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Characterization of stroma from Fuchs' endothelial dystrophy corneas.

Fuchs' endothelial dystrophy is commonly regarded as an endothelial cell disorder. In the present study we compared glycoconjugates of Fuchs' and normal corneas using FITC conjugated lectins [peanut agglutinin (PNA), castor bean agglutinin (RCA120), soybean agglutinin (SBA), and wheat germ agglutinin (WGA)]. Our results showed increased staining with RCA120 and PNA in the posterior region of the Fuchs' corneas, indicating an accumulation of terminal beta-galactose and B-D-galactose (1-3)-D-N-acetylgalactosamine residues. The stromal and epithelial regions of normal and Fuchs' corneas exhibited similar staining patterns with all lectins tested. Our collagen studies showed an increased extractability and abnormal amino acid analyses of collagen from Fuchs' corneas as compared with normals. The purified collagens did have similar banding patterns by sodium dodecyl sulfate gels. However, further characterization by 125(1) two-dimensional peptide mapping revealed that Fuchs' alpha 1-sized chains contained fingerprints that were distinctly different from normal cornea stromal collagen. These data suggest that in addition to abnormal accumulation of RCA120- and PNA-specific glycoconjugates in the posterior cornea, Fuchs' corneas contained stromal collagens with altered biochemical properties. We postulate that the characteristic deterioration of endothelial function in Fuchs' dystrophy may compromise the microenvironment of the stroma and its keratocytes, and thereby lead to an altered collagenous extracellular matrix.

Amino Acids↗

Loss of phosphine from unsealed bins of wheat at six combinations of grain temperature and grain moisture content.

Hard red winter wheat (1.4 t) at 11.1 or 13.5% moisture content (wet basis) and 20, 25, or 30 degrees C was fumigated with tablets of an aluminum phosphide formulation in unsealed, cylindrical grain bins of corrugated metal. The fumigant leakage rate was manipulated to approximate that commonly encountered in farm and commercial-scale bins of this type. Phosphine concentration profiles were recorded and phosphine loss and sorption were characterized to determine which conditions provided the greatest probability of successful fumigation in these bins. Phosphine leakage and sorption were both positively related to grain temperature and moisture content. The fumigant concentration profiles were compared with previously-published data relating temperature to the developmental rate and fumigant susceptibility of lesser grain borer eggs, which are phosphine-resistant but become less resistant as they age. The mean phosphine concentration observed at the time corresponding to one-half of the calculated egg development time was compared to the lethal concentration (LC(99)) for a 2-day exposure at each temperature-moisture combination. In the low-moisture grain at 20 degrees C, the observed fumigant concentration was below the lethal concentration, due to the long development time under these conditions. At 25 and 30 degrees C in the low-moisture wheat, the likelihood of complete kill appeared more favorable because the fumigant concentration remained above the published LC(99) for more than half of the egg development time. In the wheat with 13.5% moisture content, rapid fumigant sorption and loss resulted in phosphine concentrations below the LC(99) at one-half of the development time at 20 or 25 degrees C. At 30 degrees C, due to the very rapid development rate, the observed phosphine concentration exceeded the LC(99) half-way through the egg development period despite the rapid rate of fumigant sorption and loss. Repeated fumigation of the same grain reduced the rate at which phosphine sorbed into the grain.

Journal Article↗

Assessing potential exposure of birds to pesticide-treated seeds.

Seed treatments are widely used for crop protection and present a particular risk to granivorous birds. UK risk assessment for seed treatments has tended to focus on highly granivorous species; however, under some conditions, non-granivorous birds will take seeds. Better data is needed on which species eat seeds for which pesticide treatments are used. To identify which species will take and eat a range of crop seeds in common usage in the UK, birds visiting bait stations at which untreated seed was presented were video recorded. Information was also obtained on how much seed is taken by individual birds. The seeds tested were wheat, barley, maize, oilseed rape, grass, peas and pelleted sugar beet. For many of the species observed at the bait stations, the amounts of seed consumed during single visits were sufficient to pose a potential risk (toxicity-exposure ratio < 10) if the seed had been treated with one of the more acutely toxic seed treatments. Previous studies have shown that de-husking of seeds can substantially reduce birds' exposure. This paper provides information on which of the species recorded de-husked which seeds, in field conditions. The use of these data in pesticide risk assessment is considered.

Animals↗

Assessment of differences in ascomycete communities in the rhizosphere of field-grown wheat and potato.

To assess effects of plant crop species on rhizosphere ascomycete communities in the field, we compared a wheat monoculture and an alternating crop rotation of wheat and potato. Rhizosphere soil samples were taken at different time points during the growing season in four consecutive years (1999-2002). An ascomycete-specific primer pair (ITS5-ITS4A) was used to amplify internal transcribed spacer (ITS) sequences from total DNA extracts from rhizosphere soil. Amplified DNA was analyzed by denaturing gradient gel electrophoresis (DGGE). Individual bands from DGGE gels were sequenced and compared with known sequences from public databases. DGGE gels representing the ascomycete communities of the continuous wheat and the rotation site were compared and related to ascomycetes identified from the field. The effect of crop rotation exceeded that of the spatial heterogeneity in the field, which was evident after the first year. Significant differences between the ascomycete communities from the rhizospheres of wheat in monoculture and one year after a potato crop were found, indicating a long-term effect of potato. Sequencing of bands excised from the DGGE gels revealed the presence of ascomycetes that are common in agricultural soils.

Agriculture↗

Oats can diversify a gluten-free diet in celiac disease and dermatitis herpetiformis.

Finnish celiac disease and dermatitis herpetiformis patients have used oat-containing gluten-free diets since 1997. The aim of this study was to evaluate how the use of oats has been adopted. The use of oats and the effect of oats on symptoms and quality of life were investigated in 1,000 randomly selected members of the Celiac Society. Altogether, 710 patients responded: 423 (73%) with celiac disease and 70 (55%) with dermatitis herpetiformis were currently consuming oats. Patients appreciated the taste, the ease of use, and the low costs; 94% believed that oats diversified the gluten-free diet; 15% of celiac disease and 28% of dermatitis herpetiformis patients had stopped eating oats. The most common reasons for avoiding oats were fear of adverse effects or contamination. There is a market demand for oats, and celiac societies and dietitians should make efforts to promote the development of products free of wheat contamination.

Adolescent↗

Plant host associations of Penthaleus species and Halotydeus destructor (Acari: Penthaleidae) and implications for integrated pest management.

Integrated pest management programs seek to minimise reliance on pesticides and provide effective long-term control of pests. Cultural control strategies, such as crop rotations, trap and border crops, and weed management, require a thorough understanding of pest host associations. This paper examines the effects of different plant hosts on the persistence and reproduction of blue oat mites, Penthaleus spp., and the redlegged earth mite, Halotydeus destructor (Tucker), which are major agricultural pests in southern Australia. Field and shade-house experiments were conducted testing several crop and plant types. All species survived and reproduced from one mite season to the next when confined to pasture. Canola and a common weed, 'bristly ox-tongue', were suitable hosts for H. destructor and Penthaleus falcatus (Qin and Halliday), whereas Penthaleus sp. x and Penthaleus major (Dugés) failed to persist on these plants. A mixture of wheat and oats sustained P. sp. x and H. destructor, but not P. falcatus or P. major. Lentils were generally a poor host plant for all mite species. These findings show that earth mite species differ in their ability to persist on different plant types, highlighting the importance of distinguishing mite species before implementing control strategies. Results are discussed with respect to cultural control options for the management of these winter pests.

Animals↗

Selectable markers: antibiotic and herbicide resistance.

The low efficiencies of most plant transformation methods necessitate the use of selectable marker genes to identify those cells that successfully integrate and express transferred DNA. Genes conferring resistance to various antibiotics or herbicides are commonly used in laboratory transformation research. They encode proteins that detoxify corresponding selection agents and allow the preferential growth of transformed cells. This chapter describes the application of two selection systems on the transformation of wheat. One is based on the nptII gene and corresponding aminoglycoside antibiotics, the other is based on the bar gene and corresponding glufosinate ammonium herbicides.

Base Sequence↗

Urban-rural comparison of nutrient intake by adult women in Shaanxi Province, China.

Triplet surveys were conducted in the city of Xi' an and two villages (one in the vicinity and the other at a distance) in Shaanxi Province in China in October-November (when agricultural activities were low), 1997, to elucidate nutrient intakes with a focus on possible urban-rural differences. Total food duplicate samples were collected from non-smoking and non-habitually drinking adult healthy women (about 50 subjects per site and 149 in total). The nutrient intakes were estimated from the weight of food items in reference to national food composition tables. On average, the women took 1873 kcal energy, 54 g protein and 37 g lipid per day, with a lipid energy ratio of 18.4%. Both excess and insufficient energy intake was observed as a result of food intake analysis and body mass index determination. With regard to minor nutrient intakes, insufficiency was serious in the case of calcium, vitamin A and vitamin B2, but not with iron. Whereas dependency on plant foods for sources of energy and protein was common to the three regions, Xi' an people consumed more animal foods than those in the villages. Intake of fish and shellfish was quite low throughout the three regions. Among the four types of cereals, wheat was consumed most substantially in the three regions and in three meals (except for the village where people essentially did not take lunch in reflection of low agricultural activities), whereas rice was consumed more in Xi' an than in the two villages. Maize consumption was higher in the two villages (especially for breakfast) than in the city. In contrast, foxtail millet (although in small amounts) was taken primarily in Xi' an and only at the time of breakfast.

Adult↗

Occupational asthma in a highly industrialized region of UK: report from a local surveillance scheme.

Estimates of the incidence of occupational asthma may be derived from surveillance schemes established in several countries. SHIELD is a voluntary surveillance scheme for occupational asthma in the West Midlands, a highly industrialized region of UK. The aim of this study was to estimate the general and specific incidence of occupational asthma in the West Midlands in 1990-97. The annual incidence was 41.2/million. There was a two fold difference in the incidence by sex (male 59.6/million/yr; female 27.4/million/yr). The highest annual incidence (53.2/million) was observed in the age group 45-64 yr (male) and 45-59 yr (female). Spray painters were the occupation at the highest risk of developing occupational asthma, followed by electroplaters, rubber and plastic workers, bakery workers and moulders. Although the percentage of reported cases was low among healthcare workers, there was a raising trend. Isocyanates still remained the most common causative agents with 190 (17.3%) out of the total 1097 cases reported to the surveillance scheme in seven years. There was a decrease in the reported cases due to colophony (9.5% to 4.6%), flour & wheat (8.9% to 4.9%). There was an increase of reported cases due to latex (0.4% to 4.9%) and glutaraldehyde (1.3% to 5.6%). The serial mesurement of peak expiratory flow at and away from work was the most used method of diagnosis to confirm the occupational cause of asthma. Specific bronchial challenge test with the occupational agents were used when the serial measurement of peak expiratory flow was not able to confirm undoubtdely the diagnostic suspicion or when it was difficult to identify the possible causative agent due to multiple exposures in the workplace. Following diagnosis, 24% of the patients were moved away from exposure within the same workplace in 1997, compared to 15.8% in the previous years. Those remaining exposed to the causative agent in the same workplace decreased from 28.3% to 17.7% between 1990-97. The surveillance of occupational asthma trough this voluntary scheme has allowed to monitor the incidence of the disease in the region and to identify clusters of cases, where control measures are a priority.

Adolescent↗

[Physico-chemical properties related to gastrointestinal function of 6 sources of dietary fiber].

The physicochemical properties of the six sources of dietary fiber more commonly consumed in Mexico were studied. The knowledge of these properties may allow a prediction of the functional effect of the fibers on the GI tract. The content of dietary fiber varied from 43% in wheat bran (WB) to 87% in Psyllium plantago (PP). Glucomannan (GM) showed the highest proportion of soluble fiber (97%) followed by dried cactus from Opuntia sp (DC) (28%) and PP (13%), whereas WB and soy isolate (SI) showed the highest proportion of insoluble fiber (95 and 92%, respectively). PP and GM showed a higher water holding capacity and viscosity and a lower ion exchange capacity: we anticipate that these fibers would be more effective to regulate plasma levels of glucose and cholesterol in contrast, WB and SI with a lower water holding capacity and viscosity and a higher proportion of insoluble fiber would be more effective to increase fecal bulk. Fibers from DC and from cactus fiber isolate were intermediate in these responses.

Absorption↗

Spinal organization and steroid sensitivity of motoneurons innervating the pubococcygeus muscle in the male rat.

Male rat motoneurons innervating the pubococcygeus muscle were located in the ventral nucleus of lamina IX at the sixth lumbar (L6) and first sacral (S1) spinal cord segments. Retrograde labeling with horseradish peroxidase-wheat germ agglutinin was transported up to second-order dendrites and revealed that these motoneurons have a "U-shaped arborization" of dendrites toward the intermediolateral and intermediomedial nuclei area of lamina VII. This dendritic organization makes a wide "final common path" that probably integrates afferent information from several sources, accounting for the participation of the pubococcygeus muscle in autonomic and somatic processes, such as those related to micturition and reproduction. Castration produced a decrement in the morphometry of these motoneurons. A main effect was a decrement in dendritic length. Steroid replacement indicated that testosterone and estradiol, but not dihydrotestosterone, are able to induce a recovery of morphometric alterations. However, estrogen induced recovery after 2 weeks of treatment, whereas testosterone took 4 weeks. Thus, it is proposed that supraspinal aromatization of testosterone in the male central nervous system might be an important process for the appropriate organization of the pubococcygeus muscle motoneurons and that estradiol seems to need a shorter time of action than testosterone because of differential up-regulation and down-regulation of steroid receptors.

Animals↗

Distribution of lutein, zeaxanthin, and related geometrical isomers in fruit, vegetables, wheat, and pasta products.

Quantitative data with regard to dietary (3R,3'R,6'R)-lutein, (3R,3'R)-zeaxanthin, and their (E/Z)-geometrical isomers are scarce, and in most cases, only the combined concentrations of these two carotenoids in foods are reported. Lutein and zeaxanthin accumulate in the human macula and have been implicated in the prevention of age-related macular degeneration (AMD). The qualitative and quantitative distributions of lutein, zeaxanthin, and their (E/Z)-isomers in the extracts from some of the most commonly consumed fruits, vegetables, and pasta products were determined by HPLC employing a silica-based nitrile-bonded column. Green vegetables had the highest concentration of lutein (L) and zeaxanthin (Z), and the ratios of these carotenoids (L/Z) were in the range 12-63. The yellow-orange fruits and vegetables, with the exception of squash (butternut variety), had much lower levels of lutein in comparison to greens but contained a higher concentration of zeaxanthin. The ratio of lutein to zeaxanthin (L/Z) in two North American bread varieties of wheat (Pioneer, Catoctin) was 11 and 7.6, respectively, while in a green-harvested wheat (Freekeh) imported from Australia, the ratio was 2.5. Between the two pasta products examined, lasagne and egg noodles, the latter had a much higher concentration of lutein and zeaxanthin. The levels of the (E/Z)-geometrical isomers of lutein and zeaxanthin in these foods were also determined.

Chromatography, High Pressure Liquid↗

Silver enhancement of lectin-gold and enzyme-gold cytochemical labelling of eggs of the nematode Onchocerca gibsoni.

Wheat germ agglutinin-gold and chitinase-gold complexes were used to demonstrate the presence of chitin on the surfaces of eggs of the animal parasitic nematode Onchocerca gibsoni. The gold complexes were enhanced by silver intensification and examined by light microscopy (LM), transmission electron microscopy (TEM) and scanning electron microscopy (SEM). Distinctive labelling of the egg surfaces was obtained with both probes in all three microscope modes. The results indicate that the small colloidal gold markers (3-10 nm) commonly used for high resolution TEM studies may be silver enhanced and also used for sensitive LM and SEM studies.

Animals↗

Selective inhibition of cyclic AMP-dependent protein kinase by amphiphilic triterpenoids and related compounds.

A set of plant- and animal-derived amphiphilic triterpenoids have been shown to be potent and selective inhibitors of the catalytic subunit of rat liver cyclic AMP-dependent protein kinase (cAK). Thus plant-derived 18 alpha- and 18 beta-glycyrrhetinic acid, ursolic acid, oleanolic acid and betulin and animal-derived lithocholic acid, 5-cholenic acid and lithocholic acid methyl ester are inhibitors of cAK with IC50 values (concentrations for 50% inhibition) in the range 4-20 microM. These compounds are ineffective or relatively ineffective as inhibitors of various other eukaryote signal-regulated protein kinases namely wheat embryo Ca(2+)-dependent protein kinase (CDPK), avian calmodulin-dependent myosin light chain kinase (MLCK) and rat brain Ca(2+)- and phospholipid-dependent protein kinase C (PKC). These naturally occurring triterpenoids have a common structural motif involving polar residues located at opposite ends of an otherwise non-polar triterpenoid nucleus. A variety of triterpenoids not possessing this structural motif are relatively inactive as inhibitors of cAK and of CDPK, PKC and MLCK. The terpenoid amphiphilic compound crocetin is also a potent and relatively selective inhibitor of cAK (IC50 value for cAK 3.0 microM). 12-Hydroxystearic acid and 10-hydroxydecanoic acid do not inhibit CDPK, PKC or MLCK but are selective inhibitors of cAK (IC50 values 127 and 138 microM, respectively), consistent with a simple model for amphiphile inhibition of cAK involving two polar groups separated by a non-polar region. However, laurylgallate and 15-pentadecanolide are also potent and selective inhibitors of cAK (IC50 values 1.5 and 20 microM, respectively) although the structures of both of these compounds involve a large non-polar portion associated with only one polar region. Crocetin and the plant-derived amphiphilic triterpenoids described here are the most potent non-aromatic plant-derived inhibitors of cAK yet found.

Amino Acid Sequence↗

1H NMR and fluorescence studies of the complexation of DMPG by wheat non-specific lipid transfer protein. Global fold of the complex.

Plant non-specific lipid transfer proteins (LTPs) are proteins which transfer lipids between membranes in vitro and are believed to be involved in the transport of cutin monomers to the cuticle layer in vivo or in the plant defence against phytopathogens. The complexation of DMPG, a diacyl phospholipid, by wheat ns-LTP, a protein extracted from wheat seeds, was followed by 1H NMR and fluorescence spectroscopy. The global fold of the protein was calculated using the DIANA software package from a list of 968 distance constraints. The internal cavity volume, a feature common to all known ns-LTP structures, was estimated to be 750 A3 using the 'CAVITE' program. This model of the complex was obtained by inserting a lipid molecule in the cavity and was energy minimized. The study showed that the protein fold described for the free form was only weakly affected by the insertion of the bulky lipid. Observation of some intermolecular NOEs between the protein and the lipid glycerol moiety revealed that the cavity entrance was located between residues His35 and Arg44. The resulting solution structure was compared to the crystal structure of the maize ns-LTP/palmitate complex.

Arginine↗

Uptake and metabolic fate of [14C]-2,4-dichlorophenol and [14C]-2,4-dichloroaniline in wheat (Triticum aestivum) and soybean (Glycine max).

The uptake and metabolism of [14C]-2,4-dichlorophenol (DCP) and [14C]-2,4-dichloroaniline (DCA) were investigated in wheat and soybean. Seeds were exposed to a nutrient solution containing 50 microM of one of two radiolabeled compounds, and plant organs were harvested separately after 18 days of growth. In wheat, uptake of [14C]-2,4-DCP was 16.67 +/- 2.65 and 15.50 +/- 2.60% of [14C]-2,4-DCA. In soybean, uptake of [14C]-2,4-DCP was significantly higher than [14C]-2,4-DCA uptake, 38.39 +/- 2.56 and 18.98 +/- 1.64%, respectively. In the case of [14C]-2,4-DCP, the radioactivity absorbed by both species was found mainly associated with roots, whereas [14C]-2,4-DCA and related metabolites were associated with aerial parts, especially in soybean. In wheat, nonextractable residues represented 7.8 and 8.7% of the applied radioactivity in the case of [14C]-2,4-DCP and [14C]-2,4-DCA, respectively. In soybean, nonextractable residues amounted to 11.8 and 5.8% of the total radioactivity for [14C]-2,4-DCP and [14C]-2,4-DCA, respectively. In wheat, nonextractable residues were nearly equivalent to extractable residues for [14C]-2,4-DCP, whereas they were greater for [14C]-2,4-DCA. In soybean, the amount of extractable residues was significantly greater for both chemicals. However, in both species, nonextractable residues were mainly associated with roots. Isolation of soluble residues was next undertaken using excised shoots (wheat) or excised fully expanded leaves including petioles (soybean). Identification of metabolite structures was made by comparison with authentic standards, by enzymatic hydrolyses, and by electrospray ionization-mass spectrometric analyses. Both plant species shared a common metabolism for [14C]-2,4-DCP and [14C]-2,4-DCA since the malonylated glucoside conjugates were found as the final major metabolites.

Aniline Compounds↗

Bovine retinal rod guanyl cyclase represents a new N-glycosylated subtype of membrane-bound guanyl cyclases.

The molecular properties of retinal rod guanyl cyclase were investigated. Peptides were derived from a 112-kDa protein previously identified as the particulate bovine retinal rod guanyl cyclase. The peptides showed 100% identity to the deduced amino acid sequence of the cloned human retina-specific membrane guanyl cyclase, whereas identity to the members of the natriuretic peptide receptor guanyl cyclases was 14-59%. The 112-kDa protein was further purified by a new approach using wheat-germ agglutinin chromatography. This indicated N-linked glycosylation in retinal rod guanyl cyclase. N-glycosylation was unexpected from the sequence of the human retina-specific membrane guanyl cyclase, although it is a common property of natriuretic peptide receptors. Therefore, we further analyzed the carbohydrate composition of bovine retinal rod guanyl cyclase by lectin binding using the lectins Galanthus nivalis agglutinin, Sambucus nigra agglutinin, Maackia amurensis agglutinin, Ricinus communis agglutinin, Datura stramonium agglutinin, peanut agglutinin and by chromatography of the purified enzyme using concanavalin-A-Sepharose. The oligosaccharide side chains were of the high-mannose type or hybrid type, probably with mannose, N-acetylglucosamine and sialic acid as terminal sugars. Enzymic deglycosylation by N-glycosidase F was achieved after proteolytic digestion with endoproteinase Glu-C. Lectins neither influenced the basal nor the stimulated guanyl-cyclase activity at low calcium concentrations. Our results indicate that the particulate rod guanyl cyclase represents an unusual new subtype of membrane-bound guanyl cyclases.

Amino Acid Sequence↗

Aero-allergens in canine atopic dermatitis in southeastern Australia based on 1000 intradermal skin tests.

OBJECTIVE: To determine the most relevant aero-allergens involved in canine atopic dermatitis in southeastern Australia and provide information about these aero-allergens to the general practitioner. PROCEDURE: Dogs presented to the Animal Skin & Allergy Clinic with history and clinical signs of atopic dermatitis were injected intradermally with 38 different allergens and negative and positive control. Intradermal skin tests in 1000 dogs were retrospectively evaluated. RESULTS: One third of all patients reacted to the house dust mite Dermatophagoides farinae. Allergens reacting in more than 15% of the patients were wheat (Triticum aestivum), sweet vernal (Anthoxanthum odoratum), English couch (Agropyron repens), yellow dock (Rumex crispus), Mexican tea (Chenopodium ambrosioides), plantain (Plantago lanceolata), melaleuca (Melaleuca quinquenervia) and peppercorn (Schimus spp). CONCLUSION: House dust mites are the most common allergens in canine atopic dermatitis in southeastern Australia and D farinae is involved most frequently. However, a number of grass, weed and tree pollens also are involved regularly.

Aerosols↗