Thunderstorms, Pollen, and Severe Asthma in a Midwestern, USA, Urban Environment, 2007-2018.

BACKGROUND: Previous research has shown an association between individual thunderstorm events in the presence of high pollen, commonly called thunderstorm asthma, and acute severe asthma events, but little work has studied risk over long periods of time, using detailed measurements of storms and pollen. METHODS: We estimated change in the risk of asthma-related emergency room visits related to thunderstorm asthma events in the Minneapolis-St. Paul metropolitan area over the years 2007-2018. We defined thunderstorm asthma events as daily occurrence of two or more lightning strikes during high pollen periods interpolating weather and pollen monitor data and modeling lightning counts. We acquired daily counts of asthma-related emergency department visits from the Minnesota Hospital Association and used a quasi-Poisson time-series regression to estimate overall relative risk of emergency department visits during thunderstorm asthma events. RESULTS: We observed a 1.047 times higher risk (95% confidence interval = 1.012, 1.083) of asthma-related emergency department visits on the day of thunderstorm asthma event. Our findings are robust to adjustment for temperature, humidity, wind, precipitation, ozone, PM 2.5 , day of week, and seasonal variation in asthma cases. Occurrence of lightning alone or pollen alone showed no association with the risk of severe asthma. A two-stage analysis combining individual zip code-level results shows similar RR, and we see no evidence of spatial correlation or spatial heterogeneity of effect. DISCUSSION: Our results support an association between co-occurrence of lightning and pollen and risk of severe asthma events. Our approach incorporates lightning and pollen data and small-spatial area exposure and outcome counts.

Spatiotemporal control of axillary meristem formation by interacting transcriptional regulators.

Branching is a common feature of plant development. In seed plants, axillary meristems (AMs) initiate in leaf axils to enable lateral shoot branching. AM initiation requires a high level of expression of the meristem marker SHOOT MERISTEMLESS (STM) in the leaf axil. Here, we show that modules of interacting transcriptional regulators control STM expression and AM initiation. Two redundant AP2-type transcription factors, DORNRÖSCHEN (DRN) and DORNRÖSCHEN-LIKE (DRNL), control AM initiation by regulating STM expression. DRN and DRNL directly upregulate STM expression in leaf axil meristematic cells, as does another transcription factor, REVOLUTA (REV). The activation of STM expression by DRN/DRNL depends on REV, and vice versa. DRN/DRNL and REV have overlapping expression patterns and protein interactions in the leaf axil, which are required for the upregulation of STM expression. Furthermore, LITTLE ZIPPER3, another REV-interacting protein, is expressed in the leaf axil and interferes with the DRN/DRNL-REV interaction to negatively modulate STM expression. Our results support a model in which interacting transcriptional regulators fine-tune the expression of STM to precisely regulate AM initiation. Thus, shoot branching recruits the same conserved protein complexes used in embryogenesis and leaf polarity patterning.

Clonal sector analysis and cell ablation confirm a function for DORNROESCHEN-LIKE in founder cells and the vasculature in Arabidopsis.

MAIN CONCLUSION: Inducible lineage analysis and cell ablation via conditional toxin expression in cells expressing the DORNRÖSCHEN-LIKE transcription factor represent an effective and complementary adjunct to conventional methods of functional gene analysis. Classical methods of functional gene analysis via mutational and expression studies possess inherent limitations, and therefore, the function of a large proportion of transcription factors remains unknown. We have employed two complementary, indirect methods to obtain functional information for the AP2/ERF transcription factor DORNRÖSCHEN-LIKE (DRNL), which is dynamically expressed in flowers and marks lateral organ founder cells. An inducible, two-component Cre-Lox system was used to express beta-glucuronidase GUS in cells expressing DRNL, to perform a sector analysis that reveals lineages of cells that transiently expressed DRNL throughout plant development. In a complementary approach, an inducible system was used to ablate cells expressing DRNL using diphtheria toxin A chain, to visualise the phenotypic consequences. These complementary analyses demonstrate that DRNL functionally marks founder cells of leaves and floral organs. Clonal sectors also included the vasculature of the leaves and petals, implicating a previously unidentified role for DRNL in provasculature development, which was confirmed in cotyledons by closer analysis of drnl mutants. Our findings demonstrate that inducible gene-specific lineage analysis and cell ablation via conditional toxin expression represent an effective and informative adjunct to conventional methods of functional gene analysis.

Gene regulatory networks controlled by FLOWERING LOCUS C that confer variation in seasonal flowering and life history.

Responses to environmental cues synchronize reproduction of higher plants to the changing seasons. The genetic basis of these responses has been intensively studied in the Brassicaceae. The MADS-domain transcription factor FLOWERING LOCUS C (FLC) plays a central role in the regulatory network that controls flowering of Arabidopsis thaliana in response to seasonal cues. FLC blocks flowering until its transcription is stably repressed by extended exposure to low temperatures in autumn or winter and, therefore, FLC activity is assumed to limit flowering to spring. Recent reviews describe the complex epigenetic mechanisms responsible for FLC repression in cold. We focus on the gene regulatory networks controlled by FLC and how they influence floral transition. Genome-wide approaches determined the in vivo target genes of FLC and identified those whose transcription changes during vernalization or in flc mutants. We describe how studying FLC targets such as FLOWERING LOCUS T, SQUAMOSA PROMOTER BINDING PROTEIN-LIKE 15, and TARGET OF FLC AND SVP 1 can explain different flowering behaviours in response to vernalization and other environmental cues, and help define mechanisms by which FLC represses gene transcription. Elucidating the gene regulatory networks controlled by FLC provides access to the developmental and physiological mechanisms that regulate floral transition.

Functional dissection of the DORNRÖSCHEN-LIKE enhancer 2 during embryonic and phyllotactic patterning.

MAIN CONCLUSION: The Arabidopsis DORNRÖSCHEN-LIKE enhancer 2 comprises a high-occupancy target region in the IM periphery that integrates signals for the spiral phyllotactic pattern and cruciferous arrangement of sepals. Transcription of the DORNRÖSCHEN-LIKE (DRNL) gene marks lateral organ founder cells (LOFCs) in the peripheral zone of the inflorescence meristem (IM) and enhancer 2 (En2) in the DRNL promoter upstream region essentially contributes to this phyllotactic transcription pattern. Further analysis focused on the phylogenetically highly conserved 100-bp En2core element, which was sufficient to promote the phyllotactic pattern, but was recalcitrant to further shortening. Here, we show that En2core functions independent of orientation and create a series of mutations to study consequences on the transcription pattern. Their analysis shows that, first, in addition to in the inflorescence apex, En2core acts in the embryo; second, cis-regulatory target sequences are distributed throughout the 100-bp element, although substantial differences exist in their function between embryo and IM. Third, putative core auxin response elements (AuxREs) spatially activate or restrict DRNL expression, and fourth, according to chromatin configuration data, En2core enhancer activity in LOFCs correlates with an open chromatin structure at the DRNL transcription start. In combination, mutational and chromatin analyses imply that En2core comprises a high-occupancy target (HOT) region for transcription factors, which implements phyllotactic information for the spiral LOFC pattern in the IM periphery and coordinates the cruciferous array of floral sepals. Our data disfavor a contribution of activating auxin response factors (ARFs) but do not exclude auxin as a morphogenetic signal.

The AP2-type transcription factors DORNRÖSCHEN and DORNRÖSCHEN-LIKE promote G1/S transition.

The paralogous genes DORNRÖSCHEN (DRN) and DORNRÖSCHEN-LIKE (DRNL) encode AP2-type transcription factors that are expressed and act cell-autonomously in the central stem-cell zone or lateral organ founder cells (LOFCs) in the peripheral zone of the Arabidopsis shoot meristem (SAM), but their molecular contribution is unknown. Here, we show using the Arabidopsis thaliana MERISTEM LAYER 1 promoter that DRN and DRNL share a common function in cell cycle progression and potentially provide local competence for G1-S transitions in the SAM. Analysis of double transgenic DRN::erGFP and DRNL::erCERULEAN promoter fusion lines suggests that the trajectory of this cellular competence starts with DRN activity in the central stem-cell zone and extends locally via DRNL activity into groups of founder cells at the IM or FM periphery. Our data support the scenario that after gene duplication, DRN and DRNL acquired different transcription domains within the shoot meristem, but retained protein function that affects cell cycle progression, either centrally in stem cells or peripherally in primordial founder cells, a finding that is of general relevance for meristem function.

Founder-cell-specific transcription of the DORNRÖSCHEN-LIKE promoter and integration of the auxin response.

Transcription of the DORNRÖSCHEN (DRNL) promoter marks lateral-organ founder cells throughout Arabidopsis development, from cotyledons to flowers or floral organs. In the inflorescence apex, DRNL::GFP depicts incipient floral phyllotaxy, and organs in the four floral whorls are differentially prepatterned: the sepals unidirectionally along an abaxial-adaxial axis, the four petals and two lateral stamens in two putative morphogenetic fields, and the medial stamens subsequently in a ring-shaped domain, before two groups of carpel founder cells are specified. The dynamic DRNL transcription pattern is controlled by three enhancer elements, which redundantly and synergistically control qualitative or quantitative aspects of expression, and differentially integrate the auxin response in Arabidopsis inflorescence and floral meristems. The high sequence conservation of all three enhancer elements among the Brassicaceae is striking, which suggests that densely packed cis-regulatory elements are conserved to recruit multiple transcription factors, including auxin response factors, into higher-order enhanceosome complexes. The spatial organization of the enhancers is also conserved, by a microsynteny that extends beyond the Brassicaceae, which relates to enhancer sharing, as the distal element En1 bidirectionally serves DRNL and the upstream At1g24600 gene; the genes are transcribed in opposite directions and possibly comprise a conserved functional chromatin domain.

Phase 1, dose-ranging study of emixustat hydrochloride (ACU-4429), a novel visual cycle modulator, in healthy volunteers.

BACKGROUND: Emixustat hydrochloride (formerly ACU-4429) is a nonretinoid compound with a unique mode of action in the retinal pigment epithelium, where it modulates the biosynthesis of visual chromophore through its effect on retinal pigment epithelium-specific 65 kDa protein isomerase. This study provides clinicians with a background for understanding the pharmacokinetics and safety profile of orally administered emixustat. METHODS: This randomized, double-masked, placebo-controlled Phase 1b study evaluated the pharmacokinetics, tolerability, and safety of a 14-day course of oral emixustat (5, 10, 20, 30, or 40 mg) or placebo (3:1 ratio) once daily in healthy volunteers. RESULTS: A total of 40 subjects were enrolled (mean age, 38 years; 75% male). Emixustat (n = 30) was rapidly absorbed (median T(max), 3.0-5 hours) and readily eliminated (mean t(1/2), 4.6-7.9 hours), and mean C(max) and AUC(0-24) generally increased in proportion to dose. No significant accumulation of emixustat was observed with multiple-dose administration. Ocular adverse events occurred in 67% of the subjects who received emixustat; all were considered mild and resolved after study completion. Systemic adverse events were minimal. CONCLUSION: Oral emixustat was safe and well tolerated when administered once daily for 14 days with minimal systemic adverse events reported. These data support evaluation of emixustat in subjects with geographic atrophy associated with dry age-related macular degeneration.

Genetic integration of DORNRÖSCHEN and DORNRÖSCHEN-LIKE reveals hierarchical interactions in auxin signalling and patterning of the Arabidopsis apical embryo.

DORNRÖSCHEN (DRN) and DORNRÖSCHEN-LIKE (DRNL) encode AP2-domain transcription factors, which act redundantly in cotyledon organogenesis. A more detailed genetic study now integrates DRN and DRNL into the CUP-SHAPED COTYLEDON (CUC) regulatory network and places DRN and DRNL differentially within the auxin signalling network: DRNL function overlaps with that of PIN-FORMED1, and DRN with PINOID. DRN and DRNL act cell-autonomously and are co-expressed in the early globular embryo, whereas expression patterns diverge during later stages of embryogeny. Both genes synergize to provide essential patterning information in the apical embryo domain, to establish correct CUC, SHOOTMERISTEMLESS and WUSCHEL expression domains, which relates to the patterning of SAM anlagen to a central apical position to create two planes of bilateral symmetry in wild type Arabidopsis thaliana embryos.

Local auxin production: a small contribution to a big field.

Auxin is a plant growth regulator involved in diverse fundamental developmental responses. Much is now known about auxin transport, via influx and efflux carriers, and about auxin perception and its role in gene regulation. Many developmental processes are dependent on peaks of auxin concentration and, to date, attention has been directed at the role of polar auxin transport in generating and maintaining auxin gradients. However, surprisingly little attention has focussed on the role and significance of auxin biosynthesis, which should be expected to contribute to active auxin pools. Recent reports on the function of the YUCCA flavin monooxygenases and a tryptophan aminotransferase in Arabidopsis have caused us to look again at the importance of local biosynthesis in developmental processes. Many alternative and redundant pathways of auxin synthesis exist in many plants and it is emerging that they may function in response to environmental cues.

Auxin as compère in plant hormone crosstalk.

The architecture of many hormone perceptions and signalling pathways has been recently well established, together with an awareness that plant hormone responses are the product of networks of interactions involving multiple hormones. As growth is quantitative, so are hormone responses, which underlie a systems approach to development and response. Auxin is arguably one of the best characterised hormones in plant development, and despite many excellent reviews on auxin perception, polar transport, and signal transduction, too little attention has been given to auxin crosstalk. This review, therefore, gives a précis of recent developments in hormone crosstalk involving auxin. For decades, the literature has described the involvement of multiple hormones in particular processes, although the mechanistic bases underlying points of crosstalk have been harder to pinpoint. Crosstalk falls into different categories, such as direct, indirect, or co-regulation. One conclusion for auxin crosstalk is that crosstalk operates extensively via the metabolism of other hormones, however, microarray approaches are increasingly identifying co-regulated genes and nodes of crosstalk at shared signalling components. Auxin crosstalk is often local, and is spatially and temporally regulated to provide adaptive value to environmental conditions and fine-tuning of responses.

Histology versus phylogeny: Viewing plant embryogenesis from an evo-devo perspective.

The goal of evolutionary developmental (evo-devo) biology compares inter-organism developmental processes to infer ancestral relationships and evolutionary adaptations. Frameworks to address macroevolutionary traits such as plant embryogenesis commonly involve two complementary approaches. Historically, focus has been placed on comparative morphology and histology, but more recently, accumulating genome data from diverse taxa have elicited the construction of molecular phylogenies, which aid the identification of gene homologies and orthologies that have been adaptive and that underlie differences in form. Distinguishing between ancestral or derived traits in phyletic or cladistic-driven approaches is challenging, but relates to the broader applicability of existing developmental models such as Arabidopsis thaliana.

Cotyledon organogenesis.

The cotyledon represents one of the bases of classification within the plant kingdom, providing the name-giving difference between dicotyledonous and monocotyledonous plants. It is also a fundamental organ and there have been many reports of cotyledon mutants in many species. The use of these mutants where they have arisen in Arabidopsis has allowed us to unravel some of the complexities of embryonic patterning and cotyledon development with a high degree of resolution. The cloning of genes involved in cotyledon development from other species, together with physiological work, has supported the hypothesis that there exists a small number of orthologous gene hierarchies, particularly those involving auxin. The time is therefore appropriate for a summary of the regulation of cotyledon development gleaned from cotyledon mutants and regulatory pathways in the model species Arabidopsis and what can be inferred from cotyledon mutants in other species. There is an enormous variation in cotyledon form and development throughout the plant kingdom and this review focuses on debates about the phylogenetic relationship between mono- and dicotyledony, discusses gymnosperm cotyledon development and pleiocotyly in natural populations, and explores the limits of homology between cotyledons and leaves.

Class VIIIb APETALA2 Ethylene Response Factors in Plant Development.

The APETALA2 (AP2) transcription factor superfamily in many plant species is extremely large. In addition to well-documented roles in stress responses, some AP2 members in arabidopsis, such as those of subgroup VIIIb, which includes DORNRÖSCHEN, DORNRÖSCHEN-LIKE, PUCHI, and LEAFY PETIOLE, are also important developmental regulators throughout the plant life cycle. Information is accumulating from orthologs of these proteins in important crop species that they influence key agronomic traits, such as the release of bud-burst in woody perennials and floral meristem identity and branching in cereals, and thereby represent potential for agronomic improvement. Given the increasing recognition of their developmental significance, this review highlights the function of these proteins and addresses their phylogenetic and evolutionary relationships.

When negative is positive in functional genomics.

The post-genomic era has caused classical approaches to analyse gene function to be reviewed and refined. Conventional reverse genetic approaches to predict gene function have drawbacks in terms of genetic redundancy and being limited mainly to well-characterized genomes. The relatively recent use of dominant-negative transgenes has been successful in elaborating certain pathways and it is now possible to extend this technique to convert transcription factors into dominant repressor functions by fusion to repressor domains such as ENGRAILED from Drosophila. This methodology opens up new possibilities to overcome genetic redundancy, to identify novel gene functions and also to apply information on conserved protein domains between species to repress heterologous protein function.

Stem Cell Fate versus Differentiation: the Missing Link.

The shoot apical meristem provides a microenvironment that ensures stem cell fate and proliferation via homeostasis between WUSCHEL (WUS) activity and CLAVATA signalling. New data from maize and arabidopsis reveal that an evolutionarily conserved signal deriving from primordium cells links WUS transcription to the morphogenetic programme.

Ocular surface involvements in ectrodactyly-ectodermal dysplasia-cleft syndrome.

PURPOSE: To present the ocular manifestation of 2 cases of ectrodactyly-ectodermal dysplasia-cleft syndrome, a multiple congenital anomaly syndrome caused by a single point mutation of the p63 gene that controls epidermal development and homeostasis and to present treatment options. CASE REPORTS AND DISCUSSION: Patient 1 presented with mild signs and symptoms of dry eye and limbal stem cell deficiency with retention of 20/30 vision. Patient 2 presented with severe signs and symptoms of limbal stem cell deficiency with diffuse corneal scarring and counting fingers vision. This second patient's course was complicated by allergic conjunctivitis and advanced steroid-induced glaucoma. The cause of visual loss in ectrodactyly-ectodermal dysplasia-cleft syndrome appears to be multifactorial and likely includes inflammation of the ocular surface, tear film abnormalities, eyelid abnormalities, and limbal stem cell deficiency. Treatment modalities including lubrication, contact lenses, and limbal stem cell transplantation are reviewed. CONCLUSIONS: The ophthalmic conditions seen in ectrodactyly-ectodermal dysplasia-cleft syndrome frequently lead to vision loss. Early correct diagnosis and appropriate therapy are paramount because p63 gene mutations have a critical role in maintaining the integrity of the ocular surface in the setting of limbal stem cell deficiency, especially if there are other ocular surface insults such as lid disease, meibomian gland dysfunction and toxicity from topical medications. Patients should be monitored at regular, frequent intervals; and particular attention should be taken to avoid adverse secondary effects of these conditions and medications.

Founder cell specification.

Lateral organs arise from individual or groups of cells either on the flanks of meristems or within defined cellular positional contexts. The first event in organogenesis is founder cell specification. Auxin is one necessary signal in different organ specification contexts, but it is difficult to distinguish between correlative and causal signals and evidence is emerging that other signals exist and that the interplay between these signals is important for organ initiation. This review analyses the progress in understanding which signals contribute to founder cell specification and outlines the emerging complexities in the perception of positional information that are context-dependent and reliant on the establishment and coordination of different types of competencies.

Nanosized dendritic polyguanidilyated translocators for enhanced solubility, permeability, and delivery of gatifloxacin.

PURPOSE: Dendrimeric polyguanidilyated translocators (DPTs) are nanosized novel dendrimers that efficiently translocate molecules across biological barriers. The purpose of this study was to develop a DPT that could serve as an ophthalmic delivery vehicle for gatifloxacin and to evaluate its in vitro and in vivo delivery after topical application. METHODS: The gatifloxacin (GFX) solubility-enhancing property of a six-guanidine group-containing dendrimer (g6 DPT) was investigated as a function of pH and dendrimer concentration. Mechanisms of drug interaction with the dendrimer were investigated by using isothermal titration calorimetry (ITC), Fourier-transformed infrared spectroscopy (FTIR), and nuclear magnetic resonance spectroscopy (NMR). Permeability of the dendrimer was assessed in human corneal epithelial cells (HCECs) and across isolated bovine sclera-choroid-RPE (SCRPE). In vitro efficacy of the dendrimer formulation was evaluated with a time-to-kill assay for methicillin resistant Staphylococcus aureus (MRSA). In vivo delivery of GFX in a dendrimer eye drop formulation was studied in New Zealand White rabbits after a single dose or multiple doses over 3 weeks. Drug levels in various ocular tissues were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). RESULTS: An optimized DPT-GFX formulation (final pH 5.9, no preservative) increased GFX solubility by fourfold. The dendrimer formed isotonically stable, nanosized (346-nm) complexes with GFX via ionic bond, hydrogen bond, and hydrophobic interactions. The dendrimer gained rapid entry into the HCECs (within 5 minutes) and increased the transport of GFX by 40% across the SCRPE in 6 hours. DPT-GFX exhibited a three times faster killing rate for MRSA when compared with GFX alone. In vivo administration of DPT-GFX (1.2% wt/vol) resulted in ∼13-fold, and ∼2-fold higher areas under the curve (AUCs) for tissue concentrations in conjunctiva and cornea, respectively, when compared with GFX (0.3%) after a single dose. Further, a single dose of DPT-GFX sustained aqueous humor and vitreous humor drug levels during the 24-hour study, with a t(1/2) of 9 and 32 hours, respectively. After multiple doses, similar advantages were seen with DPT-GFX. CONCLUSIONS: The DPT forms stable complexes with GFX and enhances its solubility, permeability, anti-MRSA activity, and in vivo delivery, potentially allowing a once-daily dose regimen.

BIM1, a bHLH protein involved in brassinosteroid signalling, controls Arabidopsis embryonic patterning via interaction with DORNROSCHEN and DORNROSCHEN-LIKE.

The BIM1 protein which has been implicated in brassinosteroid (BR) signal transduction was identified from a two hybrid screen using the N-terminus, including the AP2 domain, of the transcription factors DORNROESCHEN (DRN) and DORNROESCHEN-LIKE (DRNL) which control embryonic patterning. The protein-protein interaction between BIM1 and DRN or DRNL was confirmed by co-immunoprecipitation and for DRN also in vivo by bimolecular fluorescence complementation. BIM1 can also physically interact with PHAVOLUTA (PHV), another interaction partner of DRN and DRNL. Loss of BIM1 function results in embryo patterning defects at low penetrance, including cell division defects in the hypophyseal region and apical domain defects such as cotyledon fusion and polycotyledony, in addition to polyembryony. BIM1 expression overlaps with that of DRN and DRNL from early globular embryo stages onwards. Higher order mutants between bim1, drn, drnl and phv suggest that although BIM1 may act partially redundantly with DRN in early embryo development, all genes function within the same pathway determining cotyledon development, supporting the hypothesis that they participate in a multimeric transcription factor complex. A role of BIM1 in embryonic development not only implicates a function for brassinosteroids in this process, but the interaction of BIM1 with DRN, involved with auxin signalling, represents a possible point of hormonal crosstalk in embryonic patterning and the first example of an interaction of components of the auxin and BR signalling pathways.