Developmental Genetic and Molecular Analysis of Drosophila Central Complex Lineages.

Complex behaviors are mediated by a diverse class of neurons and glia produced during development. Both neural stem cell-intrinsic and -extrinsic temporal cues regulate the appropriate number, molecular identity, and circuit assembly of neurons. The Drosophila central complex (CX) is a higher-order brain structure regulating various behaviors, including sensory-motor integration, celestial navigation, and sleep. Most neurons and glia in the adult CX are formed during larval development by 16 Type II neural stem cells (NSCs). Unlike Type I NSCs, which directly give rise to the ganglion mother cells (GMCs), Type II NSCs give rise to multiple intermediate neural progenitors (INPs), and each INP in turn generates multiple GMCs, hence fostering the generation of longer and more diverse lineages. This makes Type II NSCs a suitable model to unravel the molecular mechanisms regulating neural diversity in more complex lineages. In this review, we elaborate on the classification and identification of NSCs based on the types of division adopted and the molecular markers expressed in each type. In the end, we discuss genetic methods for lineage analysis and birthdating. We also explain the temporal expression of stem cell factors and genetic techniques to study how stem cell factors may regulate neural fate specification.

Assessment of the probability of introduction of Thaumatotibia leucotreta into the European Union with import of cut roses.

Following a request from the European Commission, the EFSA Panel on Plant Health performed a quantitative pest risk assessment to assess whether the import of cut roses provides a pathway for the introduction of Thaumatotibia leucotreta (Lepidoptera: Tortricidae) into the EU. The assessment was limited to the entry and establishment steps. A pathway model was used to assess how many T. leucotreta individuals would survive and emerge as adults from commercial or household wastes in an EU NUTS2 region climatically suitable in a specific season. This pathway model for entry consisted of three components: a cut roses distribution model, a T. leucotreta developmental model and a waste model. Four scenarios of timing from initial disposal of the cut roses until waste treatment (3, 7, 14 and 28 days) were considered. The estimated median number of adults escaping per year from imported cut roses in all the climatically suitable NUTS2 regions of the EU varied from 49,867 (90% uncertainty between 5,298 and 234,393) up to 143,689 (90% uncertainty between 21,126 and 401,458) for the 3- and 28-day scenarios. Assuming that, on average, a successful mating will happen for every 435 escaping moths, the estimated median number of T. leucotreta mated females per year from imported cut roses in all the climatically suitable NUTS2 regions of the EU would vary from 115 (90% uncertainty between 12 and 538) up to 330 (90% uncertainty between 49 and 923) for the 3- and 28-day scenarios. Due to the extreme polyphagia of T. leucotreta, host availability will not be a limiting factor for establishment. Climatic suitability assessment, using a physiologically based demographic modelling approach, identified the coastline extending from the northwest of the Iberian Peninsula through the Mediterranean as area suitable for establishment of T. leucotreta. This assessment indicates that cut roses provide a pathway for the introduction of T. leucotreta into the EU.

Elucidation of novel TRAP1-Selective inhibitors that regulate mitochondrial processes.

Hsp90 isoform-selective inhibitors represent a new paradigm for novel anti-cancer drugs as each of the four isoforms have specific cellular localization, function, and client proteins. The mitochondrial isoform, TRAP1, is the least understood member of the Hsp90 family due to the lack of small molecule tools to study its biological function. Herein, we report novel TRAP1-selective inhibitors used to interrogate TRAP1's biological function along with co-crystal structures of such compounds bound to the N-terminus of TRAP1. Solution of the co-crystal structure allowed for a structure-based approach that resulted in compound 36, which is a 40 nM inhibitor with >250-fold TRAP1 selectivity over Grp94, the isoform with the highest structural similarity to TRAP1 within the N-terminal ATP binding site. Lead compounds 35 and 36 were found to selectively induce TRAP1 client protein degradation without inducing the heat shock response or disrupting Hsp90-cytosolic clients. They were also shown to inhibit OXPHOS, alter cellular metabolism towards glycolysis, disrupt TRAP1 tetramer stability, and disrupt the mitochondrial membrane potential.

Nesting Behavior, Phenology, and Bionomics of the High Andean Leaf-Cutter Bee Megachile (Cressoniella) amparo.

Megachile amparo (González, Revista Colombiana De Entomología 32(1):93-96, 2006) is the only high Andean leaf-cutter bee reported in Colombia and is possibly endemic to the Colombian Andes. Although it is frequently observed, even in urban areas, its biology and ecology remain unknown. The present study aimed to describe detailed aspects of its bionomy. Trap-nests were installed on the Campus of the Nueva Granada University (Cajicá, Colombia) from June/2018 to March/2020. The trap-nests were wooden blocks (25 × 15 × 14 cm) with 30 cavities of Ø = 1 cm and different lengths (50 mm, 75 mm, and 100 mm) lined with waxed paper straws. During the observations, an increasing number of trap-nests were installed, increasing from 250 to 720 cavities. The trap-nests were monitored three times a week, recording both the date the start and end building by female. Most of the nest were maintained in the field to estimate the sex ratio, cell survival, and total development time under natural conditions. Thirty-two nests were removed at different times of the observation period to establish number of cells per nest, and cells built per female per day. We incubated 20 cells from different nests at 18 °C, 22 °C, 26 °C, and 32 °C to estimate the base temperature, thermal constant k (developmental time in degree days), and cell survival. Young cells of different positions were dissected and weighed to characterize food provision and brood cells. Computerized tomography-CT scans were performed in 30 brood cells to determine if diapause occurred during prepupal stage. Females nested 7- and 10-cm-long cavities and the number of cells per nest varied with cavity length. The brood cells had a length of 1.23 ± 0.12 cm and a diameter of 0.92 ± 0.05 cm. The female spends 1.17 ± 0.29 days to build a brood cell. Food provision varied according to the position of the brood cell in the nest. The adults of M. amparo present a marked seasonality being more active during dry months. Base temperature and thermal constant k were different for males and females. The sex ratio is female biased (1.9:1), and cell survival in the field was 89% with no cleptoparasites or predators recorded.

A structure activity relationship study of 3,4'-dimethoxyflavone for ArlRS inhibition in Staphylococcus aureus.

Infections caused by methicillin-resistant Staphylococcus aureus (MRSA) are difficult to treat due to their resistance to many ß-lactam antibiotics, and their highly coordinated excretion of virulence factors. One way in which MRSA accomplishes this is by responding to environmental stimuli using two-component systems (TCS). The ArlRS TCS has been identified as having a key role in regulating virulence in both systemic and local infections caused by S. aureus. We recently disclosed 3,4'-dimethoxyflavone as a selective ArlRS inhibitor. In this study we explore the structure-activity relationship (SAR) of the flavone scaffold for ArlRS inhibition and identify several compounds with increased activity compared to the parent. Additionally, we identify a compound that suppresses oxacillin resistance in MRSA, and begin to probe the mechanism of action behind this activity.

The Drivers of Diversity: Integrated genetic and hormonal cues regulate neural diversity.

Proper functioning of the nervous system relies not only on the generation of a vast repertoire of distinct neural cell types but also on the precise neural circuitry within them. How the generation of highly diverse neural populations is regulated during development remains a topic of interest. Landmark studies in Drosophila have identified the genetic and temporal cues regulating neural diversity and thus have provided valuable insights into our understanding of temporal patterning of the central nervous system. The development of the Drosophila central complex, which is mostly derived from type II neural stem cell (NSC) lineages, showcases how a small pool of NSCs can give rise to vast and distinct progeny. Similar to the human outer subventricular zone (OSVZ) neural progenitors, type II NSCs generate intermediate neural progenitors (INPs) to expand and diversify lineages that populate higher brain centers. Each type II NSC has a distinct spatial identity and timely regulated expression of many transcription factors and mRNA binding proteins. Additionally, INPs derived from them show differential expression of genes depending on their birth order. Together type II NSCs and INPs display a combinatorial temporal patterning that expands neural diversity of the central brain lineages. We cover advances in current understanding of type II NSC temporal patterning and discuss similarities and differences in temporal patterning mechanisms of various NSCs with a focus on how cell-intrinsic and extrinsic hormonal cues regulate temporal transitions in NSCs during larval development. Cell extrinsic ligands activate conserved signaling pathways and extrinsic hormonal cues act as a temporal switch that regulate temporal progression of the NSCs. We conclude by elaborating on how a progenitor's temporal code regulates the fate specification and identity of distinct neural types. At the end, we also discuss open questions in linking developmental cues to neural identity, circuits, and underlying behaviors in the adult fly.

Invasive potential of tropical fruit flies in temperate regions under climate change.

Tropical fruit flies are considered among the most economically important invasive species detected in temperate areas of the United States and the European Union. Detections often trigger quarantine and eradication programs that are conducted without a holistic understanding of the threat posed. Weather-driven physiologically-based demographic models are used to estimate the geographic range, relative abundance, and threat posed by four tropical tephritid fruit flies (Mediterranean fruit fly, melon fly, oriental fruit fly, and Mexican fruit fly) in North and Central America, and the European-Mediterranean region under extant and climate change weather (RCP8.5 and A1B scenarios). Most temperate areas under tropical fruit fly propagule pressure have not been suitable for establishment, but suitability is predicted to increase in some areas with climate change. To meet this ongoing challenge, investments are needed to collect sound biological data to develop mechanistic models to predict the geographic range and relative abundance of these and other invasive species, and to put eradication policies on a scientific basis.

Activation of gene expression by detergent-like protein domains.

The mechanisms by which transcriptional activation domains (tADs) initiate eukaryotic gene expression have been an enigma for decades because most tADs lack specificity in sequence, structure, and interactions with targets. Machine learning analysis of data sets of tAD sequences generated in vivo elucidated several functionality rules: the functional tAD sequences should (i) be devoid of or depleted with basic amino acid residues, (ii) be enriched with aromatic and acidic residues, (iii) be with aromatic residues localized mostly near the terminus of the sequence, and acidic residues localized more internally within a span of 20-30 amino acids, (iv) be with both aromatic and acidic residues preferably spread out in the sequence and not clustered, and (v) not be separated by occasional basic residues. These and other more subtle rules are not absolute, reflecting absence of a tAD consensus sequence, enormous variability, and consistent with surfactant-like tAD biochemical properties. The findings are compatible with the paradigm-shifting nucleosome detergent mechanism of gene expression activation, contributing to the development of the liquid-liquid phase separation model and the biochemistry of near-stochastic functional allosteric interactions.

The coffee agroecosystem: bio-economic analysis of coffee berry borer control (Hypothenemus hampei).

Coffee, after petroleum, is the most valuable commodity globally in terms of total value (harvest to coffee cup). Here, our bioeconomic analysis considers the multitude of factors that influence coffee production. The system model used in the analysis incorporates realistic field models based on considerable new field data and models for coffee plant growth and development, the coffee/coffee berry borer (CBB) dynamics in response to coffee berry production and the role of the CBB parasitoids and their interactions in control of CBB. Cultural control of CBB by harvesting, cleanup of abscised fruits, and chemical sprays previously considered are reexamined here to include biopesticides for control of CBB such as entomopathogenic fungi (Beauveria bassiana, Metarhizium anisopliae) and entomopathogenic nematodes (Steinernema sp., Heterorhabditis). The bioeconomic analysis estimates the potential of each control tactic singly and in combination for control of CBB. The analysis explains why frequent intensive harvesting of coffee is by far the most effective and economically viable control practice for reducing CBB infestations in Colombia and Brazil.

Small world in the real world: Long distance dispersal governs epidemic dynamics in agricultural landscapes.

Outbreaks of a plant disease in a landscape can be meaningfully modelled using networks with nodes representing individual crop-fields, and edges representing potential infection pathways between them. Their spatial structure, which resembles that of a regular lattice, makes such networks fairly robust against epidemics. Yet, it is well-known how the addition of a few shortcuts can turn robust regular lattices into vulnerable 'small world' networks. Although the relevance of this phenomenon has been shown theoretically for networks with nodes corresponding to individual host plants, its real-world implications at a larger scale (i.e. in networks with nodes representing crop fields or other plantations) remain elusive. Focusing on realistic spatial networks connecting olive orchards in Andalusia (Southern Spain), the world's leading olive producer, we show how even very small probabilities of long distance dispersal of infectious vectors result in a small-world effect that dramatically exacerbates a hypothetical outbreak of a disease targeting olive trees (loosely modelled on known epidemiological information on the bacterium Xylella fastidiosa, an important emerging threat for European agriculture). More specifically, we found that the probability of long distance vector dispersal has a disproportionately larger effect on epidemic dynamics compared to pathogen's intrinsic infectivity, increasing total infected area by up to one order of magnitude (in the absence of quarantine). Furthermore, even a very small probability of long distance dispersal increased the effort needed to halt a hypothetical outbreak through quarantine by about 50% in respect to scenarios modelling local/short distance pathogen's dispersal only. This highlights how identifying (and disrupting) long distance dispersal processes may be more efficacious to contain a plant disease epidemic than surveillance and intervention concentrated on local scale transmission processes.

Design of a Bioreactor to Assess the Effect of Passive Joint Loading in a Live Chick Embryo In Ovo.

There is increasing interest in understanding how mechanical cues (e.g., physical forces due to kicking and other movements) influence the embryological development of tissues and organs. For example, recent studies from our laboratory and others have used the chick embryo model to demonstrate that the compositional and mechanical properties of developing tendons are strongly regulated by embryo movement frequency. However, current research tools for manipulating embryological movements and in ovo (or in utero) mechanical forces are generally limited to chemical treatments that either paralyze or overstimulate muscles without allowing for precise control of physical cues. Thus, in this study, we introduce an instrument that enables application of passive, dynamic ankle flexion at prescribed amplitudes and frequencies in live, developing chick embryos. This device meets the design goals of allowing for precise (<1.5°) control of different waveforms of ankle motion at a physiologically relevant frequency (0.17 Hz) across a range of ankle angles (0-90° plantarflexion) with maintenance of embryo viability comparable to other methods. Impact Statement We describe the design and implementation of a novel bioreactor to precisely control ankle motion in a chick embryo within its physiological environment. The chick embryo has been used for decades to study mechanobiology of musculoskeletal tissue development and regeneration, but approaches have been limited to chemical treatments that either paralyze or overstimulate muscles without allowing for precise control of physical cues. Thus, this novel instrument is a major advancement over current research tools for manipulating chick embryological movements in ovo (or in utero).

Fine-scale ecological and economic assessment of climate change on olive in the Mediterranean Basin reveals winners and losers.

The Mediterranean Basin is a climate and biodiversity hot spot, and climate change threatens agro-ecosystems such as olive, an ancient drought-tolerant crop of considerable ecological and socioeconomic importance. Climate change will impact the interactions of olive and the obligate olive fruit fly (Bactrocera oleae), and alter the economics of olive culture across the Basin. We estimate the effects of climate change on the dynamics and interaction of olive and the fly using physiologically based demographic models in a geographic information system context as driven by daily climate change scenario weather. A regional climate model that includes fine-scale representation of the effects of topography and the influence of the Mediterranean Sea on regional climate was used to scale the global climate data. The system model for olive/olive fly was used as the production function in our economic analysis, replacing the commonly used production-damage control function. Climate warming will affect olive yield and fly infestation levels across the Basin, resulting in economic winners and losers at the local and regional scales. At the local scale, profitability of small olive farms in many marginal areas of Europe and elsewhere in the Basin will decrease, leading to increased abandonment. These marginal farms are critical to conserving soil, maintaining biodiversity, and reducing fire risk in these areas. Our fine-scale bioeconomic approach provides a realistic prototype for assessing climate change impacts in other Mediterranean agro-ecosystems facing extant and new invasive pests.

Eradication of invasive species: why the biology matters.

Published bi- and tri-trophic physiologically based demographic system models having similar sub components are used to assess prospectively the geographic distributions and relative abundance (a measure of invasiveness) of six invasive herbivorous insect species across the United States and Mexico. The plant hosts and insect species included in the study are: 1) cotton/pink bollworm, 2) a fruit tree host/Mediterranean fruit fly, 3) olive/olive fly, 4) a perennial host/light brown apple moth, 5) grapevine/glassy-winged sharpshooter and its two egg parasitoids, and 6) grapevine/European grapevine moth. All of these species are currently or have been targets for eradication. The goal of the analyses is to predict and explain prospectively the disparate distributions of the six species as a basis for examining eradication or containment efforts against them. The eradication of the new world screwworm is also reviewed in the discussion section because of its pivotal role in the development of the eradication paradigm. The models used are mechanistic descriptions of the weather driven biology of the species. Observed daily weather data (i.e., max-min temperatures, solar radiation) from 1,221 locations across the United States and Mexico for the period 1983-2003 were used to drive the models. Soil moisture and nutrition were assumed nonlimiting. The simulation results were mapped using GRASS GIS. The mathematical underpinnings of the modeling approach are reviewed in the appendix and in the supplemental materials.

Geographic distribution and relative abundance of the invasive glassy-winged sharpshooter: effects of temperature and egg parasitoids.

The capacity to predict the geographic distribution and relative abundance of invasive species is pivotal to developing policy for eradication or control and management. Commonly used methods fall under the ambit of ecological niche models (ENMs). These methods were reviewed and shortcomings identified. Weather-driven physiologically based demographic models (PBDMs) are proposed that resolve many of the deficiencies of ENMs. The PBDM approach is used to analyze the invasiveness of the polyphagous glassy-winged sharpshooter (Homalodisca vitripennis [Germar]), a pest native to the southeastern United States and northeastern Mexico that extended its range into California in 1989. Glassy-winged sharpshooter vectors the pathogenic bacterium, Xylella fastidiosa (Wells) that causes Pierce's disease in grape and scorch-like diseases in other plants. PBDMs for glassy-winged sharpshooter and its egg parasitoids (Gonatocerus ashmeadi Girault and G. triguttatus Girault) were developed and linked to a PBDM for grape published by Wermelinger et al. (1991). Daily weather data from 108 locations across California for the period 1995-2006 were used to drive the PBDM system, and GRASS GIS was used to map the simulation results. The geographic distribution of glassy-winged sharpshooter, as observed, is predicted to be largely restricted to the warm areas of southern California, with the action of the two egg parasitoids reducing its abundance >90%. The average indispensable mortality contributed by G. triguttatus is <1%. A temperature-dependent developmental rate model for X. fastidiosa was developed that suggests its geographic range is also limited to the warm inland areas of southern California. Biological control of glassy-winged sharpshooter further decreases the pathogen's relative range. Climate warming scenarios of +2°C and +3°C suggest that the distribution and severity of glassy-winged sharpshooter and X. fastidiosa will increase in the agriculturally rich central valley of California. The utility of holistic analyses for formulating control policy and tactics for invasive species is discussed.

Ecosocial consequences and policy implications of disease management in East African agropastoral systems.

International research and development efforts in Africa have brought ecological and social change, but analyzing the consequences of this change and developing policy to manage it for sustainable development has been difficult. This has been largely due to a lack of conceptual and analytical models to access the interacting dynamics of the different components of ecosocial systems. Here, we examine the ecological and social changes resulting from an ongoing suppression of trypanosomiasis disease in cattle in an agropastoral community in southwest Ethiopia to illustrate how such problems may be addressed. The analysis combines physiologically based demographic models of pasture, cattle, and pastoralists and a bioeconomic model that includes the demographic models as dynamic constraints in the economic objective function that maximizes the utility of individual consumption under different level of disease risk in cattle. Field data and model analysis show that suppression of trypanosomiasis leads to increased cattle and human populations and to increased agricultural development. However, in the absence of sound management, these changes will lead to a decline in pasture quality and increase the risk from tick-borne diseases in cattle and malaria in humans that would threaten system sustainability and resilience. The analysis of these conflicting outcomes of trypanosomiasis suppression is used to illustrate the need for and utility of conceptual bioeconomic models to serve as a basis for developing policy for sustainable agropastoral resource management in sub-Saharan Africa.

A regional analysis of weather mediated competition between a parasitoid and a coccinellid predator of oleander scale.

The regulation of an asexual population of the oleander scale [Aspidiotus nerii Bouchè (Hemiptera: Diaspididae)] on California bay tree [Umbellularia californica (Hopk. & Arn.) Nut.] by two natural enemies; an idiobiont, ectoparasitoid Aphytis chilensis Howard (Hymenoptera: Aphelinidae) and a coccinellid predator (Rhysobius lophanthae (Blaisd.) (Coleoptera: Coccinellidae), was examined using a general weather-driven, tri-trophic, physiologically based age-mass structured demographic model. The model is of intermediate complexity and was parameterized using extensive laboratory data and field observations from Albany, CA. Temperature-dependent physiological indices were estimated from the laboratory data and used to scale per capita growth, fecundity and survivorship rates from maximal values in a time varying environment. The tri-trophic model was integrated in a GIS (geographic information system) and the species dynamics examined across years and across the ecological zones of California. Field data and simulation results suggested the coccinellid predator was the most important regulating agent of oleander scale in the mild climate of Albany. However, multiple linear regression analysis of simulation data across all ecological zones of California shows that the parasitoid A. chilensis is the most important agent in suppressing oleander scale densities in warmer climates, while the predator R. lophanthae increases scale density an average of 9.7% across all regions.

A regional analysis of weather mediated competition between a parasitoid and a coccinellid predator of oleander scale

The regulation of an asexual population of the oleander scale [Aspidiotus nerii Bouchè (Hemiptera: Diaspididae)] on California bay tree [Umbellularia californica (Hopk. & Arn.) Nut.] by two natural enemies; an idiobiont, ectoparasitoid Aphytis chilensis Howard (Hymenoptera: Aphelinidae) and a coccinellid predator (Rhysobius lophanthae (Blaisd.) (Coleoptera: Coccinellidae), was examined using a general weather-driven, tri-trophic, physiologically based age-mass structured demographic model. The model is of intermediate complexity and was parameterized using extensive laboratory data and field observations from Albany, CA. Temperature-dependent physiological indices were estimated from the laboratory data and used to scale per capita growth, fecundity and survivorship rates from maximal values in a time varying environment. The tri-trophic model was integrated in a GIS (geographic information system) and the species dynamics examined across years and across the ecological zones of California. Field data and simulation results suggested the coccinellid predator was the most important regulating agent of oleander scale in the mild climate of Albany. However, multiple linear regression analysis of simulation data across all ecological zones of California shows that the parasitoid A. chilensis is the most important agent in suppressing oleander scale densities in warmer climates, while the predator R. lophanthae increases scale density an average of 9.7 percent across all regions.

O controle natural de uma população assexuada de cochonilha-da-espirradeira (Aspidiotus nerii Bouchè (Hemiptera: Diaspididae) em plantas do louro da California [Umbellularia californica (Hopk. & Arn.) Nut.] por dois de seus inimigos naturais [Aphytis chilensis Howard (Hymenoptera: Aphelinidae) e Rhysobius lophanthae (Blaisd.) (Coleoptera: Coccinellidae)] foi examinado em meio-ambiente com mudanças climáticas usando um modelo geral tritrófico, com base em estrutura populacional de idade e massa e acionado por condições climáticas. O modelo é de complexidade intermediária e os parâmetros foram obtidos através de extensas observações de laboratório e campo em Albany, Califórnia. índices fisiológicos dependentes de temperatura foram desenvolvidos a partir de dados de laboratório e usados para modificar o crescimento per capita, fecundidade e taxas de sobrevivência dos valores máximos. O modelo tritrófico foi integrado em GIS (sistema geográfico de informação ) e a dinâmica das espécies foi examinada através dos anos e das zonas ecológicas da Califórnia. Dados de campo e resultados de modelagem sugerem que o coccinelídeo predador é o agente mais importante no controle da cochonilha-da-espirradeira no clima ameno de Albany. Entretanto, a análise de regressão linear multivariada de dados de simulações regionais, demonstra que, sob altas temperaturas, A. chilensis é o fator mais importante suprimindo as densidades da cochonilha, enquanto o predador R. lophanthae é mais eficiente em regiões mais frias. A presença de R. lophanthae aumenta a densidade da cochonilha 9,7 por cento em média através das zonas ecológicas da Califórnia.