Dual-Labeled Single Fluorescent Probes for the Simultaneous Two-Color Visualization of Dual Organelles and for Monitoring Cell Autophagy.

Dual-labeled single fluorescent probes are powerful tools for studying autophagy on the molecular scale, yet their development has been hampered by design complexity and a lack of valid strategies. Herein, for the first time, we introduce a combinatorial regulation strategy to fabricate dual-labeled probes for studying autophagy by integrating the specific organelle-targeting group and the functional fluorescence switch into a pentacyclic pyrylium scaffold (latent dual-target scaffold). For proof of concept, we prepared a range of dual-labeled probes (TMOs) that display different emission colors in duple organelles. In these probes, TMO1 and TMO2 enabled the simultaneous two-color visualization of the lysosomes and mitochondria. The other probes (TMO3 and TMO4) discriminatively targeted lysosomes/nucleolus and lysosomes/lipid droplets (LDs) with dual-color emission characteristics, respectively. Intriguingly, by simply connecting the endoplasmic reticulum (ER) targeting group to the pentacyclic pyrylium scaffold, we created the first dual-labeled probe TMO5 for simultaneously labeling lysosomes/ER in distinctive fluorescent colors. Subsequently, using the dual-labeled probe TMO2, drug-induced mitophagy was successfully recorded by evaluating the alterations of multiple mitophagy-related parameters, and the mitophagy defects in a cellular model of Parkinson's disease (PD) were also revealed by simultaneous dual-color/dual-organelle imaging. Further, the probe TMO4 can track the movement of lysosomes and LDs in real time and monitor the dynamic process of lipophagy. Therefore, this work not only presents attractive dual-labeled probes to promote the study of organelle interactions during autophagy but also provides a promising combinatorial regulation strategy that may be generalized for designing other dual-labeled probes with multiple organelle combinations.

Genome-wide analysis of the ABC gene family in almond and functional predictions during flower development, freezing stress, and salt stress.

ABC (ATP-binding cassette) transporter proteins are one of the most extensive protein families known to date and are ubiquitously found in animals, plants, and microorganisms. ABCs have a variety of functions, such as plant tissue development regulation, hormone transport, and biotic and abiotic stress resistance. However, the gene characterization and function of the ABC gene family in almond (Prunus dulcis) have not been thoroughly studied. In this study, we identified 117 PdABC genes using the whole genome of 'Wanfeng' almond obtained by sequencing and explored their protein characterization. The PdABC family members were classified into eight subfamilies. The members of the same subfamily had conserved motifs but poorly conserved numbers of exons and introns and were unevenly distributed among the eight subfamilies and on the eight chromosomes. Expression patterns showed that PdABC family members were significantly differentially expressed during almond development, dormant freezing stress, and salt stress. We found that PdABC59 and PdABC77 had extremely high expression levels in pollen. PdABC63 and PdABC64 had high expression levels during almond petal development and multiple stages of flower development. PdABC98 was highly expressed in annual dormant branches after six temperature-freezing stress treatments. PdABC29, PdABC69, and PdABC98 were highly expressed under different concentrations of salt stress. This study preliminarily investigated the expression characteristics of ABC genes in different tissues of almond during flower development, freezing stress and salt stress, and the results will provide a reference for further in-depth research and breeding of almond in the future.

Accurate Discrimination of Benign Biliary Diseases and Cholangiocarcinoma with Serum Multiomics Revealed by High-Throughput Nanoassisted Laser Desorption Ionization Mass Spectrometry.

Cholangiocarcinoma (CCA) is an aggressive malignant tumor with a poor prognosis. Carbohydrate antigen 19-9 is an essential biomarker for CCA diagnosis, but its low sensitivity (72%) makes the diagnosis unreliable. To explore potential biomarkers for the diagnosis of CCA, a high-throughput nanoassisted laser desorption ionization mass spectrometry technique was constructed. We performed serum lipidomics and peptidomics analyses from 112 patients with CCA and 123 patients with benign biliary diseases. Lipidomics analysis showed that various lipids, such as glycerophospholipids, glycerides, and sphingolipids, were perturbed. Peptidomics analysis revealed perturbations of multiple proteins involved in the coagulation cascade, lipid transport, and so on. After data mining, 25 characteristic molecules including 20 lipids and 5 peptides were identified as potential diagnostic biomarkers. After screening various machine learning algorithms, artificial neural network was selected to construct a multiomics model for CCA diagnosis with 96.5% sensitivity and 96.4% specificity. The sensitivity and specificity of the model in the independent test cohort were 93.8 and 87.5%, respectively. Furthermore, integrated analysis with transcriptomic data in the cancer genome atlas confirmed that genes altered in CCA significantly affected multiple lipid- and protein-related pathways. Data are available via MetaboLights with the identifier MTBLS6712.

IFN-beta and EIF2AK2 are potential biomarkers for interstitial lung disease in anti-MDA5 positive dermatomyositis.

OBJECTIVE: DM with positive anti-melanoma differentiation-related gene 5 (MDA5) antibody is an autoimmune disease with multiple complications. Interstitial lung diseases (ILDs) are significantly associated with DM and are particularly related to MDA5+ DM. This article aims to explore potential molecular mechanisms and develop new diagnostic biomarkers for MDA5+ DM-ILD. METHODS: The series matrix files of DM and non-specific interstitial pneumonia (NSIP) were downloaded from the Gene Expression Omnibus (GEO) database to identify the differentially expressed genes (DEGs). Gene set enrichment analysis (GSEA) was used to screen the common enriched pathways related to DM and NSIP. Next, the co-expressed differential expressed genes (co-DEGs) between MDA5+, MDA5- and NSIP groups were identified by Venn plots, and then selected for different enrichment analyses and protein-protein interaction (PPI) network construction. The mRNA expression levels of IFN-beta and EIF2AK2 were measured by RT-qPCR. The protein expression levels of IFN-beta were measured by ELISA. RESULTS: Using GSEA, the enriched pathway 'herpes simplex virus 1 infection' was both up-regulated in DM and NSIP. Enrichment analysis in MDA5+ DM, MDA5- DM and NSIP reported that the IFN-beta signalling pathway was an important influencing factor in the MDA5+ DM-ILD. We also identified that eukaryotic translation initiation factor 2 alpha kinase 2 (EIF2AK2) was an important gene signature in the MDA5+ DM-ILD by PPI analysis. The expression levels of IFN-beta and EIF2AK2 were significantly increased in MDA5+ DM-ILD patients. CONCLUSIONS: IFN-beta and EIF2AK2 contributed to the pathogenesis of MDA5+ DM-ILD, which could be used as potential therapeutic targets.

Construction and validation of cognitive frailty risk prediction model for elderly patients with multimorbidity in Chinese community based on non-traditional factors.

BACKGROUND AND OBJECTIVES: Early identification of risk factors and timely intervention can reduce the occurrence of cognitive frailty in elderly patients with multimorbidity and improve their quality of life. To explore the risk factors, a risk prediction model is established to provide a reference for early screening and intervention of cognitive frailty in elderly patients with multimorbidity. METHODS: Nine communities were selected based on multi-stage stratified random sampling from May-June 2022. A self-designed questionnaire and three cognitive frailty rating tools [Frailty Phenotype (FP), Montreal Cognitive Assessment (MoCA), and Clinical Qualitative Rating (CDR)] were used to collect data for elderly patients with multimorbidity in the community. The nomogram prediction model for the risk of cognitive frailty was established using Stata15.0. RESULTS: A total of 1200 questionnaires were distributed in this survey, and 1182 valid questionnaires were collected, 26 non-traditional risk factors were included. According to the characteristics of community health services and patient access and the logistic regression results, 9 non-traditional risk factors were screened out. Among them, age OR = 4.499 (95%CI:3.26-6.208), marital status OR = 3.709 (95%CI:2.748-5.005), living alone OR = 4.008 (95%CI:2.873-5.005), and sleep quality OR = 3.71(95%CI:2.730-5.042). The AUC values for the modeling and validation sets in the model were 0. 9908 and 0.9897. Hosmer and Lemeshow test values for the modeling set were χ2 = 3.857, p = 0.870 and for the validation set were χ2 = 2.875, p = 0.942. CONCLUSION: The prediction model could help the community health service personnel and elderly patients with multimorbidity and their families in making early judgments and interventions on the risk of cognitive frailty.

Job burnout and its influencing factors in Chinese medical staffs under China's prevention and control strategy for the COVID-19 pandemic.

OBJECTIVE: This study aimed to investigate the influencing factors of burnout among grassroots medical staff in China so as to provide a reference for improving their physical, psychological, and social statuses under China's prevention and control strategy for the COVID-19 pandemic and ensuring the sustainable supply of high-quality medical resources. METHODS: This study was performed on medical staff in five primary hospitals in Jiangsu Province, China, from May 1, 2022, to June 1, 2022, using a general information questionnaire and Maslach Burnout Inventory Scale. SPSS 25.0 and Stata 15.0 were used for two-track data entry and analysis. The OLS regression model was established to analyze the influencing factors for the job burnout of health care personnel. RESULTS: Two hundred seventy valid questionnaires were analyzed. The total score of job burnout was (30.16 ± 10.99). The scores of emotional exhaustion, depersonalization, and self-achievement were (9.88 ± 3.839), (11.99 ± 5.68), and (8.29 ± 5.18), respectively. Feeling depressed and stressed after the pandemic, days working over the past week, and work hours per shift had a positive impact on the Maslach Burnout total score. Increased income and hours working every week had a negative impact on the Maslach Burnout total score. However, sex, age in years, degree, professional title, job category, workplace, marital status, years in practice, health status, active management of health, idea of resignation, and promotion after the pandemic did not affect the Maslach Burnout total score. CONCLUSION: The job burnout of medical staff is affected by health conditions, working conditions, the psychological consequences of a pandemic, wages and marital status. Hospital managers should formulate incentive measures according to different psychological changes in medical staff to create a good medical working environment under the normalization of COVID-19 pandemic prevention and control.

A high-quality genome of the dobsonfly Neoneuromus ignobilis reveals molecular convergences in aquatic insects.

Neoneuromus ignobilis is an archaic holometabolous aquatic predatory insect. However, a lack of genomic resources hinders the use of whole genome sequencing to explore their genetic basis and molecular mechanisms for adaptive evolution. Here, we provided a high-contiguity, chromosome-level genome assembly of N. ignobilis using high coverage Nanopore and PacBio reads with the Hi-C technique. The final assembly is 480.67 MB in size, containing 12 telomere-ended pseudochromosomes with only 17 gaps. We compared 42 hexapod species genomes including six independent lineages comprising 11 aquatic insects, and found convergent expansions of long wavelength-sensitive and blue-sensitive opsins, thermal stress response TRP channels, and sulfotransferases in aquatic insects, which may be related to their aquatic adaptation. We also detected strong nonrandom signals of convergent amino acid substitutions in aquatic insects. Collectively, our comparative genomic analysis revealed the evidence of molecular convergences in aquatic insects during both gene family evolution and convergent amino acid substitutions.

Predicting the relationship between anxiety and health-related quality of life in post-stroke patients: The role of sleep duration.

OBJECTIVES: Sleep and psychological disorders are common complaints in stroke survivors. The effectiveness of sleep duration in post-stroke on mental well-being and health outcomes has been reported recently. This study aimed to investigate the mediating effect of sleep duration on the relationship between anxiety and health-related quality of life in Chinese post-stroke; MATERIALS AND METHODS: We conducted a quantitative, cross-sectional study with participants recruited through a multistage, stratified, probability proportional to size sampling method. Anxiety, health-related quality of life, and sleep duration were measured by Zung Self-rating Anxiety Scale, World Health Organization Quality of Life Questionnaire, and a self-administered, structured questionnaire. A multiple linear regression analysis was conducted to identify the association between anxiety, sleep duration, and quality of life. The direct and indirect effects of sleep duration on health-related quality of life was assessed using the bootstrap method via Model 4 (parallel mediation) of SPSS PROCESS macro; RESULTS: A total of 856 post-stroke patients participated in the study, and incidence of anxiety symptom amongst post-stroke was 33.53%. Sleep duration mainly plays partial mediating roles in the relationship between mild-to-moderate anxiety and quality of life in physical, psychological, and environment domains, with sleep duration of 7-8h playing a major role; CONCLUSIONS: A significant relationship among anxiety, sleep duration, and quality of life in post-stroke was found in this study. The sleep duration partially mediated the association between anxiety and quality of life. Suitable prevention methods and early interventions for sleep duration may improve the quality of life for post-stroke anxiety.

Hierarchical Lotus-Seedpod-Derived Porous Activated Carbon Encapsulated with NiCo2S4 for a High-Performance All-Solid-State Asymmetric Supercapacitor.

Converting biowaste into carbon-based supercapacitor materials provides a new solution for high-performance and environmentally friendly energy storage applications. Herein, the hierarchical PAC/NiCo2S4 composite structure was fabricated through the combination of activation and sulfuration treatments. The PAC/NiCo2S4 electrode garnered advantages from its hierarchical structure and hollow architecture, resulting in a notable specific capacitance (1217.2 F g-1 at 1.25 A g-1) and superior cycling stability. Moreover, a novel all-solid-state asymmetric supercapacitor (ASC) was successfully constructed, utilizing PAC/NiCo2S4 as the cathode and PAC as the anode. The resultant device exhibited exceptionally high energy (49.7 Wh kg-1) and power density (4785.5 W kg-1), indicating the potential of this biomass-derived, hierarchical PAC/NiCo2S4 composite structure for employment in high-performance supercapacitors.

Genome-wide identification of members of the Skp1 family in almond (Prunus dulcis), cloning and expression characterization of PsdSSK1.

Skp1 (S-phase kinase-associated protein 1) is the core gene of SCF ubiquitin ligase, which mediates protein degradation, thereby regulating biological processes such as cell cycle progression, transcriptional regulation, and signal transduction. A variety of plant Skp1 gene family studies have been reported. However, the almond Skp1 gene family has not yet been studied. In this study, we identified 18 members of the Prunus dulcis PdSkp1 family that were unevenly distributed across six chromosomes of the almond genome. Phylogenetic tree analysis revealed that the PdSkp1 members can be divided into three groups: I, II, and III. PdSkp1 members in each subfamily have relatively conserved motif types and exon/intron numbers. There were three pairs of fragment duplication genes and one pair of tandem repeat genes, and their functions were highly evolutionarily conserved. Transcriptome data showed that PdSkp1 is expressed in almond flower tissues, and that its expression shows significant change during cross-pollination. Fluorescence quantitative results showed that eight PdSkp1 genes had different expression levels in five tissues of almond, i.e., branches, leaves, flower buds, flesh, and cores. In addition, we cloned a PsdSSK1 gene based on PdSkp1. The cloned PsdSSK1 showed the same protein sequence as PdSkp1-12. Results of qPCR and western blot analysis showed high expression of PsdSSK1 in almond pollen. In conclusion, we report the first clone of the key gene SSK1 that controls self-incompatibility in almonds. Our research lays a foundation for future functional research on PdSkp1 members, especially for exploring the mechanism of almond self-incompatibility. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-023-01278-9.

Dual-Mechanism-Driven Strategy for High-Coverage Detection of Serum Lipids on a Novel SALDI-MS Target.

Serum lipid metabolites have been emerging as ideal biomarkers for disease diagnosis and prediction. In the current stage, nontargeted or targeted lipidomic research mainly relies on a liquid chromatography-mass spectrometry (LC-MS) platform, but future clinical applications need more robust and high-speed platforms. Surface-assisted laser desorption ionization mass spectrometry (SALDI-MS) has shown excellent advantages in the high-speed analysis of lipid metabolites. However, the platform in the positive ion mode is more inclined to target a certain class of lipids, leading to the low coverage of lipid detection and limiting its practical translation to clinical applications. Herein, we proposed a dual-mechanism-driven strategy for high-coverage detection of serum lipids on a novel SALDI-MS target, which is a composite nanostructure comprising vertical silicon nanowires (VSiNWs) decorated with AuNPs and polydopamine (VSiNW-Au-PDA). The performance of laser desorption and ionization on the target can be enhanced by charge-driven desorption coupled with thermal-driven desorption. Simultaneous detection of 236 serum lipids (S/N ≥ 5) including neutral and polar lipids can be achieved in the positive ion mode. Among these, 107 lipid peaks were successfully identified. When combined with VSiNW-Au-PDA and VSiNW chips, 479 lipid peaks can be detected in serum samples in positive and negative ion modes, respectively. Based on the platform, serum samples from 57 hepatocellular carcinoma (HCC) patients and 76 healthy controls were analyzed. After data mining, 14 lipids containing different lipid types (TAG, CE, PC) were selected as potential lipidomic biomarkers. With the assistance of an artificial neural network, a diagnostic model with a sensitivity of 92.7% and a specificity of 96% was constructed for HCC diagnosis.

Unraveling the evolutionary history of the snakefly family Inocelliidae (Insecta: Raphidioptera) through integrative phylogenetics.

Inocelliidae is one of the two extant families of the holometabolan order Raphidioptera (snakeflies), with the modern fauna represented by seven genera and 44 species. The evolutionary history of the family is little-known. Here we present the first phylogenetic and biogeographical analyses based on a worldwide sampling of taxa and datasets combined with morphological characters and mitochondrial genomes, aiming to investigate the intergeneric phylogeny and historical biogeography of Inocelliidae. The phylogenetic inference from the combined analysis of morphological and molecular data recovered the sister-group relationship between a clade of (Negha + Indianoinocellia) + Sininocellia and a clade of Fibla + the Inocellia clade (interiorly nested by Amurinocellia and Parainocellia). Amurinocellia stat.r. and Parainocellia stat.r. et emend.n. are relegated to subgeneric status within Inocellia, whereas a newly erected subgenus of Inocellia, Epinocellia subgen.n., accommodates the former Parainocellia burmana (U. Aspöck and H. Aspöck, 1968) plus a new species Inocellia (Epinocellia) weii sp.n. Further, the Inocellia crassicornis group constitutes the nominotypical subgenus Inocellia stat.n., but the Inocellia fulvostigmata group is paraphyletic. Diversification within Inocelliidae is distinguished by an Eocene divergence leading to extant genera and a Miocene radiation of species. A biogeographical scenario depicts how the diverse inocelliid fauna from East Asia could have originated from western North America via dispersal across the Beringia during the early Tertiary, and how the Miocene ancestors of Inocellia could have accomplished long-distance dispersals via the Tibet-Himalayan corridor or eastern Palaearctic to western Palaearctic. Our results shed new light specifically on the evolution of Inocelliidae and, in general, the Raphidioptera.

Similar pattern, different paths: tracing the biogeographical history of Megaloptera (Insecta: Neuropterida) using mitochondrial phylogenomics.

The sequential breakup of the supercontinent Pangaea since the Middle Jurassic is one of the crucial factors that has driven the biogeographical patterns of terrestrial biotas. Despite decades of effort searching for concordant patterns between diversification and continental fragmentation among taxonomic groups, increasing evidence has revealed more complex and idiosyncratic scenarios resulting from a mixture of vicariance, dispersal and extinction. Aquatic insects with discreet ecological requirements, low vagility and disjunct distributions represent a valuable model for testing biogeographical hypotheses by reconstructing their distribution patterns and temporal divergences. Insects of the order Megaloptera have exclusively aquatic larvae, their adults have low vagility, and the group has a highly disjunct geographical distribution. Here we present a comprehensive phylogeny of Megaloptera based on a large-scale mitochondrial genome sequencing of 99 species representing >90% of the world genera from all major biogeographical regions. Molecular dating suggests that the deep divergence within Megaloptera pre-dates the breakup of Pangaea. Subsequently, the intergeneric divergences within Corydalinae (dobsonflies), Chauliodinae (fishflies) and Sialidae (alderflies) might have been driven by both vicariance and dispersal correlated with the shifting continent during the Cretaceous, but with strikingly different and incongruent biogeographical signals. The austral distribution of many corydalids appears to be a result of colonization from Eurasia through southward dispersal across Europe and Africa during the Cretaceous, whereas a nearly contemporaneous dispersal via northward rafting of Gondwanan landmasses may account for the colonization of extant Eurasian alderflies from the south.

Embedded growth of colorful CsPbX3(X = Cl, Br, I) nanocrystals in metal-organic frameworks at Room Temperature.

Herein, we develop a novel strategy for preparing all-inorganic cesium lead halide (CsPbX3, X = Cl, Br, I) perovskite nanocrystals (NCs)@Zn-based metal-organic framework (MOF) composites through interfacial synthesis. The successful embedding of fluorescent perovskite NCs in Zn-MOFs is due to thein situconfined growth, which is attributed to the re-nucleation of water-triggered phase transformation from Cs4PbBr6to CsPbBr3. The controllable synthesis of mixed-halide based composites with various emission wavelength can be achieved by adding the desired amount of halide (Cl or I) salts in the re-nucleation process. More importantly, the anion exchange reaction is inhibited among various composites with different halogen atoms by being trapped in MOFs. Besides, a white light-emitting diode (WLED) is produced using a blue LED chip with the green-emitting and red-emitting composites, which has a color coordinate of (0.3291, 0.3272) and a wide color gamut. This work provides a novel route to achieving perovskite NCs growth in MOFs, which also can be extended to the other NCs embedded in frames as well.

MALT1 in asthma children: A potential biomarker for monitoring exacerbation risk and Th1/Th2 imbalance-mediated inflammation.

BACKGROUND: Mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1) participates in the immune-related allergic response and inflammation flare, while its clinical role in asthma children is still unknown. Herein, this study aimed to investigate MALT1 expression, and its correlation with exacerbation risk, T helper (Th)1, Th2 cells (and their secreted cytokines), as well as inflammatory cytokines in asthma children. METHODS: Sixty children with asthma exacerbation and 60 children with remission asthma were enrolled in this study; then their blood MALT1, Th1, Th2 cells, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interferon-gamma (IFN-γ), and interleukin-4 (IL-4) were detected. Besides, blood MALT1 in another 20 health controls was also determined. RESULTS: Mucosa-associated lymphoid tissue lymphoma translocation protein 1 was highest in children with asthma exacerbation, followed by children with remission asthma, and lowest in health controls (p < 0.001). MALT1 could distinguish children with asthma exacerbation from children with remission asthma (area under the curve (AUC): 0.757, 95% CI: 0.670-0.843). In children with asthma exacerbation, MALT1 was negatively linked with IFN-γ (p = 0.002) and Th1 cells (p = 0.050), but positively related to Th2 cells (p = 0.027) and exhibited a positive correlation trend (without statistical significance) with IL-4 (p = 0.066); meanwhile, MALT1 was positively correlated with exacerbation severity (p = 0.010) and TNF-α (p = 0.003), but not linked with IL-6 (p = 0.096). In children with remission asthma, MALT1 only was negatively associated with Th1 cells (p = 0.023), but positively linked with TNF-α (p = 0.023). CONCLUSION: Mucosa-associated lymphoid tissue lymphoma translocation protein 1 serves as a potential biomarker for monitoring exacerbation risk and Th1/Th2 imbalance-mediated inflammation of asthma children.

Terahertz coded-aperture imaging for moving targets based on an incoherent detection array.

Terahertz coded-aperture imaging relies on sequential coding and sampling. Its capturing speed is limited by the switching rate of the coded-aperture antenna; therefore, it is not suitable for capturing moving targets. In this paper, we focus on imaging for moving targets using the terahertz coded-aperture imaging technique. An incoherent detection array is adopted to replace sequential sampling by spatial sampling, therefore improving the capturing speed significantly. Then, a signal processing method based on phase retrieval and image enhancement is proposed to reconstruct targets with improved quality. Numerical experimental results verify the effectiveness of the method.

Simple and eco-friendly synthesis of crude orange-peel-derived carbon nanoparticles for detection of Fe3+ and ascorbic acid.

Although fluorescence sensors based on carbon dots (CDs) have been developed widely, multicomponent detection using CDs without extra and tedious surface modification remains a challenge. Here, the crude carbon nanoparticles (CPs) as a fluorescence sensor were prepared through one-pot hydrothermal process using orange peel as the precursor. The method was simple, rapid, economical, and eco-friendly given that such extra steps as dialysis and lyophilization were not required. By adding ethanol into the reaction solvent, the fluorescence properties of orange-peel-derived CPs as well as their sensitivity of detecting Fe3+ with a limit of detection of 0.25 µM were improved. Additionally, orange-peel-derived CPs could be used as a fluorescence sensor for detection of ascorbic acid (AA) with a LOD of 5 µM. More importantly, the proposed fluorescence methods were successfully used to qualitatively and quantitatively analyze Fe3+ and AA in real samples. Recovery of Fe3+ from tap water was within the range 97.2-105.4%. Conversely, recovery of AA from vitamin C tablets and orange juices laid within the ranges 97.7-99.3% and 93.2-97.6%, respectively.

Contribution to understanding the evolution of holometaboly: transformation of internal head structures during the metamorphosis in the green lacewing Chrysopa pallens (Neuroptera: Chrysopidae).

BACKGROUND: Metamorphosis remains one of the most complicated and poorly understood processes in insects. This is particularly so for the very dynamic transformations that take place within the pupal sheath of holometabolous insects. Only few studies address these transformations especially with regard to cranial structures of those holometabolous species where the larval and adult forms have a similar diet. It thus remains unclear to what extent the internal structures undergo histolysis and rebuilding. Here, the development of the brain and skeleto-muscular system of the head of Chrysopa pallens (Rambur, 1838) is studied. This species is a predator of aphids in the larval and adult stage. RESULTS: We used micro-computed-tomography (µ-CT) to study the transformations of the larval, prepupal and pupal head within the cocoon. We first assessed the morphological differences and similarities between the stages. We then determined the point in time when the compound eyes appear and describe the re-orientation of the head capsule which transforms the prognathous larva into a hypognathous adult. The internal head muscles are distinctly more slender in larvae than adults. In addition, the adults have a significantly larger brain which is likely needed for the processing of the signals obtained by the adults vastly expanded sensory organs that are presumably needed for dispersal and mating. Our study shows that the histolysis and modification of the inner muscles and skeletal elements take place within the prepupa. The central nervous system persists throughout metamorphosis but its morphology changes significantly. CONCLUSION: Our study reveals that not only the inner structures, but also the outer morphology continues to change after the final larval moult. The adult cuticle and internal structures form gradually within the cocoon. The histolysis and rebuilding begin with the skeletal elements and is followed by changes in the central nervous system before it concludes with modifications of the musculature. This order of events is likely ancestral for Holometabola because it is also known from Hymenoptera, Diptera, Mecoptera, and Coleoptera.

Correction to: An integrative phylogenomic approach to elucidate the evolutionary history and divergence times of Neuropterida (Insecta: Holometabola).

An amendment to this paper has been published and can be accessed via the original article.

An integrative phylogenomic approach to elucidate the evolutionary history and divergence times of Neuropterida (Insecta: Holometabola).

BACKGROUND: The latest advancements in DNA sequencing technologies have facilitated the resolution of the phylogeny of insects, yet parts of the tree of Holometabola remain unresolved. The phylogeny of Neuropterida has been extensively studied, but no strong consensus exists concerning the phylogenetic relationships within the order Neuroptera. Here, we assembled a novel transcriptomic dataset to address previously unresolved issues in the phylogeny of Neuropterida and to infer divergence times within the group. We tested the robustness of our phylogenetic estimates by comparing summary coalescent and concatenation-based phylogenetic approaches and by employing different quartet-based measures of phylogenomic incongruence, combined with data permutations. RESULTS: Our results suggest that the order Raphidioptera is sister to Neuroptera + Megaloptera. Coniopterygidae is inferred as sister to all remaining neuropteran families suggesting that larval cryptonephry could be a ground plan feature of Neuroptera. A clade that includes Nevrorthidae, Osmylidae, and Sisyridae (i.e. Osmyloidea) is inferred as sister to all other Neuroptera except Coniopterygidae, and Dilaridae is placed as sister to all remaining neuropteran families. Ithonidae is inferred as the sister group of monophyletic Myrmeleontiformia. The phylogenetic affinities of Chrysopidae and Hemerobiidae were dependent on the data type analyzed, and quartet-based analyses showed only weak support for the placement of Hemerobiidae as sister to Ithonidae + Myrmeleontiformia. Our molecular dating analyses suggest that most families of Neuropterida started to diversify in the Jurassic and our ancestral character state reconstructions suggest a primarily terrestrial environment of the larvae of Neuropterida and Neuroptera. CONCLUSION: Our extensive phylogenomic analyses consolidate several key aspects in the backbone phylogeny of Neuropterida, such as the basal placement of Coniopterygidae within Neuroptera and the monophyly of Osmyloidea. Furthermore, they provide new insights into the timing of diversification of Neuropterida. Despite the vast amount of analyzed molecular data, we found that certain nodes in the tree of Neuroptera are not robustly resolved. Therefore, we emphasize the importance of integrating the results of morphological analyses with those of sequence-based phylogenomics. We also suggest that comparative analyses of genomic meta-characters should be incorporated into future phylogenomic studies of Neuropterida.