Orthoptera-specific target enrichment (OR-TE) probes resolve relationships over broad phylogenetic scales.

Phylogenomic data are revolutionizing the field of insect phylogenetics. One of the most tenable and cost-effective methods of generating phylogenomic data is target enrichment, which has resulted in novel phylogenetic hypotheses and revealed new insights into insect evolution. Orthoptera is the most diverse insect order within polyneoptera and includes many evolutionarily and ecologically interesting species. Still, the order as a whole has lagged behind other major insect orders in terms of transitioning to phylogenomics. In this study, we developed an Orthoptera-specific target enrichment (OR-TE) probe set from 80 transcriptomes across Orthoptera. The probe set targets 1828 loci from genes exhibiting a wide range of evolutionary rates. The utility of this new probe set was validated by generating phylogenomic data from 36 orthopteran species that had not previously been subjected to phylogenomic studies. The OR-TE probe set captured an average of 1037 loci across the tested taxa, resolving relationships across broad phylogenetic scales. Our detailed documentation of the probe design and bioinformatics process is intended to facilitate the widespread adoption of this tool.

Annual temperature, body size, and sexual size dimorphism in the evolution of Pyrgomorphidae.

In many animal species, larger body size is positively correlated with male mating success and female fecundity. However, in the case of insects, in high seasonality environments, natural selection favors a faster maturation that decreases the risk of pre-reproductive death. However, this advantageous adaptation comes at a tradeoff, resulting in a reduction in body size. Maturation time is influenced by environmental factors, such as temperature, season length, and food availability during the rains. The geographic variation in these parameters provides an opportunity to study their impact on the adaptive evolution of body size in Pyrgomorphidae grasshoppers. These grasshoppers exhibit remarkable variation in body size and wing development and can be found in diverse plant communities across Africa, Asia, Australia, and tropical America. In this study, we utilized a phylogenetic approach to examine the evolution of body size, considering climatic factors, and the influence of sexual selection on size differences between males and females. We found a positive correlation between mean annual temperature and sexual size dimorphism (SSD). Remarkably, species exhibiting a strong bias toward larger females were found to be adapted to regions with higher temperatures. In the Pyrgomorphidae family, an intermediate body size was identified as the ancestral trait. Additionally, winged male and female grasshoppers were observed to be larger than their wingless counterparts. Despite the potential conflicting pressures on body size in males and females, these grasshoppers adhere to Rench's Rule, suggesting that sexual selection on males' body size may explain the evolution of SSD.

Phylogeny and biogeography of the wingless orthopteran family Rhaphidophoridae.

Cave crickets (Rhaphidophoridae) are insects of an ancient and wingless lineage within Orthoptera that are distributed worldwide except in Antarctica, and each subfamily has a high level of endemicity. Here, we show the comprehensive phylogeny of cave crickets using multi-gene datasets from mitochondrial and nuclear loci, including all extant subfamilies for the first time. We reveal phylogenetic relationships between subfamilies, including the sister relationship between Anoplophilinae and Gammarotettiginae, based on which we suggest new synapomorphies. Through biogeographic analyses based on divergence time estimations and ancestral range reconstruction, we propose novel hypotheses regarding the biogeographic history of cave crickets. We suggest that Gammarotettiginae in California originated from the Asian lineage when Asia and the Americas were connected by the Bering land bridge, and the opening of the western interior seaway affected the division of Ceuthophilinae from Tropidischiinae in North America. We estimate that Rhaphidophoridae originated at 138 Mya throughout Pangea. We further hypothesize that the loss of wings in Rhaphidophoridae could be the result of their adaptation to low temperatures in the Mesozoic era.

Surface modification of polypropylene hollow fiber membranes using fluorosilane for CO2 absorption in a gas-liquid membrane contactor.

Conventional methods for improving the hydrophobicity of polypropylene (PP) membranes to prevent wetting phenomena require complex pretreatment procedures in order to activate the surface for enabling the reaction with fluorosilane (FS)-based materials. This study successfully prepared PP membrane contactors with enhanced hydrophobicity through a simple single-step dip-coating method using perfluoroether-grafted silanes for CO2 capture. The FS coating layer on the PP membrane surface was confirmed through ATR-FTIR spectroscopy, XPS, FE-SEM, and EDS. Furthermore, the evaluation of the CO2 absorption performance and long-term stability of the FS-coated PP membrane according to the variation of the gas flow rate (50, 100, 200, 400, and 800 mL/min) confirmed the superior chemical stability and durability of our membranes to those of previously reported hydrophobic membranes. The as-prepared FS-coated PP membrane expands the application scope of gas-liquid membrane contactors for CO2 capture from the flue gas of coal-fired power plants.

Differential responses to double-stranded RNA injection and feeding in Mormon cricket (Orthoptera: Tettigoniidae).

The Mormon cricket, Anabrus simplex, is a flightless katydid, one of the major devastating rangeland pests in several states of the western United States. During the past few years, their sudden and periodic outbreaks into massive migratory bands caused significant economic losses to the rangeland forage and agricultural crops, particularly grain crops. Current population management methods rely heavily on broad-spectrum chemical insecticides, which could be toxic to nontargets, and even the targeted species might develop resistance in the long run. Therefore, we assessed the potential of RNA interference (RNAi)-based alternative management strategies that could supplement the current methods. In insects, RNAi efficiency varies with the method of double-stranded RNA (dsRNA) delivery. We tested 2 different methods of dsRNA delivery: injection and oral feeding of dsRNA. The results showed that Mormon crickets are sensitive to injection of dsRNA in a dose-dependent manner, but refractory to the oral feeding of dsRNA. Further, we confirmed the high nuclease activity in the insect midgut. In order to protect the dsRNA from the dsRNase activity and facilitate its uptake in the midgut, we encapsulated dsRNA inside poly lactic-co-glycolic acid (PLGA) nanoparticles and studied its release kinetics and RNAi efficiency by oral feeding. The release kinetics clearly suggested that the PLGA nanoparticle permeates from the insect digestive system to the hemolymph; however, it failed to induce an efficient RNAi response of the targeted genes. In conclusion, our findings suggest the different responses to dsRNA delivery methods in Mormon crickets, and further investigations involving dsRNA stability and its uptake mechanism are required to use RNAi as an alternative Mormon cricket population management strategy.

Detection of Abnormal Changes on the Dorsal Tongue Surface Using Deep Learning.

Background and Objective: The tongue mucosa often changes due to various local and systemic diseases or conditions. This study aimed to investigate whether deep learning can help detect abnormal regions on the dorsal tongue surface in patients and healthy adults. Materials and Methods: The study collected 175 clinical photographic images of the dorsal tongue surface, which were divided into 7782 cropped images classified into normal, abnormal, and non-tongue regions and trained using the VGG16 deep learning model. The 80 photographic images of the entire dorsal tongue surface were used for the segmentation of abnormal regions using point mapping segmentation. Results: The F1-scores of the abnormal and normal classes were 0.960 (precision: 0.935, recall: 0.986) and 0.968 (precision: 0.987, recall: 0.950), respectively, in the prediction of the VGG16 model. As a result of evaluation using point mapping segmentation, the average F1-scores were 0.727 (precision: 0.717, recall: 0.737) and 0.645 (precision: 0.650, recall: 0.641), the average intersection of union was 0.695 and 0.590, and the average precision was 0.940 and 0.890, respectively, for abnormal and normal classes. Conclusions: The deep learning algorithm used in this study can accurately determine abnormal areas on the dorsal tongue surface, which can assist in diagnosing specific diseases or conditions of the tongue mucosa.

Reversible sulfur dioxide capture by amino acids containing a single amino group at low sulfur dioxide concentrations.

SO2, an air pollutant, is harmful to human health and causes air pollution; therefore, numerous studies have focused on the development of SO2 control technologies. Although limestone- and ammonia-based absorbents have been widely used in wet desulfurization, they are difficult to regenerate and do not enable the recycling of SO2, which is a useful resource. Recently, amino acids have attracted attention as reversible SO2 absorbents because they are eco-friendly and have excellent reactivity with SO2, as well as high regeneration performance. Glycine, L-alanine, ß-alanine, 4-aminobutyric acid, 5-aminovaleric acid, and 6-aminohexanoic acid were analyzed to investigate the relationship between SO2 absorption and the amino acid molecular structure using the simulated actual flue gas (200 ppmv SO2 + 13% CO2 in N2 balance). The SO2 absorption of amino acids (with the molecular structure of glycine and alkyl chains of various lengths) improved as the alkyl chain length increased, possibly owing to a decrease in the inductive effect in the molecular structure of the amino acid. Furthermore, 13C-nuclear magnetic resonance spectroscopy was conducted to analyze the SO2 absorption reaction mechanism (including the possible generation of irreversible species), and experiments involving a number of consecutive absorption-desorption cycles were used to confirm the reusability of the amino acids. The tested amino acids exhibited higher cyclic capacities compared to those of deep eutectic solvents and ionic liquids reported in the literature, thereby exhibiting excellent potential as SO2 absorbents. Thus, this study can guide the future design and development of eco-friendly SO2 absorbents.

The time course of behavioural phase change in the Central American locust Schistocerca piceifrons.

Locusts exhibit an extreme form of phenotypic plasticity and can exist as two alternative phenotypes, known as solitarious and gregarious phases. These phases, which can transform from one to another depending on local population density, show distinctly different behavioural characteristics. The proximate mechanisms of behavioural phase polyphenism have been well studied in the desert locust Schistocerca gregaria and the migratory locust Locusta migratoria, and what is known in these species is often treated as a general feature of locusts. However, this approach might be flawed, given that there are approximately 20 locust species that have independently evolved phase polyphenism. Using the Central American locust Schistocerca piceifrons as a study system, we characterised the time course of behavioural phase change using standard locust behavioural assays, using both a logistic regression-based model and analyses of separate behavioural variables. We found that for nymphs of S. piceifrons, solitarisation was a relatively fast, two-step process, but that gregarisation was a much slower process. Additionally, the density of the gregarisation treatment seemed to have no effect on the rate of phase change. These data are at odds with what we know about the time course of behavioural phase change in S. gregaria, suggesting that the mechanisms of locust phase polyphenism in these two species are different and may not be phylogenetically constrained. Our study represents the most in-depth study of behavioural gregarisation and solitarisation in locusts to date.

Controlled Drug Release Using Chitosan-Alginate-Gentamicin Multi-Component Beads.

This study aimed to develop improved multi-component beads with controlled, sustained delivery of antibiotics, such as gentamicin (GM). Antibiotic-loaded beads with rapid-release and the sustained-release system can be used for bone restoration. Single and multi-component beads were prepared by gelation using various combinations of chitosan and calcium chloride as cationic components and alginate and citric acid as anions. GM release was also controlled by crosslinking using citric acid. The optimum beads were obtained using 5% or 2% sodium alginate, 3% chitosan, and 0.1 mol/L citric acid. The beads were characterized by FTIR, TG-DTG, swelling behavior, and SEM. All GM-loaded beads revealed good antimicrobial activity. The rate and kinetics of release in the phosphate buffer solution were controlled by changing the amount of chitosan in the calcium chloride solution and using citric acid as the crosslinking agent. Crosslinked beads were prepared for the release of about 80% of the loaded drug within 24 h. The study concluded that the chitosan-alginate beads provided faster GM release but crosslinking with citric acid was efficient for sustained-release beads containing gentamicin.

Microstructure and Electrochemical Characterization of Ti-Sn Binary Alloys for Dental Applications.

This work investigated the microstructure and corrosion behaviors of Ti-Sn alloys with x wt% Sn (x = 5, 10, 15, and 20) for dental applications. The microstructures of commercially pure titanium (cp-Ti) and Ti-Sn alloys were characterized by X-ray diffractometry, optical microscopy, and transmission electron microscopy. The Vickers hardness of the Ti-Sn alloys was compared to that of cp-Ti. The corrosion behaviors of Ti-Sn alloys were tested in 0.9% NaCl solution at 37 °C using open circuit potential, potentiodynamic polarization, AC impedance, and galvanic corrosion tests. Ti-Sn alloys had a hexagonal close-packed structure and their microstructures were transformed from the equiaxed structure with irregular grain boundaries to the martensitic structure as the content of Sn in Ti-Sn alloys increased by over 15 wt%. Among the sample groups, Ti-15Sn and Ti-20Sn alloys exhibited better Vickers hardness values. Ti-Sn alloys had better corrosion resistance than cp-Ti. Ti-15Sn, which showed narrow martensitic bands, exhibited the highest corrosion resistance properties in AC impedance measurements due to its higher resistance and better capacitive parameters. Among the tested groups, the galvanic coupling of Ti-15Sn with cp-Ti showed higher corrosion potentials and lower current densities, which indicates that there was higher corrosion resistance.

Development of RNAi Methods for the Mormon Cricket, Anabrus simplex (Orthoptera: Tettigoniidae).

Mormon crickets are a major rangeland pest in the western United States and are currently managed by targeted applications of non-specific chemical insecticides, which can potentially have negative effects on the environment. In this study, we took the first steps toward developing RNAi methods for Mormon crickets as a potential alternative to traditional broad-spectrum insecticides. To design an effective RNAi-based insecticide, we first generated a de novo transcriptome for the Mormon cricket and developed dsRNAs that could silence the expression of seven housekeeping genes. We then characterized the RNAi efficiencies and time-course of knockdown using these dsRNAs, and assessed their ability to induce mortality. We have demonstrated that it is possible to elicit RNAi responses in the Mormon cricket by injection, but knockdown efficiencies and the time course of RNAi response varied according to target genes and tissue types. We also show that one of the reasons for the poor knockdown efficiencies could be the presence of dsRNA-degrading enzymes in the hemolymph. RNAi silencing is possible in Mormon cricket, but more work needs to be done before it can be effectively used as a population management method.

Microwave Treatment of Calcium Phosphate/Titanium Dioxide Composite to Improve Protein Adsorption.

Calcium phosphate has attracted enormous attention as a bone regenerative material in biomedical fields. In this study, we investigated the effect of microwave treatment on calcium phosphate deposited TiO2 nanoflower to improve protein adsorption. Hierarchical rutile TiO2 nanoflowers (TiNF) fabricated by a hydrothermal method were soaked in modified simulated body fluid for 3 days to induce calcium phosphate (CAP) formation, followed by exposure to microwave radiation (MW). Coating the dental implants with CAP/TiNF provides a means of improving the biological properties, as the structure, morphology, and thickness of the composites can be controlled. The composites were characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD), field emission transmission electron microscopy (TEM), and Fourier-transform infrared spectroscopy (FTIR), respectively. The composites were identified to be composed of aggregated nano-sized particles with sphere-like shapes, and the calcium phosphate demonstrated low crystallinity. The ability of bovine serum albumin (BSA) to adsorb on MW-treated CAP/TiNF composites was studied as a function of BSA concentration. The Sips isotherm was used to analyze the BSA adsorption on MW-treated CAP/TiNF composites. The MW-treated samples showed high protein adsorption capacity, thereby indicating their potential in various biomedical applications.

Auditory mechanics in the grig (Cyphoderris monstrosa): tympanal travelling waves and frequency discrimination as a precursor to inner ear tonotopy.

Ensiferan orthopterans offer a key study system for acoustic communication and the process of insect hearing. Cyphoderris monstrosa (Hagloidea) belongs to a relict ensiferan family and is often used for evolutionary comparisons between bushcrickets (Tettigoniidae) and their ancestors. Understanding how this species processes sound is therefore vital to reconstructing the evolutionary history of ensiferan hearing. Previous investigations have found a mismatch in the ear of this species, whereby neurophysiological and tympanal tuning does not match the conspecific communication frequency. However, the role of the whole tympanum in signal reception remains unknown. Using laser Doppler vibrometry, we show that the tympana are tonotopic, with higher frequencies being received more distally. The tympana use two key modalities to mechanically separate sounds into two auditory receptor populations. Frequencies below approximately 8 kHz generate a basic resonant mode in the proximal end of the tympanum, whereas frequencies above approximately 8 kHz generate travelling waves in the distal region. Micro-CT imaging of the ear and the presented data suggest that this tonotopy of the tympana drive the tonotopic mechanotransduction of the crista acustica (CA). This mechanism represents a functional intermediate between simple tuned tympana and the complex tonotopy of the bushcricket CA.

Taxonomic notes on the Australian skyhopper genus Kosciuscola Sjstedt (Orthoptera: Acrididae: Oxyinae).

The Australian skyhopper genus Kosciuscola Sjstedt consists of brachypterous species that inhabit the Australian alpine and subalpine region. The genus used to include 5 species and 1 subspecies, but according to a recent phylogenomic study, there could be as many as 14 species in the genus, that are genetically and geographically isolated from each other. This study represents the first step in describing and documenting the diversity of this interesting genus. In this study, we redefine the type species K. tristis, and elevate its subspecies K. tristis restrictus as a valid species on the basis of distinct morphological traits, geographical isolation, and phylogenomic evidence.

Studies in Mexican Grasshoppers: Four new species of Reyesacris Fontana, Buzzetti amp; Mario-Prez, 2011 (Orthoptera: Acrididae: Ommatolampidinae).

Four new species of Reyesacris Fontana, Buzzetti Mario-Prez, 2011 are described from Southern Mexico (Guerrero and Oaxaca States). Reyesacris zihua sp. nov., R. atoyacensis sp. nov., R. mephaa sp. nov. and R. tika sp. nov. A dichotomous key to species of Reyesacris and distribution map are provided together with an analysis of external and internal male genitalia to place this genus in the subtribe Vilernina within the tribe Ommatolampidini.

Albumin adsorption on microwave-treated titanium dioxide for dental implant materials.

Surface modification of biomedical implants is an established strategy to improve osseointegration. TiO2 nanoflowers (TNF) were deposited on a titanium substrate by hydrothermal technique followed by microwave radiation at 700 W for 5-15 min. Microwave-treated samples were analyzed by X-ray diffraction (XRD), scanning electron microscopy, and Raman spectroscopy to determine their physicochemical characteristics. The XRD and Raman results showed crystalline rutile TiO2. The surfaces of microwave-treated TNF were hydrophilic with split hierarchical structure. The Sips isotherm was used to analyze BSA adsorption on microwave treated TNF samples. The microwave treatment enhances protein adsorption capacity.

Motorized Treadmill and Optical Recording System for Gait Analysis of Grasshoppers.

(1) Background: Insects, which serve as model systems for many disciplines with their unique advantages, have not been extensively studied in gait research because of the lack of appropriate tools and insect models to properly study the insect gaits. (2) Methods: In this study, we present a gait analysis of grasshoppers with a closed-loop custom-designed motorized insect treadmill with an optical recording system for quantitative gait analysis. We used the eastern lubber grasshopper, a flightless and large-bodied species, as our insect model. Gait kinematics were recorded and analyzed by making three grasshoppers walk on the treadmill with various speeds from 0.1 to 1.5 m/s. (3) Results: Stance duty factor was measured as 70-95% and decreased as walking speed increased. As the walking speed increased, the number of contact legs decreased, and diagonal arrangement of contact was observed at walking speed of 1.1 cm/s. (4) Conclusions: This pilot study of gait analysis of grasshoppers using the custom-designed motorized insect treadmill with the optical recording system demonstrates the feasibility of quantitative, repeatable, and real-time insect gait analysis.

Cytogenomic analysis unveils mixed molecular evolution and recurrent chromosomal rearrangements shaping the multigene families on Schistocerca grasshopper genomes.

Multigene families are essential components of eukaryotic genomes and play key roles either structurally and functionally. Their modes of evolution remain elusive even in the era of genomics, because multiple multigene family sequences coexist in genomes, particularly in large repetitive genomes. Here, we investigate how the multigene families 18S rDNA, U2 snDNA, and H3 histone evolved in 10 species of Schistocerca grasshoppers with very large and repeat-enriched genomes. Using sequenced genomes and fluorescence in situ hybridization mapping, we find substantial differences between species, including the number of chromosomal clusters, changes in sequence abundance and nucleotide composition, pseudogenization, and association with transposable elements (TEs). The intragenomic analysis of Schistocerca gregaria using long-read sequencing and genome assembly unveils conservation for H3 histone and recurrent pseudogenization for 18S rDNA and U2 snDNA, likely promoted by association with TEs and sequence truncation. Remarkably, TEs were frequently associated with truncated copies, were also among the most abundant in the genome, and revealed signatures of recent activity. Our findings suggest a combined effect of concerted and birth-and-death models driving the evolution of multigene families in Schistocerca over the last 8 million years, and the occurrence of intra- and interchromosomal rearrangements shaping their chromosomal distribution. Despite the conserved karyotype in Schistocerca, our analysis highlights the extensive reorganization of repetitive DNAs in Schistocerca, contributing to the advance of comparative genomics for this important grasshopper genus.

Comparative analysis of phenotypic plasticity sheds light on the evolution and molecular underpinnings of locust phase polyphenism.

Locusts exhibit one of nature's most spectacular examples of complex phenotypic plasticity, in which changes in density cause solitary and cryptic individuals to transform into gregarious and conspicuous locusts forming large migrating swarms. We investigated how these coordinated alternative phenotypes might have evolved by studying the Central American locust and three closely related non-swarming grasshoppers in a comparative framework. By experimentally isolating and crowding during nymphal development, we induced density-dependent phenotypic plasticity and quantified the resulting behavioural, morphological, and molecular reaction norms. All four species exhibited clear plasticity, but the individual reaction norms varied among species and showed different magnitudes. Transcriptomic responses were species-specific, but density-responsive genes were functionally similar across species. There were modules of co-expressed genes that were highly correlated with plastic reaction norms, revealing a potential molecular basis of density-dependent phenotypic plasticity. These findings collectively highlight the importance of studying multiple reaction norms from a comparative perspective.

Molecular phylogeny of the grasshopper family Pyrgomorphidae (Caelifera, Orthoptera) reveals rampant paraphyly and convergence of traditionally used taxonomic characters.

The grasshopper family Pyrgomorphidae is one of the most colorful orthopteran lineages, and includes biologically fascinating and culturally important species. Recent attempts to reconstruct the phylogeny of this family have resulted in a large degree of conflicts between a morphology-based study and a molecular-based study, mainly due to convergent morphological traits that affected phylogenetic reconstruction. In this study, a molecular phylogeny of Pyrgomorphidae based on 32 ingroup species and mitochondrial genome data is proposed, which is used to test the monophyly of the taxonomic groupings used in the current classification scheme. Using the ancestral character state reconstruction analyses and character mapping, we demonstrate that some of the morphological characters, including the male genitalia, which were considered to be taxonomically important, have evolved convergently across the phylogeny. We discuss the discrepancies between our phylogeny and the previous studies and propose an approach to establish a natural classification scheme for Pyrgomorphidae.