Emerging Trends in Ant-Pollinator Conflict in Extrafloral Nectary-Bearing Plants.

The net outcomes of mutualisms are mediated by the trade-offs between the costs and benefits provided by both partners. Our review proposes the existence of a trade-off in ant protection mutualisms between the benefits generated by the ants' protection against the attack of herbivores and the losses caused by the disruption of pollination processes, which are commonly not quantified. This trade-off has important implications for understanding the evolution of extrafloral nectaries (EFNs), an adaptation that has repeatedly evolved throughout the flowering plant clade. We propose that the outcome of this trade-off is contingent on the specific traits of the organisms involved. We provide evidence that the protective mutualisms between ants and plants mediated by EFNs have optimal protective ant partners, represented by the optimum point of the balance between positive effects on plant protection and negative effects on pollination process. Our review also provides important details about a potential synergism of EFN functionality; that is, these structures can attract ants to protect against herbivores and/or distract them from flowers so as not to disrupt pollination processes. Finally, we argue that generalizations regarding how ants impact plants should be made with caution since ants' effects on plants vary with the identity of the ant species in their overall net outcome.

Ecosystem engineering and leaf quality together affect arthropod community structure and diversity on white oak (Quercus alba L.).

Shelter building caterpillars act as ecosystem engineers by creating and maintaining leaf shelters, which are then colonized by other arthropods. Foliage quality has been shown to influence initial colonization by shelter-building caterpillars. However, the effects of plant quality on the interactions between ecosystem engineers and their communities have yet to be studied at the whole plant level. We examined how leaf tying caterpillars, as ecosystem engineers, impact arthropod communities on Quercus alba (white oak), and the modifying effect of foliage quality on these interactions. We removed all leaf tying caterpillars and leaf ties on 35 Q. alba saplings during the season when leaf tying caterpillars were active (June-September), and compared these leaf tie removal trees to 35 control trees whose leaf ties were left intact. Removal of these ecosystem engineers had no impact on overall arthropod species richness, but reduced species diversity, and overall arthropod abundance and that of most guilds, and changed the structure of the arthropod community as the season progressed. There was an increase in plant-level species richness with increasing number of leaf ties, consistent with Habitat Diversity Hypothesis. In turn, total arthropod density, and that of both leaf tying caterpillars and free-feeding caterpillars were affected by foliar tannin and nitrogen concentrations, and leaf water content. The engineering effect was greatest on low quality plants, consistent with the Stress-Gradient Hypothesis. Our results demonstrate that interactions between ecosystem engineering and plant quality together determine community structure of arthropods on Q. alba in Missouri.

Testing the role of local plant chemical diversity on plant-herbivore interactions and plant species coexistence.

Accumulating evidence suggests that herbivorous insects influence the local composition and richness of Neotropical plant species, particularly in species-rich genera. Species richness, phylogenetic diversity, and chemical diversity all influence the ability of insect herbivores to find and utilize their hosts. The relative impact of these components of diversity on species coexistence and plant-herbivore interactions is not well understood. We constructed 60 local communities of up to 13 species of Piper (Piperaceae) in native, mature forest at a lowland wet forest location in Costa Rica. The species composition of each community was chosen such that species richness, phylogenetic diversity, and GCMS-based chemical diversity were varied independently among communities. We predicted that chemical diversity would most strongly affect the communities across time, with smaller effects of taxonomic and phylogenetic diversity. At 13 months after the experimental planting, we assessed survivorship of each cutting, measured total leaf area loss of the survivors, leaf area loss to generalist and specialist herbivorous insect species, and local extinction of species. Generalist and specialist herbivory decreased with increasing levels of species richness and phylogenetic diversity, respectively. Surprisingly, there was no independent effect of chemical diversity on any of the three measures of herbivore damage. Nevertheless, plots with a higher chemical and phylogenetic diversity showed decreased plant mortality and local species extinction. Overall, our results suggested that both chemical and phylogenetic similarity are important factors in the assembly and maintenance of tropical plant communities. The fact that chemical diversity influences plant mortality suggests that leaf herbivores, and possibly other plant natural enemies, could increase plant diversity via the selective mortality of similar chemotypes.

Climate variability and aridity modulate the role of leaf shelters for arthropods: A global experiment.

Current climate change is disrupting biotic interactions and eroding biodiversity worldwide. However, species sensitive to aridity, high temperatures, and climate variability might find shelter in microclimatic refuges, such as leaf rolls built by arthropods. To explore how the importance of leaf shelters for terrestrial arthropods changes with latitude, elevation, and climate, we conducted a distributed experiment comparing arthropods in leaf rolls versus control leaves across 52 sites along an 11,790 km latitudinal gradient. We then probed the impact of short- versus long-term climatic impacts on roll use, by comparing the relative impact of conditions during the experiment versus average, baseline conditions at the site. Leaf shelters supported larger organisms and higher arthropod biomass and species diversity than non-rolled control leaves. However, the magnitude of the leaf rolls' effect differed between long- and short-term climate conditions, metrics (species richness, biomass, and body size), and trophic groups (predators vs. herbivores). The effect of leaf rolls on predator richness was influenced only by baseline climate, increasing in magnitude in regions experiencing increased long-term aridity, regardless of latitude, elevation, and weather during the experiment. This suggests that shelter use by predators may be innate, and thus, driven by natural selection. In contrast, the effect of leaf rolls on predator biomass and predator body size decreased with increasing temperature, and increased with increasing precipitation, respectively, during the experiment. The magnitude of shelter usage by herbivores increased with the abundance of predators and decreased with increasing temperature during the experiment. Taken together, these results highlight that leaf roll use may have both proximal and ultimate causes. Projected increases in climate variability and aridity are, therefore, likely to increase the importance of biotic refugia in mitigating the effects of climate change on species persistence.

Subtle structures with not-so-subtle functions: A data set of arthropod constructs and their host plants.

The construction of shelters on plants by arthropods might influence other organisms via changes in colonization, community richness, species composition, and functionality. Arthropods, including beetles, caterpillars, sawflies, spiders, and wasps often interact with host plants via the construction of shelters, building a variety of structures such as leaf ties, tents, rolls, and bags; leaf and stem galls, and hollowed out stems. Such constructs might have both an adaptive value in terms of protection (i.e., serve as shelters) but may also exert a strong influence on terrestrial community diversity in the engineered and neighboring hosts via colonization by secondary occupants. Although different traits of the host plant (e.g., physical, chemical, and architectural features) may affect the potential for ecosystem engineering by insects, such effects have been, to a certain degree, overlooked. Further analyses of how plant traits affect the occurrence of shelters may therefore enrich our understanding of the organizing principles of plant-based communities. This data set includes more than 1000 unique records of ecosystem engineering by arthropods, in the form of structures built on plants. All records have been published in the literature, and span both natural structures (91% of the records) and structures artificially created by researchers (9% of the records). The data were gathered between 1932 and 2021, across more than 50 countries and several ecosystems, ranging from polar to tropical zones. In addition to data on host plants and engineers, we aggregated data on the type of constructs and the identity of inquilines using these structures. This data set highlights the importance of these subtle structures for the organization of terrestrial arthropod communities, enabling hypotheses testing in ecological studies addressing ecosystem engineering and facilitation mediated by constructs. There are no copyright restrictions and please cite this paper when using the data in publications.

Overcoming the Pregnane X Receptor Liability: Rational Design to Eliminate PXR-Mediated CYP Induction.

The pregnane X receptor (PXR) regulates expression of proteins responsible for all three phases required for the detoxification mechanism, which include CYP450 enzymes, phase II enzymes, and multidrug efflux pumps. Therefore, PXR is a prominent receptor that is responsible for xenobiotic excretion and drug-drug interactions. Pyrimidinone 1 is an antagonist of the calcium sensing receptor (CaSR) and a strong activator of PXR. Repeat oral administration revealed diminished exposures over time, which prohibited further progression. A medicinal chemistry campaign was initiated to understand and abolish activation of PXR in order to increase systemic exposures. Rational structure-activity relationship investigations utilizing cocrystal structures and a de novo pharmacophore model resulted in compounds devoid of PXR activation. These studies culminated in the first orally active CaSR antagonist 8 suitable for progression. Cocrystallography, the pharmacophore model employed, and additional observations reported herein supported rational elimination of PXR activation and have applicability across diverse chemical classes to help erase PXR-driven drug-drug interactions.

Ecosystem engineering in the arboreal realm: heterogeneity of wood-boring beetle cavities and their use by cavity-nesting ants.

Wood-boring beetle larvae act as ecosystem engineers by creating stem cavities that are used secondarily as nests by many arboreal ant species. Understanding the heterogeneity and distribution of available cavities and their use by ants is therefore key to understanding arboreal ant community assembly and diversity. Our goals were to quantify the abundance and diversity of beetle-produced cavity resources in a tropical canopy, reveal how ants use these resources, and determine which characteristics of the cavity resource contribute to ant use. We dissected branches from six common tree species in the Brazilian Cerrado savanna, measuring cavity characteristics and identifying the occupants. We sampled 2310 individual cavities, 576 of which were used as nests by 25 arboreal ant species. We found significant differences among tree species in the proportion of stem length bored by beetles, the number of cavities per stem length, average entrance-hole size, and the distribution of cavity volumes. The likelihood that a cavity was occupied was greater for cavities with larger entrance-hole sizes and larger volumes. In particular, there was a strong positive correlation between mean head diameters of ant species and the mean entrance-hole diameter of the cavities occupied by those ant species. Wood-boring beetles contribute to the structuring of the Cerrado ant community by differentially attacking the available tree species. In so doing, the beetles provide a wide range of entrance-hole sizes which ant species partition based on their body size, and large volume cavities that ants appear to prefer.

Revisiting ecological dominance in arboreal ants: how dominant usage of nesting resources shapes community assembly.

Ecologically dominant species can shape the assembly of ecological communities via altering competitive outcomes. Moreover, these effects may be amplified under limited niche differentiation. Nevertheless, the influences of ecological dominance and niche differentiation on assembly are rarely considered together. Here, we provide a novel examination of dominance in a diverse arboreal ant community, defining dominance by the prevalent usage of nesting resources and addressing how it influences community assembly. We first used a series of quantitative observational and experimental studies to address the natural nesting ecology, colony incidence on surveyed trees, and level of dominance over newly available nesting resources by our focal species, Cephalotes pusillus. The experimental studies were then used further to examine whether C. pusillus shapes assembly via an influence on cavity usage by co-occurring species. C. pusillus was confirmed as a dominant user of cavity nesting resources, with highly generalized nesting ecology, occupying about 50% of the trees within the focal system, and accounting for more than a third of new cavity occupation in experiments. Our experiments showed further that the presence of C. pusillus was associated with modest effects on species richness, but significant decreases in cavity-occupation levels and significant shifts in the entrance-size usage by co-occurring species. These results indicate that C. pusillus, as a dominant user of nesting resources, shapes assembly at multiple levels. Broadly, our findings highlight that complex interactions between a dominant species and the resource-usage patterns of other species can underlie species assembly in diverse ecological communities.

Correction to Identification of Quinoline-Based RIP2 Kinase Inhibitors with an Improved Therapeutic Index to the hERG Ion Channel.

[This corrects the article DOI: 10.1021/acsmedchemlett.8b00344.].

Understanding Pharmacokinetic Disconnect in Preclinical Species for 4-Aminoquinolines: Consequences of Low Permeability and High P-glycoprotein Efflux Ratio on Rat and Dog Oral Pharmacokinetics.

Receptor Interacting Protein 2 (RIP2) kinase inhibitors have been reported for therapeutic opportunities in inflammatory bowel diseases such as Ulcerative Colitis and Crohn's disease. During lead optimization, team identified 4-aminoquinoline series and several compounds from this series were investigated in rat and dog pharmacokinetic studies. While compounds such as GSKA and GSKB demonstrated acceptable pharmacokinetics in rat and dog, further progression of these compounds was halted due to adverse findings in advanced safety studies. Structurally similar analogues incorporating polarity at C-7 position of 4-aminoquinoline resulted in identification of GSKC - GSKF. Interestingly, following oral administration to rat at similar low dose, GSKC - GSKF demonstrated significantly low systemic drug exposure compared to GSKA and GSKB (3-17-fold difference). However, in dog, dose normalized oral systemic exposure for GSKC - GSKF was comparable to GSKA and GSKB (within 2-fold). A series of studies were conducted to understand the disconnect which highlighted that an intrinsic reduction in permeability and high P-glycoprotein (P-gp) efflux ratio for C-7 substituted analogues were driving pharmacokinetic disconnect between rat and dog. Oral absorption was minimally impacted in dog by P-gp mediated efflux compared to rat because the leakier gastrointestinal tract in dog likely overcomes this effect.

Discovery of Pyrazolocarboxamides as Potent and Selective Receptor Interacting Protein 2 (RIP2) Kinase Inhibitors.

Herein we report the discovery of pyrazolocarboxamides as novel, potent, and kinase selective inhibitors of receptor interacting protein 2 kinase (RIP2). Fragment based screening and design principles led to the identification of the inhibitor series, and X-ray crystallography was used to inform key structural changes. Through key substitutions about the N1 and C5 N positions on the pyrazole ring significant kinase selectivity and potency were achieved. Bridged bicyclic pyrazolocarboxamide 11 represents a selective and potent inhibitor of RIP2 and will allow for a more detailed investigation of RIP2 inhibition as a therapeutic target for autoinflammatory disorders.

Tri-trophic interactions: bridging species, communities and ecosystems.

A vast body of research demonstrates that many ecological and evolutionary processes can only be understood from a tri-trophic viewpoint, that is, one that moves beyond the pairwise interactions of neighbouring trophic levels to consider the emergent features of interactions among multiple trophic levels. Despite its unifying potential, tri-trophic research has been fragmented, following two distinct paths. One has focused on the population biology and evolutionary ecology of simple food chains of interacting species. The other has focused on bottom-up and top-down controls over the distribution of biomass across trophic levels and other ecosystem-level variables. Here, we propose pathways to bridge these two long-standing perspectives. We argue that an expanded theory of tri-trophic interactions (TTIs) can unify our understanding of biological processes across scales and levels of organisation, ranging from species evolution and pairwise interactions to community structure and ecosystem function. To do so requires addressing how community structure and ecosystem function arise as emergent properties of component TTIs, and, in turn, how species traits and TTIs are shaped by the ecosystem processes and the abiotic environment in which they are embedded. We conclude that novel insights will come from applying tri-trophic theory systematically across all levels of biological organisation.

Discovery of a First-in-Class Receptor Interacting Protein 2 (RIP2) Kinase Specific Clinical Candidate, 2-((4-(Benzo[d]thiazol-5-ylamino)-6-(tert-butylsulfonyl)quinazolin-7-yl)oxy)ethyl Dihydrogen Phosphate, for the Treatment of Inflammatory Diseases.

RIP2 kinase has been identified as a key signal transduction partner in the NOD2 pathway contributing to a variety of human pathologies, including immune-mediated inflammatory diseases. Small-molecule inhibitors of RIP2 kinase or its signaling partners on the NOD2 pathway that are suitable for advancement into the clinic have yet to be described. Herein, we report our discovery and profile of the prodrug clinical compound, inhibitor 3, currently in phase 1 clinical studies. Compound 3 potently binds to RIP2 kinase with good kinase specificity and has excellent activity in blocking many proinflammatory cytokine responses in vivo and in human IBD explant samples. The highly favorable physicochemical and ADMET properties of 3 combined with high potency led to a predicted low oral dose in humans.

Identification of a RIP1 Kinase Inhibitor Clinical Candidate (GSK3145095) for the Treatment of Pancreatic Cancer.

RIP1 regulates cell death and inflammation and is believed to play an important role in contributing to a variety of human pathologies, including immune-mediated inflammatory diseases and cancer. While small-molecule inhibitors of RIP1 kinase have been advanced to the clinic for inflammatory diseases and CNS indications, RIP1 inhibitors for oncology indications have yet to be described. Herein we report on the discovery and profile of GSK3145095 (compound 6). Compound 6 potently binds to RIP1 with exquisite kinase specificity and has excellent activity in blocking RIP1 kinase-dependent cellular responses. Highlighting its potential as a novel cancer therapy, the inhibitor was also able to promote a tumor suppressive T cell phenotype in pancreatic adenocarcinoma organ cultures. Compound 6 is currently in phase 1 clinical studies for pancreatic adenocarcinoma and other selected solid tumors.

Author Correction: Design of amidobenzimidazole STING receptor agonists with systemic activity.

Change history: In this Letter, author Ana Puhl was inadvertently omitted; this error has been corrected online.An amendment to this paper has been published and can be accessed via a link at the top of the paper.

A Schlemm Canal Microstent for Intraocular Pressure Reduction in Primary Open-Angle Glaucoma and Cataract: The HORIZON Study.

OBJECTIVE: To compare cataract surgery with implantation of a Schlemm canal microstent with cataract surgery alone for the reduction of intraocular pressure (IOP) and medication use after 24 months. DESIGN: Prospective, multicenter, single-masked, randomized controlled trial. PARTICIPANTS: Subjects with concomitant primary open-angle glaucoma (POAG), visually significant cataract, and washed-out modified diurnal IOP (MDIOP) between 22 and 34 mmHg. METHODS: Subjects were randomized 2:1 to receive a single Hydrus Microstent (Ivantis, Inc, Irvine, CA) in the Schlemm canal or no stent after uncomplicated phacoemulsification. Comprehensive eye examinations were conducted 1 day, 1 week, and 1, 3, 6, 12, 18, and 24 months postoperatively. Medication washout and MDIOP measurement were repeated at 12 and 24 months. MAIN OUTCOME MEASURES: The primary and secondary effectiveness end points were the proportion of subjects demonstrating a 20% or greater reduction in unmedicated MDIOP and change in mean MDIOP from baseline at 24 months, respectively. Hypotensive medication use was tracked throughout the course of follow-up. Safety measures included the frequency of surgical complications and adverse events. RESULTS: A total of 369 eyes were randomized after phacoemulsification to Hydrus Microstent (HMS) and 187 to no microstent (NMS). At 24 months, unmedicated MDIOP was reduced by ≥20% in 77.3% of HMS group eyes and in 57.8% of NMS group eyes (difference = 19.5%, 95% confidence interval [CI] 11.2%-27.8%, P < 0.001). The mean reduction in 24-month unmedicated MDIOP was -7.6±4.1 mmHg (mean ± standard deviation) in the HMS group and -5.3±3.9 mmHg in the NMS group (difference = -2.3 mmHg; 95% CI, -3.0 to -1.6; P < 0.001). The mean number of medications was reduced from 1.7±0.9 at baseline to 0.3±0.8 at 24 months in the HMS group and from 1.7±0.9 to 0.7±0.9 in the NMS group (difference = -0.4 medications; P < 0.001). There were no serious ocular adverse events related to the microstent, and no significant differences in safety parameters between the 2 groups. CONCLUSIONS: This 24-month multicenter randomized controlled trial demonstrated superior reduction in MDIOP and medication use among subjects with mild-to-moderate POAG who received a Schlemm canal microstent combined with phacoemulsification compared with phacoemulsification alone.

Design of amidobenzimidazole STING receptor agonists with systemic activity.

Stimulator of interferon genes (STING) is a receptor in the endoplasmic reticulum that propagates innate immune sensing of cytosolic pathogen-derived and self DNA1. The development of compounds that modulate STING has recently been the focus of intense research for the treatment of cancer and infectious diseases and as vaccine adjuvants2. To our knowledge, current efforts are focused on the development of modified cyclic dinucleotides that mimic the endogenous STING ligand cGAMP; these have progressed into clinical trials in patients with solid accessible tumours amenable to intratumoral delivery3. Here we report the discovery of a small molecule STING agonist that is not a cyclic dinucleotide and is systemically efficacious for treating tumours in mice. We developed a linking strategy to synergize the effect of two symmetry-related amidobenzimidazole (ABZI)-based compounds to create linked ABZIs (diABZIs) with enhanced binding to STING and cellular function. Intravenous administration of a diABZI STING agonist to immunocompetent mice with established syngeneic colon tumours elicited strong anti-tumour activity, with complete and lasting regression of tumours. Our findings represent a milestone in the rapidly growing field of immune-modifying cancer therapies.

Identification of Quinoline-Based RIP2 Kinase Inhibitors with an Improved Therapeutic Index to the hERG Ion Channel.

RIP2 kinase was recently identified as a therapeutic target for a variety of autoimmune diseases. We have reported previously a selective 4-aminoquinoline-based RIP2 inhibitor GSK583 and demonstrated its effectiveness in blocking downstream NOD2 signaling in cellular models, rodent in vivo models, and human ex vivo disease models. While this tool compound was valuable in validating the biological pathway, it suffered from activity at the hERG ion channel and a poor PK/PD profile thereby limiting progression of this analog. Herein, we detail our efforts to improve both this off-target liability as well as the PK/PD profile of this series of inhibitors through modulation of lipophilicity and strengthening hinge binding ability. These efforts have led to inhibitor 7, which possesses high binding affinity for the ATP pocket of RIP2 (IC50 = 1 nM) and inhibition of downstream cytokine production in human whole blood (IC50 = 10 nM) with reduced hERG activity (14 µM).

Trade-offs between growth, reproduction and defense in response to resource availability manipulations.

The Brazilian Cerrado is one of the most endangered biomes in the world. We evaluated the sustainability of leaf harvest in one of the most important Cerrado tree species, Stryphnodendron adstringens. The bark of this tree is used as a source of medicinal tannin. Harvesting bark, however, often kills the tree. In a manipulative field experiment, we tested the hypothesis that harvesting leaves, which might serve as an alternative source of tannin, would be less detrimental for tree survival, growth, reproduction, and defense than harvesting bark. In a two-way crossed experimental design, we either clipped 100% of a plant's leaves or applied NPK fertilizer to the soil. Our predictions of the experimental outcomes were based on plant resource and defense theory. Growth was determined by total leaf dry mass production, reproduction by inflorescence and fruit production traits, and defense by total phenolics, hydrolyzable tannins, and condensed tannins. Fertilization had a marginally positive effect on total leaf dry mass. Defoliation had no effect on subsequent leaf production, and most importantly, no plants died as a result of defoliation. We found high tannin amounts in leaves of S. adstringens produced both prior to and subsequent to clipping, further suggesting that leaves could serve as a sustainable alternative source of tannin. After clipping, plants invested more in tannin production and less in reproduction. Our results suggest that leaf harvest may be more sustainable than harvesting of bark in S. adstringens. We suggest the need for further investigation of the medicinal properties of leaf tannins to formulate a viable sustainable management plan for the exploitation of this plant species.

Usage of an Electronic Database and Checklist System for Improvement in Magnetic Resonance Imaging Acquisition.

PURPOSE: To determine whether implementation of an easily accessible electronic database promotes significant reporting of magnetic resonance imaging (MRI) acquisition errors. Additionally, we wanted to see if analysis of the error reports could be used to create a comprehensive checklist to avoid the most common errors. METHODS: A new, simple, and efficient electronic database reporting system was written in-house and implemented at our institution. Over the course of 4 months, the use of this database enabled collection and analysis of sufficient data for trend analysis. A simple 4-point checklist for MRI technologist use was developed based on the most commonly reported errors. Reported MRI acquisition error rates were collected and analyzed thereafter. RESULTS: By the first full month of implementation, MRI scan error reporting increased from a previous negligible baseline rate to 3.03%. The comprehensive checklist was based on the 4 most common issues reported. Verification of checklist use showed that adherence to this requirement averaged greater than 94%. Immediately following roll out of the checklist, the percentage of errors reported fell to 1.7% with a continued decline in error reports thereafter. An approximately 60% reduction in errors in the last month of the study was evident as compared to the first month of data collection. CONCLUSIONS: The use of an efficient error reporting system and implementation of a checklist based on the most common MRI acquisition errors results in a substantial decrease in the baseline MRI acquisition error rates.