Allergic Polysensitization Clusters: Newly Recognized Severity Marker in Urban Asthmatic Adults.

INTRODUCTION: While reliable, quantitative in vitro testing for sensitivity to aeroallergens has been available for decades, such information has largely been ignored in clustering analyses of asthma. Our aim is to explore allergic polysensitization as a possible marker of asthma severity and, as such, to be considered as an integral marker in future asthma clustering analyses. METHODS: We constructed a database of sensitizations to the 25 aeroallergens in our geographic area (zone 1, Northeastern US) using the ImmunoCAP® in vitro assay. We used the Scikit-Learn® machine learning library for model-based clustering to identify allergic polysensitization clusters. Clusters were compared for differences in common office-based clinical markers of asthma. RESULTS: The database consisted of 509 patients. Unbiased machine learning identified ten clusters of increasing allergic polysensitization of varying sizes (n = 1-339) characterized by significant increases in mean serum immunoglobulin E (p < 0.001), peripheral blood eosinophil count (p < 0.001), and DLCO (p = 0.02). There was a significant decline in mean age at presentation (p < 0.001), FEV1/FVC (p = 0.01), and FEF25-75 (p = 0.002) with increasing allergic polysensitization. Finally, we identified two divergent paths for the poly-atopic march, one driven by perennial and the other by seasonal allergens. CONCLUSION: This pilot study showed that allergic polysensitization, using readily available qualitative and quantitative in vitro sensitization data, largely ignored in cluster analyses to date, may add further clinical precision in cluster analyses of asthma. We suggest the methods used here can be applied and tested using larger databases and aeroallergens present in diverse geographic regions.

Festulolium and fungal endophyte associations: host status for Meloidogyne incognita and nematotoxic plant extracts.

Festulolium hybrids are forage grasses used worldwide in temperate climates. They are associated with the fungal endophyte Epichloë uncinata, which aids in nutrient uptake, drought tolerance, and production of metabolites that protect against parasites and herbivores. Epichloë uncinata produces loline alkaloids, which can deter insect pests. Festulolium has not been widely studied for susceptibility to plant-parasitic nematodes, so Festulolium lines, with and without fungal endophytes, were tested in the greenhouse for host status to the root-knot nematode Meloidogyne incognita. All were poor hosts, regardless of line or endophyte status. Pepper seedlings planted into soil following removal of the Festulolium plants were infected by nematodes, likely because of surviving nematodes from the original inoculation combined with some reproduction on Festulolium. Lolines were found in shoots and roots of all endophyte-associated lines, and some types of lolines in roots increased after nematode infection. Methanolic extracts from roots and shoots of a tested Festulolium line did not inhibit egg hatch, but killed nearly a third of second-stage juveniles whether an endophyte was present or not. Further studies would indicate whether these Festulolium lines aid in suppressing field populations of M. incognita.Festulolium hybrids are forage grasses used worldwide in temperate climates. They are associated with the fungal endophyte Epichloë uncinata, which aids in nutrient uptake, drought tolerance, and production of metabolites that protect against parasites and herbivores. Epichloë uncinata produces loline alkaloids, which can deter insect pests. Festulolium has not been widely studied for susceptibility to plant-parasitic nematodes, so Festulolium lines, with and without fungal endophytes, were tested in the greenhouse for host status to the root-knot nematode Meloidogyne incognita. All were poor hosts, regardless of line or endophyte status. Pepper seedlings planted into soil following removal of the Festulolium plants were infected by nematodes, likely because of surviving nematodes from the original inoculation combined with some reproduction on Festulolium. Lolines were found in shoots and roots of all endophyte-associated lines, and some types of lolines in roots increased after nematode infection. Methanolic extracts from roots and shoots of a tested Festulolium line did not inhibit egg hatch, but killed nearly a third of second-stage juveniles whether an endophyte was present or not. Further studies would indicate whether these Festulolium lines aid in suppressing field populations of M. incognita.

Epichloë uncinata Infection and Loline Content Protect Festulolium Grasses From Crickets (Orthoptera: Gryllidae).

Experiments with artificial diets demonstrated that black field cricket (Teleogryllus commodus (Walker)) and Lepidogryllus sp. were highly responsive to presence of lolines in their diet-quantities of diet consumed declined exponentially with increasing loline concentration. Amount consumed by black field cricket and Lepidogryllus sp. on diet containing 5,600 µg/g lolines was only 8 and 2% relative to those on loline-free diet, respectively. Additional experiments with Festulolium seeds demonstrated that both cricket species predated heavily on endophyte-free seed but largely avoided Epichloë uncinata-infected seed. By 12 h, black field cricket had destroyed 98.8% of endophyte-free but only 24.8% of E. uncinata-infected, loline-containing seed. By 36 h, Lepidogryllus sp. crickets had destroyed 40% of endophyte-free but had not fed on E. uncinata-infected, loline-containing seed. Glasshouse experiments demonstrated this aversion to lolines greatly reduces the damage potential of black field cricket in E. uncinata-infected Festulolium. When microswards were sown with E. uncinata-infected Festulolium, seedling numbers were reduced 25-26%, and yields 29-40%, by black field crickets relative to microswards sown without insect infestation. This contrasts with 70-78% reduction in seedling numbers and 67-80% reduction in yields in microswards sown to either endophyte-free Festulolium, endophyte-free perennial ryegrass, or Epichloë festucae var. lolii-infected Festulolium. Yields of mature E. uncinata-infected Festulolium plants were not adversely affected by black field crickets, irrespective of the presence of the endophyte-free standard Festulolium sown as a companion. In contrast, yields of endophyte-free Festulolium, endophyte-free perennial ryegrass, and E. festucae var. lolii-infected Festulolium plants were reduced by 56-61% by crickets.