Synthesis and Photochemical Properties of Fluorescent Metabolites Generated from Fluorinated Benzoylmenadiones in Living Cells.

This work describes the reactivity and properties of fluorinated derivatives (F-PD and F-PDO) of plasmodione (PD) and its metabolite, the plasmodione oxide (PDO). Introduction of a fluorine atom on the 2-methyl group markedly alters the redox properties of the 1,4-naphthoquinone electrophore, making the compound highly oxidizing and particularly photoreactive. A fruitful set of analytical methods (electrochemistry, absorption and emission spectrophotometry, and HRMS-ESI) have been used to highlight the products resulting from UV photoirradiation in the absence or presence of selected nucleophiles. With F-PDO and in the absence of nucleophile, photoreduction generates a highly reactive ortho-quinone methide (o-QM) capable of leading to the formation of a homodimer. In the presence of thiol nucleophiles such as ß-mercaptoethanol, which was used as a model, o-QMs are continuously regenerated in sequential photoredox reactions generating mono- or disulfanylation products as well as various unreported sulfanyl products. Besides, these photoreduced adducts derived from F-PDO are characterized by a bright yellowish emission due to an excited-state intramolecular proton transfer (ESIPT) process between the dihydronapthoquinone and benzoyl units. In order to evidence the possibility of an intramolecular coupling of the o-QM intermediate, a synthetic route to the corresponding anthrones is described. Tautomerization of the targeted anthrones occurs and affords highly fluorescent stable hydroxyl-anthraquinones. Although probable to explain the intense visible fluorescence emission also observed in tobacco BY-2 cells used as a cellular model, these coupling products have never been observed during the photochemical reactions performed in this study. Our data suggest that the observed ESIPT-induced fluorescence most likely corresponds to the generation of alkylated products through reduction species, as demonstrated with the ß-mercaptoethanol model. In conclusion, F-PDO thus acts as a novel (pro)-fluorescent probe for monitoring redox processes and protein alkylation in living cells.

Towards a Light-mediated Gene Therapy for the Eye using Caged Ethinylestradiol and the Inducible Cre/lox System.

Increasingly, retinal pathologies are being treated with virus-mediated gene therapies. To be able to target viral transgene expression specifically to the pathological regions of the retina with light, we established an in vivo photoactivated gene expression paradigm for retinal tissue. Based on the inducible Cre/lox system, we discovered that ethinylestradiol is a suitable alternative to Tamoxifen as ethinylestradiol is more amenable to modification with photosensitive protecting compounds, i.e., "caging." Identification of ethinylestradiol as a ligand for the mutated human estradiol receptor was supported by in silico binding studies showing the reduced binding of caged ethinylestradiol. Caged ethinylestradiol was injected into the eyes of double transgenic GFAP-CreERT2 mice with a Cre-dependent tdTomato reporter transgene followed by irradiation with light of 450 nm. Photoactivation significantly increased retinal tdTomato expression compared to controls. We thus demonstrated a first step towards the development of a targeted, light-mediated gene therapy for the eyes.

Photoactivatable Liposomes for Blue to Deep Red Light-Activated Surface Drug Release: Application to Controlled Delivery of the Antitumoral Drug Melphalan.

Liposome-based nanoparticles able to release, via a photolytic reaction, a payload anchored at the surface of the phospholipid bilayer were prepared. The liposome formulation strategy uses an original drug-conjugated blue light-sensitive photoactivatable coumarinyl linker. This is based on an efficient blue light-sensitive photolabile protecting group modified by a lipid anchor, which enables its incorporation into liposomes, leading to blue to green light-sensitive nanoparticles. In addition, the formulated liposomes were doped with triplet-triplet annihilation upconverting organic chromophores (red to blue light) in order to prepare red light sensitive liposomes able to release a payload, by upconversion-assisted photolysis. Those light-activatable liposomes were used to demonstrate that direct blue or green light photolysis or red light TTA-UC-assisted drug photolysis can effectively photorelease a drug payload (Melphalan) and kill tumor cells in vitro after photoactivation.

Adaptive Introgression across Semipermeable Species Boundaries between Local Helicoverpa zea and Invasive Helicoverpa armigera Moths.

Hybridization between invasive and native species has raised global concern, given the dramatic increase in species range shifts and pest outbreaks due to anthropogenic dispersal. Nevertheless, secondary contact between sister lineages of local and invasive species provides a natural laboratory to understand the factors that determine introgression and the maintenance or loss of species barriers. Here, we characterize the early evolutionary outcomes following secondary contact between invasive Helicoverpa armigera and native H. zea in Brazil. We carried out whole-genome resequencing of Helicoverpa moths from Brazil in two temporal samples: during the outbreak of H. armigera in 2013 and 2017. There is evidence for a burst of hybridization and widespread introgression from local H. zea into invasive H. armigera coinciding with H. armigera expansion in 2013. However, in H. armigera, the admixture proportion and the length of introgressed blocks were significantly reduced between 2013 and 2017, suggesting selection against admixture. In contrast to the genome-wide pattern, there was striking evidence for adaptive introgression of a single region from the invasive H. armigera into local H. zea, including an insecticide resistance allele that increased in frequency over time. In summary, despite extensive gene flow after secondary contact, the species boundaries are largely maintained except for the single introgressed region containing the insecticide-resistant locus. We document the worst-case scenario for an invasive species, in which there are now two pest species instead of one, and the native species has acquired resistance to pyrethroid insecticides through introgression.

Role of nutritional composition in the development and survival of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) on artificial diet and natural hosts.

Helicoverpa armigera is a pest of several crops causing significant economic impact. We evaluated the insect development on different vegetative and reproductive structures of cotton, maize, and soybean compared to artificial diet. One hundred individuals were evaluated per structure (cotton leaves and bolls; maize leaves, grains, and silk; soybean leaves and pods) and artificial diet. Centesimal analyses were performed on quantifiable nutrient contents in diets. The viability of immatures (eggs, larvae, and pupae) ranged from 30% on maize leaf to 74% on cotton bolls, while on the artificial diet, it was 70%. Maize, cotton, and soybean leaves provided viability of 30, 37, and 42%, respectively, revealing these leaves tissues are less favorable to the development of H. armigera immatures compared to 'reproductive tissues'. Centesimal composition of diets compared 14 common components in all diets, which correlated significantly with larval and pupal stages and/or pupal weight. Of the 12 dietary components that significantly affected larval development time, half were negatively correlated, indicating a decrease in developmental time from their increments. In general, when insects were confined separately to substrates, the artificial diet was the most suitable for H. armigera development compared to the evaluated natural diets. However, in natural conditions, the variability of available hosts must be considered. In addition, it is acceptable for moths to select more suitable hosts for oviposition, while their larvae move to other more suitable tissues of the same plant or even migrate to other plants.

On the permeation of large organic cations through the pore of ATP-gated P2X receptors.

Pore dilation is thought to be a hallmark of purinergic P2X receptors. The most commonly held view of this unusual process posits that under prolonged ATP exposure the ion pore expands in a striking manner from an initial small-cation conductive state to a dilated state, which allows the passage of larger synthetic cations, such as N-methyl-d-glucamine (NMDG+). However, this mechanism is controversial, and the identity of the natural large permeating cations remains elusive. Here, we provide evidence that, contrary to the time-dependent pore dilation model, ATP binding opens an NMDG+-permeable channel within milliseconds, with a conductance that remains stable over time. We show that the time course of NMDG+ permeability superimposes that of Na+ and demonstrate that the molecular motions leading to the permeation of NMDG+ are very similar to those that drive Na+ flow. We found, however, that NMDG+ "percolates" 10 times slower than Na+ in the open state, likely due to a conformational and orientational selection of permeating molecules. We further uncover that several P2X receptors, including those able to desensitize, are permeable not only to NMDG+ but also to spermidine, a large natural cation involved in ion channel modulation, revealing a previously unrecognized P2X-mediated signaling. Altogether, our data do not support a time-dependent dilation of the pore on its own but rather reveal that the open pore of P2X receptors is wide enough to allow the permeation of large organic cations, including natural ones. This permeation mechanism has considerable physiological significance.

The Granulate Cutworm (Lepidoptera: Noctuidae): Biological Parameters Under Controlled Conditions, Host Plants, and Distribution in the Americas.

Feltia subterranea (Fabricius), commonly known as the granulate cutworm, is a common species of owlet moths (Noctuidae) of major agricultural importance, widely distributed in Nearctic and Neotropical regions. This study was conducted to determine the species biological parameters, gather information about its larval host plants, and assess the agricultural significance of this species in the Americas. The viability of the egg, larval, pupal stages, and prepupal period was 98, 98, and 100%, respectively, under laboratory conditions. The average duration of the egg, larval, pupal stages, and prepupal period was 3, 17, 4, and 13 d, respectively. All laboratory-reared larvae developed through five instars. The growth ratio was 1.93 for females and 1.85 for males. The duration of the larval stage was significantly longer in females than in males from the fourth instar. The duration of the pupal stage was significantly shorter in females than in males. When larval and pupal stage durations were combined, there were no significant differences in total development time as a function of sex. In total, 159 botanical taxa belonging to 41 families were recorded as host species for F. subterranea. The families with the greatest number of host species were Fabaceae (22), Poaceae (19), Asteraceae (16), Brassicaceae (13), Solanaceae (12), Amaranthaceae (7), Cucurbitaceae (7), and Malvaceae (5). It is noteworthy that the large number of native weeds used by F. subterranea as host plants could represent a significant source of infestation of crops in the agricultural landscape.

First record of Utetheisa pulchella (Linnaeus, 1758) (Lepidoptera: Erebidae: Arctiinae) in Brazilian Amazon: implications for conservation.

The crimson speckled moth, Utetheisa pulchella (Linnaeus, 1758), is recorded in Brazilian Amazon for the first time. A moth (male) was collected with light trap in a multiple cropping area, in Mojuí dos Campos municipality, Pará State, Brazil. This record expands the area of occurrence and distribution of this rare species in the Neotropical Region. In addition, the occurrence of U. pulchella in the Amazon was unexpected, since this species is generally associated with open ecosystems and drier climates.

Life-History Parameters of Striacosta albicosta (Lepidoptera: Noctuidae) Under Laboratory Conditions.

Striacosta albicosta (Smith) is a key pest of maize and dry beans in North America. It has expanded its distribution from the western Great Plains of the United States to the Great Lakes region in the United States and Canada. There has been limited research on the baseline biological aspects of this insect under controlled conditions. The objective of this study was to detail the biological parameters of S. albicosta feeding on an artificial diet under laboratory conditions. Overall survival from neonate to adult at 26.6 ± 1°C was 36.72% and the total developmental time was approximately 110 d. Survival of the egg, larval, prepupal, and pupal stages were 75.71, 98.50, 51.78, and 95.10%, respectively. Average duration of the egg, larval, prepupal, and pupal stages was 4.64, 28.20, 41.50, and 25.91 d, respectively. During the larval stage, 92.50% of larvae developed through seven instars and the remaining through six instars. Larvae that developed through six and seven instars exhibited a mean growth ratio of 1.60 and 1.47, respectively; however, there was no difference in pupal weight. Eggs laid by field-mated moths showed a fertility of 75.71%, compared with 4.18% from laboratory-reared moths. These data suggest that S. albicosta develop primarily through seven instars and the most vulnerable developmental stage is the prepupa. Laboratory conditions strongly affected fertility success. Information presented here greatly expands our understanding of S. albicosta biology, which can be used to improve the efficiency of laboratory bioassays and management techniques for this critical crop pest.

Biotic Potential and Life Tables of Chrysodeixis includens (Lepidoptera: Noctuidae), Rachiplusia nu, and Trichoplusia ni on Soybean and Forage Turnip.

Loopers such as Chrysodeixis includens (Walker), Rachiplusia nu (Guenée), and Trichoplusia ni (Hübner) are important defoliators in soybean, sunflower, and crucifer crops, respectively, in countries of the Americas. The biotic potential of these polyphagous species of Plusiinae was comparatively examined considering crop rotation and succession scenarios in which crucifer crops are cultivated during or after Brazilian winter. All the species developed and reproduced on soybean (BRS 133 Embrapa) and forage turnip (Cati AL 1000, Wolf Seeds do Brasil). The development of C. includens was similar on both host plants. The survival of R. nu was lower on forage turnip than on soybean. In contrast, T. ni performance (survival, fecundity, pupal weight) was better on forage turnip than on soybean. This suggests that in crop rotation and succession scenarios of soybean after brassicacea, C. includens is likely to have a higher number of generations per year and could be potentially more harmful.

Synthesis and Characterization of Photoactivatable Doxycycline Analogues Bearing Two-Photon-Sensitive Photoremovable Groups Suitable for Light-Induced Gene Expression.

We report the synthesis and photolytic properties of caged 9-aminodoxycycline derivatives modified with 2-{4'-bis-[2-(2methoxyethoxy)ethyl]-4-nitrobiphenyl-3-yl}prop-1-oxy (EANBP) and PEG7-ylated (7-diethylamino-2-oxo-2H-chromen-4-yl)methyl (PEG7-DEACM) groups. 9-Aminodoxycycline is a tetracycline analogue capable of activating transcription through the inducible TetOn transgene expression system and can be regioselectively coupled to two-photon-sensitive photo-removable protecting groups by carbamoylation. The EANBP-based caged 9-aminodoxycycline showed complex photochemical reactions but did release 10 % of 9-aminodoxycycline. However, 9-(PEG7-DEACMamino)doxycycline exhibited excellent photolysis efficiency at 405 nm with quantitative release of 9-aminodoxycycline and a 0.21 uncaging quantum yield. Thanks to the good two-photon sensitivity of the DEACM chromophore, 9-aminodoxycycline release by two-photon photolysis is possible, with calculated action cross-sections of up to 4.0 GM at 740 nm. Therefore, 9-(PEG7-DEACMamino)doxycycline represents a very attractive tool for the development of a light-induced gene expression method in living cells.

Phenological patterns of Spodoptera Guenée, 1852 (Lepidoptera: Noctuidae) is more affected by ENSO than seasonal factors and host plant availability in a Brazilian Savanna.

The identification of factors responsible for the population dynamics is fundamental for pest management, since losses can reach 18% of annual production. Besides regular seasonal environmental factors and crop managements, additional supra-annual meteorological phenomena can also affect population dynamics, although its relevance has been rarely investigated. Among crop pests, Spodoptera stands out due to its worldwide distribution, high degree of polyphagy, thus causing damages in several crops in the world. Aiming to distinguish the relevance of different factors shaping population dynamics of Spodoptera in an ecosystem constituted of dry and rainy seasons, the current study used circular statistics to identify phenological patterns and test if its population fluctuation is driven by El Niño-Southern Oscillation (ENSO) effect, seasonal meteorological parameters, and/or host plant availability. Samplings were done in an intercropping system, in the Brazilian Savanna, during the new moon cycles between July/2013 and June/2016. Species were recorded all year round, but demonstrated differently non-uniform distribution, being concentrated in different seasons of the year. Population fluctuations were mostly affected by the ENSO intensity, despite the contrasting seasonal meteorological variation or host plant availability in a 400-m radius. Studies involving the observation of supra-annual phenomena, although rare, reach similar conclusions in relation to Neotropical insect fauna. Therefore, it is paramount to have long-term sampling studies to obtain a more precise response of the pest populations towards the agroecosystem conditions.

Optical control of an ion channel gate.

The powerful optogenetic pharmacology method allows the optical control of neuronal activity by photoswitchable ligands tethered to channels and receptors. However, this approach is technically demanding, as it requires the design of pharmacologically active ligands. The development of versatile technologies therefore represents a challenging issue. Here, we present optogating, a method in which the gating machinery of an ATP-activated P2X channel was reprogrammed to respond to light. We found that channels covalently modified by azobenzene-containing reagents at the transmembrane segments could be reversibly turned on and off by light, without the need of ATP, thus revealing an agonist-independent, light-induced gating mechanism. We demonstrate photocontrol of neuronal activity by a light-gated, ATP-insensitive P2X receptor, providing an original tool devoid of endogenous sensitivity to delineate P2X signaling in normal and pathological states. These findings open new avenues to specifically activate other ion channels independently of their natural stimulus.

Three-Dimensional Control of DNA Hybridization by Orthogonal Two-Color Two-Photon Uncaging.

We successfully introduced two-photon-sensitive photolabile groups ([7-(diethylamino)coumarin-4-yl]methyl and p-dialkylaminonitrobiphenyl) into DNA strands and demonstrated their suitability for three-dimensional photorelease. To visualize the uncaging, we used a fluorescence readout based on double-strand displacement in a hydrogel and in neurons. Orthogonal two-photon uncaging of the two cages is possible, thus enabling complex scenarios of three-dimensional control of hybridization with light.

Abdominal macrochaetae of female Hylesia oratex Dyar, 1913 (Insecta: Lepidoptera: Saturniidae): external morphology and medical significance.

The representatives of the genus Hylesia Hübner, [1820] are significant among the medically important Lepidoptera. Adult females use abdominal setae to wrap and protect the eggs that remain for months in nature. These setae, in contact with human skin, may cause allergic reactions including swelling, itching and local erythema, known as lepidopterism. The morphology of the abdominal scales and setae from the female H. oratex Dyar, 1913 is herein described and aspects related to their medical significance are discussed. Portions of each abdominal segment were examined through a scanning electron microscope. Two types of scales without medical importance, and two types of setae with medical importance, classified as "true setae" and "modified setae" were found. The true setae, which are slightly fusiform and have radially arranged lateral projections, are responsible for the allergic reactions caused by skin penetration. The modified setae, which are larger, curved, with the median enlarged and serrated margins, can be responsible for the release of chemical substances. This information provides a better understanding of the structure of the urticating setae, which are responsible for lepidopterism outbreaks in humans, and contributes towards the identification of the moth species involved.

A Polyurethane-Based Positive Photoresist.

Polyurethane (PU) monomer mixtures containing commercially available o-nitrobenzyl-based photocleavable monomers have been formulated and tested as low-cost positive tone photoresists. The photolysis reaction is studied by UV spectroscopy. Well-defined micropatterns on 2 µm thick photodegradable PU films are obtained using 365 nm light exposure. This strategy is also extended to improved formulations based on synthesized o-nitrobiphenylpropyl derivatives with enhanced photochemical properties for single photon excitation and high two-photon absorption cross-sections. Improved pattern resolution in 2D and the capability of 3D resolution using a scanning laser at 780 nm is demonstrated. This work demonstrates the potential of PUs as readily available, versatile, and easy-to-use photoresist materials for low-cost lithography applications.

Agonist trapped in ATP-binding sites of the P2X2 receptor.

ATP-gated P2X receptors are trimeric ion channels, as recently confirmed by X-ray crystallography. However, the structure was solved without ATP and even though extracellular intersubunit cavities surrounded by conserved amino acid residues previously shown to be important for ATP function were proposed to house ATP, the localization of the ATP sites remains elusive. Here we localize the ATP-binding sites by creating, through a proximity-dependent "tethering" reaction, covalent bonds between a synthesized ATP-derived thiol-reactive P2X2 agonist (NCS-ATP) and single cysteine mutants engineered in the putative binding cavities of the P2X2 receptor. By combining whole-cell and single-channel recordings, we report that NCS-ATP covalently and specifically labels two previously unidentified positions N140 and L186 from two adjacent subunits separated by about 18 Å in a P2X2 closed state homology model, suggesting the existence of at least two binding modes. Tethering reaction at both positions primes subsequent agonist binding, yet with distinct functional consequences. Labeling of one position impedes subsequent ATP function, which results in inefficient gating, whereas tethering of the other position, although failing to produce gating by itself, enhances subsequent ATP function. Our results thus define a large and dynamic intersubunit ATP-binding pocket and suggest that receptors trapped in covalently agonist-bound states differ in their ability to gate the ion channel.

Biotic potential, fertility and life table of Spodoptera albula (Walker) (Lepidoptera: Noctuidae), under controlled conditions.

This study aimed to evaluate the biotic potential, life table parameters and fertility of Spodoptera albula (Walker, 1857) under controlled conditions (25 ± 1°C, 70 ± 10% RH and 14 hour photo phase). The longevity, pre, post and oviposition periods, fecundity and fertility of 13 couples were evaluated. The longevity of females (13.500 days) was significantly higher than those of males (11.154 days). The mean durations of the pre, post and oviposition periods were 2.615, 1.769 and 9.385 days, respectively. The mean fecundity was 1.417.69 eggs and mean fertility was 1.340.401 larvae, per female. On average, females copulated 1, 231 times. A strong positive correlation was observed between the number of copulations and fecundity (r = 0.847, p <0.001), as well as a strong negative correlation between the number of copulations and the duration of the pre-oviposition period (r = -0.762, p = 0.002), and longevity (r = -0.788, p = 0.001). The biotic potential of S. albula was estimated at 8.768 x 1022 individuals / female / year. The net reproductive rate (Ro) was 353,904 times per generation and the mean generation time (T) was 37.187 days. The intrinsic rate of increase (rm) was 1,105, with a finite rate of increase (λ) of 3,019.

First report of Elaphria agrotina and Elaphria deltoides (Lepidoptera: Noctuidae: Elaphriini) feeding on maize.

This is the first report of Elaphria agrotina (Guenée, 1852) and Elaphria deltoides (Möschler, 1880) (Lepidoptera: Noctuidae) feeding on maize (Zea mays L.). The specimens were collected in maize fields during the crop season of 2012 and 2013 in three municipalities in Mato Grosso State, Brazil. Larvae were collected while feeding at the ear base, which often resulted in ears dropping to the ground. Larvae also were observed feeding on kernels in fallen ears. Ear injury often leads to reduced grain quality, and when the ears fall to the ground, reduced yield. A previous report of Striacosta albicosta (Smith, 1888) feeding on maize in Brazil was probably a misidentification of an E. agrotina male, which has wing pattern and coloration similarities with S. albicosta.

Immature stages of Spodoptera eridania (Lepidoptera: Noctuidae): developmental parameters and host plants.

This study aimed to detail the temporal and morphological parameters of the immature stages of southern armyworm Spodoptera eridania (Stoll, 1782) with larvae feed on artificial diet, under controlled conditions (25 ± 1°C, 70 ± 10% relative humidity and 14-h photophase) and gather information about their larval host plants. The viability of the egg, larval, pupal, and prepupal stages was 97.82, 93.62, 96.42, and 97.03%, respectively. The average duration of the egg, larval, pupal, and pre-pupal stages was 4.00, 16.18, 1.58, and 9.17 d, respectively. During the larval stage, 43.44% of females passed through seven instars, observing that the female's development was significant slower than males. The female larvae that developed through six and seven instars exhibited a mean growth rate of 1.52 and 1.44, respectively. Female pupae were significantly larger, exhibiting faster development than males. The rearing method proved to be adequate, providing more detailed observations of the biological cycle, especially at the larval stage, and resulting in an overall survival of almost 85%. Two hundred two plant species belonging to 58 families are listed as natural hosts for S. eridania, mainly including Asteraceae, Fabaceae, Solanaceae, Poaceae, Amaranthaceae, and Malvaceae.