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PubMed2002

Gaseous nitrogen losses from a forest site in the North Tyrolean Limestone Alps.

Microorganisms are responsible for the mineralisation of organic nitrogen in soils. NH4+ can be further oxidised to NO3- during nitrification and NO3- can be reduced to gaseous nitrogen compounds during denitrification. During both processes, nitrous oxide (N2O), which is known as greenhouse gas, can be lost from the ecosystem. The aim of this study was to quantify N2O emissions and the internal microbial N cycle including net N mineralisation and net nitrification in a montane forest ecosystem in the North Tyrolean Limestone Alps during an 18-month measurement period and to estimate the importance of these fluxes in comparison with other components of the N cycle. Gas samples were taken every 2 weeks using the closed chamber method. Additionally, CO2 emission rates were measured to estimate soil respiration activity. Net mineralisation and net nitrification rates were determined by the buried bag method every month. Ion exchange resin bags were used to determine the N availability in the root zone. Mean N2O emission rate was 0.9 kg N ha(-1) a(-1), which corresponds to 5% of the N deposited in the forest ecosystem. The main influencing factors were air and soil temperature and NO3- accumulated on the ion exchange resin bags. In the course of net ammonification, 14 kg NH4+-N ha(-1) were produced per year. About the same amount of NO3--N was formed during nitrification, indicating a rather complete nitrification going on at the site. NO3- concentrations found on the ion exchange resin bags were about 3 times as high as NO3- produced during net nitrification, indicating substantial NO3- immobilisation. The results of this study indicate significant nitrification activities taking place at the Mühleggerköpfl.

Altitude↗
PubMed1996

Effect of nitrogen dioxide and other combustion products on asthmatic subjects in a home-like environment.

Nitrogen dioxide (NO2) is one of a number of nitrogen compounds that are by-products of combustion and occur in domestic environments following the use of gas or other fuels for heating and cooking. In this study, we examined the effect of two levels of NO2 on symptoms, lung function and airway hyperresponsiveness (AHR) in asthmatic adults and children. In addition, in the same subjects, we examined the effects of the same levels of NO2 mixed with combustion by-products from a gas space heater. The subjects were nine adults, aged 19-65 yrs, and 11 children, aged 7-15 yrs, with diagnosed asthma which was severe enough to require daily medication. All subjects had demonstrable AHR to histamine. Exposures were for 1 h on five separate occasions, 1 week apart, to: 1) ambient air, drawn from outside the building; 2) 0.3 parts per million (ppm) NO2 in ambient air; 3) 0.6 ppm NO2 in ambient air; 4) ambient air+combustion by-products+NO2 to give a total of 0.3 ppm; and 5) ambient air+combustion by-products+NO2 to give a total of 0.6 ppm. Effects were measured as changes in lung function and symptoms during and 1 h after exposure, in AHR 1 h and 1 week after exposure, and in lung function and symptoms during the week following exposure. Exposure to NO2 either in ambient air or mixed with combustion by-products from a gas heater, had no significant effect on symptoms or lung function in adults or in children. There was a small, but statistically significant, increase in AHR after exposure to 0.6 ppm NO2 in ambient air. However, there was no effect of 0.6 ppm NO2 on AHR when the combustion by-products were included in the test atmosphere nor of 0.3 ppm NO2 under either exposure condition. We conclude that a 1 h exposure to 0.3 or 0.6 ppm NO2 has no clinically important effect on the airways of asthmatic adults or children, but that 0.6 ppm may cause a slight increase in airway hyperresponsiveness.

Adolescent↗
PubMed1999

On-line delta(18)O measurement of organic and inorganic substances

A method for the automated sample conversion and on-line oxygen isotope ratio (delta(18)O) determination for organic and inorganic substances is presented. The samples are pyrolytically decomposed at 1400 degrees C in the presence of nickelized graphite. With the system presented organic as well as inorganic samples such as nitrates, sulphates and phosphates of 50-100 &mgr;g O can be analyzed for their delta(18)O values with a standard deviation usually better than 0.5 per thousand. Additionally, carbon isotope ratios of organic substances and nitrogen isotope ratios of inorganic nitrogenous compounds are available in the same sample run. Data for international and some inter-laboratory reference materials are presented to show the accuracy and reliability of the method. The effect of some additives on the CO yield was checked for substances which do not pyrolyze completely. Copyright 1999 John Wiley & Sons, Ltd.

Journal Article↗
PubMed1980

Heterotrophic nitrification by intestinal microorganisms.

From studies of nitrate balance in man and analyses of fecal and ileostomy samples, the possibility that nitrite and nitrate are formed de novo in the intestine, possibly by heterotrophic nitrification has emerged. This proposition significantly alters our previous conceptions of man's exposure to nitrite and suggests that nitrite may play a role in the cause of intestinal cancer. Heterotrophic nitrification has been demonstrated in various microorganisms. Our work has shown that intestinal heterotrophic microbial isolates from man are able to oxidize nitrogenous compounds to nitrite. These isolates include both procaryotes and eucaryotes.

Actinomyces↗
PubMed1989

Cytochrome P450 activity and distribution in the human colon mucosa.

Enzymatic activity associated with the mixed-function oxidase system was determined in microsomes prepared from the mucosal cells extracted from normal human colons. A high activity toward nitrogen oxidation reactions was observed. 1,2-Dimethylhydrazine, a colon-specific carcinogen, was metabolized at a higher rate in vitro by human colon microsomes as compared with the rat, and exhibited a km ten-fold lower, 1.03 mmol/l versus 9.68 mmol/l, respectively. This activity was inhibited by classic cytochrome P450 inhibitors; 70% inhibition was achieved using 70 mmol/l metyrapone (2-methyl-1,2-di-3-pyridyl-1-propanone), 20 mmol/l; SKF-525A (diethylaminoethyl-2,-2-diphenylvalerate HCl), or 350 mumol/l n-octylamine. These data suggest the presence of a stable, active mixed-function oxidase system in the human colon mucosa which has a preferential activity toward nitrogenous compounds and provides a mechanism for the activation of carcinogens. Its distribution in the colon appears to parallel the reported incidence of human colonic carcinomas.

1,2-Dimethylhydrazine↗
PubMed1980

[Determination of intramuscular fat in beef by means of extraction with chloroform-methanol].

In a comparison on samples of muscular substance from 53 fattening bulls the extraction of intramuscular fat with a chloroform-methanol mixture yielded results about 1.23% higher than those obtained with the extraction with diethyl ether. This difference results from the extraction of additional water-soluble nitrogenous compounds by the chloroform-methanol mixture. These compounds were isolated by adding water to the chloroform-methanol extract; their analysis by means of chemical and spectroscopic methods revealed, among other compounds, creatinine and threonine. The fat values obtained for the ether extract were closely correlated with those found for the chloroform-methanol extract (r = 0.925). In a variant where water was added to the chloroform-methanol extract, the correlation was less close (r = 0.925). In a variant where water was added to the chloroform-methanol extract, the correlation was less close (r = 0.660). The chloroform-methanol extraction may be used instead of the ether extraction. A regression calculated from the present data permits, if required, to convert the values thus obtained into the values yielded by the ether extraction method. Apart from its high precision, the chloroform-methanol extraction has the special advantage that it requires relatively simple safety measures and that its energy demand is low.

Animals↗
PubMed1987

Gnotobiotes in nutritional studies.

Studies with gnotobiotic animals have demonstrated extensive synthesis of vitamins, involvement in the metabolism of nitrogenous compounds and modification of lipids among the activities of the conventional microflora. The extent to which they affect the host depends on factors such as the structure of the gut, the position of the sites of bacterial proliferation and the host's nutritional status. Thus the findings in experimental animals may not always be applicable to man. Results so far indicate that in circumstances of dietary inadequacy bacterial activity may be detrimental or beneficial. However, these effects are small, and it seems reasonable to conclude that the influence of the indigenous microflora on the host's nutrition is of relatively minor importance compared with its role as a barrier against invasion by undesirable organisms.

Animal Nutritional Physiological Phenomena↗
PubMed1988

Utilization of milk proteins and cereal starches to obtain co-extrudates.

Extrusion of the cereal mixtures with 25% addition of milk proteins induced several changes in the chemical and physico-chemical properties of starch. Coefficient of expansion was close to the coefficient of natural cereal flours, but the extrudates were characterized by higher water absorption. Starch was only slightly dextrinized, but it contained more nitrogen compounds and mineral salts, and less free lipids. Solubility and swelling power of the extrudates did not change in a way characteristic for cereal starch and flours. Attention should be given to higher content of dietary fibre, pointing to the formation of the starch-lipids and the starch-protein complexes, less susceptible to enzymatic hydrolysis used. Hence, it can be stated that the latter increase was apparent only. It can be said that a mixture of cereal starch and milk proteins can be used to modify quality of the extruded products of increased protein content. More attention should be devoted to biochemical characteristics of the protein added, as this largely determines quality of the extruded starch-protein products.

Chemical Phenomena↗
PubMed2006

Identification of novel mutations in the human ornithine transcarbamylase (OTC) gene of Korean patients with OTC deficiency and transient expression of the mutant proteins in vitro.

The urea cycle plays key roles to prevent the accumulation of toxic nitrogenous compound and synthesize arginine de novo. Ornithine transcarbamylase (OTC) deficiency is the most common inborn error of urea cycle, which is inherited in an X-linked manner. This study was undertaken to characterize molecular defects in Korean patients with OTC deficiency. With direct sequence analysis of OTC gene of 26 unrelated Korean patients with OTC deficiency, 23 different mutations were identified. Among these mutations, eleven were novel mutations. The novel mutations were p.Leu9X, p.Arg26Pro, p.Gly100Arg, p.Met205Thr, p.Lys221Asn, p.Asp249Gly, p.Phe281Ser, p.Val323Met, c.571delC, c.853delC, and c.796-805del. All the novel mutations in this study were tested in 100 normal alleles. In vitro expression study of some of novel missense mutations elucidated the correlation of genotype and phenotype of the OTC deficiency.

Adolescent↗
PubMed2004

Influence of growth conditions on the production of a bacteriocin by Lactococcus lactis subsp. lactis ST34BR, a strain isolated from barley beer.

Bacteriocin ST34BR, a small polypeptide of 2.9 kDa produced by Lactococcus lactis subsp. lactis ST34BR, inhibits the growth of Enterococcus faecalis, Escherichia coli, Lactobacillus plantarum, Lactobacillus casei, Pseudomonas aeruginosa and Staphylococcus aureus. MRS broth, adjusted to pH 6.0 yielded 6,400 AU/ml, compared to 400 AU/ml recorded in BHI broth, M17 broth, 10% (w/v) soy milk, and 8% and 10% (w/v) molasses. At pH of 4.5 only 800 AU/ml was produced. Based on comparative studies in MRS broth, without organic nitrogen, supplemented with different combinations of tryptone, meat extract and yeast extract, tryptone was identified as the stimulating nitrogen compound. Growth in the presence of 20 g/l glucose, maltose, mannose or sucrose yielded bacteriocin levels of 6,400 AU/ml, whereas the same concentration of lactose and fructose yielded 3,200 AU/ml and 1,600 AU/ml, respectively. No difference in bacteriocin ST34BR activity was recorded in MRS broth supplemented with 2 g/l K2HPO4 and 2 g/l, 5 g/l, 10 g/l or 50 g/l KH2PO4. However, 20 g/l KH2PO4 increased bacteriocin ST34BR production to 12,800 AU/ml. Glycerol at 1g/l to 10 g/l in MRS broth reduced bacteriocin activity to 3,200 AU/ml, whilst 20 g/l and 50 g/l yielded only 1,600 AU/ml. The presence of cyanocobalamin, L-ascorbic acid, thiamine and DL-6,8-thioctic acid in MRS broth at 1.0 ppm, respectively, did not result in increased activity levels.

Animals↗
PubMed1995

Proteolytic activity of Aeromonas caviae.

Aeromonas strains produce a variety of virulence factors including proteases. Studies on the kinetics of growth of Aeromonas caviae NRRL B-966 and its proteases suggest that the proteolytic activities are produced throughout the growth phase, with peak level occurring at stationary phase. A. caviae synthesize both intracellular and extracellular proteases with the latter account for major portion of the total activity. Optimum pH for the A. caviae proteolytic activity is at 7.0. A. caviae produces a thermoresistant protease, whose activity is dependent on Mg++ and Ca++ ions. Inhibition of proteolytic activity by phenyl methyl sulfonyl fluoride suggest the presence of a serine protease in A. caviae. Nitrogenous compounds enhance the proteolytic activity while carbohydrates tested in this study inhibit the activity.

Aeromonas↗
PubMed1979

Regulative influence of o-aminobenzoic acid on the biosynthesis of nourseothricin in cultures of Streptomyces noursei JA 3890b. IV. Bistability of metabolism and the mechanism of action of aminobenzoic acids.

Using the semi-continuous cultivation technique we could establish that specifically in Streptomyces noursei JA 3890b during growth on a medium supplied with D,L-alanine, NH4+, and maize starch there are two different phenotypes of the organism and stationary states of metabolism, respectively. The expression of either the metabolic state I with an enhanced capacity to oxidative deamination of alanine via the NAD+-dependent alanaine dehydrogenase or the metabolic state 2 which may be characterized by the preferred use of ammonium ions via the NADP+-dependent glutamate dehydrogenase was shown to depend strongly on the conditions of inoculum cultivation. When the amino acid permeases were derepressed by cultivating the inoculum cells on amino acid media, probably due to the defective mechanism of negative feedback control of amino acid influx in this strain an abnormously high uptake of alanine was observed that, consequently, was correlated to the enhanced oxidation of this amino acid as well as to the intensive production of ammonia within the cell. This overproduction of cellular NH4+ seems to bring about the subsequent repression of biosynthetic glutamate dehydrogenase and so on the accumulation of ammonia autocatalytically may rise up (metabolic state I). On the other hand, if the influx of alanine was kept low and the NADH oxidation was less efficient, respectively, or when there was high cellular activity of glutamate dehydrogenase the level of ammonia never did exceed the respressory limit and, accordingly, the expression of the metabolic state 2 was observed. Switching-over of metabolic flux from the state 2 towards the state 1 can be brought about either by increasing the level of nitrogen sources in the medium or by adding buffers pH greater than 7.5. In contrast, decrease of cellular level of NH4+ was shown to induce the transition of metabolic state 1 into the state 2. This can be achieved not only by limitation of nitrogen source but also by adding different aminobenzoic acids and, alternatively, effectors of membrane function (short-chain alcohols), inhibitors of cytochrome oxidases (sodium azide, potassium cyanide), heavy metal (Fe++)-chelating agents (catechol, 2,5'-dipyridyl, o-phenanthroline), beta-alanine, and buffers pH less than 7. This suggests that these effectors are capable of preventing the abnormously high influx of amino acids as well as its wasteful catabolism within the cell of S. noursei JA 3890b. Therefore, it seems likely that by this way the aminobenzoic acids and similar effectors can diminish the catabolite repression or inhibition of secondary metabolism by cellular excess of some nitrogen compounds in good agreement with its well-known stimulatory action on the biosynthesis of the antibiotic nourseothricin in this strain.

Alanine↗
PubMed1982

[Physiological and cell biological characterization of the protease producer Thermoactinomyces vulgaris during prolonged culture in a stirred fermenter].

The physiological behaviour of Thermoactinomyces vulgaris - producing a thermostable serine-protease - was analyzed during fermentation. During 38 h the consumption of nutrients and oxygen as well as the rates of macromolecular and protease synthesis were measured. The morphological and ultrastructural changes of the mycelia were also studied. The mycelia grew exponentially for about 5 hours. After a short lag and a second slower growth phase, growth continued about linearly until the end, as was indicated by a constant rate of incorporation of labelled thymidine. However, at the same time a changing portion of hyphae - up to 45% - underwent lysis. According to the changing ratio of growing and lysing material, regarding the physiological activity of the culture the fermentation process could be divided into 4 periods. The formation of the protease started at the transition to the slow growth phase and continued linearly. The ability to produce the protease was attributed to a mycelium being formed after the shift down caused by limitation of supply of utilizable nitrogen compounds. The end of protease production 10 h later was correlated to a drastic decrease of the respiratory activity of the mycelia, probably caused by exhaustion of easily utilizable carbohydrates.

Amino Acids↗
PubMed2004

An overview of polylactides as packaging materials.

Polylactide polymers have gained enormous attention as a replacement for conventional synthetic packaging materials in the last decade. By being truly biodegradable, derived from renewable resources and by providing consumers with extra end-use benefits such as avoiding paying the "green tax" in Germany or meeting environmental regulations in Japan, polylactides (PLAs) are a growing alternative as a packaging material for demanding markets. The aim of this paper is to review the production techniques for PLAs, summarize the main properties of PLA and to delineate the main advantages and disadvantages of PLA as a polymeric packaging material. PLA films have better ultraviolet light barrier properties than low density polyethylene (LDPE), but they are slightly worse than those of cellophane, polystyrene (PS) and poly(ethylene terephthalate) (PET). PLA films have mechanical properties comparable to those of PET and better than those of PS. PLA also has lower melting and glass transition temperatures than PET and PS. The glass transition temperature of PLA changes with time. Humidity between 10 and 95% and storage temperatures of 5 to 40 degrees C do not have an effect on the transition temperature of PLA, which can be explained by its low water sorption values (i.e. <100 ppm at Aw = 1). PLA seals well at temperatures below the melting temperature but an appreciable shrinking of the films has been noted when the material is sealed near its melting temperature. Solubility parameter predictions indicate that PLA will interact with nitrogen compounds, anhydrides and some alcohols and that it will not interact with aromatic hydrocarbons, ketones, esters, sulfur compounds or water. The CO2, O2 and water permeability coefficients of PLA are lower than those of PS and higher than those of PET. Its barrier to ethyl acetate and D-limonene is comparable to PET. The amount of lactic acid and its derivatives that migrate to food simulant solutions from PLA is much lower than any of the current average dietary lactic acid intake values allowed by several governmental agencies. Thus, PLA is safe for use in fabricating articles for contact with food.

Biocompatible Materials↗
PubMed1994

A new family of carbon-nitrogen hydrolases.

Using computer methods for database search and multiple alignment, statistically significant sequence similarities were identified between several nitrilases with distinct substrate specificity, cyanide hydratases, aliphatic amidases, beta-alanine synthase, and a few other proteins with unknown molecular function. All these proteins appear to be involved in the reduction of organic nitrogen compounds and ammonia production. Sequence conservation over the entire length, as well as the similarity in the reactions catalyzed by the known enzymes in this family, points to a common catalytic mechanism. The new family of enzymes is characterized by several conserved motifs, one of which contains an invariant cysteine that is part of the catalytic site in nitrilases. Another highly conserved motif includes an invariant glutamic acid that might also be involved in catalysis.

Amidohydrolases↗
PubMed2005

Identification of volatile basic components in tobacco by headspace liquid-phase microextraction coupled to matrix-assisted laser desorption/ionization with Fourier transform mass spectrometry.

A method incorporating headspace liquid-phase microextraction (HS-LPME) coupled to matrix-assisted laser desorption/ionization (MALDI) with Fourier transform mass spectrometry (FTMS) was established to analyze volatile basic components in tobacco. The sample preparation volume for MALDI-MS was compatible with the volume of the solvent microdrop in the HS-LPME procedure. The pH and the polarity of the solvent for HS-LPME were adjusted by choice of the MALDI matrix and matrix additive. Based on the elemental composition and tandem mass spectrometry information, 25 volatile nitrogenous compounds in tobacco were detected and identified. The approach is fast and sensitive, and has the potential for automation for high-throughput analysis. This approach offers an alternative method for analysis of trace volatile organic compounds in complex samples.

Gas Chromatography-Mass Spectrometry↗
PubMed2006

A 15N-aided artificial atmosphere gas flow technique for online determination of soil N2 release using the zeolite Köstrolith SX6.

N2 is one of the major gaseous nitrogen compounds released by soils due to N-transformation processes. Since it is also the major constituent of the earth's atmosphere (78.08% vol.), the determination of soil N2 release is still one of the main methodological challenges with respect to a complete evaluation of the gaseous N-loss of soils. Commonly used approaches are based either on a C2H2 inhibition technique, an artificial atmosphere or a 15N-tracer technique, and are designed either as closed systems (non-steady state) or gas flow systems (steady state). The intention of this work has been to upgrade the current gas flow technique using an artificial atmosphere for a 15N-aided determination of the soil N2 release simultaneously with N2O. A 15N-aided artificial atmosphere gas flow approach has been developed, which allows a simultaneous online determination of N2 as well as N2O fluxes from an open soil system (steady state). Fluxes of both gases can be determined continuously over long incubation periods and with high sampling frequency. The N2 selective molecular sieve Köstrolith SX6 was tested successfully for the first time for dinitrogen collection. The presented paper mainly focuses on N2 flux determination. For validation purposes soil aggregates of a Haplic Phaeozem were incubated under aerobic (21 and 6 vol.% O2) and anaerobic conditions. Significant amounts of N2 were released only during anaerobic incubation (0.4 and 640.2 pmol N2 h(-1) g(-1) dry soil). However, some N2 formation also occurred during aerobic incubation. It was also found that, during ongoing denitrification, introduced [NO3]- will be more strongly delivered to microorganisms than the original soil [NO3]-.

Journal Article↗