Sterols, fatty acids and free amino acids from two Helvella species

Sterols, fatty acids and free amino acids of Helvella crispa and H. monachella were investigated. They contained traces of ergosterol and a high amount of ergosta 5.22-dien-3β-ol. Linoleic acid and l-Dopa are the most abundant fatty acid and free amino acid, respectively. The aqueous extract of H. monachella inhibits prostaglandin release by rat peritoneal leucocyres in vitro.     

Seasonal cycling of putrescine and amino acids in relation to biological production in a stratified coastal salt pond

Seasonal cycles of concentrations and microbial uptake of dissolved free amino acids and the polyamine, putrescine, were followed during summer stratification of a coastal salt pond. Stratification began in May and was clearly seen in profiles of temperature, salinity, pH and alkalinity. Primary production exhibited a mid-August maximum and the O₂-H₂S interface shoaled at that time. POC and phytopigments roughly followed the pattern of primary production. Cycling of putresince, like the amino acids, was strongly influenced by primary production and microbial decomposition. Putrescine concentration profiles appeared to follow the pattern of primary production more closely, while amino acids appeared to follow the pattern of microbial production. The absence of production of putrescine during the decomposition of dissolved ornithine and the correlation of putrescine concentration with primary production suggest a direct source from algae in the water column.Microbial uptake of amino acids and putrescine together accounted for 60–90% of the bacterial C production measured in oxic waters and almost 300% of that measured in the anoxic bottom layer. Since other organic carbon and nitrogen compounds are also being taken up, these data suggest that tracer uptake methods as we used them may overestimate the true microbial uptake rates, or release of other organic compounds by microbes occurs at the same time. Further work on carbon and nitrogen budgets is needed to resolve the apparent imbalance between organic C and N incorporation and bacterial production.     

Diel variation in concentration,assimilation and respiration of dissolved free amino acids in relation to planktonic primary and secondary production in two eutrophic lakes

Concentration of dissolved free amino acids (DFAA) and assimilation of the 5 most abundant DFAA (glutamic acid, serine, glycine, alanine and ornithine) were measured at 3-h intervals over 27 h in two Danish, eutrophic lakes. The carbon flux of the amino acid assimilation was compared with the major routes of carbon flux, including primary production, bacterial production and zooplankton grazing. In Frederiksborg Slotssø, the mean DFAA concentration was 275 nM with distinct peaks (up to 783 nM) 3 h after sunrise. Assimilation rates of the 5 amino acids amounted on the average to 2.03 µg Cl^–1 h^–1, but high values up to 7.41 µg Cl^–1 h^–1 occurred 3 h after sunrise and at midnight. The mean turnover time of the amino acid pools was 3.2 h. In Lake Mossø, the mean DFAA concentration was 592 nM with peak of 1 161 nM at dusk. The assimilation rate averaged 0.44 µg Cl^–1 h^–1, and the mean turnover time of the amino acid pools was 39 h. In Lake Mossø, similar turnover times of glutamic acid and serine were determined from the ¹⁴C-amino acid tracer technique and Michaelis-Menten uptake kinetics, indicating that the tracer technique gave reliable values of the actual assimilation. The average respiration percentages of the assimilated amino acids were 45% in Frederiksborg Slotssø and 51% in Lake Mossø. Extracellular organic carbon (EOC) released from the phytoplankton contributed DFAA to the water. In Lake Mossø, 81% of the ambient EOC pool was <700 daltons and 9.3% of the EOC was DFAA. This corresponded to about 2.4% of the DFAA pool. Bacterial productivity, determined by means of frequency of dividing cells and ³⁵S-SO₄ dark uptake techniques gave similar results and constituted 4.5 and 3.7 µg Cl^–1 h^–1 in Frederiksborg Slotssø and Lake Mossø, respectively. The bacterial productivity suggested that DFAA were essential substrates to the bacteria, especially in Frederiksborg Slotssø. The zooplankton biomass in Frederiksborg Slotssø was six times larger than that in Lake Mossø, but cladocerans were dominant in both lakes. The zooplankton grazing probably was an important regulatory factor for the bacterial productivity.     

Determination of free amino acids in burley tobacco by high performance liquid chromatography

A reversed-phase high performance liquid chromatographic method was developed for determining free amino acids in burley tobacco. The test was done by OPA/3-mercaptopropionic acid as the pre-column derivatizing reagent. Chromatographic column was Elitte C¹⁸ column (4.6 mm × 250 mm i.d., 5 μm). Mobile phase A was 18 mol/l NaAc (pH7.2) including 0.002%(v/v) triethylamine and 0.3%(v/v) furanidine. Mobile phase B was 100 mol/l NaAc (pH7.2)–acetonitrile–methanol (v/v = 1:2:2). The column temperature was 40 °C and the flow rate was 1.0 ml/min. The fluorescence detector was used with 350 nm excitation wave length and 450 nm emission wave length. The average recoveries of the method ranged from 95.3–100.7% with the relative standard deviation of 2.32–9.24%. The method is simple, accurate and has good repeatability. The results of the determination of seventeen kinds of free amino acids in burley leaves were produced by the way of different ratios of cake fertilizer and inorganic fertilizer. The results show that Aspartic acid has the highest content however ratio of cake fertilizer and inorganic fertilizer. The contents of most of the free amino acids are increased and then gradually decreased with the increase in organic manure. The contents of most of the free amino acids are very close at 15:85% ratio and 30:70% ratio of cake fertilizer and inorganic fertilizer. The total amount of free amino acids is the highest at 30:70% ratio of cake fertilizer and inorganic fertilizer. Considering comprehensively, the quality of burley leaves is the best at 30:70% ratio of cake fertilizer and inorganic fertilizer.     

The spatio-temporal patterns and ecological significance of free amino acids and humic substances in contrasting oligotrophic and eutrophic freshwater ecosystems

The concentrations of total and individual free amino-acids (TFAA, FAA) and humic substances (HS) were measured by HPLC in the midwater column (MWC), air-water interface (AWI) and sediment pore water (SPW) in the oligotrophic, dystrophic Isle of Thorns lake and the eutrophic Lewes Brooks drainage channel in East Sussex, UK, during each of the four seasons. Pesticide levels were low, usually below detection levels in both water bodies. At the temporal level there were tendencies for the concentration of both FAA and HS to be lowest in the winter months and for the HS concentrations to be higher in the autumn in both water bodies. At the spatial level the eutrophic Lewes Brooks was characterized by having much higher FAA/HS ratios than the oligotrophic Isle of Thorns lake and it is suggested that further comparative studies be undertaken to verify whether the FAA/HS ratios may have general application in the classification of water bodies on a trophic basis. The concentrations of TFAA are higher in the SPW and AWI than in the MWC in both water bodies and the HS concentrations are much higher in the SPW than in the MCW Statistical analyses support the view that the FAA in the MWC are mainly derived from the SPW whereas the FAA in the AWI have a different origin. The data obtained is used to address the following questions: (a) How do the spatio-temporal patterns of FAA and HS in the present study areas compare with those of other water bodies? (b) What are the origins of the FAA and HS and how are their concentrations regulated? (c) Are the distributional patterns of FAA and HS of special significance to invertebrate detritivores? (d) How can the differences in the FAA and HS patterns in oligotrophic and eutrophic waters be explained? What are the conclusions and testable hypotheses arising from the present study?     

Gradient of dissolved free amino acids and phytoplankton in a shallow bay

A 3 h survey of the concentrations of individual free amino acids, chlorophyll a and phytoplankton species biomass was conducted in the surface waters of a shallow bay. Significant coefficients of correlation were found between chlorophyll a, nanoflagellates, and DFAA concentrations. Although phytoplankton biomass variations sometimes relate to DFAA concentration patterns, consideration of the physiological activity of both phytoplankton and microheterotrophs would undoubtedly explain a more significant fraction of the DFAA variation.This work is a contribution of GREPMA (Groupe Régional d'Etudes Pélagiques en Manche-Atlantique)     

An analysis of intron positions in relation to nucleotides, amino acids, and protein secondary structure

We present an analysis of intron positions in relation to nucleotides, amino acid residues, and protein secondary structure. Previous work has shown that intron sites in proteins are not randomly distributed with respect to secondary structures. Here we show that this preference can be almost totally explained by the nucleotide bias of splice site machinery, and may well not relate to protein stability or conformation at all. Each intron phase is preferentially associated with its own set of residues: phase 0 introns with lysine, glutamine, and glutamic acid before the intron, and valine after; phase 1 introns with glycine, alanine, valine, aspartic acid, and glutamic acid; and phase 2 introns with arginine, serine, lysine, and tryptophan. These preferences can be explained principally on the basis of nucleotide bias at intron locations, which is in accordance with previous literature. Although this work does not prove that introns are inserted into genomes at specific proto-splice sites, it shows that the nucleotide bias surrounding introns, however it originally occurred, explains the observed correlations between introns and protein secondary structure.     

Seasonal changes in neutral sugars and amino acids of particulate matter in Lake Nakanuma,Japan

Amino acids and neutral sugars in particulate matter were measured weekly from April in 1987 to March in 1988 in Lake Nakanuma, Japan. Changes in concentrations of total amino acids and total neutral sugars corresponded to those of chlorophylla. The composition of amino acids varied little seasonally and vertically. On the contrary, the composition of neutral sugars changed seasonally and vertically. We discuss the relationship between the changes in the biochemical components and environmental factors.     

Concentrations of endogenous free amino acids in rabbit reticulocyte lysates and the effect of incubation

Amino acid analyses show that while the free amino acids found in the rabbit reticulocyte translation system do not increase during nuclease treatment or during prolonged storage, the endogenous levels of many amino acids are so high that the choice of a radioactive; precursor for a translation should be based not only on the abundance of the amino acid in the translation product but also on its concentration in the lysate preparation. It is shown that the variation of amino acid concentrations between different lysates is sufficiently small to allow one to use the concentrations reported in this study to calculate the amount of radioactive amino acid necessary for satisfactory incorporation.     

Changes in brain temperature and free amino acids in normal and protein deprived suckling rats exposed to room temperature

Previous reports on early-induced protein-calorie malnutrition (PCM) in rats have indicated alterations in the concentration of free amino acids and of protein synthesis in the brain. Recently it was shown that early-induced protein deprivation (PD) retards the development of thermoregulation. This resulted in a failure to maintain a normal rectal temperature after short exposure to room temperature (+22°C) still at the age of 20–25 days corresponding to changes seen in normal rats at an age of 10–15 days. In the present study, 20-day old PD and normal rats where examined with regard to the effect of exposure to room temperature on brain temperature and on brain free amino acids. The results show a similar reduction in brain and rectal temperature of the PD rats occuring within 30 minutes after exposure to room temperature. The reduction was in the range of 5°C. PD rats kept in room temperature for 5 hours and then allowed to recover at 32.5°C showed a slow increase in brain and rectal temperature but normal temperatures were not reached even after 1 hour. The concentration of free amino acids in the brain was examined in rats kept for 1 hour at room temperature or at 32.5°C. In the PD rats kept at 32.5°C, free aspartate and glutamate were reduced whereas taurine, GABA and glycine were increased as compared to their corresponding control rats. As a result of the reduced brain temperature in PD rats exposed to room temperature there was a reduction in free asparagine. The lability of the pool of asparagine may be related to the low levels of aspartate and glutamate in PD rats. On the basis of the present findings it is recommended that temperature-sensitive parameters are examined in PCM rats at a normal body temperature.Special Issue dedicated to Prof. Holger Hydén     

Changes in the levels of plant total phenols and free amino acids induced by two cereal aphids and effects on aphid fecundity

Abstract:  Phenols and amino acids have previously been involved in the resistance of cereals to aphids. In the present study, the mean total concentrations of both compounds were quantitatively estimated from leaf-tissue samples collected at three growth stages (third, fifth and seventh leaf stages) from maize and barley seedlings and then correlated with the mean 7-day fecundity of Rhopalosiphum padi (L.) and Sitobion avenae (F.) (Homoptera, Aphididae) reared separately on the corresponding growth stages of each cereal. Results revealed an inverse correlation between the total concentration of each chemical in aphid-infested maize and barley plants and the fecundity of each aphid on every growth stage of the cereals. Moreover, R. padi led to an increase in total amino acids when reared on maize or barley without affecting the phenol concentration, while S. avenae induced a decrease in phenol levels but did not change the total amino acids on each growth stage of the plants. The significance of these results for understanding potential differences in requirements of cereal aphids for host selection and acceptance are discussed.     

Orientation of carp, Cyprinm carpio L., to free amino acids from Tubifex extract in an olfactometer

We analysed the effects of L-amino acid combinations, tested at the same relative concentrations as in 500 mgl¹ of a filtrated Tubifex extract, on attraction and exploratory behaviour of juvenile carp, Cyprinus curpio L., in an olfactometer. Tubifex filtrate was significantly active in the range of concentrations tested (5 mgl⁻¹, 50 mgl⁻¹, 500 mgl⁻¹, 5 gl⁻¹). Maximum attraction was obtained in response to a concentration of 500 mg l⁻¹. Soluble TCA fraction and a mixture of the 17 synthetic amino acids found in this fraction also showed significant effects on attraction and exploratory behaviour. Tests of the four chemical groups of amino acids gave the following results: acidic amino acids (aspartic and glutamic) did not produce significant activity. Basic amino acids (histidine, arginine and lysine) and polar, uncharged amino acids (glycine, serine, threonine, tyrosine, asparagine and glutamine) were ineffective as attractants but significantly increased exploration. Non-polar amino acids (alanine, valine, leucine, isoleucine, phenylalanine, and methionine) showed significant effects on both attraction and exploration. In tests where different pairs of these amino acid groups were mixed it was shown that a combination of non-polar amino acids and polar uncharged amino acids was the most effective in inducing both attraction and exploration. The simplest combination to have a significant effect on both attraction and exploration was alanine, valine and glycine.
These results are discussed in the light of recent data on the role of free amino acids in the behaviour of fishes.     

Seasonal patterns of nucleic acid concentrations and amino acid profiles of Parapenaeus longirostris (Crustacea, Decapoda): relation to growth and nutritional condition

The present work describes the seasonal changes in nucleic acid concentrations and amino acid profiles in the muscle of juvenile Parapenaeus longirostris and their relation to growth and nutritional condition. RNA content varied significantly between seasons, being the highest values attained in spring and the lowest in winter (p < 0.05). Similar results were obtained with RNA:protein and RNA:DNA ratios. In respect to total amino acid content (TAA), a significant increase from winter to spring was observed (p < 0.05) and the major essential amino acids (EAA) were arginine, histidine and leucine. Within non-essential amino acids (NEAA) glutamic acid, aspartic acid, glycine and proline were dominant. From winter to spring, a significant variation in NEAA content occurred (26.8; p < 0.05), mainly due to the significant increase of glutamic acid (79.1) and serine (66.7) (p < 0.05). EAA content did not vary significantly between seasons (p > 0.05). In opposition, during this period a significant decrease in the free amino acid content (FAA) was observed (p < 0.05); a higher percentage of decrease was attained in free non-essential (FNEAA – 42.9) in comparison to free essential amino acids (FEAA – 40.2). The significant increase in RNA and TAA contents from winter to spring may be related with protein synthesis. On the other hand, the lowest values obtained in winter may be due to a reduction in feeding activity; in this period the muscle protein must be progressively hydrolysed, which is evident with the higher FAA content. The liberated amino acids enter FAA pool and become available for energy production. In conclusion, it was evident that the seasonal cycle in activities such as feeding and growth with nucleic acids and amino acid analyses was noticed.     

Seasonal dynamics of amino acids in two small Siberian reservoirs dominated by prokaryotic and eukaryotic phytoplankton

The comparison of the dynamics of phytoplankton biomass and total amino acid composition was made for two water bodies: in one the phytoplankton were dominated by prokaryotes (i.e., there was a bloom of cyanobacteria) and by eukaryotic microalgae in the other. The dynamics of phytoplankton biomass and of total amino acid composition of water were investigated during the vegetation season. It was found that the only factor that significantly changed the percentages of amino acids in water was the bloom of cyanobacteria in the blooming water body. During the bloom of cyanobacteria, the absolute and relative content of the Leu-Glu group increased, while the contents of other acids generally dropped. Before and after the bloom, no significant variations in the total amino acid composition were recorded. In the reservoir where eukaryotic microalgae dominated, no significant variations in amino acid composition were recorded during the season.     

Changes of plasma insulin, urea, amino acids and rumen metabolites in somatotropin treated dairy cows

Summary An experiment was performed to evaluate the effects of somatotropin on plasma free amino acid, urea and insulin concentrations and rumen fermentation pattern and to assess their relationships. Four Italian Friesian dairy cows fitted with rumen cannulae were used in a switch-back design. Slow releasing recombinant bovine somatotropin (640 mg/cow) was injected every 28 days for two consecutive periods. Rumen fluid and blood samples were collected before and after feeding at 0, 7 and 21 days after rbST injection. Exogenous rbST increased plasma insulin concentration and the insulin response to feeding, and decreased plasma urea and free essential and branched chain amino acid concentrations. rbST did not affect rumen fermentation pattern. No correlation was found between rumen and plasma parameters measured after feeding. Our results are consistent with the notion that the main effect of somatotropin is post-absorptive.     

Free amino acids in the late embryogenesis and pre-hatching stage of two coregonid fishes

Developing eggs of vendace (Coregonus albula L.) and whitefish (C. lavaretus L.) were experimentally delayed in hatching by incubation at low water temperature (1–2°). Some eggs were taken during this period to a water temperature which was gradually raised up to 8° to provoke mass hatching of embryos. The pattern of free amino acids was followed in eggs incubated at both temperatures. During a 56 days period, the content of several essential amino acids significantly decreased in eggs of both species. For instance, the lysine content dropped from 703 to 270 mg/g dry matter and the arginine content from 257 to 13.3 mg/100 g dry matter in whitefish eggs. A similar pattern of decreasing level of free amino acids in embryonated ova up to hatching was characteristic for essential amino acids and serine. Methionine was exceptional; its level remained approximately the same. On the other hand, non-essential amino acids showed a significant increase in concentration during the experimental period. For instance, the glycine level increased 4.9 and 2.1 times in whitefish and vendace eggs, respectively. Transfer of eggs to 8° accelerated the decrease of nearly all free amino acids before hatching. The consequence of such amino acid metabolism for newly hatched larvae is discussed.     

Neuroactive and other free amino acids in seed and young plants of Panax ginseng

The seeds and one to three years old plants of Asian ginseng (Panax ginseng C.A. Meyer) were analyzed for their free amino acid contents. The neuro-excitatory beta-ODAP (beta-N-oxalyl-L-alpha,beta-diaminopropionic acid), suggested to be the cause of the crippling neurolathyrism, was the major component in the seed extract (70% of the total free amino acids detected) and showed the highest concentration (0.43% by wt) compared to that in the different parts of young plants. beta-ODAP concentration was higher in the shoots as compared to roots and declined in older plants. The amount of beta-ODAP in the roots may be considered as an indirect measure of age and quality. Another neuro-active non-protein amino acid, GABA (gamma-aminobutyric acid), increased dramatically after germination and reached highest concentration in different parts of 3 year-old plants. Glutamine and arginine were the two major free proteinogenic amino acids in the ginseng plants and together they constituted over 50% of all the free amino acids detected in the root.     

Changes in amino acids in Cucumis melo in relation to life-history traits and flight propensity of Bemisia tabaci

Phloem amino acids in Cucumis melo L. were measured to determine whether changes in nitrogen availability might affect life-history traits and flight activity of Bemisia tabaci Gennadius (Homoptera: Aleyrodidae). During plant development, nineteen of the twenty common amino acids, plus hydroxyproline, citrulline, ornithine and -aminobutyric acid (GABA) were identified. For most essential amino acids, there were two peaks observed: an initial large peak associated with young plants, and a later small peak associated with senescing plants. For histidine, ornithine and citrulline, medium to large peaks in relative abundance were observed in mature plants. Arginine peaked during the first few weeks of development and was no longer detectable after wk 7. Serine and glutamine/glutamic acid were the only amino acids that peaked during plant senescence. Factor analysis was used to create a reduced number of orthogonal factors, which corresponded well with the trends that were observed for the various groups of amino acids. No single or combination of factors explained a significant amount of the variability in oviposition. For both males and females, factor 1 (predominantly essential amino acids) was the single most important predictor of adult weight. As the relative concentrations of essential amino acids decreased, whitefly weights decreased. Factors 1 and 3 (predominantly histidine and ornithine) were the most important predictors of developmental time. As these amino acids decreased in relative concentration, developmental time increased. Percent emergence was positively associated with factor 1 and negatively associated with factor 6 (predominantly aspartic acid). The distributions of flight duration for whiteflies emerging from young, mature and senescing melon plants were compared and they were always skewed towards short flights; however, the frequency of long-duration flights increased when melon plants began to senesce. Whiteflies from all plant-age categories were capable of flying for more than 2 h with fully developed eggs, but the presence of more than four mature eggs was associated with flights of reduced duration.     

Effects of branched chain amino acids,pyruvate, or ketone bodies on the free amino acid pool and release from brain cortex slices of normal and streptozotocindiabetic rats

Brain cortex slices from diabetic rats incubated in Krebs-Ringer-bicarbonate (KRB)-glucose medium show, compared to the normals, a 75% higher glutamine content. Branched chain amino acids (BCAA) added, at 0.5mM each, to this medium increase (53%) the glutamine content in the normal slices but have no effect on the glutamine content in the slices from diabetic rats. When the incubation medium is KRB-pyruvate, glutamine and glutamate contents are lower than in the KRB-glucose medium. The addition of BCAA in the KRB-pyruvate medium partially restores the contents of glutamine in the normal and of glutamine plus glutamate in the diabetic. Keto acids or BCAA added to the incubation medium of normal slices decrease the pool of most of the neutral and acidic amino acids but they do not affect this pool in slices from the diabetic rats. In addition keto acids increase the ratio glutamate in the tissue: glutamate in the medium.Abbreviations used BCAA branched chain amino acids - 3-OHB d,l-3-hydroxybutyrate - AcAc acetoacetate - KRB Krebs-Ringer-bicarbonate