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Genetic and molecular basis of grass cell-wall biosynthesis and degradability. III. Towards a forage grass ideotype.

Lignification of cell walls is the major factor controlling the digestibility of forage grasses. Thus far, from QTL analysis, about 15 locations involved in cell-wall lignification or digestibility have been identified in the maize genome, many of which colocalise with QTLs involved in corn borer susceptibility. Genetic diversity for enhancing cell-wall digestibility in maize must be identified in novel germplasm, but genetic engineering is also a relevant way both to design specific cell-wall characteristics for improved digestibility and to identify genes involved in these traits for further discovery of alleles of interest in grass germplasm.

Animal Feed↗

Measurement of IgE antibodies against purified grass-pollen allergens (Phl p 1, 2, 3, 4, 5, 6, 7, 11, and 12) in sera of patients allergic to grass pollen.

BACKGROUND: Current allergy diagnosis is performed with allergen extracts which contain a variety of allergenic and nonallergenic components. The availability of highly purified and well-characterized allergen molecules seems to be an advantage of component-based diagnosis. METHODS: With the immunoenzymatic CAP FEIA System, we measured specific IgE levels to the recombinant allergens rPhl p 1, rPhl p 2, rPhl p 5, rPhl p 6, rPhl p 7, rPhl p 11, rPhl p 12, and native Phl p 4 in 77 sera of patients allergic to grass pollen, in order to evaluate the IgE-binding frequency to these purified grass-pollen allergens and their relationship to rBet v 4, rBet v 2, and other allergens. RESULTS: The frequency of sensitization was as follows: rPhl p 1=93.5%; rPhl p 2=67.5%; rPhl p 5=72.7%; rPhl p 6=68.8%; rPhl p 7=7.8%; rPhl p 11=53.2%; rPhl p 12=35.1%; and native Phl p 4=88.3%. As expected, rPhl p 7 and rPhl p 12 had a very good correlation (Spearman's r) with Bet v 4 (r=0.95%, P<0.05) and rBet v 2 (r=0.99, P<0.05), respectively. Good correlations of rPhl p 12 with papain (r=0.93, P<0.05), latex (r=0.92, P<0.05), and bromelain (r=0.86, P<0.05) were found. Highly variable individual sensitization patterns were observed. CONCLUSIONS: A new clinical approach has allowed the determination of specific allergograms for the different patients and may therefore be of great importance for more specific diagnosis. The use of component-resolved diagnostics may be useful to evaluate the allergen content of an extract for immunotherapy by monitoring patient's IgE and IgG directed to relevant allergens.

Adolescent↗

A comparison of the clinical and immunological effects of an alum-precipitated five-grass extract with a conjugated two-grass extract in the desensitization of hay fever.

Thirty-nine patients with pollen hay fever were randomly allocated to receive either an alum-precipitated five-grass extract (Allpyral) or a two-grass conjugated extract (Conjuvac) prior to the 1982 pollen season. Efficacy was assessed by means of symptom scores, drug usage and specific IgE and IgG antibody response. Both nasal and eye symptom scores were significantly higher during the pollen season in those patients receiving Allpyral. Specific IgE and IgG measured after the pollen season showed similar rises in each group. Seven patients showed a marked elevation in specific IgE and nine a marked elevation in specific IgG, but no patient showed both. Those with a high specific IgE level tended to be younger. Symptom scores bore no relation to either specific IgE or IgG antibodies. Side-effects were mainly local and were equal in both groups.

Adolescent↗

Immunogenicity in guinea pigs and tolerance in grass pollen-sensitive volunteers of enteric-coated grass pollen allergens.

An acid-insoluble, methacrylic acid, methyl methacrylate copolymer (Eudragit L-100) was used to give an enteric coating to a grass pollen extract in order to protect it against gastric degradation. Substantial protection against the degradative effects of simulated gastric secretion was demonstrated using this preparation which was well tolerated by grass pollen-allergic volunteers. The enteric-coated allergen induced a greater secondary antibody response than did an aqueous presentation when administered orally to guinea pigs which had been primed previously by subcutaneous injection. This result indicates that an effective hyposensitisation regimen could consist of a short series of initial parenteral injections, followed by an oral course of the protected allergen.

Acrylic Resins↗

Rumen fermentation and nutrient flows for cows fed grass and grass supplemented with molassed beet pulp pellets.

An experiment was carried out to determine the effect of a grass diet and a concentrate supplement on rumen fermentation and nutrient flows to the duodenum. Perennial ryegrass was cut and fed indoors to eight rumen- and duodenum-cannulated Friesian cows with or without 3 kg/d of molassed beet pulp in a randomized design experiment. The dry matter intake of grass was significantly lower for cows fed the concentrate supplement (13.6 vs. 11.5 kg of dry matter/d), but total dry matter and organic matter (OM) intakes were similar for cows fed both diets. Cows fed the supplement had higher mean concentrations of total volatile fatty acids (108 vs. 89 mmol/L) and a higher percentage of butyrate in total volatile fatty acids (13.5 vs. 11.6 mol/100 mol). There were no differences between the diets in the flow of OM to the duodenum or in the extent of OM digestion in the rumen. Flows of nonammonia N, microbial N, and amino acids to the duodenum tended to be higher for cows fed the supplemented diet than for those fed ryegrass only. The efficiency of microbial protein synthesis also tended to be higher for cows fed the supplemented diet (42 vs. 37.7 g/kg of OM apparently digested in the rumen and 28.2 vs. 26 g/kg of OM truly digested in the rumen). Overall, there were indications that the supplement caused better capture of N in the rumen and increased the efficiency of microbial protein synthesis.

Amino Acids↗

Bahia grass pollen, a significant aeroallergen: evidence for the lack of clinical cross-reactivity with timothy grass pollen.

Thirty-eight subjects were challenged (25 nasal, 13 bronchial) with Bahia grass, Paspalum notatum, pollen extract. A positive Bahia intradermal skin test predicted a positive challenge to Bahia in all (11/11) of the nasal challenges and 75% (6/8) of the bronchial challenges. All 19 subjects with negative Bahia intradermal skin tests had negative challenges with Bahia. Specific IgE antibodies to Bahia pollen were detected by conventional RAST (greater than or equal to 2+) in 82% (14/17) of subjects with positive challenges and in 5% (1/20) of subjects with negative challenges. Eight subjects had positive intradermal skin tests to either Bahia (three) or timothy, Phleum pratense (five). Seven of the eight subjects reacted exclusively to either Bahia or timothy nasal challenge as predicted by their skin tests. Bahia grass is a significant aeroallergen, which in some subjects can be demonstrated not to cross-react with timothy.

Allergens↗

Interaction between the endophytic fungus Epichloe bromicola and the grass bromus erectus: effects of endophyte infection, fungal concentration and environment on grass growth and flowering

Epichloe bromicola is an endophytic fungal species that systemically and perennially colonizes intercellular spaces of leaf blades, leaf sheaths and culms of Bromus grass species. E. bromicola causes choke disease in B. erectus, suppressing maturation of most, if not all, host inflorescences. In an investigation of the interaction between fungus and host, we used a quantitative polymerase chain reaction technique to estimate the amount of fungal DNA, and thereby fungal concentration, in host plants. Fungal concentration was directly correlated with vegetative vigour of the plant, as measured by longest leaf length, number of tillers and vegetative above-ground biomass, suggesting that, during vegetative growth, the endophytic fungus is most beneficial for the plant when present in high concentrations. In contrast, the reproduction of the plant, as measured by the number of functional inflorescences, was inversely correlated with fungal concentration: the majority of infected plants, and all that were associated with high concentrations of fungi, were diseased. Thus, the benefit of endophyte infection for the plant is coupled with the disadvantages of infertility. Fungal concentration was shown to be at least in part genetically determined because fungal concentration differed significantly in different plant-endophyte genotype combinations (symbiotum). In a field experiment with normal and CO2-enriched environments, elevated CO2 levels favoured fungal reproductive vigour over host reproductive vigour, suggesting that these plant endophytes would be at a selective advantage in a corresponding environmental-change scenario. We conclude that a dynamic and complex relationship between fungal endophyte infection, fungal concentration, genotype and environment affects growth and fecundity of B. erectus and should contribute to the evolution of these plant-fungal interactions.

Journal Article↗

[Perloline content in fodder grasses. 2. Perloline content in selected grass types and its variability in tall fescue and intergeneric tall fescue hybrids].

The perloline content of fodder grasses is genetically determined and strongly influenced by environmental factors. Tall fescue and meadow fescue show high, perennial and Italian ryegrass low concentrations of perloline. The perloline content of tall fescue can be diminished by bastardization to a level harmless in cattle feeding.

Alkaloids↗

A comparative study of an aqueous grass pollen extract and glutaraldehyde-treated grass pollen-tyrosine adsorbate in the treatment of pollenosis.

Hyposensitization therapy gave a subjective improvement in 84% of grass pollen-sensitive patients receiving an eighteen dose course of aqueous pollen extract (SDV) and in 69% of patients receiving a three dose course of glutaraldehyde-treated pollentyrosine adsorbate (Pollinex). Systemic reactions to treatment were considerably more frequent with the aqueous course, but virtually absent with the short adsorbate course, which seems therefore more convenient for the patient. The SDV treatment induced a more pronounced IgG antibody response both in patients and in guinea-pigs receiving the clinical dosage schedule. The Pollinex treatment showed a tendency to enhance the IgE antibody response in new patients.

Adult↗

Grass pollen hyposensitization versus placebo therapy. I. Clinical effectiveness and methodological aspects of a pre-seasonal course of desensitization with a four-grass pollen extract.

The present work reports the results of a double-blind clinical trial, comparing the effects of hyposensitization treatment versus placebo in 33 patients with allergic rhinitis, sensitive to a crude extract of the pollen of four different grasses (Dactylis glomerata, Lolium perenne, Secale cereale, and Phleum pratense). The distribution of these patients in the two groups was done randomly and gave two comparable groups, as far as clinical and biological features are concerned. The treatment course included five low doses of the aqueous extract followed by 12 injections of Al(OH)3-adsorbed aliquots of the same extract. Evaluation of the clinical scores was based on diary cards on which symptoms and medications were recorded. A reevaluation of the significance of the symptom and medication scores is presented and the link between both scores is studied. Particular attention is given to the methodological and statistical problems raised during this study. The non-parametric tests reveal a significant difference (P less or equal to than 0.03) in the total clinical score between the treated and the placebo groups for the second half of the observation period, when the pathology was most intense.

Adolescent↗

Characterization of cogon grass (Imperata cylindrica) pollen extract and preliminary analysis of grass group 1, 4 and 5 homologues using monoclonal antibodies to Phleum pratense.

BACKGROUND: Previous studies have established the role of Imperata cylindrica (Ic) pollen in type I allergic disorders. However, no systematic information is available on the allergen composition of Ic pollen extract. OBJECTIVES: To characterize the IgE-binding proteins of Ic pollen extract and to detect the presence of grass group 1, 4 and 5 allergen homologues, if any. METHODS: Pollen extract of Ic was analyzed by in vivo and in vitro procedures such as intradermal tests (ID), enzyme-linked immunosorbent assay (ELISA), ELISA-inhibition, thin-layer isoelectric focusing (TLIEF), sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting. Dot blot assay was carried out to check the presence of well-known group 1, 4, and 5 allergen homologues in Ic pollen extract. RESULTS: Out of 303 respiratory allergies patients skin-tested, 27 showed sensitivity to Ic pollen extract. Specific IgE levels were elevated in all 15 serum samples tested. The extract prepared for this study was found to be highly potent since it required only 400 ng of homologous proteins for 50% inhibition of binding in ELISA inhibition assays. TLIEF of Ic pollen extract showed 44 silver-stained bands (pI 3.5-7.0) while SDS-PAGE resolved it into 24 Coomassie-Brilliant-Blue-stained bands (MW 100-10 kD). Immunoblotting with individual patient sera recognized 7 major IgE-binding bands (MW 85, 62, 57, 43, 40, 28 and 16 kD) in Ic pollen extract. A panel of monoclonal antibodies, specific to group 1, 4 and 5 allergens from Phleum pratense pollen extract identified group 5 and group 4 homologues in Ic pollen extract. CONCLUSION: Ic pollen extract was characterized for the protein profile by TLIEF and SDS-PAGE. IgE reactivity was determined by ELISA and immunoblot. Monoclonal antibodies to group 5 and group 4 allergens reacted weakly showing that this pollen contains group 5 and group 4 homologous allergens.

Adolescent↗

Chemical modification of crude timothy grass pollen extract. II. Class and specificity of antibodies induced by chemically modified timothy grass pollen extract.

The effects have been studied of three different chemical modifications of timothy grass pollen extract on various immunological properties. The ability to induce IgG antibody with specificity for native antigen was least affected by glutaraldehyde treatment; IgE antibody production was reduced to a similar extent by all three modifications; there was no increase in IgM production; delayed reactions were reduced. New antigenic determinants were introduced by all the modifications, but the effect was minimal following glutaraldehyde treatment. The significance of these results and the potential application of modified allergen in hyposensitisation therapy are discussed.

Acetoacetates↗

Allergens of Kentucky Blue Grass pollen. II. Isolation of hapten-like components from Kentucky Blue Grass pollen by preparative isoelectrofocussing.

Components with hapten-like properties were isolated from the nondialyzable fraction, i.e. the retentate (R) and the dialyzable fractions of the aqueous extract of Kentucky Blue Grass pollen (KBC aq.ext.), by preparative isoelectrofocussing on Sephadex G-100 gel. These haptenic components could not elicit the passive cutaneous anaphylaxis (PCA) reactions in rats passively sensitized with a murine reaginic antiserum to R, but could inhibit completely and specifically the PCA reaction which is normally elicitable with R. It was concluded that the specificity of the murine IgE antibodies was directed to a determinant(s) which was common to either allergenic or haptenic fractions. Moreover, by employing a pool of human sera from individuals allergic to KBG pollen in the radioallergosorbent test procedure, it was apparent that most of the haptenic fractions lacked some of the specificities present on allergenic components of R that were recognized by the human IgE antibodies. Evidence was obtained to suggest that the electrophoretic heterogeneity of allergenic components present in various fractions of KBG aq.ext. may be due primarily to differences in their net charge, rather than to differences in their allergenic specificity.

Absorption↗

Characterization of the allergen group VI in timothy grass pollen (Phl p 6). II. cDNA cloning of Phl p 6 and structural comparison to grass group V.

The complete primary structure of the major allergen Phl p 6 was determined by cDNA cloning. A cDNA library of timothy grass pollen was screened by a Phl p 6 directed mouse antiserum. Six clones were obtained. From the cDNA data we deduced a protein sequence of 110 amino acids. While the size of the leader sequences varied considerably between the different clones, only one amino acid exchange affected the mature proteins. Sequence comparison of group VI and V allergens showed a high degree of homology in the N- and C-terminal regions. The structural data strongly indicate that group VI and V are independent groups of allergens, which have derived from a common precursor gene.

Allergens↗