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Cerumen composition by flash pyrolysis-gas chromatography/mass spectrometry.

OBJECTIVE: To assess the chemical composition of cerumen by flash pyrolysis-gas chromatography/mass spectrometry. STUDY DESIGN: Collected earwax specimens were fractionated into residue and supernatant by means of deoxycholate. This natural bile acid produces significantly better disintegration of earwax in vitro than do presently available ceruminolytic preparations, and also has demonstrated excellent clinical results in vivo to date. PATIENTS: The sample for analysis was obtained from a patient with clinical earwax impaction. RESULTS: The supernatant is composed of simple aromatic hydrocarbons, C5-C17 straight-chain hydrocarbons, a complex mixture of compounds tentatively identified as diterpenoids, and steroids, in particular cholesterol. The residue, on the other hand, produced simple aromatic compounds (including benzenes, phenols, and benzonitriles), C5-C25 straight-chain hydrocarbons, greater relative quantities of nitrogen compounds and phenol, and lesser importance of the (tentatively identified) diterpenoids. CONCLUSIONS: Through the use of the detergent deoxycholate, squalene and a tentatively identified diterpenoid were revealed to be present in a free, unbound state, whereas some steroids and hydrocarbons appeared to be bound to a macromolecular structure by nitrogen linkages or other bonds. Additionally, this study reintroduces detergents as a viable method of earwax removal, specifically the bile acids.

Carbon↗

Relationships between structures and mutagenic potencies of 16 heterocyclic nitrogen mustards (ICR compounds) in Salmonella typhimurium.

16 heterocyclic nitrogen mustards (ICR compounds), which were synthesized for use as possible antitumor agents by Creech and coworkers, were tested for mutagenicity in Salmonella typhimurium strains TA1535, TA1536, TA1537, TA1538, TA98 and TA100. The compounds were incorporated into the top agar at 5 doses: 0.5, 1, 2.5, 5 and 10 micrograms/plate. All of the compounds were negative in TA1535 except ICR 449, which was positive in all 6 strains. The other 15 compounds were positive in the remaining strains with the following exceptions: ICR 371 and 355 were negative in TA100; ICR 445 was negative in TA98 and TA100; and ICR 360 was negative in TA1537, TA1538, TA98 and TA100. Good qualitative agreement was observed between the mutagenic and antitumor activities of the 16 compounds, and between the mutagenic and carcinogenic activities of the 5 compounds that have been tested for carcinogenicity by Peck and coworkers. However, no significant correlation was found between mutagenic potency in Salmonella and antitumor potency in mice for the 16 compounds. Also, for the 5 compounds that have been tested for carcinogenicity, no significant correlation was found between their mutagenic potency in Salmonella and their carcinogenic potency in mice. In Salmonella, the secondary (2 degrees) amines generally were more mutagenic than their tertiary (3 degrees) amine homologs, although the opposite result has been reported in certain eukaryotes. Relationships between structures and potencies for the different nuclei of the 16 ICR compounds are discussed, as are similarities and differences in strain sensitivities. We conclude that the Salmonella his reversion test is not a good predictor of the antitumor and carcinogenic potencies of these ICR compounds.

Adenoma↗

Atmospheric deposition of nitrogen and sulphur compounds in the Czech Republic.

Estimates of dry and wet deposition of nitrogen and sulphur compounds in the Czech Republic for the years 1994 and 1998 are presented. Deposition has been estimated from monitored and modeled concentrations in the atmosphere and in precipitation, where the most important acidifying compounds are sulphur dioxide, nitrogen oxides, ammonia, and their reaction products. Measured atmospheric concentrations of SO2, NOx, NH3, and aerosol particles (SO4(2-), NO3-, and NH4+), along with measured concentrations of SO4(2-), NO3-, and NH4+ in precipitation, weighted by precipitation amounts, were interpolated with Kriging technique on a 10- x 10-km grid covering the whole Czech Republic. Wet deposition was derived from concentration values for SO4(2-), NO3-, and NH4+ in precipitation and from precipitation amounts. Dry deposition was derived from concentrations of gaseous components and aerosol in the air, and from their deposition velocities. A multiple resistance model was used for calculation of SO2, NOx, and NH3 deposition velocities. Deposition velocities of particles were parameterized. It was estimated that the annual average deposition of SOx in the Czech Republic decreased from 1384 to 1027 mol H + ha(-1) a(-1) between 1994 and 1998. The annual average NOy deposition was estimated to be 972 and 919 mol H + ha(-1) a(-1) in 1994 and 1998, respectively. The annual average NHx deposition was estimated to be 887 mol H+ ha(-1) a(-1) and 779 mol H + ha(-1) a(-1) in 1994 and 1998, respectively. It was estimated that the annual average of the total potential acid deposition decreased from 3243 to 2725 mol H + ha(-1) a(-1) between 1994 and 1998. Sulphur compounds (SOx) contributed about 38%, oxidized nitrogen species (NOy) 34%, and reduced nitrogen species (NHx) 28% to the total potential acid deposition in 1998. The wet deposition contributed 42% to the total potential acid deposition in 1998.

Algorithms↗

Removal of carbonaceous and nitrogenous pollutants from a synthetic wastewater using a membrane-coupled bioreactor.

Two modified Ludzack-Ettinger (MLE)-type membrane-coupled bioreactors (MBRs) were investigated in this study for the purpose of removing both nitrogenous and carbonaceous pollutants from a synthetic wastewater. During the first MBR experiment, removal efficiencies were high (>90%) for chemical oxygen demand (COD) and ammonia, but total nitrogenous pollutant removal efficiency was poor (approximately 25%). Bacterial community analysis of ammonia oxidizing bacteria (AOB) by a nested PCR-DGGE approach detected two Nitrosomonas-like populations and one Nitrosospira-like population. During the initial portion of the second MBR experiment, COD and ammonia removal efficiencies were similar to the first MBR experiment until the COD of the influent wastewater was increased to provide additional electron donors to support denitrification. Total nitrogen removal efficiencies eventually exceeded 90%, with a hydraulic residence time (HRT) of 24 h and a recirculation ratio of 8. When the HRT of the MBR experiment was decreased to 12 h, however, ammonia removal efficiency was adversely affected. A subsequent increase in the HRT to 18 h helped improve removal efficiencies for both ammonia (>85%) and total nitrogenous compounds (approximately 70%). Our research demonstrates that MBRs can be effectively designed to remove both carbonaceous and nitrogenous pollutants. The ability of the microbial community to switch between anoxic (denitrifying) and oxic (nitrifying) conditions, however, represents a critical process constraint for the application of MLE-type MBR systems, such that little benefit is gained compared to conventional designs.

Aerobiosis↗

Nutritional requirements of mycelial growth of Cordyceps sinensis in submerged culture.

AIMS: The nutritional requirements for mycelial growth of Cordyceps sinensis in semi-synthetic liquid media were investigated. The results provide a basis for further physiological study and industrial fermentation of the fungus. METHODS AND RESULTS: Nutritional requirements, including 17 carbohydrates, 16 nitrogen compounds, nine vitamins, four macro-elements, four trace-elements and eight ratios of carbon to nitrogen, were studied for their effects on the mycelial growth in submerged cultures of C. sinensis by using one-factor-at-a-time and orthogonal matrix methods. Among these variables, sucrose, peptone, folic acid, calcium, zinc and a carbon to nitrogen ratio 12 : 1 were identified as the requirements for the optimum mycelial growth. The concentrations of sucrose, peptone and yeast extract were optimized and the effects of medium composition on mycelial growth were found to be in the order sucrose > yeast extract > peptone. The optimal concentration for mycelial growth was determined as 50 g l(-1) sucrose, 10 g l(-1) peptone and 3 g l(-1) yeast extract. CONCLUSIONS: Under optimal culture conditions, over 22 g l(-1) of mycelial biomass could be obtained after 40 days in submerged cultures. SIGNIFICANCE AND IMPACT OF THE STUDY: Cordyceps sinensis, one of the most valued medicinal fungi, is shown to grow in axenic culture. This is the first report on nutritional requirements and design of a simplified semi-synthetic medium for mycelial growth of this psychrophilic species, which grows slowly below 20 degrees C. The results of this study will facilitate research on mass production of the fungus under defined culture conditions.

Carbohydrates↗

Airborne sulfur and nitrogen in Finland-trends and exposure in relation to air transport sector.

We present the concentration trends and the atmospheric exposure of sulfur and nitrogen compounds in different transport sectors during the period 1981-2000, based on the air quality data of the Finnish Meteorological Institute. Sulfur and nitrogen concentrations in air and precipitation were assessed from background stations covering the whole country. A significant decrease of more than 60% was observed in the concentrations of all sulfur compounds throughout the country during 1981-2000. In the 1990's, significant trends were detected in all sulfur time series, with the exception of sulfate in precipitation at the northernmost stations. On the other hand, the concentrations of oxidized nitrogen compounds in air and precipitation have declined only slightly or not at all, especially in the 1990's. This is attributed to the fairly small domestic and European NO(x) emission reductions after 1981. The ammonium concentration in precipitation has declined by more than 50% during 1981-2000. The atmospheric sulfur exposure was found to be dominated by transport from the West and South-West sectors during summer, whereas in winter all the southward sectors were important. The largest decline in the sectoral sulfur concentrations has taken place already before 1991, with a more pronounced reduction during winter than during summer. The atmospheric nitrogen exposure was mainly dominated by transport from the sectors between west and south, but in the latter half of the study period there seems to have been a slight shift towards increased transport from the eastern and northern sectors.

Air Movements↗

Characterization of Gordonia sp. strain F.5.25.8 capable of dibenzothiophene desulfurization and carbazole utilization.

A dibenzothiophene (DBT)-degrading bacterial strain able to utilize carbazole as the only source of nitrogen was identified as Gordonia sp. F.5.25.8 due to its 16S rRNA gene sequence and phenotypic characteristics. Gas chromatography (GC) and GC-mass spectroscopy analyses showed that strain F.5.25.8 transformed DBT into 2-hydroxybiphenyl (2-HBP). This strain was also able to grow using various organic sulfur or nitrogen compounds as the sole sulfur or nitrogen sources. Resting-cell studies indicated that desulfurization occurs either in cell-associated or in cell-free extracts of F.5.25.8. The biological responses of F.5.25.8 to a series of mutagens and environmental agents were also characterized. The results revealed that this strain is highly tolerant to DNA damage and also refractory to induced mutagenesis. Strain F.5.25.8 was also characterized genetically. Results showed that genes involved in desulfurization (dsz) are located in the chromosome, and PCR amplification was observed with primers dszA and dszB designed based on Rhodococcus genes. However, no amplification product was observed with the primer based on dszC.

Biotechnology↗

Effects of carbon and nitrogen sources, carbon-to-nitrogen ratio, and initial pH on the growth of nematophagous fungus Pochonia chlamydosporia in liquid culture.

The effects of carbon and nitrogen sources, carbon-to-nitrogen ratio (C:N) and initial pH value on the growth and sporulation of the nematophagous fungus Pochonia chlamydosporia in liquid culture were examined. Among the 21 carbon sources and 15 nitrogen compounds tested, the optimal carbon and nitrogen sources for mycelial growth were sweet potato and L: -tyrosine, and for sporulation were sweet potato and casein peptone. A C:N ratio of 10:1 at pH 3.7 gave the maximum yield of conidia and a C:N ratio of 40:1 at pH 6.8 gave the maximum biomass. The initial pH value had a significant effect on mycelial growth and conidial production, with the optimal ranges being 3.5-4.5 for sporulation and 5-6 for growth. Maximum conidial production was obtained at an initial pH of 4.0 and the maximum biomass at pH 6.0. The results also showed that the final pH after 7 days cultivation was always higher than the initial value. The variability in growth and sporulation of seven strains of P. chlamydosporia in liquid culture was also compared and discussed.

Biomass↗

Simultaneous sulfur dioxide and nitrogen dioxide removal by calcium hydroxide and calcium silicate solids.

At conditions typical of a bag filter exposed to a coal-fired flue gas that has been adiabatically cooled with water, calcium hydroxide and calcium silicate solids were exposed to a dilute, humidified gas stream of nitrogen dioxide (NO2) and sulfur dioxide (SO2) in a packed-bed reactor. A prior study found that NO2 reacted readily with surface water of alkaline and non-alkaline solids to produce nitrate, nitrite, and nitric oxide (NO). With SO2 present in the gas stream, NO2 also reacted with S(IV), a product of SO2 removal, on the exterior of an alkaline solid. The oxidation of S(IV) to S(VI) by oxygen reduced the availability of S(IV) and lowered removal of NO2. Subsequent acidification of the sorbent by the removal of NO2 and SO2 facilitated the production of NO. However, the conversion of nitrous acid to sulfur-nitrogen compounds reduced NO production and enhanced SO2 removal. A reactor model based on empirical and semi-empirical rate expressions predicted rates of SO2 removal, NO2 removal, and NO production by calcium silicate solids. Rate expressions from the reactor model were inserted into a second program, which predicted the removal of SO2 and NOx by a continuous process, such as the collection of alkaline solids in a baghouse. The continuous process model, depending upon inlet conditions, predicted 30-40% removal for NOx and 50-90% removal for SO2. These results are relevant to dry scrubbing technology for combined SO2 and NOx removal that first oxidizes NO to NO2 by the addition of methanol into the flue duct.

Air Pollutants, Occupational↗

Nutritional treatment of liver cirrhosis by branched-chain amino acid-enriched nutrient mixture.

The effects of branched-chain amino acid (BCAA)-enriched nutrient mixture (nutrient-mixture) on the nitrogen metabolism and nutritional state were clinically investigated in 10 patients with liver cirrhosis. Nutrient-mixture-supplemented diet was prepared by adding 150 g nutrient-mixture daily to low-protein diet, and comparisons were made with a regular diet (control diet). Each diet supplied 2,100 kcal energy and 80 g protein per day. Patients were given control diet for 2 weeks and thereafter treated successively with nutrient-mixture-supplemented and control diet each for 2 weeks. Nitrogen balance improvement and positive balance were observed during the feeding of nutrient-mixture-supplemented diet. The composition of nitrogen compounds in urine and the fecal nitrogen excretion did not alter during the test period. Plasma aromatic amino acid (AAA) concentrations decreased and BCAA/AAA molar ratios increased significantly during the 1st and 2nd week of nutrient-mixture-supplemented diet administration. Plasma methionine concentration also decreased in the 1st week. Plasma pre-albumin levels rose significantly during the 1st and 2nd week of nutrient-mixture-supplemented diet administration, and the number connection test improved significantly following the supplemented diet. These results suggest that the use of nutrient-mixture in the nutritional treatment of liver cirrhosis had no deleterious effects on nitrogen metabolism and is useful for the improvement of plasma amino acid imbalance and protein-energy malnutrition.

Adult↗

Results for 74 substances tested for interference with determination of plasma catecholamines by "high-performance" liquid chromatography with electrochemical detection.

Various catecholamine metabolites, catecholamine-related compounds, catechols, drugs, amines, and other nitrogen compounds were injected onto an HPLC system ("ClinRep Catecholamine-Plasma" assay kit with a reversed-phase C18 column) used for measuring catecholamines. None of the 74 substances tested co-eluted with any of the catecholamines--norepinephrine, epinephrine, or dopamine--or with the internal standard, 3,4-dihydroxybenzylamine.

Catecholamines↗

Ammonium uptake and metabolism by mitrogen fixing bacteria. II. Klebsiella pneumoniae.

The primary steps of N2, ammonia and nitrate metabolism in Klebsiella pneumoniae grown in a continuous culture are regulated by the kind and supply of the nitrogenous compound. Cultures growing on N2 as the only nitrogen source have high activities of nitrogenase, unadenylated glutamine synthetase and glutamate synthase and low levels of glutamate dehydrogenase. If small amounts of ammonium salts are added continuously, initially only part of it is absorbed by the organisms. After 2-3 h complete absorption of ammonia against an ammonium gradient coinciding with an increased growth rate of the bacteria is observed. The change in the extracellular ammonium level is paralleled by the intracellular glutamine concentration which in turn regulates the glutamine synthesis and an induction of glutamate dehydrogenase synthesis. Upon deadenylation these events are reversed.--Addition of dinitrophenol causes transient leakage of intracellular ammonium into the medium.

Biological Transport↗

Block of heart potassium channels by clofilium and its tertiary analogs: relationship between drug structure and type of channel blocked.

The whole-cell arrangement of the patch clamp was used to study delayed rectifier and inward rectifier K channels in isolated guinea pig ventricular cells. Block of these channels by an externally applied quaternary nitrogen compound, clofilium, and two of its tertiary nitrogen structural analogs (LY97241 and LY97119) were investigated. Clofilium reduced delayed rectifier current but had little effect on inward rectifier currents in concentrations as high as 100 microM. The block of delayed rectifier did not reverse upon washout. In contrast, lower concentrations of the tertiary analogs blocked both delayed rectifier and inward rectifier K currents. Onset of block of delayed rectifier was fast and block was reversible. The onset of block of inward rectifier by the tertiary compounds was slower than for delayed rectifier current and more difficult to reverse. We conclude from this work that tertiary, but not quaternary, clofilium blocks inward as well as delayed rectifier channels in these cells. Block of inward rectifier current is presumably caused by access to a receptor for the molecule that is gained by the tertiary, but not the quaternary, forms of the drug.

Animals↗

Nutritional control of sexuality in Chlamydomonas reinhardi.

1. Cells of Chlamydomonas reinhardi grown in the light or dark on standard medium require an additional exposure to light in the absence of a nitrogen source, in order to become sexually active. As the culture ages, the light requirement decreases. 2. This light requirement is a function of nitrogen depletion, as shown by the observation that cells from cultures grown to maturity on a low nitrogen medium in the light or in the dark, have no additional light requirement for zygote formation. The withholding of no other component of the medium has this effect. 3. In cells requiring light for zygote formation, the light can be supplied before the mating types are mixed, indicating that light is required, not for mating per se, but for the conversion of vegetative cells to gametes. 4. Gametes can be dedifferentiated to the vegetative state by any nitrogen compound which the cells can use for growth; and by further exposure to light in the absence of a nitrogen source, these vegetative cells can again become gametic. 5. Cells grown at different nitrogen levels become gametic at widely different cell concentrations of nitrogen and carbon and C/N ratios. 6. It is postulated that the role of light in gametic differentiation is indirect, providing by photosynthesis, energy for the mating process and carbohydrates to tie up excess nitrogenous reserves; and that the concentration of some particular nitrogen fraction or compound determines whether or not gametic differentiation is initiated.

Carbon↗

[Microflora in swine slurry as a parameter in determining the efficiency of deodorizing treatment. II. Treatment with ammonium peroxydisulfate].

Fresh and stored liquid manure from the pig could be deodorized by addition of 2% (w/v) Ammonperoxodisulfate (= APS). This effect was accompanied by a rapid increase of the redox potential up to +400 mV, fluctuating oxygen contents of maximal 0.8 mg/l, a significant decrease of pH below 7.0, a complete decomposition of organic nitrogen compounds and a considerable increase of total nitrogen caused by APS-addition. After 20 days, odour production could be noted again in the fresh slurry sample. It is assumed, that this effect is due to a population of yeasts, vigorously grown up towards the end of the test. The stored slurry sample was shown to be stable with respect to deodorization effect and yeasts could be found in portions lesser than 1%. In the fresh as well as in the stored slurry samples, organisms of the genus Bifidobacterium dominated towards the end of manure handling. In spite of an oxidised substrate it is assumed, that these anaerobic to aerotolerant microorganisms found a microbiotope with a reduced environment inside the flocks, built up by APS-addition. The facultatively anaerobic microorganisms, for the most part of the genus Bacillus, which mainly belong to producers of odorous compounds almost could be eliminated within 20 days at the latest by APS-treatment. It should be focused to prevent a secondary odour production by H2S-forming yeasts. This may be managed in future by an odour-reducing minimal APS-addition under a 2%-application. In addition, the costs of APS-treatment of liquid piggery waste could be reduced considerably.

Ammonium Sulfate↗

Molecular cloning of the cDNA encoding aspartate aminotransferase from bean root nodules and determination of its role in nodule nitrogen metabolism.

A cDNA clone encoding aspartate aminotransferase (PVAAT-2) (EC 2.6.1.1) was isolated from the common bean Phaseolus vulgaris nodule cDNA library. The nucleotide sequence analysis of the full-length cDNA allowed its identification by comparison with sequence databases. The amino acid sequence of the bean PvAAT-2 showed high similarity with the AAT-2 isoforms described in other leguminous plants. The amino-terminal region of the PvAAT-2 contains a sequence, which shares common features of plastid transit peptides. Southern blot analysis showed that the PvAAT-2 clone is encoded by a single gene in the P. vulgaris genome. Analysis of the PvAAT-2 mRNA levels suggests that the expression of this gene is nodule enhanced. The PvAAT-2 transcript is more abundant in nodules with increased synthesis of amides and is down-regulated in conditions where ureides accumulate. When plants were supplemented with ureides or with amides, PvAAT-2 expression was reduced, while it was not affected when plants were treated with allopurinol, an inhibitor of ureide synthesis. On the other hand, the expression of asparagine synthetase (another enzyme involved in the synthesis of amides) is not affected either by ureides or amides. These data suggest a role for AAT-2 in the mechanism involved in the synthesis of nitrogen compounds in bean nodules.

Acetylene↗

Developing seasonal ammonia emission estimates with an inverse modeling technique.

Significant uncertainty exists in magnitude and variability of ammonia (NH3) emissions, which are needed for air quality modeling of aerosols and deposition of nitrogen compounds. Approximately 85% of NH3 emissions are estimated to come from agricultural nonpoint sources. We suspect a strong seasonal pattern in NH 3 emissions; however, current NH3 emission inventories lack intra-annual variability. Annually averaged NH 3 emissions could significantly affect model-predicted concentrations and wet and dry deposition of nitrogen-containing compounds. We apply a Kalman filter inverse modeling technique to deduce monthly NH3 emissions for the eastern U.S. Final products of this research will include monthly emissions estimates from each season. Results for January and June 1990 are currently available and are presented here. The U.S. Environmental Protection Agency (USEPA) Community Multiscale Air Quality (CMAQ) model and ammonium (NH4+) wet concentration data from the National Atmospheric Deposition Program (NADP) network are used. The inverse modeling technique estimates the emission adjustments that provide optimal modeled results with respect to wet NH4+ concentrations, observational data error, and emission uncertainty. Our results suggest that annual average NH 3 emissions estimates should be decreased by 64% for January 1990 and increased by 25% for June 1990. These results illustrate the strong differences that are anticipated for NH3 emissions.

Agriculture↗