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Eosinophilic esophagitis in adults--no clinical relevance of wheat and rye sensitizations.

BACKGROUND: Eosinophilic esophagitis (EE) is often associated with concomitant atopic diseases. In children with EE in whom food allergens have been identified as causative factors, elemental and elimination diets result in an improvement or resolution of symptoms. Most adult EE patients are sensitized to aeroallergens, which cross-react with plant-derived food allergens, most commonly to grass pollen and cereals. AIMS OF THE STUDY: To investigate the clinical relevance of the sensitization to wheat and rye, and the efficacy of an allergen-specific elimination diet in adult EE patients. METHODS: Six patients (five men, one women) with permanently active EE sensitized to grass pollen and the cereals wheat and rye underwent a double-blind placebo-controlled food challenge and were kept on an elimination diet avoiding wheat and rye for 6 weeks. RESULTS: The challenge tests with wheat and rye did not provoke any EE symptoms in all patients. The elimination diet failed in reducing disease activity. Although one patient noticed an improvement of symptoms, endoscopic and histopathologic findings remained unchanged. CONCLUSIONS: In adult EE patients, sensitization to wheat and rye does not seem causative for EE. Elimination diet is not a reliable and efficient therapeutic measure in EE patients sensitized to wheat and rye. Low specific immunoglobulin-E levels to wheat and rye may be a consequence of the underlying grass pollen allergy.

Adult↗

Synthesis and degradation of unassembled polypeptides of the coupling factor of photophosphorylation CF1 in 70S ribosome-deficient rye leaves.

The formation of polypeptides of the coupling factor CF1 was investigated in 70S ribosome-deficient rye leaves generated by growing the plants at a non-permissive elevated temperature of 32 degrees C, in order to analyse mechanisms coordinating subunit accumulation. Antibodies were raised in rabbits against total CF1 as well as against its five individual subunits purified from chloroplast thylakoids from rye leaves. Several immunological techniques applying these antibodies (immunoprecipitation, immunoblotting, antibody affinity chromatography) were unable to detect the presence of any of the CF1 subunits in heat-treated 70S ribosome-deficient leaves. After in vivo labeling with L-[35S]methionine and subsequent immunoprecipitation, however, radioactivity was found to be incorporated into the subunits gamma and delta, but not into alpha, beta and epsilon, in 70S ribosome-deficient leaves, demonstrating the cytoplasmic synthesis of CF1-gamma and CF1-delta. Chase experiments after in vivo labeling with L-[35S]methionine indicated that the unassembled subunits gamma and delta were rapidly and preferentially degraded, while they were stabilized when integrated into the complete CF1 complex in normal green leaves from permissive growth conditions. The apparent half-times of the unassembled subunits were 2 h for CF1-gamma and 4 h for CF1-delta in 32 degrees C-grown leaves. Several other, stromal, plastid proteins of cytoplasmic origin were stable in 32 degrees C-grown leaves during the period of chase. In etiolated leaves total CF1, including all subunits, appeared to be less stable than in green leaves grown under permissive temperature conditions in light. Rapid degradation of the excess of unassembled subunits is regarded as an important mechanism ensuring a constant stoichiometry and apparently synchronous development of CF1 subunits.

Edible Grain↗

Rye inhibitors of animal alpha-amylases show different specificities, aggregative properties and IgE-binding capacities than their homologues from wheat and barley.

Three new members of the alpha-amylase/trypsin-inhibitor family of cereal endosperm have been isolated from rye. N-terminal amino acid sequence comparison revealed that two of the purified proteins were the rye homologues of the barley monomeric inhibitor (BMAI-1) previously described, while the other rye protein corresponded to one of the subunits of the barley and wheat heterotetrameric inhibitors. However, the inhibitory specificities (active against human salivary alpha-amylase), aggregative behaviours (mainly as dimeric forms) and IgE-binding capacities (not recognized by sera from allergic patients) of the rye inhibitors were clearly different from those of their wheat and barley counterparts. These results indicate that homologous inhibitors may have distinctive properties in different cereal species.

Amino Acid Sequence↗

Different consequences of incorporating chloroplast ribosomal proteins L12 and S18 into the bacterial ribosomes of Escherichia coli.

We have incorporated chloroplast ribosomal proteins (R-proteins) L12 and S18 into Escherichia coli ribosomes and examined the hybrid ribosomes for their ability to form polysomes in vivo and perform poly(U)-dependent poly(Phe) synthesis in vitro. The rye chloroplast S18 used for the experiment is a highly divergent protein (170 amino acid residues; E. coil S18, 74 residues), containing a repeating, chloroplast-specific, heptapeptide motif, and has amino acid sequence identity of only 35% to E. coli S18. When expressed in E. coli, chloroplast S18 was assembled in E. coli ribosomes. The latter formed polysomes in vivo at about the same rate as the host ribosomes, indicating that the replacement of E. coli S18 with its chloroplast homologue has only a minor, if any, effect on function. The L12 protein is much more conserved in sequence and chain length, and is known to have a very important function. The Arabidopsis chloroplast L12 used in the experiment was incorporated into E. coli 50S subunits that associated with the 30S subunits to form ribosomes, but the latter were unable to form polysomes. This result indicates functional inactivation of E. coil ribosomes by a chloroplast R-protein. To further confirm this result, we overproduced chloroplast L12 through the use of a secretion vector and purified the protein to homogeneity. Chloroplast L12 could be efficiently incorporated in vitro into L7/12-lacking E. coli ribosomes, but the hybrid ribosomes were totally inactive in poly(U)-dependent poly(Phe) synthesis. Computer modeling of the spatial structure of all known chloroplast L12 proteins (using E. coli L12 coordinates) indicated a 'chloroplast loop' present only in chloroplast L12. The presence of this loop might have a role in the observed inactivation. Taken together with previously reported results (summarized in this paper), it would appear that the features of chloroplast R-proteins concerned with specific functions are more divergent than their assembly properties. We have previously described methods suitable for overproduction and purification of chloroplast R-proteins that are encoded in organellar DNA (approximately 20), but that gave poor yield for those encoded in the nuclear DNA (approximately 45). Here we describe a method that overcomes this problem and allows the purification of nucleus-encoded chloroplast R-proteins in milligram quantities.

Amino Acid Sequence↗

Passage through alternative hosts changes the fitness of Fusarium graminearum and Fusarium pseudograminearum.

Species of the necrotrophic fungal pathogen Fusarium that cause head blight and crown rot of cereals including wheat also infect a number of alternative host plants. This raises the prospect of more damaging pathogen strains originating and persisting as highly successful saprophytes on hosts other than wheat. The immediate impact on pathogenic (aggressiveness) and saprophytic (growth rate and fecundity) behaviour of six isolates with low, moderate or high initial aggressiveness was examined in two species of Fusarium after their passage through 10 alternative plant hosts. One passage through alternative hosts significantly reduced the pathogenic fitness of most isolates, but this change was not associated with a concomitant change in their overall saprophytic behaviour. The overall weak association between aggressiveness, fecundity and growth rate both before and after passage through the alternative hosts indicate that pathogenic and saprophytic fitness traits may be independently controlled in both Fusarium species. Thus, there was no trade-off between pathogenic and saprophytic fitness in these necrotrophic plant pathogens.

Edible Grain↗

Treatment of chronic prostatitis and prostatodynia with pollen extract.

Chronic abacterial prostatitis and prostatodynia are notoriously difficult both to diagnose and to treat. These patients tend to have received several courses of antibiotics, antiinflammatory agents or adrenergic blockade and various other therapeutic manoeuvres with little success. The pollen extract, Cernilton, is reported to be effective in the treatment of this condition and we present the results of an open trial with Cernilton in a group of 15 patients with chronic prostatitis and prostatodynia. In 13 patients there was either complete and lasting relief of symptoms or a marked improvement; 2 patients failed to respond. Cernilton was found to be effective in the treatment of chronic prostatitis and prostatodynia. Its precise mode of action is not known, although experimental studies suggest that it has anti-inflammatory and anti-androgenic properties.

Adult↗

In vitro evaluation of the pollen extract, cernitin T-60, in the regulation of prostate cell growth.

Nine human-derived cancer and non-cancer continuous cell lines were employed to evaluate the relative in vitro activity of the pollen extract, Cernitin T-60. Responses of the cell lines to the drug were assessed by measuring growth and cell survival as determined by cell count. The results demonstrated that of the 9 continuous cell lines tested, only those derived from the human prostate were growth inhibited by the pollen extract, whereas the non-prostate derived cells exhibited variable degrees of resistance to the T-60. The selectivity of the drug for the prostate cell lines was even more pronounced in the hormone-independent models, suggesting that there might be a place for the pollen extract in the control of abnormal growth in hormone-insensitive cells.

3-Oxo-5-alpha-Steroid 4-Dehydrogenase↗

Treatment of outflow tract obstruction due to benign prostatic hyperplasia with the pollen extract, cernilton. A double-blind, placebo-controlled study.

Whilst prostatectomy remains the "gold standard" for the treatment of outflow tract obstruction due to benign prostatic hyperplasia, medical treatment--if only for symptomatic relief--appears to be an attractive alternative. Most of the pharmacological agents in use block the hormonal or the sympathetic neurological pathways that influence prostate growth and function. All of these drugs are known to have side effects. Sixty patients with outflow obstruction due to benign prostatic hyperplasia (BPH) were entered into a double-blind, placebo-controlled study to evaluate the effect of a 6-month course of the pollen extract, Cernilton. There was a statistically significant subjective improvement with Cernilton (69% of the patients) compared with placebo (30%). There was a significant decrease in residual urine in the patients treated with Cernilton and in the antero-posterior (A-P) diameter of the prostate on ultrasound. However, differences in respect of flow rate and voided volume were not statistically significant. It is concluded that Cernilton has a beneficial effect in BPH and may have a place in the treatment of patients with mild or moderate symptoms of outflow obstruction.

Aged↗

Results of treatment with pollen extract (Cernilton N) in chronic prostatitis and prostatodynia.

We report the results of a prospective study with the pollen extract, Cernilton N, in a dose of 1 tablet tid for 6 months for the treatment of chronic prostatitis syndrome in 90 patients. The factors documented before and after 3 and 6 months' treatment were digital rectal examination (DRE) of the prostate, uroflowmetry, bacterial studies, leucocyte counts in urine and measurement of complement C3/coeruloplasmin in the seminal fluid. The patients were divided into 2 groups: those without associated complicating factors (CFs) (n = 72) and those with complicating factors, i.e. urethral strictures, prostatic calculi, bladder neck sclerosis (n = 18). In the group without CFs, 56 (78%) had a favorable response; 26 (36%) were cured of their symptoms and signs and 30 (42%) improved significantly with an increase in flow rate, a reduction in leukocyturia in the post-prostate massage urine (VB3) and a decrease in complement C3/coeruloplasmin in the ejaculate. In the patients with CFs only 1 patient showed a response. Complicating factors should be considered in patients who fail to respond to treatment within 3 months. Cernilton N was well tolerated by 97% of patients.

Adult↗

Associated mycoflora of rye bread.

Penicillium roqueforti (27%), P. corylophilum (20%) and Eurotium species (15%) made up the important mycoflora associated with rye bread on 3425 identified fungi isolates. These fungi were dominant as spoilers of packaged rye bread in almost every month of a 4 year investigation. Penicillium decumbens (3%), Paecilomyces variotti (8%) and Aspergillus flavus (5%) were found more rarely, but were the major species found over periods of a few months. Penicillium commune (5%), P. solitum (4%), A. niger (4%) and Mucor species (2%) were a constant, but small, part of the total mycoflora of rye bread. Identification of the fungi in the production environment in a rye bread factory showed the locality of potential contamination sources. Elimination of the contamination sources by improved cleaning and disinfection procedures quickly resulted in a significant reduction in the frequency of mould growth in the packaged rye bread.

Aspergillus flavus↗

Effects of variously processed starch on pH of human dental plaque.

The aim of this investigation was to study the effect of starch in various cereal products on pH of human dental plaque in vivo as compared to availability to the alpha-amylase of starch in vitro. Three separate experimental series were performed including: 1) rye and wheat products, 2) wheat products processed under both mild and severe conditions, and 3) wheat products with and without added sucrose. The pH drop in dental plaque was studied after a mouthrinse in groups of 10 subjects. Solutions of glucose or sucrose were used as references. No differences were found between boiled rye and boiled wheat, nor between rye bread and wheat bread. Severely popped wheat resulted in the greatest pH drop, followed, in order, by drum-dried and extrusion-cooked products. Mildly steam-flaked and mildly dry-autoclaved products induced comparable small pH falls. Sucrose-sweetened wheat products resulted in lower plaque pH values compared to unsweetened products. This effect was most pronounced with a mildly processed sample. In all three series, the glucose and sucrose reference solutions showed the greatest pH drop. In conclusion, all processing methods increased the fermentability of starch in human dental plaque in vivo. Thus, the more severe the processing conditions the more prominent the pH falls. The pH drop in plaque in vivo correlated (r = 0.96 in Series 1 and r = 0.92 in Series 2) with the rate of starch hydrolysis with alpha-amylase in vitro.

Biological Availability↗

Effect of rye A and B chromosomes on meiotic association of Hordeum marinum ssp. gussoneanum (4x) chromosomes in intergeneric hybrids.

Chromosome association at metaphase I was studied in PMCs of eight H. marinum ssp. gussoneanum (4x) x rye hybrids. Differences in the levels of association separated six hybrids with 2n = 23 including 14 Hordeum, 7 rye A and 2 rye B chromosomes into two groups of three plants each, a "low association" group with means of 0.03III + 4.43II (1.55 rings + 2.88 rods) + 5.10I and 6.03 chiasmata/cell, and a "high association" group with means of 0.01IV + 0.03III + 6.40II (3.55 rings + 2.85 rods) + 1.13I and 10.04 chiasmata/cell. The low number of plants studied prevents a safe estimate of the number of genes involved, but the significant difference between groups suggests the presence in the rye genome of two major genes, or two genotypes, for control of meiotic chromosome association. In two additional hybrids with 2n = 25, one of each above-mentioned group, the presence of two extra rye B chromosomes raised chiasma frequencies by ca 1.5, indicating a promoting effect on chromosome association. The level of Hordeum chromosome association in the "high association" group and the observation of up to 7 Hordeum ring bivalents in some cells agree with an autoploid origin of H. marinum ssp. gussoneanum (4x). Hordeum and rye chromosomes formed a few heterogenomic bi- and trivalents. Most rye A chromosomes formed univalents, but 2.7% were included in associations. Rye B chromosomes generally formed rod bivalents. The use of genome analysis in its traditional sense is discussed.

Chromosomes↗

Sources of wheat powdery mildew resistance from wheat-rye and wheat-Leymus hybrids.

Introgression of germplasm from rye and Leymus mollis has been used in the objective of broadening mildew resistance in wheat. The mildew resistant wheat lines were isolated from crosses of three hexaploid triticale lines, a mildew resistant double wheat-rye substitution line and a mildew resistant hexaploid wheat-L. mollis hybrid with three mildew susceptible breadwheats. Three different mildew isolates with a broad virulence have been used and lines with different resistance patterns, including resistance to all three mildew isolates, were selected. From a total of about 13,000 lines, eight lines from the triticale crosses, 58 from the double wheat-rye substitution line crosses and six from the L. mollis-hybrid crosses showed homozygous resistance to all three isolates. A differential tester set of wheat lines with known mildew resistance genes including earlier known resistance genes from rye showed a similarity between IRS-cultivars and the largest category in the triticale populations, indicating a higher frequency of IR chromosomes. The results are discussed in relation to the differential tester set and in relation to broadening the genetic base of resistance in wheat by interspecific hybridization.

Chimera↗

Phylogenetic relationships among Portuguese rye based on isozyme, RAPD and ISSR markers.

The phylogenetic relationships of 10 rye landraces and cultivars from the north of Portugal and from Brazil were analysed using 20 isozyme loci, and a total of 511 PCR markers (342 ISSRs and 169 RAPDs). The isozymes were analysed in at least 100 plants of each population/cultivar and, therefore, we have data about intra and inter population/cultivar genetic variability. However, the analyses with ISSRs and RAPDs were obtained using a mix of 25 plants of each population. Therefore, each population/cultivar was reduced to one tube and we have no data about intra genetic variability. As expected in a cross pollinated crop we found genetic diversity and a larger variation within than among the populations using isozymes. Somewhat unexpectedly, however, we found that the breeding cultivars have the same level of heterozygosity as the landraces. The phylogenetic relationships obtained using isozymes among the landraces, synthetic cultivar and the cultivars from breeding programs do not reflect their origin. Moreover, the cultivar from Brazil is not separated from the remaining populations/cultivars studied. However, the data observed using RAPDs and ISSRs are in agreement with their known origin. The populations maintained by the farmers in the north of Portugal are grouped in a cluster in the phenogram and the C902591 (from Brazil), the Alvão (synthetic variety) and Larouco (a hybrid between Montalegre and Brazil) are in a different cluster. The ISSRs and RAPDs provide a rapid method for the production of polymorphic markers, which appear to correspond to known pedigree information.

Alleles↗