Short term physiological implications of NBPT application on the N metabolism of Pisum sativum and Spinacea oleracea

The application of urease inhibitors in conjunction with urea fertilizers as a means of reducing N loss due to ammonia volatilization requires an in-depth study of the physiological effects of these inhibitors on plants. The aim of this study was to determine how the urease inhibitor N-(n-butyl) thiophosphoric triamide (NBPT) affects N metabolism in pea and spinach. Plants were cultivated in pure hydroponic culture with urea as the sole N source. After 2 weeks of growth for pea, and 3 weeks for spinach, half of the plants received NBPT in their nutrient solution. Urease activity, urea and ammonium content, free amino acid composition and soluble protein were determined in leaves and roots at days 0, 1, 2, 4, 7 and 9, and the NBPT content in these tissues was determined 48 h after inhibitor application. The results suggest that the effects of NBPT on spinach and pea urease activity differ, with pea being most affected by this treatment, and that the NBPT absorbed by the plant caused a clear inhibition of the urease activity in pea leaf and roots. The high urea concentration observed in leaves was associated with the development of necrotic leaf margins, and was further evidence of NBPT inhibition in these plants. A decrease in the ammonium content in roots, where N assimilation mainly takes place, was also observed. Consequently, total amino acid contents were drastically reduced upon NBPT treatment, indicating a strong alteration of the N metabolism. Furthermore, the amino acid profile showed that amidic amino acids were major components of the reduced pool of amino acids. In contrast, NBPT was absorbed to a much lesser degree by spinach plants than pea plants (35% less) and did not produce a clear inhibition of urease activity in this species.     

Structure Elucidation of a New n-Pentyl Fructofuranoside in Dendranthema morifolium(Ramat.) Tzvel.

A new compound (1) and eleven known compounds were isolated from the polaric fractions of Dendranthema morifolium (Ramat.) Tzvel. By means of chemical and spectral analyses, compound 1 was established as n-pentyl-β-D-fruetofuranoside. Two known compounds n-butyl and ethyl eaffeate were strong antagonists of 5-]ipoxygenase.     

Chemical constituents of Bulbophyllum wendlandianum (Kraenzl.) Dammer and their chemotaxonomic significance

Phytochemical investigation of Bulbophyllum wendlandianum (Kraenzl.) Dammer led to the isolation of twenty-three compounds 1–23 (flavanthrinin 1, coelonin 2, lusianthridin 3, densiflorol B 4, plicatol B 5, batatasin-lll 6, gigantol 7, 5-hydroxy-3,3′-dimethoxy-2-(p-hydroxybenzyl) bibenzyl 8, 2,2-dimethyl-5-hydroxy-6-carboxy-7-(2-phenylethyl) 9, tristin 10, p-hydroxybenzyl ethyl ether 11, p-hydroxybenzaldehyde 12, hydroquinone 13, coniferaldehyde 14, p-hydroxybenzyl alcohol 15, 3,4-dihydroxy benzaldehyde 16, stigmasterol 17, β-sitosterol 18, ergosterol peroxide 19, (+)pinoresinol 20, n-butyl sulfoxide 21, tridec-4E-en-l-yl acetate 22, ethyl linolate 23) including five phenanthrenes 1–5, five bibenzyls 6–10, six phenols 11–16, three sterols 17–19, one lignan 20, one n-butyl sulfoxide 21 and two fatty acids 22–23. The structures of these compounds were elucidated by spectroscopic analyses. This is the first report of isolation of compounds 1–23 from Bulbophyllum wendlandianum and compounds 8–9, 11, 13, 15–16 and 19–23 within genus Bulbophyllum. Compound 21 is a new natural product, isolated from a natural source for the first time. Furthermore, the chemotaxonomic significance of the isolates was also discussed.     

Inhibition of Jack Bean Urease by N-(n-butyl)thiophosphorictriamide and N-(n-butyl)phosphorictriamide: Determination of the Inhibition Mechanism

N-(n-butyl)thiophosphorictriamide (NBPT) and its oxygen analogue N-(n-butyl)phosphorictriamide (NBPTO) were studied as inhibitors of jack bean urease. NBPTO was obtained by spontaneous conversion of NBPT into NBPTO. The conversion under laboratory conditions was slow and did not affect NBPT studies. The mechanisms of NBPT and NBPTO inhibition were determined by analysis of the reaction progress curves in the presence of different inhibitor concentrations. The obtained plots were time-dependent and characteristic of slow-binding inhibition. The effects of different concentration of NBPT and NBPTO on the initial and steady-state velocities as well as the apparent first-order velocity constants obeyed the relationships for a one-step enzyme-inhibitor interaction, qualified as mechanism A. The inhibition constants of urease by NBPT and NBPTO were found to be 0.15 μM and 2.1 nM, respectively. The inhibition constant for NBPT was also calculated by steady-state analysis and was found to be 0.13 μM. NBPTO was found to be a very strong inhibitor of urease in contrast to NBPT.     

Production of alcohol from sugar beet molasses without heat or filter sterilization

Among three esters of p-hydroxybenzoate, n-butyl p-hydroxybenzoate was selected as the best antimicrobial substance. Molasses medium sterilized by this ester was used as a substrate for ethanol production. n-Butyl p-hydroxybenzoate (0.15% w/v) completely inhibited the growth of free yeast cell inoculum, Ca-alginate immobilized yeast inoculum and bacterial contaminants. Immobilization of the yeast cell inoculum in Ca-alginate with castor oil (6% v/v) offered a yeast cell protection against the inhibitory effect of n-butyl p-hydroxybenzoate. The presence of castor oil in this immobilization system did not affect the metabolic activity of the yeast in beads compared to the cells immobilized without castor oil. The yeast cell beads in this system completely utilized up to 25% molasses sugar with an ethanol yield of 10.58%, equal to 83% of its theoretical value. The beads were stable and could be used successfully for seven cycles of batch fermentation. The optimum fermentation temperature using this system was 35°C. Received 21 January 1997/ Accepted in revised form 05 May 1997     

Agronomic assessment and 15N recovery of urea amended with the urease inhibitor nBTPT (N-(n-butyl) thiophosphoric triamide) for temperate grassland

Three field experiments were undertaken concurrently at one site to evaluate a range of surface-applied nBTPT-amended urea products (0.01, 0.05, 0.1, 0.25 and 0.5% nBTPT w/w) on NH₃ volatilization, grass yield and ¹⁵N recovery in the plant-soil system. Each experiment was repeated on five separate occasions over the 1992 growing season to cover a range of weather conditions. Total NH₃ loss from unamended-urea ranged from 5.5% in early May to 20.8% in June. The inhibitor was highly effective in reducing ammonia volatilization and delaying the time at which maximum rate of NH₃ loss occurred. Over all time periods the % inhibition was 50.4, 82.8, 89.0, 96.5 and 97.0% at the 0.01, 0.05, 0.1, 0.25 and 0.5% nBTPT levels respectively. There was no significant difference in the overall % inhibition in ammonia loss at different times suggesting that the effectiveness of the inhibitor was not dependent on climatic conditions.Over all times incorporation of nBTPT at the 0.05% level increased dry-matter yield by 9% compared to urea alone and increased the shoot recovery of N from 66.7% to 80.9%. Nitrogen saved from volatilization was taken up by the plant, however, the subsequent translation into dry-matter yield appeared to be adversely affected at the high inhibitor rates.There was no significant effect of inhibitor on ¹⁵N recovery in soil at any depth down to 15 cms. nBTPT significantly increased (p < 0.001) the % N derived from fertilizer (% N dff) in the shoot compared to unamended-urea and increased (p < 0.01) the shoot recovery of ¹⁵N from 32% up to 39%. Total ¹⁵N recovery in the soil-plant system was increased by up to 17% by amending urea with nBTPT. This urease inhibitor has been shown to improve the efficiency of urea however, its potential for the European market will be dependent on economic factors.Faculty of Agriculture and Food Science, The Queen's University of Belfast     

Modified polynucleotides. V. Slow-down of nuclease action by 5-alkyluracil-containing DNAs

Poly d/[³H]A-r⁵U/ type of synthetic models of bacteriophage DNAs containing thymine analogues were prepared by DNA polymerase and tested for stability against nucleases /$\text{r}$ was a n-alkyl group from methyl to pentyl/. The 5-pentyluracil-containing copolymer was found to be most stable: 50 % degradation with pancreatic DNase, spleen DNase, snake venom phosphodiesterase or micrococcal nuclease required 3–15 times as much time as that of poly d/A-T/.     

Nano carriers that enable co-delivery of chemotherapy and RNAi agents for treatment of drug-resistant cancers

Tumor cells exhibit drug resistant phenotypes that decrease the efficacy of chemotherapeutic treatments. The drug resistance has a genetic basis that is caused by an abnormal gene expression. There are several types of drug resistance: efflux pumps reducing the cellular concentration of the drug, alterations in membrane lipids that reduce cellular uptake, increased or altered drug targets, metabolic alteration of the drug, inhibition of apoptosis, repair of the damaged DNA, and alteration of the cell cycle checkpoints ( and ). siRNA is used to silence the drug resistant phenotype and prevent this drug resistance response. Of the listed types of drug resistance, pump-type resistance (e.g., high expression of ATP-binding cassette transporter proteins such as P-glycoproteins (Pgp; also known as multi-drug resistance protein 1 or MDR1, encoded by the ATP-Binding Cassette Sub-Family B Member 1 (ABCB1) gene)) and apoptosis inhibition (e.g., expression of anti-apoptotic proteins such as Bcl-2) are the most frequently targeted for gene silencing. The co-delivery of siRNA and chemotherapeutic drugs has a synergistic effect, but many of the current projects do not control the drug release from the nanocarrier. This means that the drug payload is released before the drug resistance proteins have degraded and the drug resistance phenotype has been silenced. Current research focuses on cross-linking the carrier's polymers to prevent premature drug release, but these carriers still rely on environmental cues to release the drug payload, and the drug may be released too early. In this review, we studied the release kinetics of siRNA and chemotherapeutic drugs from a broad range of carriers. We also give examples of carriers used to co-deliver siRNA and drugs to drug-resistant tumor cells, and we examine how modifications to the carrier affect the delivery. Lastly, we give our recommendations for the future directions of the co-delivery of siRNA and chemotherapeutic drug treatments.     

Single administration of di(n-butyl) phthalate delays spermatogenesis in prepubertal rats

Morphological alterations in seminiferous tubules caused by single administration of di(n-butyl) phthalate (DBP) in 3-week-old rats were investigated throughout the first wave of spermatogenesis. Single administration of DBP (500 mg/kg) showed progressive detachment and displacement of spermatogenic cells and disappearance of tubular lumen at 3 h after treatment, and then showed thin seminiferous epithelia and wide tubular lumen at day 1 (D1). At D1, quite significant numbers of apoptotic spermatogenic cells were detected, and then they gradually decreased in accordance with the passage of time. In contrast, the testes revealed lower weight gain, even after completion of first wave of spermatogenesis in the DBP-treated group, compared to the control. In order to clarify whether spermatogenic cells differentiate into mature spermatids in the DBP-treated rats, immunohistochemical staining for Hsc 70t, a specific marker for elongate spermatids, was carried out. As a result, the decrease in mature spermatids in the DBP-treated testes, compared to the control, was demonstrated. For example, at D20 (41-day-old) after treatment, the most advanced spermatids in the tubules from rats in the DBP-treated groups were steps 2-4, while those of the control were steps 12-13. Moreover, in some tubules, pachytene spermatocytes were the most advanced spermatogenic cell. At D30 (51-day-old) after treatment, maturation of spermatogenic cells in the DBP-treated rats proceeded further, and the most advanced spermatids in tubules were steps 8-9, while those of the control were steps 15-19. These results lead us to the postulation that a single administration of DBP to prepubertal rats delays maturation of spermatogenic cells, even after completion of first wave of spermatogenesis.     

羽摇蚊幼虫和霍甫水丝蚓的生态毒理学研究

本文以羽摇蚊幼虫和霍甫水丝蚓为对象,进行了丁基磺毒杀作用的试验。结果表明,丁基磺不仅污染环境,而且对这两种水生生物均有较高的毒杀作用;这两种水生生物对其毒性都分别产生不同的症状;在低浓度下,亦产生不同效应;不同浓度丁基磺对摇蚊幼虫24小时毒理试验LC₅₀为256.2ppm,对水丝蚓LC₅₀为25.8ppm;不同季节出现的毒性效应均有差异。在试验结果的基础上,对水丝蚓中毒后出现了不同类型的症状;温度、酸碱度等不同生态因素对丁基磺毒性的影响以及加强水丝蚓生态毒理学工作的必要性等问题进行了讨论。