Acetylation of 5-amino-1H-[1,2,4]triazole revisited

The products of the acetylation reactions of the common herbicide 5-amino-1H-[1,2,4]triazole were investigated using HPLC, GC-MS, 1H NMR, and FTIR spectroscopy. The conventional annular monoacetylation procedures with acetyl chloride are not regioselective and furnish a mixture of isomers. Traditional diacetylation in neat acetic anhydride under reflux produces a mixture of di-, mono-, and triacetylated derivatives. By using equivalent amounts of acetic anhydride in a dimethylformamide solution, a rapid and selective annular monoacetylation of 5-amino-1H-[1,2,4]triazole was achieved. The monoacetylation proceeds via the formation of the intermediate, 1-acetyl-3-amino-1H-[1,2,4]triazole, which had not been observed previously and which undergoes transformation into the known 1-acetyl-5-amino-1H-[1,2,4]triazole. Neat acetic anhydride at room temperature affords the diacetylated derivative, 1-acetyl-3-(acetylamino)-1H-[1,2,4]triazole both from 5-amino-1H-[1,2,4]triazole itself and from either 1-acetyl-5-amino-1H-[1,2,4]triazole or 5-(acetylamino)-1H-[1,2,4]triazole. The atypical product of the second acetylation, 1-acetyl-5-(acetylamino)-1H-[1,2,4]triazole, has been identified. These results may be useful in the development of effective and selective preparative procedures for the acetylation of 5-amino-1H-[1,2,4]triazole.     

Cycloate,an inhibitor of fatty acid elongase,modulates the metabolism of very‐long‐side‐chain alkylresorcinols in rye seedlings

BACKGROUND: Cycloate inhibits the biosynthesis of very‐long‐chain fatty acids, the essential constituents of plant waxes and suberin. Fatty acids also serve as precursors of aliphatic carbon chains in resorcinolic lipids, which play a fundamental role in the plant defence system against fungal pathogens. In this study, the effect of cycloate on the biosynthesis of 5‐n‐alkylresorcinols in rye seedlings (Secale cereale L.) grown under various light and thermal conditions was examined. RESULTS: The content of alkylresorcinols biosynthesised in rye was generally increased by the herbicide in both green and etiolated plants. The presence of cycloate also affected patterns of alkylresorcinol homologues in plants grown at 15 and 22 °C; very‐long‐side‐chain compounds were less abundant, whereas both short‐chain saturated and unsaturated homologues were generally accumulated. No cycloate‐related effects caused by homologue pattern modifications were observed at elevated temperature. CONCLUSION: This study extends present understanding of the mode of action of thiocarbamate herbicides. Cycloate markedly affected the biosynthesis of very‐long‐side‐chain resorcinolic lipids in rye seedlings, confirming the existence of parallels in both fatty acid and alkylresorcinol biosynthetic pathways. The observed cycloate‐driven accumulation of 5‐n‐alkylresorcinols may improve the resistance of cereals to infections caused by microbial pathogens. Copyright © 2009 Society of Chemical Industry     

Tracing Pharmaceuticals in the Unsaturated Zone (10 pp)

Background, Aim and Scope   One of the first occurrences of pharmaceutically active compounds in groundwater was reported from the sewage irrigation farms south of Berlin. At these sewage irrigation farms treated sewage effluent passed the soil and unsaturated zone before reaching the aquifer. Clofibric acid was detected in pore water from soils of those sewage irrigation farms in concentrations between 65 ng/L and 1430 ng/L. The aim of this study was to investigate the transport behavior of regularly detected clofibric acid, diclofenac, ibuprofen, and propyphenazone under conditions comparable to those at the sewage irrigation farms in a multiple compound sand column laboratory experiment. Materials and Methods   Sediment column experiments were conducted to study the transport of pharmaceuticals in the unsaturated zone. The migration was measured in fine to medium grained sand and leaching solution containing 1 mg/L of pharmaceutically active compounds and 61 mg/L of the tracer lithium chloride (LiCl). For the analysis of the pharmaceutical compounds the water samples were adjusted to a pH value of 2 and then extracted by solid-phase extraction (SPE). Before extraction, the samples were spiked with a surrogate standard for analytical quality control. The sample extracts were analyzed by capillary gas chromatography-mass spectrometry (GC-MS) with selected ion monitoring (SIM). Depending on the sample volume (100 to 200 mL) and the matrix, the limits of detection were between 1 and 10 ng/L, and the limits of quantitation were between 5 and 25 ng/L. Analysis for calcium, magnesium and lithium were carried out using a 'Trace Scan' ICP-AES from Thermo Jarrel Ash. Sodium, potassium, iron and manganese were analyzed using a Philips PU 9400 flame AAS. Analysis of anions was performed on a Dionex ion chromatograph DX 120. Results   At the sewage irrigation farms the average concentrations of clofibric acid in the unsaturated zone declined from higher values near ground surface (480 ng/L) to lower values near the groundwater table (65 ng/L). From the pharmaceuticals analyzed only clofibric acid, primidone and propyphenazone could be analyzed in the first (upper) aquifer at the sewage irrigation farms. All other pharmaceuticals could neither be detected in the first aquifer nor in the deeper aquifers. Breakthrough curves from soil column experiments revealed no transformation and no retardation for clofibric acid, whereas transformation of diclofenac was so high (79%) that no retardation factor could be calculated. Ibuprofen was significantly transformed (37%), transformation of propyphenazone (17%) was quite low and retardation (Rf = 2.05) was in the range of previously conducted column experiments. Discussion   The results confirm previously conducted experiments with clofibric acid where this compound was identified as highly mobile and persistent. The results that diclofenac and ibuprofen are significantly transformed where unexpected as other studies exhibited much lower transformation under saturated conditions at least for diclofenac. However, lower pH values and higher oxygen contents in the unsaturated zone compared to the aquifer may explain this observed high transformation of these compounds at the column experiments. Conclusions   We conclude that irrigation with sewage effluent containing the compounds used in our experiments will lead to an input into groundwater of clofibric acid, whereas diclofenac and ibuprofen will most likely be transformed during the passage. Propyphenazone will be retarded but will most likely occur in groundwater. These results from the column experiments coincide very well with the occurrence of the pharmaceuticals clofibric acid, primidone, and propyphenazone in the first aquifer. Recommendations and Perspective  : The results underline the need to study the sorption of pharmaceuticals on various materials. e.g. organic matter, surfaces at pH values occurring in the unsaturated zone. Future field studies will also include the investigation of desorption behavior in the unsaturated zone.     

Bioefficacy of Hen Feather Keratin Hydrolysate and Compost on Vegetable Plant Growth

ABSTRACT

The potential of keratin wastes originating from poultry farms in practical application as a valuable organic fertilizer, gives rise to the need for intensive study on their effect on plants. In this study, for the first time there has been examined the influence of hen feather keratin bio-hydrolysate (FKH) and hen feather keratin compost (FKC) on plant growth, and the following features that indicate the plant condition: the leaf chlorophyll content, the activity of phenylalanine ammonia lyase (PAL) and guaiacol peroxidase (GPX) enzymes. The results of pot experiments showed a potential plant growth promoting effect of FKH, applied as a leaf treatment and of FKC manure. Fertilization of tested plants with FKC resulted in significant increase of their fresh weight. There was no effect on plant biomass after FKH treatment. The stimulating effect on plant physiology was expressed by decreased PAL activity after FKC treatment, and enhanced GPX activity, after FKH and FKC treatment. The application of FKC as a manure gave better effects for the plant condition expressed by the activity of PAL and GPX in comparison with FKH spraying. The chlorophyll content did not prove to be an efficient parameter to evaluate the impact of FKH or FKC on white cabbage, tomato, and maize plant. However, a significant increase in the leaf chlorophyll a, b, and a+b concentration was observed in cucumber plant after FKC treatment. Among the tested plants, the cucumber has shown the most profitable effect of feather compost on plant growth.     

Characterization of genetic differences between Mycobacterium avium subsp. avium strains of diverse virulence with a focus on the glycopeptidolipid biosynthesis cluster

The Mycobacterium avium complex (MAC) encompasses important pathogens in both animals and humans, yet little information is available on the factors required for MAC virulence. An animal isolate, M. avium strain 724 was found to be considerably more virulent in Balb/c mice than a human isolate, M. avium strain A5. To identify the genetic basis of this difference subtractive hybridization was applied, which resulted in the isolation of six DNA fragments unique to strain 724. BLAST searches showed that three sequences belonged to a large gene cluster responsible for biosynthesis of M. avium glycopeptidolipids (GPLs). To reveal the nature of variation between strains in the GPL cluster 27.5kb of a clone containing the A5 serotype-specific GPL (ssGPL) cluster was isolated, sequenced and compared to the corresponding region in other M. avium strains. The ssGPL cluster was highly conserved in the 5' region between all strains and serotypes tested; the 3' region reflects extensive divergence among serotypes including whole gene deletions and insertions of sequences containing open reading frames but lacking identity to any known genes.     

Exogenous leptin advances puberty in domestic hen

The present study was undertaken to examine the effect of recombinant chicken leptin administered to fed ad libitum and feed-restricted immature chickens of a layer strain on ovarian development and the timing of sexual maturity. In the first experiment 11-week-old pullets (77 days of age) fed ad libitum were injected daily with leptin at four dose levels (4, 16, 64 and 256 microg/kg body weight) until sexual maturity (lay of the first egg). Leptin treatment at the highest dose significantly (P<0.05) advanced the onset of puberty (day 116.3+/-1.0) in comparison to controls (day 121.3+/-1.2). The rises of luteinizing hormone, estradiol and progesterone in blood plasma were also advanced by leptin treatment. In the second experiment, both full-fed and feed-restricted pullets (79 days of age) were injected daily with leptin (256 microg/kg body weight). In birds fed ad libitum, exogenous leptin again significantly (P<0.05) advanced first ovipostion (day 118.4+/-1.4 versus day 124.4+/-1.7), while abolishing the significant (P<0.05) delay caused by feed restriction (day 131.5+/-1.6) and restoring the normal onset of sexual maturity (day 125.7+/-1.6). Analysis of the ovaries in 106-day-old pullets revealed that leptin injections advanced follicular development, particularly in birds fed ad libitum, and significantly (P<0.01) reduced follicular apoptosis both in full-fed and feed-restricted birds. In conclusion, we have shown that in female chickens exogenous leptin advances the onset of puberty by attenuation of ovarian apoptosis and enhancement of folliculogenesis.     

Involvement of mammalian Mus81 in genome integrity and tumor suppression

Mus81-Eme1 endonuclease has been implicated in the rescue of stalled replication forks and the resolution of meiotic recombination intermediates in yeast. We used gene targeting to study the physiological requirements of Mus81 in mammals. Mus81-/- mice are viable and fertile, which indicates that mammalian Mus81 is not essential for recombination processes associated with meiosis. Mus81-deficient mice and cells were hypersensitive to the DNA cross-linking agent mitomycin C but not to gamma-irradiation. Remarkably, both homozygous Mus81-/- and heterozygous Mus81+/- mice exhibited a similar susceptibility to spontaneous chromosomal damage and a profound and equivalent predisposition to lymphomas and other cancers. These studies demonstrate a critical role for the proper biallelic expression of the mammalian Mus81 in the maintenance of genomic integrity and tumor suppression.     

Evaluation of Deammonification Process by Response Surface Models

The influence of the operational variables on the Anammox process has been generally researched considering each variable separately. However, the optimization of the process also requires the identification of the more significant variables and their possible interactions. Response surface models were successfully applied to evaluate the performance of the Anammox process in a deammonification system (i.e., one-stage biofilm Anammox process) taking into account the combined effects caused by two sets of three variables. Specific Anammox activity was measured by a manometric method and used as the response variable. The obtained models pointed out that the significant variables were the temperature, the value of pH, and the ratio between the unionized species of the substrates (free ammonia and free nitrous acid (FA/FNA)). There were interactions among them caused by chemical equilibriums. Total nitrogen concentration and ammonium concentration were found to be not significant in the tested range. According to the models, the optimum values of temperature, pH, and free ammonia to free nitrous acid ratio within the test ranges were, respectively, 30°C, 7.0, and 0.3. Further research at higher temperatures and lower values of pH and FA/FNA ratios would be necessary in order to find the absolute optimum conditions for the process. The obtained model can be also useful in order to develop control strategies that take into account the significant variables and their optimum ranges. A strategy to control deammonification reactors has been proposed, according to the results of the modeling.     

Impact of Maillard reaction on immunoreactivity and allergenicity of the hazelnut allergen Cor a 11

Few studies exist on the influence of processing methods on structural changes and allergenic potential of hazelnut proteins. This study focused on the effect of glycation (Maillard reaction) on the immunoreactivity and degranulation capacity of the purified hazelnut 7S globulin, Cor a 11. After heating, the extent of the Maillard reaction, sensitivity to proteolysis, binding of human IgE or rabbit IgG, and degranulation capacity were analyzed. Changes in electrophoretic mobility, amount of free amino groups, and contents of bound sugar and fructosamine indicated that glycation of Cor a 11 occurred at all conditions. Glycation at 37 °C did not influence the specific IgG or IgE binding and was decreased after heating at 60 and 145 °C. Heating, with or without glucose, at 145 °C increased basophil degranulation capacity. The results suggest that glycation of Cor a 11 at 60 and 145 °C may decrease the IgE/IgG binding properties but not the degranulation capacity of basophils. This is possibly related to aggregation of the proteins as a result of the Maillard reaction.