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PubMed1994

Environment and development of respiratory allergy. II. Indoors.

Even if it is difficult to obtain correct epidemiological evidence, there is a body of evidence which suggests that the frequency of allergic respiratory diseases is increasing. The majority of atopic patients, in particular in childhood and adolescence, develop immunoglobulin E (IgE) antibodies with clinical symptoms to aeroallergens, such as those derived from house dust mites, pollens and pets. Since, in the economically-developed countries individuals spend most of their time indoors (home, school and workplace), indoor pollutants (tobacco smoke etc.) and allergens (house dust mite, cats, etc.) are the most important source of exposure. Indoor allergens are associated with a wide variety of particles in a broad size range, only some of which are microscopically identifiable, culturable, or detectable with existing immunoassay. Evaluation of indoor allergens requires both air and source sampling, and several different analytical techniques. It is likely that the increased prevalence of allergy and asthma may be caused in the indoor environment by the synergistic action of air pollution (in particular derived from tobacco smoking) with allergic sensitization. Passive smoking in infancy has also been involved in the airways allergic sensitization to common aeroallergens.

Air Pollution, Indoor↗
PubMed2004

Using genetic markers to estimate the pollen dispersal curve.

Pollen dispersal is a critical process that shapes genetic diversity in natural populations of plants. Estimating the pollen dispersal curve can provide insight into the evolutionary dynamics of populations and is essential background for making predictions about changes induced by perturbations. Specifically, we would like to know whether the dispersal curve is exponential, thin-tailed (decreasing faster than exponential), or fat-tailed (decreasing slower than the exponential). In the latter case, rare events of long-distance dispersal will be much more likely. Here we generalize the previously developed TWOGENER method, assuming that the pollen dispersal curve belongs to particular one- or two-parameter families of dispersal curves and estimating simultaneously the parameters of the dispersal curve and the effective density of reproducing individuals in the population. We tested this method on simulated data, using an exponential power distribution, under thin-tailed, exponential and fat-tailed conditions. We find that even if our estimates show some bias and large mean squared error (MSE), we are able to estimate correctly the general trend of the curve - thin-tailed or fat-tailed - and the effective density. Moreover, the mean distance of dispersal can be correctly estimated with low bias and MSE, even if another family of dispersal curve is used for the estimation. Finally, we consider three case studies based on forest tree species. We find that dispersal is fat-tailed in all cases, and that the effective density estimated by our model is below the measured density in two of the cases. This latter result may reflect the difficulty of estimating two parameters, or it may be a biological consequence of variance in reproductive success of males in the population. Both the simulated and empirical findings demonstrate the strong potential of TWOGENER for evaluating the shape of the dispersal curve and the effective density of the population (d(e)).

Computer Simulation↗
PubMed2000

Female reproductive tissues are the primary target of Agrobacterium-mediated transformation by the Arabidopsis floral-dip method.

The floral-dip method for Agrobacterium-mediated transformation of Arabidopsis allows efficient plant transformation without need for tissue culture. To facilitate use with other plant species, we investigated the mechanisms that underlie this method. In manual outcrossing experiments, application of Agrobacterium tumefaciens to pollen donor plants did not produce any transformed progeny, whereas application of Agrobacterium to pollen recipient plants yielded transformants at a rate of 0.48%. Agrobacterium strains with T-DNA carrying gusA (encoding beta-glucuronidase [GUS]) under the control of 35S, LAT52, or ACT11 promoters revealed delivery of GUS activity to developing ovules, whereas no GUS staining of pollen or pollen tubes was observed. Transformants derived from the same seed pod contained independent T-DNA integration events. In Arabidopsis flowers, the gynoecium develops as an open, vase-like structure that fuses to form closed locules roughly 3 d prior to anthesis. In correlation with this fact, we found that the timing of Agrobacterium infection was critical. Transformants were obtained and GUS staining of ovules and embryo sacs was observed only if the Agrobacterium were applied 5 d or more prior to anthesis. A 6-fold higher rate of transformation was obtained with a CRABS-CLAW mutant that maintains an open gynoecium. Our results suggest that ovules are the site of productive transformation in the floral-dip method, and further suggest that Agrobacterium must be delivered to the interior of the developing gynoecium prior to locule closure if efficient transformation is to be achieved.

Arabidopsis↗
PubMed1996

Proteinases from pollen and pests.

An examination of the proteinases present in two very different systems is described, in order to illustrate the diversity in function of this class of enzymes. In the first case we have noted the importance of gut proteinases from the fire ant Solenopsis invicta in relation to the nutritional requirements of the entire colony. In the second we have investigated the properties of endoproteases from both ragweed and mesquite pollen, relative to their role in the development of allergies and asthma. If the function of each type of enzyme(s) is correct, then it is clear that addition of exogenous inhibitors might be useful in a) controlling the infestation associated with the fire ant, and b) reducing the deleterious effects associated with the development of asthma.

Animals↗
PubMed1997

Expression of an Aspartate Kinase Homoserine Dehydrogenase Gene Is Subject to Specific Spatial and Temporal Regulation in Vegetative Tissues, Flowers, and Developing Seeds.

Although the regulation of amino acid synthesis has been studied extensively at the biochemical level, it is still not known how genes encoding amino acid biosynthesis enzymes are regulated during plant development. In the present report, we have used the [beta]-glucuronidase (GUS) reporter gene to study the regulation of expression of an Arabidopsis thaliana aspartate kinase-homoserine dehydrogenase (AK/HSD) gene in transgenic tobacco plants. The polypeptide encoded by the AK/HSD gene comprises two linked key enzymes in the biosynthesis of aspartate-family amino acids. AK/HSD-GUS gene expression was highly stimulated in apical and lateral meristems, lateral buds, young leaves, trichomes, vascular and cortical tissues of growing stems, tapetum and other tissues of anthers, pollen grains, various parts of the developing gynoecium, developing seeds, and, in some transgenic plants, also in stem and leaf epidermal trichomes. AK/HSD-GUS gene expression gradually dimished upon maturation of leaves, stems, floral tissues, and embryos. GUS expression was relatively low in roots. During seed development, expression of the AK/HSD gene in the embryo was coordinated with the initiation and onset of storage protein synthesis, whereas in the endosperm it was coordinated with the onset of seed desiccation. Upon germination, AK/HSD-GUS gene expression in the hypocotyl and the cotyledons was significantly affected by light. The expression pattern of the A. thaliana AK/HSD-GUS reporter gene positively correlated with the levels of aspartate-family amino acids and was also very similar to the expression pattern of the endogenous tobacco AK/HSD mRNA as determined by in situ hybridization.

Journal Article↗
PubMed1985

Exposure of hay fever subjects to an indoor environmental grass pollen challenge system.

A new indoor nasal allergen challenge system has been developed which allows subjects with hay fever to be exposed to pre-determined concentration of grass pollen grains in the immediate environment. Thirteen subjects were exposed during the winter months to an atmosphere containing approximately 1000 grains/m3 for 30 min. No changes in nasal airway patency or symptoms of nasal congestion were recorded. It was necessary to increase the pollen concentration to approximately 9000 and 35 000 grains/m3 in two subjects and six subjects respectively, before a positive response was recorded. This response was reproducible in all subjects. Administration of intra-nasal sodium cromoglycate before exposure significantly inhibited the decrease in nasal airway patency and when administered 1 min before exposure totally abolished symptoms of nasal congestion. This new challenge system may therefore be used as an in vivo screen to assess the potential therapeutic value of drugs in hay fever under controlled conditions.

Allergens↗
PubMed2002

Seasonal atopic dermatitis in dogs sensitive to a major allergen of Japanese cedar (Cryptomeria japonica) pollen.

Three dogs were examined because of episodes of recurrent pruritic dermatitis in the spring, the season of Japanese cedar (Cryptomeria japonica, CJ) pollination in Japan. The dogs were shown to be sensitive to CJ pollen allergen using intradermal testing and antigen-specific IgE measurement. Fluorometric enzyme-linked immunosorbant assay (ELISA) showed increased concentrations of IgE specific to Cry j 1 and a negative result for Cry j 2 in the three dogs. The concentrations of IgE specific to Cry j 1 during the season of CJ pollination were higher than the concentrations found during the off-season in all the dogs, and the variation in the concentrations correlated with the variation in clinical signs. Peripheral blood mononuclear cells showed apparent proliferative responses to crude CJ pollen antigen and Cry j 1 during CJ pollination season. These findings indicated that Cry j 1 was the major allergen recognized by IgE and lymphocytes and resulted in the development of type I hypersensitivity to CJ pollen allergen in these atopic dogs.

Allergens↗
PubMed1996

Generative cells of Lilium longiflorum possess translatable mRNA and functional protein synthesis machinery.

Metabolic labelling with [35S]-methionine demonstrated that generative cells of Lilium longiflorum possess their own set of mRNA and are capable of synthesising proteins independently from the vegetative cell. The isolated generative cells synthesised ten proteins, of which six were unique to these specialised cells. Isolation of generative cells from pollen grains after [35S]-methionine labelling resulted in an identical protein profile, therefore the synthesis of these proteins was not due to isolation shock. Addition of cycloheximide, abolished TCA-precipitable counts, whilst actinomycin D had no qualitative effect on the observed protein profile, indicating active translation of pre-existing mRNAs by the generative cells.

Cell Separation↗
PubMed1997

Biochemical and immunological characterization of recombinant allergen Lol p 1.

Pollen from perennial rye grass (Lolium perenne), a major cause of type-I allergy worldwide, contains a complex mixture of allergenic proteins among which Lol p 1 is one of the most important. We describe the expression, purification and characterization of a recombinant Lol p 1 overproduced in Escherichia coli. The recombinant allergen, expressed in high yields and purified in milligram amounts, bound to specific IgE antibodies from human sera, induced histamine release from sensitized human basophils, and elicited rabbit antisera that recognize specifically recombinant Lol p 1 and natural Lol p 1 of pollen extract. Recombinant Lol p 1 was used to develop ImmunoCAP assays for analysis of 150 sera that were Radioallergosorbent test positive to L. perenne pollen. In 130 of them (87%) the assay detected a significant level of IgE antibodies to Lol p 1, reaching on average 37% of the level obtained with a test for IgE to the whole grass pollen extract. To map epitopes on Lol p 1, we produced three deletion mutants [des-(116-240)-Lol p 1, des-(1-88)-Lol p 1 and des-(133-189)-Lol p 1], which were efficiently expressed in bacteria. These all showed a strong reactivity with the specific rabbit IgG antibodies, but lacked most or all the allergenic properties of recombinant Lol p 1. A study of the antigenic structure of Lol p 1 was performed using the three deletion mutants and a set of 17-18-residue overlapping synthetic peptides covering the whole allergen sequence. The results indicate that human IgE and rabbit IgG antibodies bind to distinct regions of Lol p 1, and that at least some important IgE epitopes are mainly conformational. The findings suggest that recombinant allergens constitute useful reagents for further development of serological diagnosis of allergy, and that it should be possible to produce immunogenic fragments of allergenic proteins without allergenic properties.

Allergens↗
PubMedDate not supplied

Anaphylaxis after laboratory rat bite: an occupational hazard.

Workers exposed to laboratory animals are at risk of developing asthma, rhinitis, angioedema, conjunctivitis, and urticaria. Approximately one in five scientists and technicians handling small animals will develop laboratory animal allergy symptoms within three years of employment, many of whom will have severe symptoms requiring a change of occupation. Individuals suffering from allergy to environmental allergens, such as pollen and ragweed, are more likely to develop allergic reactions to animals, and are more likely to develop asthma. We report a case of life-threatening anaphylaxis secondary to a rat bite in a laboratory research director with known allergies to rat urinary protein. While rodent bites are common in research settings, such severe reactions are extremely rare.

Accidents, Occupational↗
PubMed2003

Absence of microspore polarity, symmetric divisions and pollen cell fate in Brachiaria decumbens (Gramineae).

Meiotic division and male gametophyte development were analyzed in one tetraploid (2n = 4x = 36) accession of Brachiaria decumbens cv. Basilisk that showed some pollen sterility. Meiotic process was typical of polyploids in that it consisted of multiple chromosome associations. Precocious chromosome migration to the poles, laggards, and micronucleus formation were abundant in both meiosis I and II and resulted in tetrads with micronuclei. After callose dissolution, microspores were released into the anther locule and had the semblance of being normal. Although each microspore initiated its differentiation by pollen mitosis, in 43.24% of the microspores, nuclear polarization was not observed and the typical hemispherical cell plate was not detected. Division was symmetric and microspores lacked differentiation between the vegetative and the generative cell. Both nuclei were of equal size, presented equal chromatin condensation, and had a spherical shape. After the first pollen mitosis and cytokinesis, each cell underwent a new symmetric mitosis without nuclear polarization. At the end of the second pollen mitosis, four equal nuclei were observed in each pollen grain. After the second cytokinesis, the cells gave rise to four equal-sized pollen grains with a similar tetrad configuration that initially remained together. Sterile pollen grains resulted from abnormal pollen mitosis. This anomaly may be explained by a mutation, probably affecting microtubule cytoskeleton formation. The importance of this male-sterile mutation for Brachiaria breeding programs is discussed.

Brachiaria↗
PubMed1998

Genetic analysis of female gametophyte development and function.

The female gametophyte is an absolutely essential structure for angiosperm reproduction. It produces the egg cell and central cell (which give rise to the embryo and endosperm, respectively) and mediates several reproductive processes including pollen tube guidance, fertilization, the induction of seed development, and perhaps also maternal control of embryo development. Although much has been learned about these processes at the cytological level, specific molecules mediating and controlling megagametogenesis and female gametophyte function have not been identified. A genetic approach to the identification of such molecules has been initiated in Arabidopsis and maize. Although genetic analyses are still in their infancy, mutations affecting female gametophyte function and specific steps of megagametogenesis have already been identified. Large-scale genetic screens aimed at identifying mutants affecting every step of megagametogenesis and female gametophyte function are in progress; the characterization of genes identified in these screens should go a long way toward defining the molecules that are required for female gametophyte development and function.

Arabidopsis↗
PubMedDate not supplied

IgE and the allergy-asthma connection in the 23-year follow-up of Brown University students.

The Hygiene Hypothesis helps to explain the increased epidemiology of atopy, especially asthma and hay fever. This hypothesis depends on two major immunological pathways, the Th1 and the Th2 pathways, which are mutually inhibitory, with the Th2 pathway being the dominant one in fetal life and the newborn. The Th1 leads to a cellular delayed hypersensitive response while the Th2 pathway leads to increased IgE, eosinophilia, atopy, and airway/hyperresponsiveness. The ever-increasing vaccines for immunization against viral and bacterial microorganisms together with better public health hygiene procedures introduce a bias in favor of the inhibition of the Th1 pathway, thereby allowing the Th2 pathway, with its IgE hypersensitivity, to predominate. We have attempted to correlate this new hypothesis with data from our Brown University college student longitudinal study. In this study, our data have demonstrated that allergen sensitization (positive pollen skin tests reactions) leads to an increased risk factor for developing asthma. Most of our asthmatic patients in our longitudinal study had positive allergy skin tests. Also, students born in months with high concentrations of atmospheric ragweed pollen had an increased risk of developing sensitization to ragweed and later to develop hay fever, which may lead to asthma. There is a strong association of asthma with hay fever (a classic IgE disease). Also, hay fever patients have three times the risk for developing asthma than controls. There appear to be several factors needed to express the phenotype of allergic asthma: elevated IgE, eosinophilia, airway hyperresponsiveness, exposure to allergens, and the predominance of the Th2 pathway of immunologic reactions.

Allergens↗
PubMed2002

The ROP2 GTPase controls the formation of cortical fine F-actin and the early phase of directional cell expansion during Arabidopsis organogenesis.

Polar cell expansion in differentiating tissues is critical for the development and morphogenesis of plant organs and is modulated by hormonal and developmental signals, yet little is known about signaling in this fundamental process in plants. In contrast to tip-growing cells, such as pollen tubes and root hairs, cells in developing tissues are thought to expand by diffuse growth. In this study, we provide evidence that these cells expand in two phases with distinct mechanisms. In the early phase, cell expansion can occur in both longitudinal and radial or lateral directions and is mediated by Rop GTPase signaling, a mechanism known to control tip growth. The expression of a dominant-negative mutant for ROP2 (DN-rop2) inhibited polar cell expansion, whereas the expression of a constitutively active mutant (CA-rop2) caused isotropic expansion in the early phase. In the late phase, expansion occurs only in the longitudinal direction and is not affected by DN-rop2 or CA-rop2 expression. The transition from the early to the late phase coincides with the reorientation of cortical microtubules from random to transverse arrangements. Thus, cell expansion in the late phase is consistent with polar diffuse growth, in which polarity probably is defined by transverse cortical microtubules. We show that the direction of cell expansion in the early phase is associated with the localization of diffuse fine cortical F-actin in leaf epidermal cells. DN-rop2 expression specifically inhibited the formation of this F-actin, but not actin cables, whereas CA-rop2 expression caused delocalized distribution of this fine F-actin throughout the cell cortex. Furthermore, green fluorescent protein-ROP2 was localized preferentially to the cortical region of the cell, where expansion apparently occurs. These observations suggest that ROP2 control of the polar expansion of cells within tissues is analogous to the Rop control of tip growth and of tip-localized F-actin in pollen tubes and root hairs and that the tip growth mechanism also may modulate polar cell expansion in differentiating tissues.

Actins↗
PubMed2003

Injection immunotherapy with different airborne allergens did not prevent de novo sensitization to ragweed and birch pollen north of Milan.

BACKGROUND: Previous studies suggest that allergen-specific immunotherapy (SIT) with a single allergen may prevent sensitization to other airborne allergens. OBJECTIVE: To assess the effectiveness of injection SIT in preventing sensitization to birch and ragweed pollen north of Milan, a geographical area that was recently colonized by these two airborne allergens. METHODS: 691 adult patients monosensitized to grass (n = 342), pellitory (n = 76), ragweed (n = 66), birch (n = 112), and house dust mite (n = 95) were studied. 284 (41%) of them received injection SIT as part of routine outpatient care; the remaining 407 (59%) not submitted to SIT, served as controls. All patients underwent follow-up visits after no less than 2 years in order to detect possible new sensitization to birch and/or ragweed. RESULTS: 227 (33%) patients became sensitized to birch and/or ragweed pollen during the follow-up period. Surprisingly, the prevalence of new sensitizations was significantly higher among subjects receiving SIT (132/284; 46%) than among those not receiving SIT (95/407; 23%; p < 0.001). 27 subjects developed new sensitization to birch and/or ragweed pollen while undergoing SIT. These findings were consistent in all subgroups with different airborne allergies. CONCLUSION: In this study, injection SIT did not exert any preventive effect against denovosensitization to the two novel airborne allergens, birch and ragweed pollen, in adult monosensitized patients.

Adult↗