A Deep Reinforcement Learning Approach to Droplet Routing for Erroneous Digital Microfluidic Biochips.

Digital microfluidic biochips (DMFBs), which are used in various fields like DNA analysis, clinical diagnosis, and PCR testing, have made biochemical experiments more compact, efficient, and user-friendly than the previous methods. However, their reliability is often compromised by their inability to adapt to all kinds of errors. Errors in biochips can be categorized into two types: known errors, and unknown errors. Known errors are detectable before the start of the routing process using sensors or cameras. Unknown errors, in contrast, only become apparent during the routing process and remain undetected by sensors or cameras, which can unexpectedly stop the routing process and diminish the reliability of biochips. This paper introduces a deep reinforcement learning-based routing algorithm, designed to manage not only known errors but also unknown errors. Our experiments demonstrated that our algorithm outperformed the previous ones in terms of the success rate of the routing, in the scenarios including both known errors and unknown errors. Additionally, our algorithm contributed to detecting unknown errors during the routing process, identifying the most efficient routing path with a high probability.

Efficacy of scalp acupuncture combined with conventional therapy in the intervention of post-stroke depression: A systematic review and meta-analysis.

BACKGROUND: Post-stroke depression (PSD) is a common complication following a stroke, significantly impacting patients' quality of life and mental well-being. Currently, two primary approaches are employed to treat PSD: drug therapy and non-drug therapy. Among these, acupuncture, specifically scalp acupuncture (SA), has gained attention due to its cost-effectiveness and broad social benefits. SA is a precise and direct form of acupuncture that has been utilized in the treatment of PSD. Although several randomized controlled trials (RCTs) have demonstrated the efficacy of SA in treating PSD, there is a lack of comprehensive systematic reviews. Given the limitations of existing evidence, we conducted a systematic evaluation to assess the effectiveness of SA in combination with conventional therapy (CT) for intervening in PSD. METHODS: We systematically searched five databases for articles published up until May 31, 2023, pertaining to SA treatment of PSD. A team of researchers meticulously screened and assessed these articles to identify the final included studies. After extracting relevant information and outcome indicators from the selected articles, we employed RevMan5.3 software to evaluate their quality and perform statistical analysis. Throughout our research, we strictly adhered to the PRISMA 2020 guidelines. RESULTS: A total of 11 articles were included, and a meta-analysis was conducted to evaluate the effectiveness of SA combined with CT for treating PSD. The results revealed that SA combined with CT can effectively improve the treatment's success rate for PSD and reduce the severity of depressive symptoms measured by the Self-Rating Depression Scale. However, SA combined with CT did not show significant reductions in depressive symptoms assessed by the Hamilton Rating Scale for Depression, which may be related to the inclusion of high heterogeneity articles. Importantly, the combination treatment did not lead to an increase in adverse reactions among PSD patients. CONCLUSION: While the effectiveness of SA combined with CT in treating PSD still requires further validation through rigorous randomized double-blind trials, this study provides a comprehensive collection of studies that meet the criteria for SA combined with CT in PSD treatment. It objectively and systematically evaluated the impact of SA combined with CT on PSD. Consequently, the findings of this study hold certain clinical significance.

Bacteria-mediated RNAi for managing fall webworm, Hyphantria cunea: screening target genes and analyzing lethal effect.

BACKGROUND: The fall webworm, Hyphantria cunea, an invasive forest pest found worldwide, causes serious ecological and economic damage. Currently, the application of chemical pesticides is the most widely used strategy for H. cunea management. However, long-term pesticide use leads to pest resistance, phytotoxicity, human poisoning, and environmental deterioration. RNA interference (RNAi) technology may provide an environmentally friendly and cost-effective option for H. cunea control. However, effective RNAi targets and application methods for H. cunea are lacking. RESULTS: We screened and obtained two highly effective RNAi targets, vATPase A (V-type proton ATPase catalytic subunit A) and Rop (Ras opposite), from 23 candidate genes, using initial and repeat screening tests with the double-stranded RNA (dsRNA) injection method. RNAi against these two genes was effective in suppressing each target messenger RNA level and interfering with larval growth, leading to significant larval mortality and pupal abnormality. For massive production of dsRNA and practical application of RNAi technology in H. cunea, transformed bacteria expressing dsRNAs of these two genes were prepared using the L4440 expression vector and HT115 strain of Escherichia coli. Oral administration of bacterially expressed dsRNA targeting vATPase A and Rop genes showed high mortality and the same malformed phenotype as the injection treatment. To further investigate the lethal effects of targeting these two genes on larval development, transcriptome sequencing (RNA-seq) was performed on RNAi samples. The results demonstrated disorders in multiple metabolic pathways, and the expression levels of most genes related to insect cuticle metabolism were significantly different, which may directly threaten insect survival. In addition, some new findings were obtained via RNA-seq analysis; for example, the progesterone-mediated oocyte maturation and oocyte meiosis processes were significantly different after silencing vATPase A, and the insect olfactory protein-related genes were significantly downregulated after dsHcRop treatment. CONCLUSION: vATPase A and Rop are two highly effective RNAi-mediated lethal genes in H. cunea that regulate insect growth via multiple metabolic pathways. Oral delivery of bacterially expressed dsRNA specific to vATPase A and Rop can potentially be used for RNAi-based H. cunea management. This is the first study to apply bacteria-mediated RNAi for the control of this invasive pest, which is a major step forward in the application of the RNAi technology in H. cunea. © 2022 Society of Chemical Industry.

Performance of two Ips bark beetles and their associated pathogenic fungi on hosts reflects a species-specific association in the beetle-fungus complex.

When Ips bark beetles invade and colonize the host plants, their associated pathogenic fungal partners are carried into the phloem of the host trees. Host trees are lethally attacked by the beetle-fungus complex and the collective damage severely limits forestry production worldwide. It is of great importance to verify whether bark beetles and their associated fungi show concordant performance in terms of biology, physiology, and biochemistry on host trees. In this study, the two Ips bark beetles Ips typographus and Ips subelongatus (Coleoptera: Curculionidae, Scolytinae), their respective associated pathogenic fungi Endoconidiophora polonica and Endoconidiophora fujiensis, and their respective host plants Picea jezoensis and Larix olgensis were selected as test material. Cross-inoculation experiments were conducted indoors and outdoors to investigate the differences in reproduction and development of two beetles and infectivity of two fungi on two plants, as well as the differences in physiological responses of two plants to two fungal infections. The results showed that I. typographus and E. polonica had excellent host performance on P. jezoensis; however, neither successfully colonized and infected L. olgensis. In contrast, I. subelongatus and E. fujiensis showed strong host suitability on L. olgensis and some degree of suitability on P. jezoensis, although the host suitability of P. jezoensis for E. polonica was significantly higher than that for E. fujiensis. In addition, we found that the absolute amount of ergosterol accumulated on the lesion was positively correlated with lesion area. The ergosterol amount and lesion area were both strongly correlated with the release of host monoterpenes, but had no obvious correlation with the concentration of fungi-induced phenols on the lesion area and the side-chain oxidation of lignin in the xylem of the infected sites. Based on these results, we confirmed that "I. typographus-E. polonica" and "I. subelongatus-E. fujiensis" complexes both showed the most suitable consistent performances on their own traditional hosts, establishing a stable species-specific association relationship in these two beetle-fungus complexes, with the "I. subelongatus-E. fujiensis" complex showing broader host suitability. From the perspective of physiological responses of plants to fungal infections, monoterpenes are an important indicator of host suitability.

RNAi Efficiency through dsRNA Injection Is Enhanced by Knockdown of dsRNA Nucleases in the Fall Webworm, Hyphantria cunea (Lepidoptera: Arctiidae).

RNA interference (RNAi) technology is a promising approach used in pest control. The efficiency of RNAi varies considerably among different insect species, and growing evidence suggests that degradation of double-stranded RNA (dsRNA) prior to uptake is an important factor that limits RNAi efficiency in insects. Our recent work on fall webworm (Hyphantria cunea), an important invasive pest in China, showed a relatively low silencing efficiency of RNAi through dsRNA injection, which is considered the most feasible dsRNA delivery method for inducing RNAi, and the factors involved in the mechanism remain unknown. Herein, we first detected the dsRNA-degrading activity in the hemolymph and gut content of H. cunea in ex vivo assays and observed rapid degradation of dsRNA, especially in the hemolymph, which was complete within only 10 min. To determine whether dsRNA degradation could contribute to the low effectiveness of RNAi in H. cunea, four dsRNA nuclease (dsRNase) genes, HcdsRNase1, HcdsRNase2, HcdsRNase3, and HcdsRNase4, were identified by homology searching against the H. cunea transcriptome database, and their transcript levels were subsequently investigated in different tissues, developmental stages, and after dsRNA injection. Our results show that HcdsRNases are highly expressed mainly in gut tissues and hemolymph, and the expression of HcdsRNase3 and HcdsRNase4 were significantly upregulated by dsGFP induction. RNAi-of-RNAi studies, using HcCht5 as a reporter gene, demonstrated that silencing HcdsRNase3 and HcdsRNase4 significantly increases RNAi efficacy via dsHcCht5 injection, and co-silencing these two HcdsRNase genes results in a more significant improvement in efficacy. These results confirm that the RNAi efficacy in H. cunea through dsRNA injection is certainly impaired by dsRNase activity, and that blocking HcdsRNases could potentially improve RNAi, providing a reference for related studies on insects where RNAi has low efficiency.

Minimization of MEDA Biochip-Size in Droplet Routing.

With the increasing demand for fast, accurate, and reliable biological sensor systems, miniaturized systems have been aimed at droplet-based sensor systems and have been promising. A micro-electrode dot array (MEDA) biochip, which is one kind of the miniaturized systems for biochemical protocols such as dispensing, dilutions, mixing, and so on, has become widespread due to enabling dynamical control of the droplets in microfluidic manipulations. In MEDA biochips, the electrowetting-on-dielectric (EWOD) technique stands out since it can actuate droplets with nano/picoliter volumes. Microelectrode cells on MEDA actuate multiple droplets simultaneously to route locations for the purpose of the biochemical operations. Taking advantage of the feature, droplets are often routed in parallel to achieve high-throughput outcomes. Regarding parallel manipulation of multiple droplets, however, the droplets are known to be initially placed at a distant position to avoid undesirable mixing. The droplets thus result in traveling a long way for a manipulation, and the required biochip size for routing is also enlarged. This paper proposes a routing method for droplets to reduce the biochip size on a MEDA biochip with the allowance of splitting during routing operations. We mathematically derive the routing problem, and the experiments demonstrate that our proposal can significantly reduce the biochip size by 70.8% on average, compared to the state-of-the-art method.

Research Progress on the Dendrolimus spp. Pheromone: From Identification to Molecular Recognition.

Dendrolimus species (Lepidoptera, Lasiocampidae), are the most serious phytophagous pests of coniferous forests worldwide. Dendrolimus feed intensively on needles, leading to considerable economic loss and ecological damage. Notably, the outbreak of Dendrolimus is a somewhat periodic pattern, and those outbreaks cause rapid and large-scale destruction of pine forests, with those forests observed to look like "Fire without smoke". Sex pheromones play an important role during insect mating and reproduction, and there has been extensive research into the pheromone of Dendrolimus. The pheromone components of several Dendrolimus have been identified, and functions of two most important pheromone recognition genes, pheromone-binding proteins (PBPs) and pheromone receptors (PRs), were clarified. The evolution of PBP gene sequences is in good agreement with the trends in structural changes of the sex pheromone components in several Dendrolimus species, and it is interesting that PRs of Dendrolimus spp. occupy a novel lineage of PRs tuned to Type I pheromones in Lepidoptera. We present the current state of research into the sex pheromone of these important forest pests and highlight the emerging topics, to clarify future urgent work into Dendrolimus.

Pix2Pix-Based Monocular Depth Estimation for Drones with Optical Flow on AirSim.

In this work, we propose a method for estimating depth for an image of a monocular camera in order to avoid a collision for the autonomous flight of a drone. The highest flight speed of a drone is generally approximate 22.2 m/s, and long-distant depth information is crucial for autonomous flights since if the long-distance information is not available, the drone flying at high speeds is prone to collisions. However, long-range, measurable depth cameras are too heavy to be equipped on a drone. This work applies Pix2Pix, which is a kind of Conditional Generative Adversarial Nets (CGAN). Pix2Pix generates depth images from a monocular camera. Additionally, this work applies optical flow to enhance the accuracy of depth estimation. In this work, we propose a highly accurate depth estimation method that effectively embeds an optical flow map into a monocular image. The models are trained with taking advantage of AirSim, which is one of the flight simulators. AirSim can take both monocular and depth images over a hundred meter in the virtual environment, and our model generates a depth image that provides the long-distance information than images captured by a common depth camera. We evaluate accuracy and error of our proposed method using test images in AirSim. In addition, the proposed method is utilized for flight simulation to evaluate the effectiveness to collision avoidance. As a result, our proposed method is higher accuracy and lower error than a state of work. Moreover, our proposed method is lower collision than a state of work.

Insights into the Divergence of Chinese Ips Bark Beetles during Evolutionary Adaptation.

Many bark beetles of the genus Ips are economically important insect pests that cause severe damage to conifer forests worldwide. In this study, sequencing the mitochondrial genome and restriction site-associated DNA of Ips bark beetles helps us understand their phylogenetic relationships, biogeographic history, and evolution of ecological traits (e.g., pheromones and host plants). Our results show that the same topology in phylogenetic trees constructed in different ways (ML/MP/BI) and with different data (mtDNA/SNP) helps us to clarify the phylogenetic relationships between Chinese Ips bark beetle populations and Euramerican species and their higher order clades; Ips bark beetles are polyphyletic. The structure of the mitochondrial genome of Ips bark beetles is similar and conserved to some extent, especially in the sibling species Ips typographus and Ips nitidus. Genetic differences among Ips species are mainly related to their geographic distribution and different hosts. The evolutionary pattern of aggregation pheromones of Ips species reflects their adaptations to the environment and differences among hosts in their evolutionary process. The evolution of Ips species is closely related to the uplift of the Qinghai-Tibet Plateau and host switching. Our study addresses the evolutionary trend and phylogenetic relationships of Ips bark beetles in China, and also provides a new perspective on the evolution of bark beetles and their relationships with host plants and pheromones.

Five Visual and Olfactory Target Genes for RNAi in Agrilus Planipennis.

RNA interference (RNAi) is a widely used technique for gene function researches and recently pest controls. It had been applied in emerald ash borer (EAB Agrilus planipennis) larvae and adults, and achieved significant interference effects, whether by ingesting or microinjecting. Feeding in the phloem and cambial regions, the larvae of A. planipennis are difficult to be controlled by conventional insecticides, so adult stage is the critical stage for EAB control. However, the target genes of adult stage of A. planipennis need to be further screened. Here, we preliminarily screened five potential target genes of vision and olfaction for RNAi in A. planipennis. Three odorant binding proteins (OBPs) and three opsins, which expressed significantly different between newly emerged and sexually mature EABs (OBP5, OBP7, OBP10, LW opsin 1 and UV opsin 2) or highly in sexually mature male EAB (UV opsin 3), were selected as targets to design primers for gene silencing. After dsRNA injection, the gene expression levels were determined by real-time quantitative PCR. We found that the expression levels of five genes were significantly down-regulated, during the 4 days after dsRNA injection. Among these genes, the expression of LW opsin 1 was down-regulated the most, causing a reduction of 99.1% compared with the control treated with EGFP dsRNA, followed by UV opsin 3 (97.4%), UV opsin 2 (97.0%), OBP7 (96.2%), and OBP10 (88.7%). This study provides a basis for further RNAi-based new controlling method development of A. planipennis at adult stage.

Multi-class deep learning segmentation and automated measurements in periodontal sonograms of a porcine model.

OBJECTIVES: Ultrasound emerges as a complement to cone-beam computed tomography in dentistry, but struggles with artifacts like reverberation and shadowing. This study seeks to help novice users recognize soft tissue, bone, and crown of a dental sonogram, and automate soft tissue height (STH) measurement using deep learning. METHODS: In this retrospective study, 627 frames from 111 independent cine loops of mandibular and maxillary premolar and incisors collected from our porcine model (N = 8) were labeled by a reader. 274 premolar sonograms, including data augmentation, were used to train a multi class segmentation model. The model was evaluated against several test sets, including premolar of the same breed (n = 74, Yucatan) and premolar of a different breed (n = 120, Sinclair). We further proposed a rule-based algorithm to automate STH measurements using predicted segmentation masks. RESULTS: The model reached a Dice similarity coefficient of 90.7±4.39%, 89.4±4.63%, and 83.7±10.5% for soft tissue, bone, and crown segmentation, respectively on the first test set (n = 74), and 90.0±7.16%, 78.6±13.2%, and 62.6±17.7% on the second test set (n = 120). The automated STH measurements have a mean difference (95% confidence interval) of -0.22 mm (-1.4, 0.95), a limit of agreement of 1.2 mm, and a minimum ICC of 0.915 (0.857, 0.948) when compared to expert annotation. CONCLUSION: This work demonstrates the potential use of deep learning in identifying periodontal structures on sonograms and obtaining diagnostic periodontal dimensions.

Stereospecific synthesis of S-(-)-trans-verbenol and its antipode by inversion of sterically hindered alcohols.

S-(-)-trans-Verbenol (1) and its antipode, R-(+)-trans-verbenol (1') have been confirmed as the critical pheromone components of bark beetles. Synthesis of these two active secondary alcohols (1 and 1') from commercially available starting materials S-α-pinene and R-α-pinene was reported. The key steps were mainly depended on the effective SN2 stereo-inversion of the hydroxy group of sterically hindered alcohols (3 and 3'), using Mitsunobu reaction or hydrolysis of mesylate ester, alternatively. Our results provide a new and stereo-selectivity way to obtain optically active insect pheromones.

Comparative Analysis of Eight Mitogenomes of Bark Beetles and Their Phylogenetic Implications.

Many bark beetles of the subfamily Scolytinae are the most economically important insect pests of coniferous forests worldwide. In this study, we sequenced the mitochondrial genomes of eight bark beetle species, including Dendroctonus micans, Orthotomicus erosus, Polygraphus poligraphus, Dryocoetes hectographus, Ips nitidus, Ips typographus, Ips subelongatus, and Ips hauseri, to examine their structural characteristics and determine their phylogenetic relationships. We also used previously published mitochondrial genome sequence data from other Scolytinae species to identify and localize the eight species studied within the bark beetle phylogeny. Their gene arrangement matched the presumed ancestral pattern of these bark beetles. Start and stop codon usage, amino acid abundance, and the relative codon usage frequencies were conserved among bark beetles. Genetic distances between species ranged from 0.037 to 0.418, and evolutionary rates of protein-coding genes ranged from 0.07 for COI to 0.69 for ND2. Our results shed light on the phylogenetic relationships and taxonomic status of several bark beetles in the subfamily Scolytinae and highlight the need for further sequencing analyses and taxonomic revisions in additional bark beetle species.

RNAi-Mediated Silencing of the Chitinase 5 Gene for Fall Webworm (Hyphantria cunea) Can Inhibit Larval Molting Depending on the Timing of dsRNA Injection.

Chitinases, which are crucial enzymes required for chitin degradation and reconstruction, are often selectively considered to be effective molecular targets for pest control due to their critical roles in insect development. Although the Hyphantria cunea chitinase gene has been reported previously, its sequence characteristics, gene function, and feasibility as a potential target for pest management were absent. In the present study, we characterized the H. cunea chitinase gene and designated it HcCht5. Phylogenic and domain structure analysis suggested that HcCht5 contained the typical chitinase features and was clustered into chitinase group I. Tissue-specific and developmental expression pattern analysis with Real-Time Quantitative PCR (RT-qPCR) showed that HcCht5 was mainly expressed in the integument tissues and that the transcript levels peaked during molting. RNA interference (RNAi)-mediated silencing of HcCht5 caused 33.3% (2 ug) and 66.7% (4 ug) mortality rates after double-stranded RNA (dsRNA) injection. Importantly, the interference efficiency of HcCht5 depended on the injection time of double-stranded RNA (dsRNA), as the pre-molting treatment achieved molt arrest more effectively. In addition, transcriptome sequencing (RNA-seq) analysis of RNAi samples demonstrated silencing of the down-regulated HcCht5 genes related to chitin metabolism and molting hormone signaling, as well as genes related to detoxification metabolism. Our results indicate the essential role of HcCht5 in H. cunea development and detail the involvement of its gene function in the larval molting process.

Sensilla on antennae and mouthparts of adult spruce bark beetle Ips typographus (Coleoptera: Curculionidae).

The spruce bark beetle, Ips typographus, is an invasive forest pest that occurs across Eurasia. To further understand the mechanism of insect-host chemical communication and the feeding choices of adult I. typographus, we examined the sensilla on the antennae and mouthparts of I. typographus using scanning electron microscopy. No sexual dimorphism in sensilla type or distribution was observed. Ten subtypes of antennal sensilla were identified: sensilla trichodea Types I-III (TR1 , TR2 , and TR3 ); sensilla chaetica Types I and II (CH1 and CH2 ); sensilla basiconica Types I-III (BA1 , BA2 , and BA3 ); sensilla coeloconica (CO); and Böhm sensilla (BS). BA2 were the most abundant among the antennal sensilla in both sexes. Thirteen mouthpart sensilla subtypes were observed: sensilla trichodea Types I-IV (TR1 , TR2 , TR3 , and TR4 ); sensilla chaetica Types I-III (CH1 , CH2 , and CH3 ); sensilla digitiformia (DI); sensilla basiconica Types I-II (BA1 and BA2 ); and sensilla styloconica Types I-III (ST1 , ST2 , and ST3 ). Among these, TR4 were the most abundant in both sexes. The apex of each maxillary and labial palp carried the same sensilla (BA2 , ST1 , ST2 , and ST3 ), although the apex of the maxillary palp contained more total sensilla. The functional roles of each sensilla type are discussed based on their external structure and distribution. The results provide a theoretical basis for further studies on the olfactory and feeding behaviors and electrophysiology of adult I. typographus.

Mini Percutaneous Nephrolithotomy Is a Noninferior Modality to Standard Percutaneous Nephrolithotomy for the Management of 20-40mm Renal Calculi: A Multicenter Randomized Controlled Trial.

BACKGROUND: High quality of evidence comparing mini percutaneous nephrolithotomy (mPNL) with standard percutaneous nephrolithotomy (sPNL) for the treatment of larger-sized renal stones is lacking. OBJECTIVE: To compare the efficacy and safety of mPNL and sPNL for the treatment of 20-40mm renal stones. DESIGN, SETTING, AND PARTICIPANTS: A parallel, open-label, and noninferior randomized controlled trial was performed at 20 Chinese centers (2016-2019). The inclusion criteria were patients 18-70 yr old, with normal renal function, and 20-40mm renal stones. INTERVENTION: Percutaneous nephrolithotomy PNL was performed using either 18 F or 24 F percutaneous nephrostomy tracts. OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: The primary outcome was the one-session stone-free rate (SFR). The secondary outcomes included operating time, visual analog pain scale (VAS) score, blood loss, complications as per the Clavien-Dindo grading system, and length of hospitalization. RESULTS AND LIMITATIONS: The 1980 intention-to-treat patients were randomized. The mPNL group achieved a noninferior one-session SFR to the sPNL group by the one-side noninferiority test (0.5% [difference], p <  0.001). The transfusion and embolization rates were comparable; however, the sPNL group had a higher hemoglobin drop (5.2 g/l, p <  0.001). The sPNL yielded shorter operating time (-2.2 min, p = 0.008) but a higher VAS score (0.8, p <  0.001). Patients in the sPNL group also had longer hospitalization (0.6 d, p <  0.001). There was no statistically significant difference in fever or urosepsis occurrences. The study's main limitation was that only 18F or 24F tract sizes were used. CONCLUSIONS: Mini mPNL achieves noninferior SFR outcomes to sPNL, but with reduced bleeding, less postoperative pain, and shorter hospitalization. PATIENT SUMMARY: We evaluated the surgical outcomes of percutaneous nephrolithotomy using two different sizes of nephrostomy tracts in a large population. We found that the smaller tract might be a sensible alternative for patients with 20-40mm renal stones.

Synthesis and bioactivity of (13Z,15E)-octadecadienal: A sex pheromone component from Micromelalopha siversi Staudinger (Lepidoptera: Notodontidae).

BACKGROUND: Micromelalopha siversi (Staudinger) (Lepidoptera: Notodontidae) is a defoliator of poplar trees, Populus spp. (Salicaceae). In our previous study, 13,15-octadecadienal has been conformed as a female-produced candidate sex pheromone component of M. siversi, but the Z/E stereochemistry of the 1,3-diene system has not been identified so far. RESULTS: Four unsaturated aliphatic aldehydes, Z13,E15-18:Ald, Z13,Z15-18:Ald, E13,E15-18:Ald, and E13,Z15-18:Ald, were synthesized from the commercially available 12-bromo-1-decanol mainly by alkylation of lithium alkyne, normal Wittig or Wittig-Schlosser olefination, and hydroboration-protonolysis. According to gas chromatography (GC) analysis of pheromone gland extracts, Z13,E15-18:Ald was the main component, and a small amount of Z13,Z15-18:Ald was also detected, with a ratio of approximately 7:3. However, the results of GC-electroantennographic detection (GC-EAD) showed that Z13,E15-18:Ald was the only compound with electrophysiological activity, whereas Z13,Z15-18:Ald elicited no activity. In the field, traps baited with only Z13,E15-18:Ald resulted in much superior results to those with Z13,Z15-18:Ald as well as the Z13,E15-18:Ald and Z13,Z15-18:Ald binary mixture. CONCLUSIONS: Based on geometrically selective synthesis and bioactivity tests, the active sex pheromone component of M. siversi has been identified as Z13,E15-18:Ald, the pheromone component that has not been identified in Lepidoptera before. The synthetic component was attractive to male moths in preliminary field traps, which provides novel technologies to monitor and control this pest.

New records and a confirmation of three perlodid species in China, with additional notes and images of Rauserodes epiproctalis (Zwick, 1997)(Plecoptera: Perlodidae).

Three species of the family Perlodidae are newly reported or confirmed for China, Isoperla asiatica Rauser from Arxan, Inner Mongolia Autonomous Region, Isoperla eximia Zapekina-Dulkeit from Changbai Mountain, Jilin Province, and Mesoperlina capnoptera (McLachlan, 1886) from Xinjiang Uygur Autonomous Region. Based on new material, additional taxonomic notes and images of another perlodid stonefly, Rauserodes epiproctalis (Zwick, 1997) is provided.

Differences in Gut Bacterial Communities of Ips typographus (Coleoptera: Curculionidae) Induced by Enantiomer-Specific α-Pinene.

The spruce bark beetle (Ips typographus L.) is a destructive pest of Eurasian spruce forests. Although the gut bacteria of this insect are considered to play important roles in its lifecycle, the relationship between I. typographus and its gut bacterial community is poorly characterized. In this study, 16S rRNA gene sequencing was used to determine gut bacterial community composition across successive I. typographus life stages. Responses of the gut bacteria to α-pinene enantiomers were also explored. Ips typographus gut bacterial populations were dominated by the phyla Proteobacteria, Firmicutes, Bacteroidetes, and Actinobacteria, and the relative abundance of these phyla varied across different developmental stages of the beetle. Bacterial species diversity and richness indices increased with developmental stage progression. Relative abundances of the dominant genera, Erwinia (Enterobacteriales: Enterobacteriaceae), Pseudoxanthomonas (Xanthomonadales: Xanthomonadaceae), Serratia (Enterobacteriales: Enterobacteriaceae), and Romboutsia (Clostridiales: Peptostreptococcaceae), also varied across successive I. typographus life stages. Large disparities in the gut bacterial community of male adults were observed when the beetles were treated with S-(-)-α-pinene and R-(+)-α-pinene. The relative abundances of Lactococcus (Lactobacillales: Streptococcaceae) and Lelliottia (Enterobacteriales: Enterobacteriaceae) increased drastically with R-(+)-α-pinene and S-(-)-α-pinene treatment, respectively. This indicated a distinct enantiomer-specific effect of α-pinene on the I. typographus gut bacteria. This study demonstrated the plasticity of gut bacteria during I. typographus development, when α-pinene host monoterpenes are encountered. This study provides new insights into the relationship between 'I. typographus-gut bacteria' symbionts and host trees.

Coding and Non-coding RNAs: Molecular Basis of Forest-Insect Outbreaks.

Insect population dynamics are closely related to 'human' ecological and economic environments, and a central focus of research is outbreaks. However, the lack of molecular-based investigations restricts our understanding of the intrinsic mechanisms responsible for insect outbreaks. In this context, the moth Dendrolimus punctatus Walker can serve as an ideal model species for insect population dynamics research because it undergoes periodic outbreaks. Here, high-throughput whole-transcriptome sequencing was performed using D. punctatus, sampled during latent and outbreak periods, to systemically explore the molecular basis of insect outbreaks and to identify the involved non-coding RNA (ncRNA) regulators, namely microRNAs, long non-coding RNAs, and circular RNAs. Differentially expressed mRNAs of D. punctatus from different outbreak periods were involved in developmental, reproductive, immune, and chemosensory processes; results that were consistent with the physiological differences in D. punctatus during differing outbreak periods. Targets analysis of the non-coding RNAs indicated that long non-coding RNAs could be the primary ncRNA regulators of D. punctatus outbreaks, while circular RNAs mainly regulated synapses and cell junctions. The target genes of differentially expressed microRNAs mainly regulated the metabolic and reproductive pathways during the D. punctatus outbreaks. Developmental, multi-organismal, and reproductive processes, as well as biological adhesion, characterized the competing endogenous RNA network. Chemosensory and immune genes closely related to the outbreak of D. punctatus were further analyzed in detail: from their ncRNA regulators' analysis, we deduce that both lncRNA and miRNA may play significant roles. This is the first report to examine the molecular basis of coding and non-coding RNAs' roles in insect outbreaks. The results provide potential biomarkers for control targets in forest insect management, as well as fresh insights into underlying outbreak-related mechanisms, which could be used for improving insect control strategies in the future.