covers the whole inner plexiform layer, whereas tyrosine hydroxylase-immunoreactive processes show a tendency to arborize in a stratified manner. Tyrosine-hydroxylase-immunolabeled fibers can occasionally be observed in the optic nerve head of larval stages. During ontogenesis and larval development, the distribution of transmitter-expressing cells changes and their number decreases, but no general degeneration of the visual system is detectable. Adult Ichthyophis still have retinal transmitters, indicating that the eyes, although obviously playing a minor role in a subterranean ecological niche, retain all the elements of functioning sense organs.
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  • 70
    ISSN: 1432-0878
    Keywords: Key words: Intracardiac neurones ; Innervation ; Heart ; Neuropeptide Y ; Immunocytochemistry ; Electron microscopy ; Rat (Sprague Dawley) ; Guinea-pig (Dunkin Hartley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Intrinsic neuropeptide Y-containing neurones in rat and guinea-pig hearts were studied at the ultrastructural level by the pre-embedding peroxidase-antiperoxidase immunocytochemical technique. Intracardiac neuronal cell bodies were often weakly or moderately immunostained, and the labelling was usually pronounced in the Golgi complex, multivesicular bodies, some cisterns of granular endoplasmic reticulum and large granular vesicles. Neuropeptide Y-immunoreactive nerve fibres were also observed in association with intracardiac neurones. A subpopulation of neuropeptide Y-immunoreactive granule-containing cells in the rat heart are described for the first time and were very heavily labelled; other granule-containing cells were non-immunoreactive, but were contacted by neuropeptide Y-containing nerves. Preterminal regions of nerve fibres that were located in nerve bundles were only weakly neuropeptide Y-immunoreactive, in contrast to the heavy labelling observed in varicosities that contained many synaptic vesicles. Many neuropeptide Y-immunoreactive nerve fibres were associated with the coronary vasculature and were particularly prominent in the walls of small arteries and arterioles where labelled nerve varicosities were present close to the smooth muscle cells. Immunoreactive nerves were also seen in the myocardium, usually near to capillaries. In axonal varicosities, the central core of large granular vesicles was immunolabelled, and electron-dense immunoreactive material outlined the membranes of small and large clear vesicles. The significance of neuropeptide Y-immunoreactive intracardiac neurones and granule-containing cells and the origin of associated labelled nerve fibres in the heart are discussed.
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  • 71
    ISSN: 1432-0878
    Keywords: Key words Histamine ; Immunocytochemistry ; Mast cells ; Melanocytes ; Nerves ; Rana esculenta (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  The number, distribution, and ultrastructural characteristics of mast cells were assessed in the tongue, heart, and kidney of the frog Rana esculenta. The density of tongue mast cells (253±45 mast cells/mm2) was significantly higher than that of the heart (5.3±0.4/mm2) and kidney (15.3±1.4 /mm2). A striking feature of this study was the remarkable association of frog mast cells to nerves. The ultrastructural study of the mast cell/nerve association demonstrated that mast cells were closely apposed to or even embedded in nerves. Mast cells were also physically associated with melanocytes even in the heart. Mast cells were Alcian blue+/safranin+ in the tongue and in the peritoneum, whereas in the heart and in the kidney they were Alcian blue–/safranin+. The mast cells in the lamina propria of the gastrointestinal tract were Alcian blue+/safranin–. The cytoplasm of frog mast cells was packed with numerous heterogeneous, membrane-bound granules. The ultrastructure of these cytoplasmic granules was unique, being totally unlike any other previously described granules in other animal species as well as in man. The histamine content/frog mast cell (≈0.1 pg/cell) was approximately 30 times lower than that of human mast cells isolated from different tissues (≈3 pg/cell). A monoclonal anti-histamine antibody was used to confirm the ultrastructural localization of histamine in secretory granules in frog mast cells.
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  • 72
    ISSN: 1432-0878
    Keywords: Key words Pancreas ; Stomach ; Duodenum ; Ribonuclease protection assay ; Immunocytochemistry ; Protease ; Rat ; (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Prior studies have revealed the presence of chymotrypsinlike protease in peripheral organs, although no definitive evidence for the synthesis of this enzyme in tissue other than the pancreas is available. In an attempt to detect chymotrypsinogen mRNA in peripheral organs, a fragment of the pancreatic chymotrypsin mRNA from rat was amplified using PCR. The sequence was identified as a portion of the rat chymotrypsin B gene overlapping exon 5 through exon 7. It was subcloned into the pGEM-4Z vector and used as a template for the vitro transcription of an antisense riboprobe. Using ribonuclease protection and Northern blot analyses, chymotrypsin mRNA was detected in the rat pancreas, stomach, duodenum, ovary, and spleen. Monoclonal and polyclonal antisera against chymotrypsin detected chymotrypsinlike immunoreactivity in rat and human pancreas, rat stomach, duodenum and jejunum. Electrophoresis and immunoblotting revealed chymotrypsin-chymotrypsinogen bands (25–29 kDa) in the stomach and duodenum. Synthesis of a potent protease such as chymotrypsin in tissue other than pancreas is significant, suggesting a potential physiological and/or pathological role in these tissues.
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  • 73
    ISSN: 1432-0878
    Keywords: Key words Acrosome ; Epididymal maturation ; Monoclonal antibody ; Immunocytochemistry ; Spermatozoa ; Guinea pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  We have previously shown that a 90-kDa intra-acrosomal antigen, MN7, is restricted to the anterior acrosomal region of mouse, rat, and hamster spermatozoa. The present study has examined the localization and the behavior of MN7 during sperm maturation in the epididymis of the guinea pig by immunoelectron microscopy. MN7 showed not only a specific localization in the apical segment of the guinea pig sperm acrosome, but also a distinct alteration during maturation, as follows. MN7 was exclusively found both at the dorsal matrix and on the outer acrosome membrane (OAM)/matrix-associated materials in the apical segment. MN7 was initially distributed throughout the electron-lucent dorsal matrix in immature sperm but, during maturation, became more restricted to the spherical bodies within the electron-lucent area. MN7 on OAM/matrix-associated materials was first distributed along the ventral margin and the small area posterior to the dorsal matrix but, during maturation, disappeared from the ventral margin and became restricted to the dorsal region. These results indicate that MN7 is a good tool for studying the stepwise maturation of epididymal spermatozoa.
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  • 74
    ISSN: 1432-0878
    Keywords: Key words: Epididymis ; Efferent ducts ; Cell culture ; Immunocytochemistry ; Immunoprecipitation ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The way in which the human epididymis modifies spermatozoa during their sojourn in this structure might be clarified by knowledge of the nature of its secretions. We have examined the presence of several lysosomal hydrolases in human epididymal tissue and fluids, and their synthesis and secretion by monolayer cultures. Tissues were obtained from men undergoing orchidectomy for prostatic carcinoma. The enzymes cathepsin D and acid α-glucosidase were localised in the lysosomes of epithelial cells from the corpus epididymidis, by an immunocytochemical technique. Cathepsin D was also found in epithelial cells of the efferent ducts within lysosomes, apical vesicles and multivesicular bodies. No immunolocalisation of acid glucosidase in the efferent ducts or on the microvilli of the corpus was demonstrable. Cathepsin D, β-hexosaminidase (N-acetylglucosaminidase) and α-glucosidase were measurable in the luminal fluid from the human corpus epididymidis; β-hexosaminidase was secreted into the culture medium by confluent monolayers of epididymal and efferent duct cells. Immunoprecipitation of cell extracts and culture medium of these cultures incubated with 35S-methionine revealed that the precursors of cathepsin D and β-hexosaminidase were synthesized and secreted by such monolayers. Thus, active lytic enzymes are secreted by the human epididymis and could modify sperm membranes.
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  • 75
    ISSN: 1432-0878
    Keywords: Compound eye ; Photoreceptor cells ; Ion pumps ; Polarity ; Immunocytochemistry ; Manduca sexta (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Immunohistochemical and physiological studies on various insect photoreceptors have demonstrated that the Na,K-ATPase (sodium pump) is restricted to the nonreceptive nonmicrovillar area of the plasma membrane. Here, we examined the distribution of the Na,K-ATPase in photoreceptor cells of the superposition-type compound eye in the moth Manduca sexta. Using immunofluorescent and immunogold cytochemistry, we show that the Na,K-ATPase is localized to both the nonmicrovillar and the microvillar parts of the plasma membrane. Manduca photoreceptors thus deviate from the common concept that the sodium pump and the molecular components of the photoreceptive machinery reside on different domains of the plasma membrane.
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  • 76
    ISSN: 1432-0878
    Keywords: Neurohemal areas ; Neuropeptides ; Monoamines ; Immunocytochemistry ; Nervous system, insect ; Gryllus bimaculatus (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The morphology and position of putative neurohemal areas in the peripheral nervous system (ventral nerve cord and retrocerebral complex) of the cricket Gryllus bimaculatus are described. By using antisera to the amines dopamine, histamine, octopamine, and serotonin, and the neuropeptides crustacean cardioactive peptide, FMRFamide, leucokinin 1, and proctolin, an extensive system of varicose fibers has been detected throughout the nerves of all neuromeres, except for nerve 2 of the prothoracic ganglion. Immunoreactive varicose fibers occur mainly in a superficial position at the neurilemma, indicating neurosecretory storage and release of neuroactive compounds. The varicose fibers are projections from central or peripheral neurons that may extend over more than one segment. The peripheral fiber varicosities show segment-specific arrangements for each of the substances investigated. Immunoreactivity to histamine and octopamine is mainly found in the nerves of abdominal segments, whereas serotonin immunoreactivity is concentrated in subesophageal and terminal ganglion nerves. Immunoreactivity to FMRFamide and crustacean cardioactive peptide is widespread throughout all segments. Structures immunoreactive to leucokinin 1 are present in abdominal nerves, and proctolin immunostaining is found in the terminal ganglion and thoracic nerves. Codistribution of peripheral varicose fiber plexuses is regularly seen for amines and peptides, whereas the colocalization of substances in neurons has not been detected for any of the neuroactive compounds investigated. The varicose fiber system is regarded as complementary to the classical neurohemal organs.
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  • 77
    ISSN: 1432-0878
    Keywords: Interleukin ; Stellate reticulum ; Immunocytochemistry ; Epidermal growth factor ; Interleukin-1 receptor type I messenger RNA ; Tooth eruption ; Rat (Sprague Dawley)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Immunolocalization of interleukin-1α in the first mandibular molars of rats from day 0–12 postnatally showed that the protein was localized in the epithelial stellate reticulum adjacent to the dental follicle. Staining of the stellate reticulum was most prominent in the early days postnatally and was absent by postnatal day 11. Injection of epidermal growth factor into rats at day 0 greatly increased the intensity of the staining for interleukin-1α in the stellate reticulum. Epidermal growth factor (EGF) enhanced the gene expression of interleukin-1α in stellate reticulum cells in vitro, and this study suggests there is enhanced translation of interleukin-1α messenger RNA in the stellate reticulum following EGF injection. In turn, the interleukin-1α may exert its effect on the dental follicle cells adjacent to the stellate reticulum because EGF also enhanced expression of the interleukin-1 receptor type I messenger RNA in cultured dental follicle cells as well as enhancing its expression in vivo. In view of the fact that injection of EGF will stimulate precocious eruption of teeth, its stimulus of interleukin-1α synthesis in the stellate reticulum may be the mechanism by which EGF initiates a cascade of molecular events to signal the onset of tooth eruption.
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  • 78
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    Cell & tissue research 285 (1996), S. 165-169 
    ISSN: 1432-0878
    Keywords: Key words: Ciliary axoneme ; Cones ; Cytoskeleton ; Immunocytochemistry ; Microtubules ; Outer segments ; Photoreceptor cells ; Xenopus laevis (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The ciliary axoneme in photoreceptors from the retina of Xenopus laevis was examined by immunofluorescent staining of tubulin throughout the light/dark cycle. The immunofluorescent axoneme extended along only part of the length of rod outer segments but the entire length of cone outer segments. Both the cone axoneme and outer segment elongated during the day and shortened (presumably by shedding) during the night. Fragments of immunofluorescent axonemes were found within packets of outer segment material breaking off from cone tips. These findings show that the ciliary axoneme in the Xenopus retina is replaced during the renewal of outer segments in cones. No evidence has been found for renewal of the axoneme in rods, and thus the stability of the ciliary axoneme may differ in rod and cone photoreceptors.
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  • 79
    ISSN: 1432-0878
    Keywords: Key words: Galectin ; β-Galactoside-binding lectin ; Human ; Skin ; Immunocytochemistry ; Immunohistochemistry ; Hybridization ; in situ ; Langerhans cell ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The localization of an endogenous 14-kDa β-galactoside-binding lectin (galectin) and its pattern of gene expression were examined in normal human skin by light- and electron microscopy. Under the light microscope, immunostaining of 14-kDa galectin was observed in the cell membrane of cells in the basal and spinous layers of the epidermis. Galectin was also found in the Langerhans cells, as shown by double labeling using anti-14-kDa galectin and anti-CD1a antibodi es. In the dermis, immunostaining for the 14-kDa galectin was positive in the extracellular matrix and fibroblasts. At the electron-microscopic level of resolution, galectin was located primarily along the plasma membrane of keratinocytes, and in both the cytoplasm and nucleus of Langerhans cells in the epidermis, whereas in the dermis it was detected in the extracellular matrix and in both the nucleus and cytoplasm of fibroblasts. The gene expression of 14-kDa galectin was visualized by the HRP-staining me thod following in situ hybridization techniques. The expression was detected in the cytoplasm of cells in the basal and spinous layers of the epidermis; whereas, in the dermis, it was detected in the cytoplasm of fibroblasts. Moreover, SDS-polyacrylamide gel electrophoresis and lectin-blot analysis revealed that this galectin bound to glycoproteins of approximately 17, 62, and 72 kDa in the epidermis and to those of 29, 54, and 220 kDa in the dermis. The present study indicates that 1) normal human skin produces the β-galactoside-binding 14-kDa galectin, and 2) this galectin is located in both the epidermis, particularly in the keratinocytes and Langerhans cells, and in the dermis. These results suggest that galectin is important for cell-cell contact and/or adhesion in the epidermis and for cell-extracellular matrix interaction in the dermis.
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  • 80
    ISSN: 1432-0878
    Keywords: Key words: Diuretic hormone ; Endocrine cells ; Midgut ; Malpighian tubules ; Bioassay ; Immunocytochemistry ; Locusta migratoria (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. This is an investigation of an endocrine cell type in the midgut of the migratory locust Locusta migratoria. This cell type is found in the posterior region of the midgut and is especially common in the ampullae through which Malpighian tubules drain into the gut at the midgut-hindgut junction. Strong Locusta diuretic hormone-like immunoreactivity in these cells was colocalized with FMRFamide- and substance P-like immunoreactivities. At the ultrastructural level, immunoreactivity for Locusta diuretic hormone was found in spherical granules (mean diameter of 450 nm), the contents of which showed variable electron density. Fractionation of a methanolic extract of the ampullae by reversed-phase high performance liquid chromatography revealed the presence of two peaks of Locusta diuretic hormone-like immunoreactive material, both of which stimulate cyclic AMP production by isolated Malpighian tubules. The more hydrophobic material is most likely Locusta diuretic hormone, which has the same retention time when chromatographed under identical conditions.
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  • 81
    ISSN: 1432-0878
    Keywords: Galectin ; β-Galactoside-binding lectin ; Human ; Skin ; Immunocytochemistry ; Immunohistochemistry ; Hybridization, in situ ; Langerhans cell ; Man
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The localization of an endogenous 14-kDa β-galactoside-binding lectin (galectin) and its pattern of gene expression were examined in normal human skin by light- and electron microscopy. Under the light microscope, immunostaining of 14-kDa galectin was observed in the cell membrane of cells in the basal and spinous layers of the epidermis. Galectin was also found in the Langerhans cells, as shown by double labeling using anti-14-kDa galectin and anti-CD1a antibodies. In the dermis, immunostaining for the 14-kDa galectin was positive in the extracellular matrix and fibroblasts. At the electron-microscopic level of resolution, galectin was located primarily along the plasma membrane of keratinocytes, and in both the cytoplasm and nucleus of Langerhans cells in the epidermis, whereas in the dermis it was detected in the extracellular matrix and in both the nucleus and cytoplasm of fibroblasts. The gene expression of 14-kDa galectin was visualized by the HRP-staining method following in situ hybridization techniques. The expression was detected in the cytoplasm of cells in the basal and spinous layers of the epidermis; whereas, in the dermis, it was detected in the cytoplasm of fibroblasts. Moreover, SDS-polyacrylamide gel electrophoresis and lectin-blot analysis revealed that this galectin bound to glycoproteins of approximately 17, 62, and 72 kDa in the epidermis and to those of 29, 54, and 220 kDa in the dermis. The present study indicates that 1) normal human skin produces the β-galactoside-binding 14-kDa galectin, and 2) this galectin is located in both the epidermis, particularly in the keratinocytes and Langerhans cells, and in the dermis. These results suggest that galectin is important for cell-cell contact and/or adhesion in the epidermis and for cell-extracellular matrix interaction in the dermis.
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  • 82
    ISSN: 1432-0878
    Keywords: Defence mechanisms ; Encapsulation ; Granulocytes ; Immunocytochemistry ; Parasitism ; Perkinsus sp. (Protozoa) ; Tapes semidecussatus (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Parasitosis by the trophozoite protozoan Perkinsus sp. (Apicomplexa, Perkinsea) induces in the gill filaments of the clam Tapes semidecussatus (Mollusca, Bivalvia) a cellular reaction, which is constituted by infiltrated granulocytes. This cellular reaction has characteristics of those of a holocrine gland, since the parasites are encapsulated by the secretion product of the granulocytes after cell death. An enriched fraction of prezoosporangia and their associated capsule was obtained after culture of the parasitized gills in fluid thioglycollate medium. Specific polypeptides from this fraction were separated by SDS-PAGE and isolated for rabbit immunizations. The serum obtained against an Mr 225 kDa polypeptide, revealed its exclusive localization in the capsule and in the granules of the infiltrated granulocytes, thus indicating that this polypeptide is synthesized by these cells and secreted, in a polarized way, around the trophozoites resulting in their encapsulation. Selective deglycosylation of the polypeptide, by Endo H and alkaline β-elimination, did not show an effect on its molecular weight or antibody recognition. Furthermore, the absence of the 225 kDa band in the Western-blots of non-parasitized gills indicated the specific association of this polypeptide with the parasitosis. Finally, this is the first tissue-specific factor described in molluscs in relation to defence mechanisms.
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  • 83
    ISSN: 1432-0878
    Keywords: Oocyte ; Yolk granules ; Ribonuclease ; Immunocytochemistry ; Bullfrog, Rana catesbeiana (Anura)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract To determine the localization of the pyrimidine-guanine sequence-specific ribonuclease in Rana catesbeiana (bullfrog) oocytes, the RNase was first isolated and used to prepare a specific rabbit antiserum. Only one protein of similar molecular size to the RNase was immunoprecipitated from ovary homogenate by the antiserum, but two bands were observed by Western blotting analysis. These two proteins were shown by further purification of antibody and Western blotting analysis to have similar antigenicity. Immunoprecipitation and Western blotting of tissue homogenates showed that the RNase was found predominantly in the ovary, but not in other tissues. The specific localization of the RNase was determined by immuno-electron microscopy of oocyte sections incubated with the specific antiserum; the yolk granules, but not other organelles, were found to contain the RNase. Most of the RNase was evenly distributed in the lateral amorphous area of the yolk granule but not in the central yolk crystal area which contains stored vitellogenin proteins. Our results indicate that the RNase is compartmentalized in the yolk granules of oocytes, which might prevent damage to cellular RNAs.
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  • 84
    ISSN: 1432-0878
    Keywords: Esophagus ; Epithelial cells ; Intestinal lectin, L-36 ; RI-H fragment ; Immunocytochemistry ; Rat (Wistar)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Using an affinity purified antibody raised against the RI-H fragment of rat intestinal lectin L-36, the latter protein has been identified within the esophageal epithelium by means of ultracryotomy followed by immunogold labeling. The epithelium consists of 4 morphologically distinct cell-types, namely, the basal, spiny, granular and squamous cells, and each of these exhibits a different immunolabeling pattern. The basal cells form a layer on the basal lamina, and in these a diffuse cytoplasmic staining is observed. This basal cell layer is overlaid by spiny cells that extend many cell processes into wide intercellular spaces. In these cells, immunogold particles are found only on small granular inclusions consisting of an electron-lucent homogeneous substance. The granular cells from a third layer over the spiny cells, and are characterized by a number of large granular inclusions with an electron-dense core rimmed by a less electron-dense substance. Immunogold labeling is found on these granules, both on the core and peripheral region. Squamous cell-types constitute the most superficial layer of the epithelium. They are without granular inclusions, and immunogold labeling is confined to the cytoplasmic surface of the thickened plasma membrane. These findings suggest that L-36 is produced in the basal cells as free cytosolic protein, then becomes progressively aggregated into the granular inclusions of the spiny and granular cells, and is eventually transferred onto the cytoplasmic surface of the squamous cell plasma membrane where it may interact with complementary glycoconjugate(s) located at this site. The membrane lining substance thus formed may play a role in stabilizing the squamous cell membranes, thereby maintaining the structural integrity of the epithelium against mechanical stress coming from the esophageal lumen.
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  • 85
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    Cell & tissue research 281 (1995), S. 101-108 
    ISSN: 1432-0878
    Keywords: Somite ; Intergin ; Extracellular matrix, structures ; Embryo ; Laminin ; Immunocytochemistry ; Macaca fascicularis (Primates)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The distribution of integrin subunits α6 and β1, and the α6β1 integrin ligand, laminin, was examined during somitogenesis in developmental stages 11, 13, and 16 in the long-tailed macaque, using peroxidase immunocytochemistry. Within differentiating somites in stage 11, α6 expression was observed in the sclerotome, basal surface of dermamyotomal cells adjacent to the basal lamina and on scattered cells throughout the dermamyotome. In further advanced somites in stages 13 and 16, α6 immunoreactivity become restricted to the myotome, α6 was expressed on mesenchymal core cells within the myocele of undifferentiated epitheliod somites and the ventromedial wall of somites commencing differentiation at each stage. β1 distribution resembled that of α6 in stage 11 somitic tissue, however, it remained present on myotome and sclerotome cells in the later stages, and was also expressed on dermatomal cells in stage 16. Laminin immunoreactivity, while more intense and prevalent than α6 and β1 in each stage examined, occurred on the same somite cell populations as the 2 integrin subunits. These results show a defined distribution of α6 on somitic tissue, and suggest this integrin is involved in somite differentiation. They also support a possible role for α6 in myoblast formation and migration. Overlapping of β1 and laminin immunoreactivity with that of α6 further suggests that α6 paris with β1 as a functional heterodimer for laminin in defined somitic regions.
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  • 86
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    Cell & tissue research 286 (1996), S. 69-80 
    ISSN: 1432-0878
    Keywords: Key words: Growth hormone ; Growth hormone receptors ; Growth hormone-binding proteins ; Immune system ; Immunocytochemistry ; Domestic fowl (Aves ; Phasianiformes)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. It is well established that the activity and proliferation of lymphoid cells and lymphoid organs are stimulated by growth hormone. These actions on lymphoid cells may be direct or mediated by actions on the epithelial and non-immune tissue cells that regulate immune function. The occurrence and cellular localization of growth hormone receptors in immune tissues has therefore been investigated to determine the target-sites of growth hormone action. Growth hormone receptor mRNA was first detected by Northern blotting in the spleen, bursa of Fabricius, and thymus of domestic fowl. In addition to the 4.4-kb transcript thought to encode the full-length growth hormone receptor, smaller transcripts of 2.8 kb and 1.0 kb, which may encode growth hormone-binding proteins, were also occasionally observed. Further analysis using the polymerase chain reaction revealed that mRNA sequences encoding the extracellular and intracellular domains of the growth hormone receptor were present in all tissues and highly homologous with hepatic transcripts. Translation of these transcripts also occurs in immune tissues, since immunoreactive growth hormone-binding proteins or growth hormone receptors of approximately 56 kDa were detected in hepatic, splenic, thymic, and bursal extracts. Immunocytochemistry of these tissues subsequently revealed that macrophages probably contain the bulk of this immunoreactivity, although some thymic medullary epithelial cells (including Hassall’s corpuscles) and splenic ellipsoids and interdigitating cells were also immunoreactive. This immunoreactivity is present in immune tissues of newly hatched and adult chickens. Importantly, B-lymphocytes were rarely, if ever, immunoreactive, and T-lymphocytes containing growth hormone receptors or binding proteins were not observed. These results suggest that a number of primary (thymus and bursa) and secondary (spleen) lymphoid tissues in the chicken contain growth hormone receptors and are thus target-sites for growth hormone action. The distribution of growth hormone receptor/growth hormone-binding protein immunoreactivity in these tissues would further suggest that growth hormone plays a major role in macrophage proliferation and/or activity and may indirectly affect lymphocyte maturation and storage via effects on thymic and splenic stromal cells.
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  • 87
    ISSN: 1432-0878
    Keywords: Key words: GABA receptor ; RDL subunit ; Nervous system ; Immunocytochemistry ; Drosophila melanogaster (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. An antibody raised against a peptide based on the C-terminal derived amino acid sequence from a cloned Drosophila melanogaster (fruit fly) gene, Rdl (resistant to dieldrin), was used to investigate localization of a GABA receptor subunit in adult male D. melanogaster. Many regions in the brain and thoracic ganglia were stained with this antibody. For example, staining was detected in the medulla, lobula and lobular plate optic neurpiles. Also stained were the antennal lobe glomeruli, the ellipsoid body of the central complex and the mushroom bodies. These results suggest possible roles for an RDL-like GABA receptor subunit in the processing of olfactory, visual and mechanosensory information in the nervous system of D. melanogaster.
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  • 88
    ISSN: 1432-0878
    Keywords: Key words: Gap junction ; Intercellular junction ; Insect ; Arthropod ; Immunocytochemistry ; Heliothis virescens (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Gap junction-enriched fractions were prepared from larvae of the tobacco budworm Heliothis virescens using the NaOH procedure in the presence or absence of protease inhibitors and were analyzed by SDS-PAGE, immunoblotting and EM immunocytochemistry. Protease inhibitor fractions contained a 48-kDa protein in addition to the ∼10 proteins in fractions with and without inhibitors. Three polyclonal antibodies were used as probes for gap junction plaques and proteins: R16, against an ∼40-kDa candidate gap junction protein from Drosophila melanogaster; R17, against the 40-kDa candidate gap junction protein from H. virescens; and R18AP, an affinity purified antibody against a consensus sequence of N-terminal amino acids 2–21 of the H. virescens 40-kDa protein. R16, R17, and R18AP stain the 40- and 48-kDa proteins, R16 and R18AP stain a 64-kDa protein, and R16 stains an ∼30-kDa protein in the absence of inhibitors. Inclusion of protease inhibitors had no effect on gap junction ultrastructure. R16 and R17 label gap junction plaques in crude membrane and NaOH fractions, whereas R18AP exhibits only a low level of reactivity with gap junctions in crude membrane fractions and none with gap junctions in NaOH fractions. The results show that the 30-, 40-, 48- and 64-kDa proteins are immunologically related and are associated with gap junctions in H. virescens, the N-terminus of the 40-kDa protein is relatively inaccessible or easily lost, and the 48-kDa protein is protease-sensitive.
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  • 89
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    Cell & tissue research 281 (1995), S. 507-515 
    ISSN: 1432-0878
    Keywords: Microtubules ; Isoforms ; Nervous system ; Locomotion ; Cilia ; Immunocytochemistry ; Western blotting ; Lymnaea stagnalis (Mollusca)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Immunocytochemistry and Western blotting techniques demonstrated that the nervous system and foot of the pond snail Lymnaea stagnalis are rich sources of tubulin, which can be extracted and assembled in vitro in the presence of taxol. Various broad-spectrum antibodies raised against α-tubulin and β-tubulin yielded qualitatively similar results. One monoclonal antibody to trypanosome α-tubulin, however, labelled α-tubulin more strongly on both probed sections and Western blots. Cytochemistry and immunoblotting revealed that tyrosinated tubulin constitutes a large proportion of total α-tubulin in locomotor cilia of the foot and in axons of the nervous system. Detyrosinated tubulin also appeared to be abundant in the foot cilia but only a very faint band of detyrosinated tubulin was found on protein blots extracted from the central ganglia, and staining was barely detectable in central ganglia or peripheral nerves. Similarly, acetylated tubulin appeared to be abundant in foot cilia, but Western blotting indicated only low levels of acetylated tubulin in the nervous system. Immunocytochemistry indicated that, while most neurons possessed little or no acetylated tubulin, a small number of axons contained significant amounts of this isoform. Thus, while a large amount of tubulin was expected in the nervous system and locomotor cilia of L. stagnalis, the observed distribution of isoforms was unanticipated. Specifically, neurons of other organisms have generally been reported to contain substantial amounts of both detyrosinated α-tubulin and acetylated α-tubulin. Our results indicate that such findings cannot be generalized across all species. L. stagnalis, with its well studied nervous system and unusual distribution of tubulin isoforms, may prove to be particularly useful for studying the roles of tubulin isoforms in microtubule function and cell activity.
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  • 90
    ISSN: 1432-0878
    Keywords: Circadian rhythm ; Colocalization ; Immunocytochemistry ; Brain (CNS), invertebrate ; Optic lobe ; Pigment-dispersing hormone, insect ; Leucophaea maderae (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Several lines of evidence suggest that pigment-dispersing hormone-immunoreactive neurons with ramifications in the accessory medulla are involved in the circadian system of insects. The present study provides a detailed analysis of the anatomical and neurochemical organization of the accessory medulla in the brain of the cockroach Leucophaea maderae. We show that the accessory medulla is compartmentalized into central dense nodular neuropil surrounded by a shell of coarse fibers. It is innervated by neurons immunoreactive to antisera against serotonin and the neuropeptides allatostatin 7, allatotropin, corazonin, gastrin/cholecystokinin, FMRFamide, leucokinin I, and pigment-dispersing hormone. Some of the immunostained neurons appear to be local neurons of the accessory medulla, whereas others connect this neuropil to various brain areas, including the lamina, the contralateral optic lobe, the posterior optic tubercles, and the superior protocerebrum. Double-label experiments show the colocalization of immunoreactivity against pigment-dispersing hormone with compounds related to FMRFamide, serotonin, and leucokinin I. The neuronal and neurochemical organization of the accessory medulla is consistent with the current hypothesis for a role of this brain area as a circadian pacemaking center in the insect brain.
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  • 91
    ISSN: 1432-0878
    Keywords: Neuropeptide Y ; Gastroenteropancreatic (GEP) endocrine system ; Development, ontogenetic ; Vitellointestinal duct ; Pancreas, exocrine ; Pancreas, endocrine ; Immunocytochemistry ; Scyliorhinus torazame (Elasmobranchii)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract This immunocytochemical study was carried out to elucidate the ontogenetic development of neuropeptide Y-like-immunoreactive cells in the gastroenteropancreatic endocrine system of the cloudy dogfish, Scyliorhinus torazame. Immunostained cells first appeared in the pancreas of the embryo at the 15-mm stage, and were also detected in the vitellointestinal duct of the yolk stalk at the 20-mm stage. These cells were polymorphic, with occasional processes that were sometimes directed toward the vascular wall or into the cavity of the vitellointestinal duct. At the 34-mm stage, immunostained cells could also be found in the proximal part of the spiral intestine and, by the 74-mm stage, immunopositive cells were present in the gastric mucosa. In the gut and pancreas, the cells gradually increased in number with development, whereas in the vitellointestinal duct and internal yolk sac, they decreased and seemed to disappear following hatching. Thus, in juveniles, the distribution of the neuropeptide Y-like-immunoreactive cells in the gastroenteropancreatic endocrine system had attained that of adults. Electron-microscopic immunocytochemistry demonstrated that, in the labeled cells of the vitellointestinal duct, the neuropeptide Y-like antigen was located in cytoplasmic granules, as in the cells of the gut and pancreas.
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  • 92
    ISSN: 1432-0878
    Keywords: Key words: Circadian rhythm ; Colocalization ; Immunocytochemistry ; Brain (CNS) ; invertebrate ; Optic lobe ; Pigment-dispersing hormone ; insect ; Leucophaea maderae (Insecta)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Several lines of evidence suggest that pigment-dispersing hormone-immunoreactive neurons with ramifications in the accessory medulla are involved in the circadian system of insects. The present study provides a detailed analysis of the anatomical and neurochemical organization of the accessory medulla in the brain of the cockroach Leucophaea maderae. We show that the accessory medulla is compartmentalized into central dense nodular neuropil surrounded by a shell of coarse fibers. It is innervated by neurons immunoreactive to antisera against serotonin and the neuropeptides allatostatin 7, allatotropin, corazonin, gastrin/cholecystokinin, FMRFamide, leucokinin I, and pigment-dispersing hormone. Some of the immunostained neurons appear to be local neurons of the accessory medulla, whereas others connect this neuropil to various brain areas, including the lamina, the contralateral optic lobe, the posterior optic tubercles, and the superior protocerebrum. Double-label experiments show the colocalization of immunoreactivity against pigment-dispersing hormone with compounds related to FMRFamide, serotonin, and leucokinin I. The neuronal and neurochemical organization of the accessory medulla is consistent with the current hypothesis for a role of this brain area as a circadian pacemaking center in the insect brain.
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  • 93
    ISSN: 1432-0878
    Keywords: Key words: Quinolinic acid ; Interferon-γ ; Kynurenine ; Electron microscopy ; Immunocytochemistry ; Excitotoxicity ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract.  Quinolinate (QUIN), a tryptophan-derived excitotoxin, was localized ultrastructurally in human peripheral blood monocytes/macrophages (MØ) by immuno-electron microscopy. A combined carbodiimide/glutaraldehyde/paraformaldehyde-based fixation procedure was developed for optimal retention of QUIN in the cell as well as minimal loss of ultrastructure; a silver-enhanced colloidal gold detection system was used for electron-microscopic analysis. Gold particles representing QUIN immunoreactivity were associated with the inner side of the plasma membrane in normal MØ. The number of gold particles increased significantly when QUIN levels were elevated by treatment with its precursor kynurenine, but location of the gold particles remained essentially the same under this condition. Treatment with interferon-γ increased the number of Golgi bodies, vacuoles and pseudopodia, reflecting the activated state of the cell. Significantly increased numbers of gold particles representing QUIN were detectable in approximately the same location as in the case of kynurenine treatment. Combined treatment with kynurenine and interferon-γ maximally increased the number of gold particles at the periphery of the cell. The pseudopodia were intensely stained with gold particles, while they were not detectable in the inner part of the cytoplasm or in any other organelle even under this activated condition. The significance of the specific location of QUIN revealed in the present study and its relation to the release and subsequent actions of QUIN are discussed.
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  • 94
    ISSN: 1432-0878
    Keywords: Key words: Subcommissural organ ; Secretory glycoproteins ; Antibodies ; Immunochemistry ; Immunocytochemistry ; Lectins ; Chick embryo (White Leghorn)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The subcommissural organ is an ependymal brain gland that secretes, into the ventricular cerebrospinal fluid, high molecular weight glycoproteins that form Reissner’s fiber. Precursor and processed forms of secretion have been demonstrated by immunoblotting in the subcommissural organ of mammals and fish. In the chicken only a processed form has as yet been identified. In the present report, we have studied the subcommissural organ of 13-day-old chick embryos using (1) an antiserum against bovine Reissner’s fiber, and (2) the lectins, concanavalin A and Limax flavus agglutinin. Paraffin sections of the subcommissural organ and blots of subcommissural organ extracts have been analyzed. The ependymal cells of sectioned subcommissural organ are strongly stained with the antiserum. Concanavalin A binds to materials in all cytoplasmatic regions, whereas Limax flavus agglutinin identifies materials confined to the apex of the ependymal cells. In the blots, a band of 540 kDa is immunostained. This band is positive for concanavalin A positive but negative for Limax flavus agglutinin and is thereby regarded as representing a precursor form of the secretion.
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  • 95
    ISSN: 1432-0878
    Keywords: Key words: Vasotocin ; Sex dimorphism ; Bed nucleus of the stria terminalis ; Dehydration ; Immunocytochemistry ; In situ hybridization ; Domestic fowl
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. A strong sex dimorphism in the distribution of immunoreactive arginine-vasotocin (AVT) and AVT mRNA was observed in telencephalic and dorsal diencephalic areas of the domestic fowl using immunocytochemistry and in situ hybridization. Two subgroups of immunoreactive parvocellular perikarya surrounded by dense plexus of immunoreactive fibres were found within the bed nucleus of the stria terminalis and the dorsal part of the diencephalic paraventricular region of males. No signs of immunoreactivity were observed within corresponding regions of the female brain. Instead, in females a few scattered weakly stained perikarya were observed rostrally to the level of the anterior commissure, juxtapositioned to the nucleus accumbens and the floor of the lateral ventricle. The distribution of AVT mRNA containing cell profiles fully confirmed the immunocytochemical findings. Osmotic stress induced by water deprivation for 48 h had no influence on the number of immunoreactive or AVT mRNA containing parvocellular cell bodies. However, it resulted in an increase of immunoreactive cell area in the bed nucleus of the stria terminalis and dorsal diencephalon of 5.9 and 11.7%, respectively. We suggest that the sexually dimorphic vasotocinergic circuit may be involved in the co-ordination of behavioural and autonomic functions in response to environmental stress.
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  • 96
    ISSN: 1432-0878
    Keywords: Key words: NO/cGMP pathway ; Testis ; Leydig cells ; Immunocytochemistry ; RIA ; Cell culture ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. In this study we sought to determine whether the main components of the nitric oxide (NO) pathway are localized within the Leydig cells of the human testis and whether the soluble guanylyl cyclase (sGC), the enzyme that accounts for NO effects, is functionally active in these cells. Using an amplified immunocytochemical technique, immunoreactivity for nitric oxide synthase (NOS-I), sGC and cyclic guanosine monophosphate (cGMP) was detected within the cytoplasm of human Leydig cells. Distinct differences in staining intensity were found between individual Leydig cells, between cell groups and between Leydig cells of different patients. By means of a specific cGMP-RIA, a concentration-dependent increase in the quantity of cGMP was measured in primary cultures of human Leydig cells following exposure to the NO donor sodium nitroprusside. In addition, NOS-I immunoreactivity was seen in Sertoli cells, whereas cGMP and sGC immunoreactivity was found in Sertoli cells, some apically situated spermatids and residual bodies of seminiferous tubules. Dual-labelling studies and the staining of consecutive sections showed that there are several populations of Leydig cells in the human testis. Most cells were immunoreactive for NOS-I, sGC and cGMP, but smaller numbers of cells were unlabelled by any of the antibodies used, or labelled for NOS-I or cGMP alone, for sGC and cGMP, or for NOS-I and sGC. These results show that the Leydig cells possess both the enzyme by which NO is produced and the active enzyme which mediates the NO effects. There are different Leydig cell populations that probably reflect variations in their functional (steroidogenic) activity.
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  • 97
    ISSN: 1432-0878
    Keywords: Key words: Urothelium ; Tartrate-resistant acid phosphatase ; Nitric oxide synthase I ; Superoxide dismutase ; Immunocytochemistry ; Free radicals ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Three enzymes, viz., tartrate-resistant acid phosphatase (TRAP), nitric oxide synthase I (NOS-I), and superoxide dismutase (SOD), involved in the production and metabolism of free radicals or radical equivalents, were demonstrated by immunocytochemistry in the urothelium of the ureters of six patients of various ages. Two of these enzymes (TRAP and NOS-I) were colocalized in the most apical and lateral border of the superficial cells of the urothelium. In contrast, SOD showed a patchy or granular distribution within the supranuclear region of these cells. Intra- and subepithelial macrophages exhibited a weak TRAP, but no NOS-I or SOD, immune reaction. On the basis of the immunocytochemical findings, arguments in favor of a cytotoxic function of the superficial cells of the human urothelium are presented.
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  • 98
    ISSN: 1432-0878
    Keywords: Key words: Oxytocin ; Immunocytochemistry ; Paraventricular nucleus ; Superior cervical ganglion ; Spinal cord ; Sympathetic nervous system ; Retrograde tracing ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The paraventricular nucleus of the hypothalamus is a major integrative nucleus for relaying information from the suprachiasmatic nucleus to the autonomic system. The precise pathway by which this information can influence autonomic functions, such as melatonin synthesis in the pineal gland, is not clear. In the present study, we used a retrograde tracer injected in the superior cervical ganglion to identify spinal preganglionic neurons. One of the main neurotransmitters present in descending projections of the paraventricular nucleus of the hypothalamus, oxytocin, was detected with immunocytochemistry to visualise possible contacts with the neurons located in the intermediolateral column of the spinal cord and projecting to the superior cervical ganglion. Although many appositions could be seen at the light-microscopic level, this abundance could not be confirmed at the electron-microscopic level. The implications of these observations for the overall timing message received by the spinal preganglionic neurons are discussed.
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  • 99
    ISSN: 1432-0878
    Keywords: Key words Retina ; NOS ; Immunocytochemistry ; Synaptic connectivity ; Guinea pig
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Immunocytochemical methods with an antiserum against neuronal nitric oxide synthase (NOS) were applied to identify the morphology and synaptic connectivity of NOS-like immunoreactive neurons in the guinea pig retina. In the present study, two types of amacrine cells were labeled with anti-NOS antisera. Type 1 cells had large somata located in the inner nuclear layer (INL) with long, sparsely branched processes ramifying mainly in stratum 3 of the inner plexiform layer (IPL). The somata of type 2 cells (smaller diameters) were located in the INL. Some displaced amacrine cells in the ganglion cell layer were labeled. The soma size of the displaced amacrine cells was similar to that of the type 2 amacrine cells. However, processes originating from type 2 amacrine cells and displaced amacrine cells stratified mainly in strata 1 and 5, respectively. Some cone bipolar cells were weakly NOS-immunoreactive. The synaptic connectivity of NOS-like immunoreactive amacrine cells was identified in the IPL by electron microscopy. NOS-labeled amacrine cell processes received synaptic input from other amacrine cell processes and bipolar cell axon terminals in all strata of the IPL. The most frequent postsynaptic targets of NOS-immunoreactive amacrine cells were other amacrine cell processes. Cone bipolar cells were postsynaptic to NOS-labeled amacrine cells in all strata of the IPL. Labeled amacrine cells synapsing onto ganglion cells were found only in sublamina b. A few synaptic contacts were observed between labeled cell processes. In the outer plexiform layer, dendrites of labeled bipolar cells made basal contact with cone pedicles or formed a synaptic triad opposed to a synaptic ribbon of cone pedicles.
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  • 100
    ISSN: 1432-0878
    Keywords: Key words: Crustacean hyperglycaemic hormone ; Immunocytochemistry ; Neurohaemal organ ; Daphnia magna ; Artemia salina (Crustacea)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Crustacean hyperglycaemic hormone-immunoreactive neuronal systems are detected in the central and peripheral nervous systems of two entomostracan crustaceans, Daphnia magna and Artemia salina, by immunocytochemistry using specific antisera against crustacean hyperglycaemic hormones of the decapod crustaceans Orconectes limosus and Carcinus maenas. In D. magna, four small putative interneurones are detected in the brain. In the thorax, ten bipolar peripheral neurones are stained by both antisera. They are obviously segmental homologues with centrally projecting axons that form interdigitating varicose fibres and terminals in putative neurohaemal areas next to the surface of the anterior part of the thoracic ganglia. Similar immunopositive neurones occur both in the central and peripheral nervous systems of A. salina. A total of five groups of neurones occur in the protocerebrum, the deutocerebrum and the mandibular ganglion. Some of the protocerebral neurones are bipolar and project to the dorsal frontal organ. A single pair of peripheral multipolar neurones in the maxillary segment projects centrally into the ventral nerve cord and innervates unidentified somatic muscles and tissues in the maxillary and the first appendage segments. None of the brain neurones in both species show similarities to decapod X-organ sinus gland neurosecretory neurones. Chromatography of brain extracts of D. magna combined with immunodot blotting revealed two strongly immunoreactive fractions at retention times close to that of the crustacean hyperglycaemic hormone of crayfish. Moreover, preabsorption controls suggest that the cross-reacting peptides of D. magna and A. salina are structurally closely related to those of decapods.
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