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PubMed1987

Isotypic and antigenic restriction of the blocking antibody response to ryegrass pollen: correlation of rye group I antigen-specific IgG1 with clinical response.

To investigate the role of blocking antibodies in allergen immunotherapy (IT), we analyzed IgE, IgG, and IgG subclass 1 to 4 antibody responses to ryegrass group I antigen (RGGI) in a prospective double-blind, heterologous allergen, allergen-controlled trial of grass-pollen IT in 18 adults with seasonal rhinitis and asthma. Serum was assayed preseasonally before starting IT and again in midseason at time of documented highest natural exposure. Antibodies were measured by ELISA, and immunogenic specificities of ryegrass extract were examined by Western immunoblots. Nine subjects receiving grass-pollen IT and nine control subjects had similar clinical and immunologic status before IT. RGGI-specific IgE antibodies (sIgE) did not change from pretreatment levels in actively treated subjects but increased in control subjects (p less than 0.002). RGGI sIgG increased approximately thirteen-fold with active IT versus threefold during natural seasonal exposure (p less than 0.0005). The IgG-blocking response to RGGI was restricted to IgG1 and IgG4. Ten nonatopic subjects had similar RGGI sIgG1 but lower or undetectable sIgE and sIgG4 than the 18 atopic study subjects. Active IT dramatically increased RGGI sIgG4 (p less than 0.001) and to a lesser extent RGGI sIgG1 (p less than 0.01). Immunoblots demonstrated eight IgE-binding ryegrass-polypeptide allergens, with RGGI ubiquitous, and 11 IgG-binding polypeptides, including all eight allergens. A negative correlation between seasonal rhinitis symptom-medication scores and RGGI sIgG1 levels was found (r = -0.62, p less than 0.01), but no other immunologic parameters assayed were related to clinical improvement. Although RGGI sIgG4 predominates in the blocking response and is a useful marker of effective IT, early beneficial biologic effects may involve IgG1 antibodies.

Allergens↗
PubMed1989

Identification of two distinct allergenic sites on ryegrass-pollen allergen, Lol p IV.

Lol p IV is an important allergen of ryegrass pollen. For the immunochemical identification of antigenic and/or allergenic site(s), murine monoclonal antibodies (MAbs) were prepared against Lol p IV. The hybridoma cell-culture supernatants were screened for anti-Lol p IV antibodies by a combination of ELISA and Western immunoblot analyses. The MAbs were finally purified from ascites on a Mono Q ion-exchange column. In a competitive radioimmunoassay with Lol p IV as the solid phase and 125I-labeled MAbs, it was established that MAbs 90, 91, 92, 93, and 94, although they differed in their relative affinities, recognized in common with one another an epitope designated as antigenic site A, whereas MAb 12 recognized a different epitope referred to as site B. Sites A and B were also demonstrated to constitute allergenic determinants of Lol p IV. Differences in the repertoire of specificities of the human IgE antibodies directed to Lol p IV were also demonstrated. Interestingly, it was found that sera from both allergic as well as from nonatopic individuals had IgG antibodies to sites A and/or B.

Allergens↗
PubMedDate not supplied

Clinical evaluation of long-term treatment using cernitin pollen extract in patients with benign prostatic hyperplasia.

Seventy-nine patients with benign prostatic hyperplasia (BPH) were treated with cernitin pollen extract. Patient ages ranged from 62 to 89 years (mean, 68 years). Mean baseline prostatic volume was 33.2 cm3. Cernitin pollen extract was administered in a dosage of 126 mg (2 tablets, 63 mg each), three times a day, for more than 12 weeks. Symptom scores, based on a modified Boyarsky scoring scale, uroflowmetry, prostatic volume, residual urine volume, and urinalysis results were examined before and after administration of cernitin pollen extract. Symptom scores significantly decreased from baseline, and the favorable results continued during the treatment period. Urine maximum flow rate and average flow rate increased significantly from 9.3 mL/s to 11 mL/s and from 5.1 mL/s to 6 mL/s, respectively. Residual urine volume decreased significantly from 54.2 mL to less than 30 mL. There was no change in prostatic volume. However, 28 patients treated for more than 1 year showed a mean decrease of prostatic volume to 26.5 cm3. No adverse reactions were observed. Clinical efficacy at 12 weeks was rated excellent, good, satisfactory, and poor in 11%, 39%, 35%, and 15% of patients, respectively. Overall clinical efficacy was 85%. In conclusion, cernitin pollen extract showed a mild beneficial effect on prostatic volume and urination variables in patients with symptomatic BPH.

Administration, Oral↗
PubMed1988

Anti-idiotypic antibodies as probes for the determination of idiotopes shared by human and monoclonal murine antibodies.

Murine monoclonal antibodies (MAbs) against rye grass Group I (rye I) allergen were previously produced (290A-167, 348A-6 and 539A-6) and are further characterized herein. By competitive binding to polystyrene-bound allergen, it was shown that the three MAbs are directed against three different non-overlapping epitopes on rye I. Human rye I-specific IgG isolated from the serum of one rye grass sensitive patient could recognize the same or closely related epitopes than those recognized by MAbs. Antisera were then raised in rabbits against purified F(ab')2 fragments of human rye I-specific IgG and F(ab')2 fragments of 539A-6 MAb. The antisera were rendered idiotype specific by adsorption with insolubilized non-specific human IgG from the same donor, insolubilized normal murine immunoglobulins and finally with rye I. As shown by competitive binding to polystyrene-bound allergen, rabbit anti-idiotypic antibodies (anti-ID antibodies) produced against F(ab')2 fragments of human rye I specific IgG could inhibit the reaction between two MAbs (290A-167 and 539A-6) and the relevant allergen. Furthermore, human rye I specific IgG could inhibit the binding of 125I-labelled 539A-6 MAb to its specific anti-ID antibody to a significant degree. Human autoanti-idiotypic antibodies were also shown to inhibit the reaction between the three anti-rye I MAbs and the antigen. These observations suggest that cross-reactivity exists between idiotypic determinants of human rye I-specific IgG and the three murine MAbs, which implies structural similarity in the V genes coding for the variable region of the antibody from these two different species.

Allergens↗
PubMed1989

Mapping of Lol p I allergenic epitopes by using murine monoclonal antibodies.

Murine monoclonal antibodies (MAbs) against three non-overlapping epitopes of Lol p I allergen were previously produced and subsequently used for purification of the allergen. In the present study, these MAbs were further characterized, and the biological activity of the purified allergen assessed. The three MAbs were of the IgG isotype and carried a kappa light chain. Their affinity constants were in the range of 7.4-15.1 x 10(-9) mol/l. Purified Lol p I kept its biological activity, as shown by its ability to induce histamine release by basophils of Lol p I-sensitive patients. The profiles of histamine release induced by either Lol p I or crude Lolium perenne extracts were comparable. This observation suggests that human IgE bound to basophils are polyspecific which has been confirmed by immunoblot and inhibition assay. Our data indicated also that Lol p I possesses a major allergenic epitope recognized by all human serum IgE tested. This epitope seems to be partially shared by those recognized by the three MAbs. Finally, preincubation of Lol p I with either one of the Mabs did not affect significantly the basophil-histamine release induced by the purified allergen. This suggests that Lol p I possesses allergenic sites other than the one shared by MAbs and IgE Abs.

Allergens↗
PubMed1995

Analysis of the primary structure of the chloroplast isozyme of triosephosphate isomerase from rye leaves by protein and cDNA sequencing indicates a eukaryotic origin of its gene.

The primary structure of the chloroplast isozyme of triosephosphate isomerase from rye leaves was identified by protein and cDNA sequencing and compared to the deduced amino acid sequence of a cDNA for the cytosolic isozyme. The mature cytosolic and chloroplast isozyme proteins share 64% amino acid sequence identity. The cDNA for the chloroplast isozyme codes for a precursor protein consisting of an N-terminal transit peptide of Mr 4351 and a mature subunit of Mr 27,282. Southern blot analysis indicates that the two rye isozymes are encoded by two independent single genes. Amino acid residues or sequence regions of basic functional relevance in known triosephosphate isomerases are strictly conserved in the chloroplast isozyme. The chloroplast isozyme contains 6 cysteine residues, instead of 4 in the cytosolic isozyme. A cysteine at position 143 of the chloroplast isozyme appears to be modified. Phylogenetic trees constructed on the basis of sequence comparisons for triosephosphate isomerases from different species of all major taxonomic groups indicate that the chloroplast isozyme is much more closely related to eukaryotic cytosolic enzymes than to eubacterial enzymes. The results indicate that the nuclear gene for the chloroplast isozyme originated with that for the cytosolic isozyme through duplication of an ancestral eukaryotic gene, rather than through gene transfer from a prokaryotic endosymbiont.

Amino Acid Sequence↗
PubMed1996

Identification of major rye secalins as coeliac immunoreactive proteins.

Six distinct gamma- and omega-type secalins, together with two new low molecular mass glycoproteins, have been identified as the major coeliac immunoreactive proteins from a chloroform/methanol soluble extract from rye endosperm. These components were characterized by a combination of reverse-phase high-performance liquid chromatography, immunoblotting using a coeliac serum and microsequencing analysis. This allowed the identification of a group of secalins with different molecular masses according to their N-terminal amino-acid sequence: one omega-type secalin of 40 kDa (omega 1-40); three gamma-type secalins, one of 70 kDa (gamma-70) and two of 35 kDa (gamma-35); as well as two low molecular mass glycoproteins of 15 and 18 kDa, all exhibiting coeliac serum antigenicity. Moreover, four additional rye components, including two low molecular mass proteins, which did not react with coeliac sera, have also been identified. Analysis by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) of the three main purified coeliac immunogenic secalins, gamma-70, gamma-35 and omega 1-40, indicated molecular masses of 71457, 32240 and 39117 Da, respectively. The omega 1-40 secalin displays a significant absorption in the visible region which could be related to its peculiar low capacity to bind both coeliac sera antibodies and Coomassie brilliant blue dye.

Amino Acid Sequence↗
PubMed2004

Occurrence of proteinaceous endoxylanase inhibitors in cereals.

Cereals contain proteinaceous inhibitors of endoxylanases, which affect the efficiency and functionality of these enzymes in cereal processing. This review relates their first discovery in wheat and the subsequent purification of two distinct classes of endoxylanase inhibitors, namely Triticum aestivum xylanase inhibitor (TAXI)-type and xylanase inhibitor protein (XIP)-type inhibitors in cereals. Both inhibitor classes occur in monocots as multi-isoform families. The reported data provide an overview of the relative quantitative and qualitative variation of these inhibitors in cereals. Wheat and rye are particularly rich in TAXI-type and XIP-type inhibitors with the latter inhibitors being more abundant. Lower inhibitor levels are present in durum wheat and barley, while maize contains solely XIP-type inhibitors. No inhibitors have been isolated from rice, oats and buckwheat.

Bacteria↗
PubMed2005

Nitrogen budgets for three cropping systems fertilised with cattle manure.

A field plot experiment was set up on a sandy loam soil of SW England in order to determine the efficiency of nitrogen use from different cattle manures. The manure treatments were low and high dry matter cattle slurries and one farmyard manure applied at a target rate of 200 kg total N ha(-1)year(-1), and an untreated control. There were three different cropping systems: ryegrass/clover mixture, maize/rye and maize/bare soil, which were evaluated during 1998/99 and 1999/00. Measurements were made of N losses, N uptake and herbage DM yields. Result showed that manure type had a significant effect on N utilisation only for maize. N balances were negative in maize (approximately -247 to -10 kg N) compared to grass (approximately 5-158 kg N). Agronomic management was more important than manure type in influencing N losses, where soil cultivation appeared to be a key factor when comparing maize and grass systems.

Animals↗
PubMed2005

Dilute acid pretreatment of rye straw and bermudagrass for ethanol production.

Ethanol production from lignocellulosic materials provides an alternative energy production system. Rye and bermudagrass that are used in hog farms for nutrient uptake from swine wastewater have the potential for fuel ethanol production because they have a relative high cellulose and hemicellulose content. Dilute sulfuric acid pretreatment of rye straw and bermudagrass before enzymatic hydrolysis of cellulose was investigated in this study. The biomass at a solid loading rate of 10% was pretreated at 121 degrees C with different sulfuric acid concentrations (0.6, 0.9, 1.2 and 1.5%, w/w) and residence times (30, 60, and 90 min). Total reducing sugars, arabinose, galactose, glucose, and xylose in the prehydrolyzate were analyzed. In addition, the solid residues were hydrolyzed by cellulases to investigate the enzymatic digestibility. With the increasing acid concentration and residence time, the amount of arabinose and galactose in the filtrates increased. The glucose concentration in the prehydrolyzate of rye straw was not significantly influenced by the sulfuric acid concentration and residence time, but it increased in the prehydrolyzate of bermudagrass with the increase of pretreatment severity. The xylose concentration in the filtrates increased with the increase of sulfuric acid concentration and residence time. Most of the arabinan, galactan and xylan in the biomass were hydrolyzed during the acid pretreatment. Cellulose remaining in the pretreated feedstock was highly digestible by cellulases from Trichoderma reesei.

Bioelectric Energy Sources↗
PubMed2006

Quantitative determination of five ergot alkaloids in rye flour by liquid chromatography-electrospray ionisation tandem mass spectrometry.

A confirmatory method for detecting five ergot alkaloids, ergocristine, ergotamine, ergonovine, ergocornine and alpha-ergokryptine, in rye flour is described using high performance liquid chromatography coupled to tandem mass spectrometry detection by monitoring two transition reactions per analyte. The procedure entails a liquid-liquid extraction followed by a clean-up step using a C18 solid-phase extraction (SPE) cartridge. An analogue compound, methysergide hydrogen maleinate, was used to assess both repeatability sample preparation and potential MS response fluctuations. The method was fully validated according to the European Union (EU) criteria. Detection and quantification limits of all analytes were calculated ranging from 7 to 11 microg/kg and from 23 to 37 microg/kg, respectively. Fifteen rye flour samples were investigated with the newly developed method, and none of them were above the current Swiss limits of 200mg/kg for total ergot alkaloids.

Chromatography, High Pressure Liquid↗
PubMed2005

Phytoremediation of aquaculture wastewater for water recycling and production of fish feed.

Five plants were examined for their ability to remove nutrients from aquaculture wastewater and suitability as fish feed: alfalfa, white clover, oat, fall rye, barley. The seeds were first germinated in water in a hydroponic system, and the plants were fed wastewater from Tilapia production facility. Clover and alfalfa seeds were infected with fungus shortly after germination, and their roots were completely destroyed by day 14. Oat, rye and barley had the fastest growth and showed greater tolerance to fungal disease compared with alfalfa and clover. Although substantial amounts of soluble and insoluble substances were released by the seeds during the germination period, the plants were able to remove all the pollutants in wastewater and significant portions of those released substances. The total reductions in total solids, COD, NO3-N, NO2-N, phosphate and potassium ranged from 54.7% to 91.0%, 56.0% to 91.5%, 82.9% to 98.1%, 95.9% to 99.5%, 54.5% to 93.6% and 99.6% to 99.8%, respectively. Oat, barley and rye grow well in this type of hydroponic system and can be used as a fish feed after being supplemented with fat, Ca, Na, Mn and Fe. Oil seeds and the chlorides of these elements could be added to these plants when formulating the fish feed. For a continuous operation, a two-unit system could be configured to allow for one week germination and one week cleaning and startup in one unit while the other unit is in operation.

Animals↗
PubMed2005

Study of calcium-dependent lead-tolerance on plants differing in their level of Ca-deficiency tolerance.

The main aim of the study was to determine the role of calcium in the amelioration of lead toxic effects in plants with accordingly high/low level of Pb-tolerance and high/low Ca-deficiency tolerance. The study was performed on maize, rye, tomato and mustard. Plants were cultivated in modified Knop's solution. They were subjected to Ca-deficiency, and to lead nitrate administered in the presence of four calcium nitrate concentrations 3.0, 2.4, 1.2, 0.3mM. Lead-tolerance and tolerance to Ca-deficiency were determined, as were concentration of the studied elements in plant tissues, and the Pb deposition pattern at the ultrastructural level (electron microscopy study, X-ray microanalysis). In all studied plants, lead toxicity increased as medium calcium content decreased, however, only in the Ca-deficiency sensitive mustard with low Pb-tolerance was it accompanied by a rise in tissue lead concentration. In contrast, lead root and shoot levels did not increase in the highly Ca-deficiency tolerant tomato, mustard and rye with high Pb-tolerance irrespective of the Ca(2+) regimens applied. Thus, in these plants, lead's unfavourable effects resulted only from the higher toxicity of the same amount of lead in tissues at low calcium in the medium. Of particular relevance is the finding by electron microscopy and X-ray microanalysis, that under low calcium in both highly Ca-deficiency tolerant and Ca-deficiency sensitive plants, less efficient Pb(2+) detoxification was accompanied by the restriction of the formation of large lead deposits in cell walls. Obtained results are novel in demonstrating calcium involvement in the lead deposition in the cell wall, thus in the regulation of the internal lead detoxification.

Air Pollutants↗
PubMed2006

Impact of sourdough on the texture of bread.

Sourdough has been used since ancient times and its ability to improve the quality and increase the shelf-life of bread has been widely described. During sourdough fermentation, lactic acid bacteria (LAB) produce a number of metabolites which have been shown to have a positive effect on the texture and staling of bread, e.g. organic acids, exopolysaccharides (EPS) and/or enzymes. EPS produced by LAB have the potential to replace more expensive hydrocolloids used as bread improvers. Organic acids affect the protein and starch fractions of flour. Additionally, the drop in pH associated with acid production causes an increase in the proteases and amylases activity of the flour, thus leading to a reduction in staling. While improving the textural qualities of bread, sourdough fermentation also results in increased mineral bioavailability and reduced phytate content. In this review we will be discussing the effect of sourdough on wheat and rye bread as well as the potential of sourdough to improve the quality of gluten-free bread.

Bread↗
PubMed2006

Bran fermentation as a means to enhance technological properties and bioactivity of rye.

Response surface methodology was applied to study the effects of fermentation on the levels of phytochemicals (folates, phenolic compounds, alkylresorcinols) and on the solubilization of pentosans in rye bran from native and peeled grains. Furthermore, the microbial composition of the brans before and after fermentation was studied. Peeling reduced the microbial load and lower microbial counts were detected in the fermentation experiments carried out with the bran from peeled grains. High temperature and long fermentation time favoured the growth of indigenous lactic acid bacteria (LAB), and a diverse microbial community was detected. The brans contained low levels of aerobic spore-forming bacteria, but their number was not increased during the fermentations. Fermentation of both brans increased the levels of folates, easily extractable total phenolics and free ferulic acid. During fermentation of bran from native grains, the levels of alkylresorcinols slightly increased but during fermentation of bran from peeled grains they decreased. Significant increase in soluble pentosans was established in both types of rye bran fermentations. Enhanced bioactivity and solubilization of pentosans with limited microbial growth were obtained after 12-14 h fermentation at 25 degrees C. The results suggest that fermentation is a potential bioprocessing technology for improved technological properties and bioactivity of rye bran.

Bread↗
PubMed2005

Effects of yeasts and bacteria on the levels of folates in rye sourdoughs.

Fermentation of rye dough is often accompanied with an increase in folate content. In this study, three sourdough yeasts, Candida milleri CBS 8195, Saccharomyces cerevisiae TS 146, and Torulaspora delbrueckii TS 207; a control, baker's yeast S. cerevisiae ALKO 743; and four Lactobacillus spp., L. acidophilus TSB 262, L. brevis TSB 307, L. plantarum TSB 304, and L. sanfranciscensis TSB 299 originally isolated from rye sourdough were examined for their abilities to produce or consume folates. The microorganisms were grown in yeast extract-peptone-d-glucose medium as well as in small-scale fermentations that modelled the sourdough fermentation step used in rye baking. Total folate contents were determined using Lactobacillus rhamnosus (ATCC 7469) as the growth indicator organism. The microorganisms studied did not excrete folates into the media in significant amounts. Yeasts increased the folate contents of sterilised rye flour-water mixtures from 6.5 microg/100 g to between 15 and 23 microg/100 g after 19-h fermentation, whereas lactic acid bacteria decreased it to between 2.9 and 4.2 microg/100 g. Strains of Lactobacillus bulgaricus, L. casei, L. curvatus, L. fermentum, L. helveticus, Pediococcus spp., and Streptococcus thermophilus that were also tested gave folate contents after fermentation that varied between 2 and 10.4 microg/100 g. Although the four Lactobacillus spp. from sourdough consumed folates their effect on folate contents in co-cultivations was minimal. It was concluded that the increase of folate content during fermentation was mainly due to folate synthesis by yeasts. Fermentation of non-sterilised flour-water mixtures as such resulted in three-fold increases in the folate contents. Two folate producing bacteria were isolated from the non-sterilised flour and identified as Enterobacter cowanii and Pantoea agglomerans.

Bread↗
PubMed2004

Identification of cereal alkylresorcinol metabolites in human urine-potential biomarkers of wholegrain wheat and rye intake.

Alkylresorcinols, phenolic lipids present in high amounts in wholegrain wheat and rye, are of interest as potential biomarkers of the intake of these cereals. Alkylresorcinols are known to be absorbed by humans and animals, but little is known about their metabolism or resulting metabolites. A preliminary human study was carried out to identify alkylresorcinol metabolites in human urine. Urine samples, collected before and after a wheat-bran based meal, were deconjugated with beta-glucuronidase/sulphatase and then extracted with ethyl acetate. Extracts were separated by thin-layer chromatography, and fractions containing alkylresorcinols and possible metabolites were identified by retention on the plate compared to standard compounds, and staining with fast blue B. These fractions were further analysed by gas chromatography-mass spectrometry. Deconjugated human urine after the wheat-bran based meal contained two alkylresorcinol metabolites, 3,5-dihydroxybenzoic acid and 3-(3,5-dihydroxyphenyl)-1-propanoic acid, as well as smaller amounts of unchanged alkylresorcinols, confirming the hypothesis that alkylresorcinols are metabolised in humans via beta-oxidation of their alkyl chain.

Biomarkers↗
PubMed2004

Ergot of rye-the first specific for migraine.

Over the past decade the various triptan derivatives have been accepted as the most effective available agents for relieving migraine attacks. Prior to that, for a period of half a century, ergotamine was the only 'specific' available for this purpose. In 1918, Stoll had isolated it from the various alkaloids present in extracts of the sclerotia of the fungus Claviceps purpurea (ergot), which grow on rye and, to a lesser extent, on other grasses. By 1925 ergotamine was beginning to be used to treat migraine attacks. However, as ergotamine was present in extracts of ergot, which had been used to treat migraine first in Italy in 1862, and then by Edward Woakes (1868) in England, and after him by Albert Eulenburg in Germany (1883), the drug had actually come into unrecognised use for the disorder more than half a century before ergotamine itself was known to exist. Unfortunately, because of ergotamine's chemical and pharmacokinetic properties, extracts of ergot of rye were incapable of producing consistent therapeutic results, so that general acceptance that the first specific substance for migraine treatment existed had to wait until pure ergotamine was available for administration.

Claviceps↗