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PubMed2006

Catalytic characteristics of peroxidase from wheat grass.

The crude enzyme extract of wheat grass was heated at 60 degrees C for 30 min, followed by ammonium sulfate fractionation and isoelectric chromatofocusing on Polybuffer exchanger (PBE 94) for purification. The purified peroxidase was then characterized for its catalytic characteristics. It was found that AgNO3 at a concentration of 0.25 mM and MnSO4 and EDTA at concentrations of 5 mM significantly inhibited the activity of wheat grass peroxidase. However, KCl, NaCl, CuCl2, CaCl2, ZnCl2, and MgCl2 at concentrations of 5.0 mM and HgCl2 at a concentration of 0.25 mM enhanced enzyme activity. Chemical modification significantly influenced the activity of wheat grass peroxidase. Particularly, N-bromosuccinimide (5 mM) inhibited 16% of the enzyme activity, whereas N-acetylimidazole (2.5 mM), diethyl pyrocarbonate (2.5 mM), and phenylmethanesulfonyl fluoride (2.5 mM) enhanced by 18-29% of the enzyme activity. Such results implied that tryptophan, histidine, tyrosine, and serine residues are related to enzyme activity. The pH optima for wheat grass peroxidase to catalyze the oxidation of o-phenylenediamine (OPD), catechol, pyrogallol, and guaiacol were 5.0, 4.5, 6.5, and 5.0, respectively. The apparent Km values for OPD, catechol, pyrogallol, and guaiacol were 2.9, 18.2, 2.5, and 3.8 mM, respectively. Under optimal reaction conditions, wheat grass peroxidase catalyzed the oxidation of OPD (an aromatic amine substrate) 3-11 times more rapidly than guaiacol, catechol, and pyrogallol (phenolic substrates containing one to three hydroxy groups in the benzene ring).

Buffers↗

PubMed1967

Comparative studies on the activity of carboxylases and other enzymes in relation to the new pathway of photosynthetic carbon dioxide fixation in tropical grasses.

1. The activity per unit of chlorophyll of certain carboxylases, and of other enzymes involved in photosynthesis, was determined in leaf extracts of the tropical grasses, sugar-cane, maize and sorghum, and compared with the activities for wheat, oat and silver-beet. Maximum rates of photosynthetic carbon dioxide uptake were also measured for comparison with enzyme activities. 2. Phosphopyruvate carboxylase activity was about 60 times greater in the tropical grasses than in wheat, oat and silver-beet and was severalfold higher than the rates of photosynthetic carbon dioxide uptake. Most of the enzyme was located in the chloroplast fraction of cell extracts. 3. Phosphopyruvate carboxylase was apparently the major photosynthetic carbon dioxide-fixing enzyme in the tropical grasses, although malic enzyme may contribute to a lesser extent. 4. Tropical grasses contained less than one-tenth of the ribulose diphosphate carboxylase activity present in wheat, oat and silver-beet. For the tropical grasses this activity, determined with a saturating concentration of bicarbonate, was approx. 10% of the rate of photosynthesis. 5. The fraction-1 protein content of leaf extracts paralleled the ribulose diphosphate carboxylase activity. 6. In contrast, the activity of several other enzymes of the Calvin cycle was similar in the different species examined.

Acid Phosphatase↗

PubMed1999

Relationship between allergic status and specificity of IgG antibody to inhaled allergens: the grass pollen model.

BACKGROUND: We have previously reported that IgG antibodies from healthy individuals and patients suffering from non-seasonal mite allergy bind to different sets of epitopes on Der p 1, allowing almost complete discrimination of the populations. OBJECTIVES: To confirm this observation in a seasonal allergy model where a clear relationship between allergic symptoms and exposure to the offending agent is established. To investigate whether the pattern of modified specificity is related to the differences in IgG subclass hierarchy usually exhibited by nonallergic and allergic populations. METHODS: The capacity of individual sera from patients allergic to grass pollen and healthy individuals, including grass pollen-sensitized subjects, to prevent the binding of pooled IgG, IgG1, and IgG4 fractions from grass pollen-allergic patients and healthy individuals to solid-phase bound grass pollen antigen was evaluated in enzyme-linked immunosorbent assay (ELISA) using streptavidin-biotin technology. Specificity controls were performed using sera from patients allergic to cat dander and house dust mite. RESULTS: The capacity of sera to prevent the antigen binding of allergic IgG averaged 84 +/- 5% for allergic sera and 53 +/- 6% for healthy sera (P < 0.001 by one-way anova). Conversely, using the antigen-binding capacity of healthy control IgG as reference, percentage inhibitions averaged 46 +/- 9% in grass pollen-allergic subjects compared with 80 +/- 4%, 82 +/- 2% in healthy individuals, and mite- and cat-allergic patients, respectively, resulting in two well-separated populations (P < 0.0001 by one-way anova). Similar results were found regardless of whether pooled IgG1 or IgG4 were used. CONCLUSION: Together with previous data, our results define a new type of humoral signature in the immune response to inhaled allergens. Allergic and healthy status differ not only in the presence or absence of specific IgE antibody but also in the preferential expression of distinct IgG specificities that are better correlated with clinical manifestations and are unrelated to subclass distribution.

Adult↗

PubMed1999

The prevalence of immunoglobulin E antibodies to the proteins of rubber (Hevea brasiliensis) latex and grass (Phleum pratense) pollen in sera of British blood donors.

BACKGROUND: Although there have been many studies of the prevalence of latex allergy in populations deemed to be at risk, little is known of the potential allergic susceptibility to latex products prevailing in the general population. OBJECTIVE: To assess the possible prevalence of allergy to latex goods in a population of blood donors by measurement of specific antilatex immunoglobulin (Ig) E in blood, to relate this to prevalence of antigrass IgE in the blood donations, and to assess the prevalence of antibodies to grass, house dust mite and cat allergens in those donors having antilatex IgE antibodies. METHODS: Sera from two groups of donations obtained in the English West Midlands were assayed. A group of 2000 donations obtained in midwinter was assayed for antilatex and antigrass pollen IgE. A group of 5000 midsummer donations was assayed for total IgE, and antilatex IgE and the sera giving a positive reaction, assayed for antigrass pollen, antihouse dust mite and anticat IgE. The nature of the principal latex and grass pollen polypeptides reacting with IgE in the sera was assessed by immunoblotting. RESULTS: Anti-latex IgE was detected in approximately 4% of the winter and 7% of the summer donations. The prevalence of antigrass IgE in the winter donations was approximately 20% and amongst the latex-positive sera approximately 84% contained antigrass IgE. Of the summer donations of latex-positive sera, 96% contained antigrass, 48.6% antimite IgE and 34% anticat IgE. The prevalence of both antilatex and antigrass IgE was age and sex related. Inhibition studies indicated cross-reactivity of IgE with latex and grass pollen proteins. CONCLUSIONS: Whilst 4-7% of the population may have serum IgE reacting with latex, the levels are low compared with those reacting with the aeroallergens studied. The apparent cross-reactivity of some serum IgE with both latex and grass pollen taken with other evidence suggests that, in some individuals, allergy to latex may arise from an initial sensitization by grass pollen.

Adolescent↗

PubMed2000

Upregulation of nasal mucosal eotaxin in patients with allergic rhinitis during grass pollen season: effect of a local glucocorticoid.

BACKGROUND: Allergic rhinitis is a common disease characterized by infiltration of eosinophils into the nasal mucosa during the periods of symptoms. Among chemokines, which attract cells to the site of inflammation, eotaxin is relatively specific for eosinophils. OBJECTIVE: We examined the influence of grass pollen season on nasal eotaxin expression in patients with seasonal allergic rhinitis, as well as the effect of a nasal glucocorticoid on this eotaxin expression. METHODS: Nineteen patients with allergic rhinitis received treatment with either nasal beclomethasone (400 microgram/day) or placebo over a grass pollen season. In these patients, nasal biopsies were taken prior to and during the peak of the pollen season and stained immunohistochemically for eotaxin and EG2 + eosinophils. Five healthy subjects served as controls and gave nasal biopsies once prior to the pollen season. RESULTS: Prior to pollen season, there was no significant difference in nasal eotaxin expression between patients with allergic rhinitis and healthy subjects. Grass pollen season induced significant increase in eotaxin expression in placebo-treated (P = 0.04; n = 9) but not in beclomethasone-treated rhinitis patients (P = 0.8; n = 10). During peak grass pollen season, the eotaxin expression in placebo-treated patients was significantly higher compared with healthy subjects outside season (P = 0.03). There was no significant correlation between the expression of eotaxin and the number of EG2 + eosinophils in nasal mucosa. The serum levels of eotaxin in rhinitis patients remained stable over the pollen season. CONCLUSION: Expression of eotaxin in nasal mucosa of grass-pollen allergic rhinitis patients is upregulated during pollen season and treatment with a nasal glucocorticoid protects against this upregulation.

Administration, Topical↗

PubMed2003

Skin test with a timothy grass (Phleum pratense) pollen extract vs. IgE to a timothy extract vs. IgE to rPhl p 1, rPhl p 2, nPhl p 4, rPhl p 5, rPhl p 6, rPhl p 7, rPhl p 11, and rPhl p 12: epidemiological and diagnostic data.

BACKGROUND: The diagnostic approach to grass pollen allergy is now possible by detecting specific IgE to its allergenic components. OBJECTIVE: To compare the IgE reactivity to a timothy grass pollen extract with the IgE reactivity to eight allergenic components from the same source (Phl p 1, 2, 4, 5, 6, 7, 11, 12). Both were compared with the skin test reactivity to a timothy grass extract. METHODS: A population survey was carried out by means of the skin test to identify grass-allergic subjects, and to characterize them in terms of demographic and allergological parameters. Seven hundred and forty-nine sera were available for IgE detection to a timothy extract, to the recombinant Phl p 1, 2, 5, 6, 7, 11, 12, and to native Phl p 4 and bromelain. Results were stratified by means of demographic and allergy parameters. RESULTS: Ninety-five per cent of the sera had detectable IgE to the timothy extract. Prevalence of IgE reactivity increased from 86.8% to 93.3% as the number of combined reactive molecules rose from 2 to 8. Adjusted prevalences for each allergen were: rPhl p 1 = 83%, rPhl p 2 = 55%, nPhl p 4 = 70%, rPhl p 5 = 50%, rPhl p 6 = 44%, rPhl p 7 = 7%, rPhl p11 = 43%, rPhl p 12 = 15%. Isolated reactivity to rPhl p 1 was 6%, whereas it was negligible for the remaining molecules. IgE reactivity prevalence and mean values differed when patients were stratified on the basis of their associated pollen reactivity and their skin test reactivity grade. No differences were found when age, symptom type and duration were considered. Up to eight-fold higher IgE concentrations were found when the sum of IgE to molecules was compared with IgE to the extract. Testing for the IgE reactivity to the glycan of the native Phl p 4 allergen showed a possible interference with prevalence and value estimation. Higher prevalence values were found in previously immunotherapy-treated patients. CONCLUSIONS: The use of a complete panel of grass allergenic molecules can mimic the current use of allergenic extracts, but new relevant information, such as individual pattern of reactivity, adjusted prevalence, correct specific IgE concentration, can be achieved only by means of discrete allergenic molecules.

Adolescent↗

PubMed1999

A role for glutathione transferases functioning as glutathione peroxidases in resistance to multiple herbicides in black-grass.

Black-grass (Alopecurus myosuroides) is a major weed of wheat in Europe, with several populations having acquired resistance to multiple herbicides of differing modes of action. As compared with herbicide-susceptible black-grass, populations showing herbicide cross-resistance contained greatly elevated levels of a specific type I glutathione transferase (GST), termed AmGST2, but similar levels of a type III GST termed AmGST1. Following cloning and expression of the respective cDNAs, AmGST2 differed from AmGST1 in showing limited activity in detoxifying herbicides but high activities as a glutathione peroxidase (GPOX) capable of reducing organic hydroperoxides. In contrast to AmGST2, other GPOXs were not enhanced in the herbicide-resistant populations. Treatment with a range of herbicides used to control grass weeds in wheat resulted in increased levels of hydroperoxides in herbicide-susceptible populations but not in herbicide-resistant plants, consistent with AmGST2 functioning to prevent oxidative injury caused as a primary or secondary effect of herbicide action. Increased AmGST2 expression in black-grass was associated with partial tolerance to the peroxidizing herbicide paraquat. The selective enhancement of AmGST2 expression resulted from a constitutively high expression of the respective gene, which was activated in herbicide-susceptible black-grass in response to herbicide safeners, dehydration and chemical treatments imposing oxidative stress. Our results provide strong evidence that GSTs can contribute to resistance to multiple herbicides by playing a role in oxidative stress tolerance in addition to detoxifying herbicides by catalysing their conjugation with glutathione.

Amino Acid Sequence↗

PubMed2000

Expulsion of allergen-containing materials from hydrated rye grass (Lolium perenne) pollen revealed by using immunogold field emission scanning and transmission electron microscopy.

BACKGROUND: Several studies demonstrated episodes of grass pollen-induced allergic asthma after heavy rainfalls. It has been hypothesized that these asthma attacks might be due to the release of respirable allergen-bearing particles from pollen cytoplasm. OBJECTIVE: In this study we investigated the release mechanism of the most potent and frequently recognized grass pollen allergens, group 1 and group 5, from freshly harvested and subsequently hydrated rye grass pollen at the ultrastructural level. METHODS: Rabbit antisera against purified recombinant group 1 and group 5 allergens were used to investigate, by using field emission scanning and transmission immunogold electron microscopy, the allergen release from rye grass pollen grains into isotonic aqueous solutions or water. RESULTS: Pollen grains exposed to isotonic aqueous solutions remained intact and released allergens by means of diffusion. However, pollen grains hydrated in distilled water or rainwater expelled starch grains and cytoplasmic debris of respirable size. Group 1 and group 5 allergens were observed on and within these materials. CONCLUSIONS: Exposure of rye grass pollen to water leads to an expulsion of subcellular allergen-containing pollen components of respirable size. Our ultrastructural data thus support the idea that this release of allergen-containing respirable pollen materials may be a cause of asthma attacks after heavy rainfalls.

Allergens↗

PubMed2002

Utility and distribution of conserved noncoding sequences in the grasses.

Control of gene expression requires cis-acting regulatory DNA sequences. Historically these sequences have been difficult to identify. Conserved noncoding sequences (CNSs) have recently been identified in mammalian genes through cross-species genomic DNA comparisons, and some have been shown to be regulatory sequences. Using sequence alignment algorithms, we compared genomic noncoding DNA sequences of the liguleless1 (lg1) genes in two grasses, maize and rice, and found several CNSs in lg1. These CNSs are present in multiple grass species that represent phylogenetically disparate lineages. Six other maize/rice genes were compared and five contained CNSs. Based on nucleotide substitution rates, these CNSs exist because they have biological functions. Our analysis suggests that grass CNSs are smaller and far less frequent than those identified in mammalian genes and that mammalian gene regulation may be more complex than that of grasses. CNSs make excellent pan-grass PCR-based genetic mapping tools. They should be useful as characters in phylogenetic studies and as monitors of gene regulatory complexity.

Animals↗

PubMed2000

Establishing a grassland signature in veins: 18O in the leaf water of C3 and C4 grasses.

We show that 18O evaporative enrichment of bulk leaf water in grass species can be significantly more enriched than predicted by the Craig-Gordon model, with C4 grasses considerably more enriched than C3 grasses. Our results suggest that the unanticipated 18O leaf water enrichment of grasses is attributable to the progressive evaporative enrichment along parallel veins (a function of both leaf length and interveinal distance), a pattern that does not occur in Dicotyledonous species. We propose that the differential 18O enrichment of grasses will result in distinct C18O16O biospheric signals from grassland and forest ecosystems, allowing for further partitioning of terrestrial carbon fluxes.

Journal Article↗

PubMed1985

Insulin action and glucose metabolism in sheep fed on dried-grass or ground, maize-based diets.

The effect of an exogenous supply of glucose, provided by the digestion of maize starch in the small intestine, on endogenous glucose metabolism and insulin action was studied in sheep using the euglycaemic insulin clamp procedure. Insulin was infused intravenously at rates of 0.2, 0.5, 1.0 and 6.0 mU/min per kg live weight for four consecutive periods in each of four sheep fed on dried-grass and maize-based diets. Glucose was also infused intravenously at a variable rate, sufficient to maintain the plasma glucose concentration at basal levels. Whole-body rates of glucose metabolism were determined using a continuous infusion of [6-3H]glucose. From the resulting insulin dose-response curves, it was observed that, when the sheep were fed on the dried-grass diet, the responsiveness of glucose metabolism to insulin was less than that reported for non-ruminants. When fed the maize-based diet, the glucose metabolic clearance rates (MCR) observed during insulin infusions were significantly greater (P less than 0.05) than those observed for the dried-grass diet. However, after correcting for the non-insulin-mediated glucose disposal, differences between diets were not significant. The sensitivity of glucose utilization to insulin was not affected by diet. The plasma insulin concentrations causing half-maximal insulin-mediated glucose MCR were 103 (SE 21) and 85 (SE 11) mU/l for the dried-grass and maize-based diets respectively. The sensitivity of endogenous glucose production to insulin was also unaffected by diet. The plasma insulin concentrations resulting in the suppression of endogenous glucose production to half the basal level were 80 (SE 26) and 89 (SE 29) mU/l for the dried-grass and maize-based diets respectively. It is concluded that the observed increase in glucose utilization on the maize-based diet was due partly to a slight change in responsiveness to insulin and also partly to a change in the rate of non-insulin-mediated glucose disposal.

Animal Feed↗

PubMed1993

[Effect of bovine growth hormone on performance criteria of dairy cows during summer feeding with grass. 1. Feed intake and protein and energy supply].

The influence of long-term administration of bovine growth hormone (bGH) with prolonged release (Somidibove from Lilly Germany GmbH, section Elanco, 640 mg/28 d) in six consecutive injection periods on feed intake under summer feeding conditions with grass feeding ad libitum was investigated. After a pretreatment period of 22 days 30 dairy cows were divided into 15 blocks with two cows each in consideration of milk yield, days postpartum and number of calving. In each block the bGH-treated animal received the same concentrate supply as the corresponding control animal which was supplemented in order to requirement. In a 42 day posttreatment period without bGH-injection possible effects of bGH-treatment on feed intake (grass silage and maize silage) was investigated. The first injection was administrated 75 days postpartum, respectively. During all six injection periods feed intake corrected to the pretreatment period was increased by 0.63 kg DM/animal.d resulting from a higher grass intake in the bGH-treated dairy cows compared to the control group. This difference in feed intake raises from 0.40 kg DM/animal.d in the first injection period up to 0.84 kg DM/animal.d in injection period six and continued in the posttreatment period without bGH-injection (+0.89 kg DM/animal.d). The higher feed intake of grass in the injection periods by the bGH-treated dairy cows resulted in an increased energy intake sufficient on average for the production of 1.2 kg milk/animal.d. Possibilities and limits of a sufficient nutrient supply in bGH-treated dairy cows by an increasing intake of grass are discussed.

Animals↗

PubMed1995

Influence of grass source on net flux of nutrients across splanchnic tissues in sheep with restricted intake.

Crossbred wethers (22 months old; 46 +/- 1.3 kg body weight), with catheters in a hepatic vein, the portal vein and a mesenteric vein and artery, consumed warm (W; bermudagrass hay) or cool season grass hay (C; ryegrass-wheat) at 1.6% body weight (dry matter basis) in a crossover design experiment. Warm and cool season grasses were 13.6 and 9.9% crude protein, 77 and 66% neutral detergent fibre and 4.6 and 4.0% acid detergent lignin, respectively. Neutral detergent fibre digestibility (70.3 and 77.4%) and digestible energy intake (8.5 and 9.3 mJ/d) were greater (P < 0.02) for C than for W, and digestible nitrogen intake (11.5 and 8.0 g/d for W and C, respectively) was greater (P < 0.01) for W. Ruminal fluid concentrations of ammonia nitrogen and total volatile fatty acids were not altered by grass source, and acetate: propionate was greater (P < 0.02) for W (3.80) than for the C (3.54). Portal-drained viscera blood flow (118 and 119 l/h; SE 8.0), oxygen consumption (141 and 142 mM/h; SE 3.7), alpha-amino nitrogen release (13.4 and 13.1 mM/h; SE 3.42), urea nitrogen uptake (22.8 and 22.5 mM/h; SE 4.97), ammonia nitrogen release (14.9 and 15.7 mM/h; SE 3.36), glucose uptake (10.0 and 6.5 mM/h; SE 1.30), propionate release (14.5 and 16.4 mM/h; SE 1.88), lactate release (4.64 and 5.03 mM/h; SE 1.908) and acetate release (54.8 and 55.4 mM/h for W and C, respectively; SE 8.82) were similar between grasses. Energy consumption by the portal-drained viscera accounted for a slightly greater (P < 0.01) percentage of digestible energy intake with W vs C (18.8 vs 17.0%; SE 0.10). In conclusion, with restricted consumption of W or C by mature sheep, grass source had little impact on net flux of oxygen and nutrients across the portal-drained viscera and splanchnic bed.

Animal Feed↗

PubMed1996

[Feed intake and performance of dairy cows during feeding of fodder beets supplementing grass silage].

In a feeding experiment 3 x 12 dairy cows (German Simmental x Red Holstein Friesian) were used to investigate the influence of different amounts of fodder beets on feed intake, milk yield and milk composition. The fodder beets (variety Kyros) were fed in exchange for concentrate. The amounts of fodder beets were either 22 kg cow-1 d-1 or 44 kg cow-1 d-1 or zero (control group). All groups received 4 kg hay cow-1 d-1, grass silage ad libitum, and concentrate corresponding to the milk yield. In order to balance the rations the concentrate was either rich in protein or rich in energy. Feeding 22 kg or 44 kg fodder beets increased forage intake (including fodder beets) significantly. However, the total feed intake diminished on average from 18.3 kg DM cow-1 d-1 (control group) to 17.2 kg DM cow-1 d-1 (fodder beet groups). In each treatment the intake of hay was 3.3 kg DM cow-1 d-1. However, the intake of grass silage offered ad libitum was significantly suppressed by the fodder beets. Considering the different amounts of concentrate the quantity of suppression of grass silage by feeding fodder beets was calculated at -0.8 kg DM of grass silage per kg DM of fodder beet intake. The milk yield decreased linearly with increasing fodder beets on average up to 4 kg cow-1 d-1. However, the milk composition was scarcely influenced. The fat content increased a little, the mean alterations of the protein and lactose content were below 0.1 percentage points. Therefore, the change in daily excretion of milk fat, protein or lactose were similar to the change in milk yield. In conclusion, feeding fodder beets in combination to grass silage causes a positive effect on forage intake (including fodder beets). However, the amount of concentrate should not be reduced equivalently to the energy contents of the added fodder beets.

Animal Feed↗

PubMedDate not supplied

Responses of loblolly pine, sweetgum and crab grass roots to localized increases in nitrogen in two watering regimes.

Root responses to differences in availability of nitrogen and soil water were studied in loblolly pine (Pinus taeda L.) seedlings grown in monoculture and in competition with sweetgum (Liquidambar styraciflua L.) or crab grass (Digitaria spp.). Rhizotron cells were maintained at high soil water availability (approximately -0.1 MPa) or subjected to three dry-down cycles to low soil water availability (approximately -1.0 MPa), over two growing seasons. Localized increases in nitrogen availability were created by adding nitrogen in solution to root ingress cores placed in each rhizotron cell. Presence of competitors reduced loblolly pine root growth regardless of the nitrogen or soil water treatment. On average, both total root length density and root surface area were reduced 60% when loblolly pine seedlings were grown with crab grass and 31% when grown with sweetgum. Low water availability reduced loblolly pine root length density and root surface area by 25 and 28%, respectively, compared with well-watered seedlings. Sweetgum root surface area was reduced 18% by the low water availability treatment, whereas crab grass root surface area was unaffected by this treatment. At all soil depths, loblolly pine root surface area and root length density were increased in localized areas of increased nitrogen availability. Sweetgum and crab grass root surface areas were also greater in areas of increased nitrogen availability. In the high soil water availability treatment, loblolly pine root surface area increased 128% in localized areas of increased nitrogen in all competition treatments. In the low soil water availability treatment, loblolly pine roots responded to increased nitrogen only in the absence of competitors. In general, loblolly pine and sweetgum roots responded to increases in resource availability similarly, whereas crab grass roots were relatively less affected.

Journal Article↗

PubMed1978

Cyclic adenosine 3':5'-monophosphate in axenic rye grass endosperm cell cultures.

Cyclic adenosine 3':5'-monophosphate (cAMP) was extensively purified from rye grass (Lolium multiflorum) endosperm cells grown in axenic suspension culture. The cAMP was purified by neutral alumina and anion and cation exchange chromatography. The cAMP was quantitated by means of a radiochemical saturation assay using a beef heart cAMP-binding protein and also by an assay involving activation of beef heart protein kinase. The cAMP levels found (corrected for recovery of tracer cyclic 3',5'-[8-(3)H]AMP included from the point of sample extraction) ranged from 2 to 12 pmol/g fresh weight. The material purified from rye grass cultures was indistinguishable from authentic cAMP with respect to chromatography in two cellulose thin layer systems, behavior on dilution in both the saturation and protein kinase activation assays, and rates of degradation by a mammalian cAMP phosphodiesterase. The cAMP from rye grass cultures was completely degraded by a mammalian cAMP phosphodiesterase, and 1-methyl-3-isobutylxanthine inhibited such degradation. The protein kinase activation and saturation assays gave essentially the same values for the cAMP content of axenic rye grass culture extracts. Material satisfying the above criteria for identity with cAMP was also isolated from the culture medium. The increase observed in medium cAMP levels during culture growth provides evidence for the synthesis and secretion of cAMP by rye grass endosperm cells in suspension culture.

Journal Article↗

PubMed1980

Variations in K(m)(CO(2)) of Ribulose-1,5-bisphosphate Carboxylase among Grasses.

A survey of the K(m)(CO(2)) values of ribulose-1,5-bisphosphate carboxylase from 60 grass species shows that enzyme from C(3) grasses consistently exhibits lower K(m)(CO(2)) than does that from C(4) grasses. Systematically ordered variation in K(m)(CO(2)) of ribulose-1,5-bisphosphate carboxylases from C(3) and C(4) grasses is also apparent and, among C(4) grasses, this shows some correlation with C(4) types.

Journal Article↗

PubMed1990

Evidence of grass-pollen allergenic activity in the smaller micronic atmospheric aerosol fraction.

In June 1988, during the grass-pollen season in Leiden, The Netherlands, outdoor airborne particulate matter was collected and separated into fractions according to aerodynamic sizes (greater than or equal to 10 microns, 4.9-10 microns, 2.7-4.9 microns, 1.3-2.7 microns, 0.6-1.3 microns, less than or equal to 0.6 microns), with a cascade impactor mounted on top of a high volume sampler. The different fractions were tested for the presence of grass-pollen allergenic activity using a RAST-inhibition assay: specific IgE-antibody-containing patient serum was applied on the particle-loaded impaction strips, and the serum was recovered by descending elution for further analysis in the RAST. Simultaneously, continuous measurements were made of the airborne grass-pollen concentration using a volumetric pollen trap. Sampling observations lasting 7-9 hr during a period with relatively high airborne grass-pollen concentrations showed reliably detectable amounts of grass-pollen allergen, not only in the first impaction stage where intact pollen were collected, but also in the lower stages collecting the smaller, paucimicronic and submicron atmospheric aerosol fraction. It is evident that this result has serious implications for the understanding of the bronchial symptoms frequently seen in hay fever patients on days with high pollen concentrations in the air.

Air↗