Phenol oxidases from Rhodnius prolixus: Temporal and tissue expression pattern and regulation by ecdysone

Rhodnius prolixus 5th instar nymphs have significant PO enzymatic activity in the anterior midgut, fat body and hemolymph. The tissue with the major amount of PO activity is the anterior midgut while those with higher specific activities are the fat body and hemolymph. In this work the temporal pattern of PO enzymatic activity in different tissues was investigated. In fat body, PO peaks occur at 7, 12 and 16 days after a blood meal. In hemolymph, PO diminishes until day 7, and then recovers by day 14. In the anterior midgut tissue, PO peaks on day 9 and just before ecdysis; a similar pattern was observed in the anterior midgut contents. Some of these activities are dependent on the release of ecdysone, as feeding blood meal containing azadirachtin suppresses them and ecdysone treatment counteracts this effect. These results suggest that during the development of the 5th instar, the insect has natural regulating cycles of basal PO expression and activation, which could be related to the occurrence of natural infections. The differences in temporal patterns of activity and the effects of azadirachtin and ecdysone in each organ suggest that, at least in R. prolixus, different tissues are expressing different PO genes.     

Phytochrome control of oscillating levels of phenylalanine ammonia lyase in Hordeum vulgare shoots

Five-day-old etiolated barley shoots respond to brief illumination with red light by increasing their level of PAL ca 50% within 5 hr. When assayed s     

Ascaris lumbricoides: succinate and tiglate in hemolymph

When hemolymph is taken from Ascaris lumbricoides at the time the worm is collected from pigs, it contains acetic, propionic, 2-methylbutyric, n-valeric, 2-methylvaleric, and succinic acid radicals; tiglic acid is absent.     

Identification of amino compounds synthesized and translocated in symbiotic Parasponia

We investigated the synthesis and translocation of amino compounds in Parasponia, a genus of the Ulmaceae that represents the only non-legumes known to form a root nodule symbiosis with rhizohia. In the xylem sap of P. andersonii we identified asparagine. aspartate. glutamine, glutamated significant quantities of a non-protein amino acid. 4-methylglutamte(2-amino-4-methylpentanedioic acid). This identification was confirmed by two methods, capillary gas chromatography (GC) electron ionization (El) mass spectrometry (MS) and reverse phase high pressure liquid chromatography (HPLC) analysis of derivatized compounds. In leaf, root and nodule samples from P. andersonii and P. parviflora we also identified the related compounds 4-methyleneglutamate and 4-methyleneglulamine. Using ¹⁵N₂ labelling and GC-Ms analysis of root nodule extracts we followed N₂ fixation and ammonia assimilation in P. andersonii root nodules and observed Label initially in glutamine and subsequently in glutamate, suggesting operation of the glutamine synthetase/glutamine:2-oxoglutarate aminotransferase (GS/GOGAT) pathway. Importantly, we observed the incorporation of significant quantities of ¹⁵N into 4-methylglutamate in nodules, demonstrating the de nova synthesis of this non protein amino acid and suggesting a role in the translation of N in symbioticParasponia.     

Effects of glyphosate on phenolic metabolism in yellow nutsedge leaves

The action of the herbicide glyphosate [N-(phosphonomethyl)-glycine] on phenolic metabolism and phenylalanine ammonia lyase (PAL; EC 4.3.1.5) activity was investigated in yellow nutsedge (Cyperus esculentus L.). Glyphosate caused significant increases in the amount of total soluble hydroxyphenolics in the three fractions studied (neutral, acid and residual). Qualitative and quantitative differences in relation to these fractions and the amount of applied glyphosate were observed. Most of the phenolic compounds which increased after glyphosate treatment were benzoic acids (gentisic. p -OH-benzoic, salicylic and vanillic). Gentisic acid showed the greatest increase in neutral and acid fractions, being twenty- and four-fold, respectively, of the amount found in the control. PAL activity was not affected by the lowest doses of glyphosate (10−⁴and 10−³ M) , but a dramatic decrease in PAL activity was observed after 10−² M treatment. These findings, together with the low levels of cinnamic acids measured in treated yellow nutsedge plants, suggest that PAL activity is only marginally involved in glyphosate action. Since the herbicidal action probably takes place at 5-enol-pyruvylshikimate-3-P synthase (EC 2.5.1.19), an alternative pathway to PAL in phenolic biosynthesis should be activated yielding benzoic acids.     

Model of ammonia volatilization from calcareous soils

A quantitative model of ammonia volatilization from the calcareous soil uppermost 1-cm layer was developed and tested. The model accounts for the following processes: ammonium-ammonia equilibration in the soil solution, cation exchange between calcium and ammonium which results in ammonium distribution between soil liquid and solid phases, nitrification of dissolved ammonium, distribution of ammonia between liquid and gaseous phases and diffusion of gaseous ammonia in the soil air. The combined effect of various characteristics such as soil pH, cation exchange capacity, water capacity and nitrification rate on ammonia losses from various soil types have been studied. The model was validated against experimental results of ammonia losses from different soils for its use as a predicting tool. The model shows that most of ammonia losses can be explained by the interactive effect of high soil pH and low cation exchange capacity. Computations show increased ammonia volatilization with decreasing soil water capacity. Increasing fertilizer application rate has a small effect on percentage of ammonia losses. Increased nitrification rate and shorter “lag” period of nitrification reduce ammonia losses considerably. Good agreement was obtained between model calculations and experimental results of ammonia volatilization from 13 soils.     

Effects of medium glutamine,glutamate, and ammonia on glutamine synthetase activity in cultured mouse astroglial cells

Mouse astroglial cells were grown during the last week of culture in either glutamine-free or glutamine-containing medium. The addition of cortisol to the glutamine-containing medium resulted in a doubling of astroglial glutamine synthetase (GS) activity. Withdrawal of glutamine from the medium resulted in a 50% elevation of GS and addition of cortisol to such a medium resulted in a further increase in GS which was not additive to glutamine withdrawal. Both in glutamine-free and glutamine-containing medium, the addition of glutamate resulted in a depression of both basal and cortisol induced GS activity. The simultaneous addition of ammonia plus glutamate to the culture medium ameliorated the glutamate mediated depressive effects on cortisol induced but not basal GS activity. Glutamine withdrawal from the culture medium resulted in an astroglial protein deficit. The addition of ammonia to the medium considerably reduced this deficit and the addition of glutamate completely eliminated this protein deficit.     

Peat,zeolite and basalt as adsorbents of ammoniacal nitrogen during manure decomposition

The effectiveness of sphagnum peat, zeolite (clinoptilonite) and basalt in reducing ammonia losses during aerobic manure decomposition was determined in an incubation experiment. Peat placed in the spent air-stream adsorbed all of the ammonia volatilized during the first 8 days of decomposition, and reduced overall ammonia losses by 59%. Zeolite reduced total ammonia losses by 16%, and basalt by 6%.All adsorbents were considerably less effective in reducing ammonia losses when mixed with the manure. Reductions in ammonia losses of 24% and 1.5% were obtained with the peat and zeolite, respectively. The addition of basalt increased losses.Ammonia and ammonium adsorption isotherms were determined for the three materials. The adsorption capacities and affinity terms of the adsorbents calculated from the isotherms, reflected their ability to reduce ammonia losses in the incubation experiment. Zeolite had both the highest affinity for ammonium and the highest ammonium adsorption capacity. The peat had a very high affinity for ammonia and a high adsorption capacity (23.4 mg NH₃–N g^–1), whereas zeolite and basalt had a much lower adsorption capacity (1.8 and 0.05 mg NH₃–N g^–1, respectively) compared with their capacity to adsorb ammonium (18.1 and 0.18 mg NH₄–N g^–1).