Short-term urea feeding decreases in vitro hatching of bovine blastocysts

Cows fed high-protein diets may have impaired reproductive performance. Although the pathogenesis has not been completely elucidated, it appears that not only the uterus, but also the follicle and oocyte, are affected by excessive plasma urea nitrogen (PUN) concentrations. Thus, the objective was to determine the effects of short-term urea feeding on the competence of bovine oocytes. Forty crossbred heifers (Bos indicus vs Bos taurus) were allocated to two groups, namely CONTROL (maintenance diet) and UREA (maintenance diet supplemented with 75 g of urea/day), following a cross-over design. Heifers received their respective diets for 6 d (without adaptation). On the sixth day, blood samples were harvested both before and 3 h after feeding, and cumulus oocyte complexes (COCs) were collected by ovum pick-up. Although PUN concentrations were higher in UREA than CONTROL heifers (31.31 mg/dL +/- 1.13 vs 22.12 mg/dL +/- 0.86; mean +/- SEM), neither the number of COCs recovered (8.8 +/- 1.0 vs 9.2 +/- 0.8, UREA vs CONTROL, respectively) nor their quality (based on morphology) differed significantly between groups. Next, oocytes were fertilized and cultured in vitro to assess developmental rates. There was an absence of significant differences between groups for rates of cleavage (Day 3) or blastocyst formation (Days 6, 7 and 9), but the hatched blastocyst rate on Day 11 after fertilization was lower (P < 0.05) in the UREA than the CONTROL groups (64.3 vs 83.5%). Therefore, we inferred that the effects of urea were only manifest later in development. In conclusion, high PUN concentrations decreased oocyte competence in heifers, reinforcing the hypothesis that poor reproductive performance in cows with high PUN was due, at least in part, to a deleterious effect on oocytes. (C) 2011 Elsevier Inc. All rights reserved.

Fate of urea nitrogen applied to a banana crop in the wet tropics of Queensland

This paper reports a study in the wet tropics of Queensland on the fate of urea applied to a dry or wet soil surface under banana plants. The transformations of urea were followed in cylindrical microplots (10.3 cm diameter x 23 cm long), a nitrogen (N) balance was conducted in macroplots (3.85 m x 2.0 m) with N-15 labelled urea, and ammonia volatilization was determined with a mass balance micrometeorological method. Most of the urea was hydrolysed within 4 days irrespective of whether the urea was applied onto dry or wet soil. The nitrification rate was slow at the beginning when the soil was dry, but increased greatly after small amounts of rain; in the 9 days after rain 20% of the N applied was converted to nitrate. In the 40 days between urea application and harvesting, the macroplots the banana plants absorbed only 15% of the applied N; at harvest the largest amounts were found in the leaves (3.4%), pseudostem (3.3%) and fruit (2.8%). Only 1% of the applied N was present in the roots. Sixty percent of the applied N was recovered in the soil and 25% was lost from the plant-soil system by either ammonia volatilization, leaching or denitrification. Direct measurements of ammonia volatilization showed that when urea was applied to dry soil, and only small amounts of rain were received, little ammonia was lost (3.2% of applied N). In contrast, when urea was applied onto wet soil, urea hydrolysis occurred immediately, ammonia was volatilized on day zero, and 17.2% of the applied N was lost by the ninth day after that application. In the latter study, although rain fell every day, the extensive canopy of banana plants reduced the rainfall reaching the fertilized area under the bananas to less than half. Thus even though 90 mm of rain fell during the volatilization study, the fertilized area did not receive sufficient water to wash the urea into the soil and prevent ammonia loss. Losses by leaching and denitrification combined amounted to 5% of the applied N.

Significance of gaseous nitrogen loss from a tropical dairy pasture fertilised with urea

This paper reports a study in the wet tropics of Queensland on the fate of urea applied to a dairy pasture in the absence of grazing animals. A nitrogen balance was conducted in cylindrical plots with N-15-labelled urea, and ammonia volatilisation was determined using a mass balance micrometeorological method. The pasture plants took up 42% of the applied nitrogen in the 98 days between fertiliser application and harvest. At harvest 18% of the applied nitrogen was found in the soil, and 40% was lost from the plant-soil system. The micrometeorological study showed that 20% of the unrecovered nitrogen was lost by ammonia volatilisation. As there was no evidence for leaching or runoff losses it was concluded that the remaining 20% of the applied nitrogen was lost by denitrification. It is evident from these results that fertiliser nitrogen is not being used efficiently on dairy pastures, and that practices need to be changed to conserve fertiliser nitrogen and reduce contamination of the environment.

Effect of leucaena supplementation level, and provision of urea, on utilisation by sheep of Rhodes grass hay

Tarramba leucaena (Leucaena leucocephala cv. Tarramba) foliage had per kilogram dry matter, 169 g protein and 29.8 g condensed tannins. Its value as a supplement, given either with or without urea, to sheep given a low-quality Callide Rhodes grass (Chloris gayana cv. Callide) hay was studied. Six rumen fistulated sheep (mean +/- s.d. liveweight, 34 +/- 1.4 kg) were used to compare 6 dietary treatments in an incomplete latin square design. Rhodes grass hay was given ad libitum either alone, or with urea 7 g/day (U), or with leucaena 150 g/day (L150), or leucaena with urea (L150U), or leucaena 300 g/day (L300), or leucaena with urea (L300U). Digestible organic matter intake was increased significantly by leucaena supplementation although digestibility of the whole diet did not alter. Rumen fluid ammonia-N was not altered by leucaena supplementation, but was increased by urea. This suggests that Tarramba foliage protein has some resistance to ruminal degradation. Liquid and solids passage rates were not affected by the treatments. Microbial nitrogen supply to the intestine (g/day), and the efficiency of microbial nitrogen synthesis (g/kg organic matter apparently digested in the rumen), were increased by leucaena supplementation (P

Syntheses of polycationic dendrimers on lipophilic peptide core for complexation and transport of oligonucleotides

Synthesis of novel polycationic lipophilic peptide core(s) was accomplished and these agents successfully transfected human retinal pigment epithelium cells with ODN1 upon complexation with the oligonucleotide. The level of transfection was indirectly measured by the decreased production of the protein hVEGF (human vascular endothelial growth factor) in comparison to the transfection agent cytofectin GSV(TM). (C) 2002 Elsevier Science Ltd. All rights reserved.

Synthesis and complexation properties of some novel lariat-crown ethers

Several new lariat-crown ethers bearing either bridged bisdioxine or tetraoxaadamantane units as chiral substituents are prepared by reacting the corresponding amino-crown ether derivatives with the dimeric alpha-oxoketene, the latter obtained by flash vacuum pyrolysis of a furan-2,3-dione precursor. Complexation properties towards differently charged metal ions are investigated by H-1 NMR titration to obtain complexation constants (K-c-values for potassium/ sodium rhodanides: 480-1100 mol dm(-3)), as well as extraction experiments to explore the metal ion transportation abilities of the new lariat crown derivatives. In particular, a significantly increased ability to transport metal ions from water into chloroform was found with spherical tetraoxaadamantyl derivatives when compared with the free amino-benzocrown ethers.

Urease inhibitor (NBPT) and efficiency of single or split application of urea in wheat crop

NBPT (N-(n-butyl) thiophosphoric triamide), a urease inhibitor, has been reported as one of the most promising compounds to maximize urea nitrogen use in agricultural systems. The objective of this study was to evaluate the performance of irrigated wheat fertilized with urea or urea + NBPT as single or split application. The experiment was conducted from June to October 2006 in Viçosa, MG, Brazil. The experimental design followed a 2×2 factorial scheme, in which urea or urea + NBPT were combined with two modes of application: full dose at sowing (60kg ha-1) or split (20kg ha-1 at sowing + 40kg ha-1 as topdressing at tillering), in randomized blocks with ten replications. The split application of nitrogen fertilization does not improve the yield wheat under used conditions. The use of urease inhibitor improves the grain yield of wheat crop when urea is applied in topdressing at tillering, but its use does not promote difference when urea is applied in the furrow at planting.

Development of a biosensor for urea assay based on amidase inhibition, using an ion-selective electrode

A biosensor for urea has been developed based on the observation that urea is a powerful active-site inhibitor of amidase, which catalyzes the hydrolysis of amides such as acetamide to produce ammonia and the corresponding organic acid. Cell-free extract from Pseudomonas aeruginosa was the source of amidase (acylamide hydrolase, EC 3.5.1.4) which was immobilized on a polyethersulfone membrane in the presence of glutaraldehyde; anion-selective electrode for ammonium ions was used for biosensor development. Analysis of variance was used for optimization of the biosensorresponse and showed that 30 mu L of cell-free extract containing 7.47 mg protein mL(-1), 2 mu L of glutaraldehyde (5%, v/v) and 10 mu L of gelatin (15%, w/v) exhibited the highest response. Optimization of other parameters showed that pH 7.2 and 30 min incubation time were optimum for incubation ofmembranes in urea. The biosensor exhibited a linear response in the range of 4.0-10.0 mu M urea, a detection limit of 2.0 mu M for urea, a response timeof 20 s, a sensitivity of 58.245 % per mu M urea and a storage stability of over 4 months. It was successfully used for quantification of urea in samples such as wine and milk; recovery experiments were carried out which revealed an average substrate recovery of 94.9%. The urea analogs hydroxyurea, methylurea and thiourea inhibited amidase activity by about 90%, 10% and 0%, respectively, compared with urea inhibition.

Tuneable micro- and nano-periodic structures in urethane/urea networks

Micro- and nano-patterned materials are of great importance for the design of new nanoscale electronic, optical and mechanical devices, ranging from sensors to displays. A prospective system that can support a designed functionality is elastomeric polyurethane thin films with nano- or micromodulated surface structures ("wrinkles"). These wrinkles can be induced on different lengthscales by mechanically stretching the films, without the need for any sophisticated lithographic techniques. In the present article we focus on the experimental control of the wrinkling process. A simple model for wrinkle formation is also discussed, and some preliminary results reported. Hierarchical assembly of these tunable structures paves the way for the development of a new class of materials with a wide range of applications, from electronics to biomedicine.

A multiresidue method for the analysis of carbamate and urea pesticides from soils by microwave-assisted extraction and liquid chromatography with photodiode array detection

A multiresidue approach using microwave-assisted extraction and liquid chromatography with photodiode array detection was investigated for the determination of butylate, carbaryl, carbofuran, chlorpropham, ethiofencarb, linuron,metobromuron, and monolinuron in soils. The critical parameters of the developed methodology were studied. Method validation was performed by analyzing freshly and aged spiked soil samples. The recoveries and relative standard deviations reached using the optimized conditions were between 77.0 ± 0.46% and 120 ± 2.9% except for ethiofencarb (46.4 ± 4.4% to 105 ± 1.6%) and butylate (22.1 ± 7.6% to 49.2 ± 11%). Soil samples from five locations of Portugal were analysed.

Determination of carbamate and urea pesticide residues in fresh vegetables using microwave-assisted extraction and liquid chromatography

An analytical multiresidue method for the simultaneous determination of seven pesticides in fresh vegetable samples, namely, courgette (Cucurbita pepo), cucumber (Cucumis sativus), lettuce (Lactuca sativa, Romaine and Iceberg varieties) and peppers (Capsicum sp.) is described. The procedure, based on microwave-assisted extraction (MAE) and analysis by liquid chromatography– photodiode array (LC–PDA) detection was applied to four carbamates (carbofuran, carbaryl, chlorpropham and EPTC) and three urea pesticides (monolinuron, metobromuron and linuron). Extraction solvent and the addition of anhydrous sodium sulphate to fresh vegetable homogenate before MAE were the parameters optimised for each commodity. Recovery studies were performed using spiked samples in the range 250–403 µgkg- 1 in each pesticide. The pesticide residues were extracted using 20mL acetonitrile at 60 ºC, for 10 min. Acceptable recoveries and RSDs were attained (overall average recovery of 77.2% and RSDs are lower than 11%). Detection limits ranged between 5.8 µgkg- 1 for carbaryl to 12.3 µgkg- 1 for carbofuran. The analytical protocol was applied for quality control of 41 fresh vegetable samples bought in Oporto Metropolitan Area (North Portugal). None of the samples contained any detectable amounts of the studied compounds.

A new fluorescent double-cavity calix[4]arene: synthesis and complexation studies toward nitroanilines

Agência Financiadora: Fundação para a Ciência e a Tecnologia/MCTES (Portugal) - PEst-OE/EQB/UI0702/2012

Alternative chelating agents: Evaluation of the ready biodegradability and complexation properties

The ready biodegradability of four chelating agents, N,N -(S,S)-bis[1-carboxy-2-(imidazol-4-yl)ethyl]ethylenediamine (BCIEE), N - ethylenedi-L-cysteine (EC), N,N -bis (4-imidazolymethyl)ethylenediamine (EMI) and 2,6-pyridine dicarboxylic acid (PDA), was tested according to the OECD guideline for testing of chemicals. PDA proved to be a readily biodegradable substance. However, none of the other three compounds were degraded during the 28 days of the test. Chemical simulations were performed for the four compounds in order to understand their ability to complex with some metal ions (Ca, Cd, Co, Cu, Fe, Mg, Mn, Ni, Pb, Zn) and discuss possible applications of these chelating agents. Two different conditions were simulated: (i) in the presence of the chelating agent and one metal ion, and (ii) in the simultaneous presence of the chelating agent and all metal ions with an excess of Ca. For those compounds that were revealed not to be readily biodegradable (BCIEE, EC and EMI), applications were evaluated where this property was not fundamental or even not required. Chemical simulations pointed out that possible applications for these chelating agents are: food fortification, food process, fertilizers, biocides, soil remediation and treatment of metal poisoning. Additionally, chemical simulations also predicted that PDA is an efficient chelating agent for Ca incrustations removal, detergents and for pulp metal ions removal process.

VALIDATION OF 14C-UREA BREATH TEST FOR DIAGNOSIS OF Helicobacter pylori

The aim of this study was to validate the 14C-urea breath test for use in diagnosis of Helicobacter pylori infection. Thirty H. pylori positive patients, based on histologic test and thirty H. pylori negative patients by histology and anti-H. pylori IgG entered the study. Fasting patients drank 5 uCi of 14C-urea in 20 ml of water. Breath samples were collected at 0, 5, 10, 15, 20 and 30 min. The difference of cpm values between the two groups was significant at all the time intervals, besides time 0 (p<0.0001). At 20 min, the test gave 100% sensitivity and specificity with a cut-off value of 562 cpm. Females were higher expirers than males (p=0.005). 14C-urea breath test is highly accurate for Helicobacter pylori diagnosis. It is fast, simple and should be the non-invasive test used after treating Helicobacter pylori infection.