Cytotoxic effects of peroxynitrite, polymorphonuclear neutrophils, free-radical scavengers, inhibitors of myeloperoxidase, and inhibitors of nitric oxide synthase on bovine mammary secretory epithelial cells

OBJECTIVE: To determine cytotoxic effects of activated polymorphonuclear neutrophils (PMN) and peroxynitrite on bovine mammary secretory epithelial cells before and after addition of nitric oxide synthase inhibitors, myeloperoxidase (MPO) inhibitors, and free-radical scavengers. SAMPLE POPULATION: Polymorphonuclear neutrophils from 3 lactating cows. PROCEDURE: Cells from the bovine mammary epithelial cell line MAC-T were cultured. Monolayers were treated with activated bovine PMN, lipopolysaccharide (LPS), phorbol 12-myristate 13-acetate (PMA), 3-morpholino-sydnonimine (SIN-1), 4-amino-benzoic acid hydrazide (ABAH), NG-monomethyl-L-arginine, histidine, and superoxide dismutase (SOD). At 24 hours, activity of lactate dehydrogenase in culture medium was used as a relative index of cell death. Tyrosine nitration of proteins in MAC-T cell lysates was determined by visual examination of immunoblots. RESULTS: Lipopolysaccharide, PMA, and < or = 0.1 mM SIN-1 were not toxic to MAC-T cells. Activated PMN, > or = 6 mg of histidine/ml, and 0.5 mM SIN-1 were toxic. Together, histidine and 500,000 activated PMN/ml also were toxic. NG-monomethyl-L-arginine did not have an effect, but ABAH decreased PMN-mediated cytotoxicity. Ten and 50 U of SOD/ml protected MAC-T cells from cytotoxic effects of 0.5 mM SIN-1. Compared with control samples, nitration of MAC-T tyrosine residues decreased after addition of 500,000 PMN/ml or > or = 6 mg of histidine/ml. Superoxide dismutase increased and SIN-1 decreased tyrosine nitration of MAC-T cell proteins in a dose-responsive manner. CONCLUSIONS AND CLINICAL RELEVANCE: Peroxynitrite, MPO, and histidine are toxic to mammary secretory epithelial cells. Superoxide dismutase and inhibition of MPO activity mitigate these effects. Nitration of MAC-T cell tyrosine residues may be positively associated with viability.     

Fertilizer-induced Changes in Rhizosphere Electrical Conductivity: Relation to Forest Tree Seedling Root System Growth and Function

Fertilization is standard practice in forest tree seedling nursery culture. Additionally, fertilization at outplanting has potential to facilitate nutrient uptake and reduce transplant shock. Fertilization, however, may dramatically alter rhizosphere chemical properties such as pH, ion availability, and electrical conductivity (EC). These changes may inhibit root system growth and function by reducing soil osmotic potential and creating specific ion toxicities. The risk of root damage associated with high EC levels appears to be dependent on species, age of root system, and soil moisture availability. Root inhibition in container nursery culture of conifers is likely to occur above 2.5 dS m⁻¹, though threshold EC levels for bareroot culture and field plantings are largely unavailable. Fertilization at outplanting has the added risk that drought conditions may prevent leaching of excess fertilizer salts, which can increase rhizosphere EC beyond safe levels and ultimately impair root uptake of water or nutrients. For fertilization programs to be successful, a critical threshold balance must be maintained between optimizing seedling nutrient availability in the rhizosphere, while minimizing potential for root damage. Future research is needed to identify optimal EC levels for a range of species across all stages of the reforestation process, from nursery culture through plantation establishment.     

Influence of nursery soil amendments on water relations,root architectural development,and field performance of Douglas-fir transplants

This experiment evaluated the influence of manure, peat, and vermiculite incorporated at low and high rates (0.0118 and 0.0236 m³/m²) and under two soil moisture regimes on Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) seedling (1+0 for 1+1) xylem water potential (_xylem), whole-plant growth, root architectural development, and subsequent field performance under fertilized and non-fertilized conditions. Trends in soil moisture retention were observed (high manure > high peat > control) but there were no differences in _xylem. Root length in the wetter soil moisture experiment was initially (three months) greatest for seedlings in high vermiculite and least in high manure but there were no differences among treatments at lifting (eight months). Mean height was greatest for seedlings grown in vermiculite and peat (wetter nursery experiment) after two field seasons. Field fertilization (35 g/seedling) with controlled-release fertilizer in the planting hole stimulated height growth initially, but decreased height and diameter growth during the second growing season. Dramatic improvements associated with the use of nursery soil amendments were not realized, but the failure to identify negative effects, a potential reduction in disease incidence, and improvement of nursery soil physical and chemical properties may justify their use.     

Organic Matter Added to Bareroot Nursery Beds Influences Soil Properties and Morphology of Fraxinus pennsylvanica and Quercus rubra Seedlings

Bareroot hardwood seedling production involves intensive soil management. To increase soil organic matter (OM), nurseries commonly grow a cover crop for 1 year after every 1–2 year of seedling production. Raising soil OM levels can also be achieved through addition of soil amendments. We studied the influence of chicken manure (CM) and composted leaf, tree, and lawn trimmings (Cp) on soil properties and morphology of green ash (Fraxinus pennsylvanica Marsh.) and northern red oak (Quercus rubra L.) seedlings. CM was applied at 725, 1450, or 2900 kg ha⁻¹ (CM725, CM1450, and CM2900, respectively) and Cp was applied at 200 m³ ha⁻¹. Addition of CM and Cp significantly raised soil OM levels and altered soil chemical properties compared to the control (Ctrl). Root-collar diameter increased with addition of CM1450, CM2900, or Cp compared to CM725 or Ctrl plots for northern red oak, but was largest in soils amended with CM2900 for green ash. Conversely, height was greatest with addition of CM725 for northern red oak, but green ash seedlings were shorter in Ctrl plots than in all amendments except for CM725. Root volume of green ash and northern red oak seedlings was positively influenced by addition of CM or Cp. Seedling responses to nursery soil amendments vary with different forms and amounts of OM. Benefits to seedling growth through application of appropriate materials in the proper balance can improve seedling morphological quality and positively influence soil chemical properties.     

Nitrogen fertilization of black walnut (Juglans nigra L.) during plantation establishment. Physiology of production

Physiological mechanisms by which nitrogen (N) fertilization affects growth and development in temperate deciduous forest trees are not clearly understood, especially under intensive silvicultural systems. Grafted, Tippecanoe 1 cultivar black walnut (Juglans nigra L.) trees were grown in an intensively managed plantation in west-central Spain and subjected to six, fixed-nutrient-ratio complete fertilizer treatments (defined as 0, 25, 50, 75, 150, and 300 g N tree^?1) delivered via daily fertigation. Leaf chemistry and morphology were evaluated from June to September, and gas exchange was measured in July. Specific leaf mass, leaflet nitrogen (N), and chlorophyll concentrations varied over the course of the growing season, yet consistently increased with increasing fertilization. Net photosynthesis at ambient (A _net) and light-saturated (A _max) conditions increased from the unfertilized control to lowest treatment (25 g N) but did not increase at higher fertilizer rates. Photosynthetic N and chlorophyll use efficiencies decreased with increasing fertilization, but photosynthetic phosphorus and water use efficiencies increased. Transpiration rates and dark respiration were not significantly affected by treatment. Overall, the lowest fertilizer treatment (25 g N) had the greatest photosynthetic efficiency. Interactions between N and other nutrients with increasing fertilizer application suggested potential for nutrient imbalances at high fertilization rates. Our results provide a physiological justification for the use of low-to-moderate fertilization as an efficient strategy to promote black walnut plantation establishment under intensive cultural systems.     

Nitrogen form and concentration interact to affect the performance of two ecologically distinct Mediterranean forest trees

Most studies examining inorganic N form effects on growth and nutrition of forest trees have been conducted on single species from boreal or temperate environments, while comparative studies with species from other biomes are scarce. We evaluated the response of two Mediterranean trees of contrasting ecology, Quercus ilex L. and Pinus halepensis Mill., to cultivation with distinct inorganic N forms. Seedlings were fertilized with different NH₄ ⁺/NO₃ ^? proportion at either 1 or 10 mM N. In both species, N forms had small effects at low N concentration, but at high N concentration they markedly affected the plant performance. A greater proportion of NH₄ ⁺ in the fertilizer at high N caused toxicity as it reduced growth and caused seedling death, with the effect being greater in Q. ilex than in P. halepensis. An increase in the proportion of NO₃ ^? at high N strongly enhanced growth relative to low N plants in P. halepensis but had minor effects in Q. ilex. Relatively more NH₄ ⁺ in the fertilizer enhanced plant P concentration but reduced K concentration in both species, while the opposite effect occurred with NO₃ ^?, and these effects were enhanced under high N concentration. We conclude that species responses to inorganic N forms were related to their ecology. P. halepensis, a pioneer tree, had improved performance with NO₃ ^? at high N concentration and showed strong plasticity to changes in N supply. Q. ilex, a late successional tree, had low responsiveness to N form or concentration.