Does the use of chlorantraniliprole during queen development adversely impact health and viability?

The queen is the sole reproductive individual and the maturing brood replenishes the shorter-lived worker bees. Production of many crops relies on both pesticides and bee pollination to improve crop quantity and quality. Despite the certain knowledge on chemical pesticides caused damage to worker bee physiology and behavior, our understanding of the relationship between honeybee queen development and chemical pesticides remains weak. Here, we comprehensive investigate the effects of the widely used insecticide chlorantraniliprole on the growth, hormone levels, and detoxifying enzyme activity of queen larvae. It has been determined that chlorantraniliprole present a chronic toxic effect on queen larvae and also reduced the fitness of queen, and that these effects are positively correlated with pesticide levels. It has been found that queen larvae began to show reduced capping and emergence rates when exposed to 2 ng/larva of chlorantraniliprole. At 20 ng/larva, queen capping and emergence rates were the lowest, and there were significant reductions in larval hormone level. Chlorantraniliprole have an effect on detoxification enzyme activity and hormone levels in queen larvae. In conclusion, chlorantraniliprole can adversely affect the growth and development of queen larvae. Our findings may guide the scientifically sound use of chemical pesticides to reduce potential risks to queen larvae.

Risk assessment of honeybee larvae exposure to pyrethroid insecticides in beebread and honey.

Honeybee is an essential pollinator to crops, evaluation to the risk assessment of honeybee larvae exposure to pesticides residue in the bee bread and honey is an important strategy to protect the bee colony due to the mixture of these two matrices is main food for 3-day-old honeybee larvae. In this study, a continuous survey to the residue of five pyrethroid insecticides in bee bread and honey between 2018 and 2020 from 17 major cultivation provinces which can be determined as Northeast, Northwest, Eastern, Central, Southwest, and Southern of China, there was at least one type II pyrethroid insecticide was detected in 54.7 % of the bee bread samples and 43.4 % of the honey. Then, we assayed the acute toxicity of type II pyrethroid insecticides based on the detection results, the LD50 value was 0.2201 µg/larva (beta-cyhalothrin), 0.4507 µg/larva (bifenthrin), 2.0840 µg/larva (fenvalerate), 0.0530 µg/larva (deltamethrin), and 0.1640 µg/larva (beta-cypermethrin), respectively. Finally, the hazard quotient was calculated as larval oral ranged from 0.046 × 10-3 to 2.128 × 10-3. Together, these empirical findings provide further insight into the accurate contamination of honey bee colonies caused by chemical pesticides, which can be used as a valuable guidance for the beekeeping industry and pesticide regulation.

Tumor-Derived Membrane Vesicles Restrain Migration in Gliomas By Altering Collective Polarization.

Mechanobiology is a cornerstone in physiology. However, its role in biomedical applications remains considerably undermined. In this study, we employed cell membrane vesicles (CMVs), which are currently being used as nanodrug carriers, as tactile cues for mechano-regulation of collective cell behaviors. Gliomas, which are among the most resilient brain tumors and have a low patient survival rate, were used as the cell model. We observed that mechanical responses due to the application of glioma- or microglia-derived CMVs resulted in the doubling of the traction stress of glioma cell collectives with a 10-fold increase in the CMV concentration. Glioma-CMVs constrained cell protrusions and hindered their collective migration, with the migration speed of such cells declining by almost 40% compared to the untreated cells. We speculated that the alteration of collective polarization leads to migration speed changes, and this phenomenon was elucidated using the cellular Potts model. In addition to intracellular force modulation and cytoskeletal reorganization, glioma-CMVs altered drug diffusion within glioma spheroids by downregulating the mechano-signaling protein YAP-1 while also marginally enhancing the associated apoptotic events. Our results suggest that glioma-CMVs can be applied as an adjuvant to current treatment approaches to restrict tumor invasion and enhance the penetration of reagents within tumors. Considering the broad impact of mechano-transduction on cell functions, the regulation of cell mechanics through CMVs can provide a foundation for alternative therapeutic strategies.

Exposure to sublethal concentrations of thiacloprid insecticide modulated the expression of microRNAs in honeybees (Apis mellifera L.).

This study aimed to investigate the sublethal effects of thiacloprid on microRNA (miRNA) expression in honeybees (Apis mellifera L.) and the role of a specific miRNA, ame-miR-283-5p, in thiacloprid resistance. The high-throughput small RNA-sequencing was used to analyze global miRNA expression profile changes in honeybees orally exposed to sublethal concentrations of thiacloprid (20 mg/L and 4 mg/L) for 72 h. Thiacloprid at 20 mg/L had no observed adverse effects. In addition, bees were fed with miRNA mimics or inhibitors to increase or decrease ame-miR-283-5p expression, and its effects on P450 gene expression (CYP9Q2 and CYP9Q3) were examined. Thiacloprid susceptibility was also detected. The results showed that treatment with thiacloprid at 20 mg/L and 4 mg/L induced 11 and five differentially expressed miRNAs (DEMs), respectively. Bioinformatic analysis suggested that the DEMs are mainly involved in the regulation of growth and development, metabolism, nerve conduction, and behavior. ame-miR-283-5p was downregulated by both concentrations, which was validated using quantitative real-time reverse transcription PCR analysis. Enhancing ame-miR-283-5p expression significantly inhibited CYP9Q2 mRNA and protein expression, and increased thiacloprid toxicity, while reducing ame-miR-283-5p expression significantly promoted CYP9Q2 expression, and decreased thiacloprid susceptibility. Our study revealed a novel role of miRNAs in insecticide resistance in honeybees.

Digestion dynamics of acetamiprid during royal jelly formation and exposure risk assessment to honeybee larva based on processing factor.

Previous studies to the exposure effects of acetamiprid on honeybees were based on the analysis of bee pollen and honey sacs from field trials or of beebread and honey in the hive, which overestimate or underestimate the risk of exposure to pesticide residues. It was believed that the processing factor (PF) is an important variable to determine the final pesticide residue during royal jelly formation and the actual risk to honeybee larva. Hence, a QuEChERS method to determine acetamiprid contents in honeybee samples was established in this study. Then, the PFs for acetamiprid in beebread fermentation, honey brewing, and royal jelly formation were determined to be 0.85, 0.76, and 0.16, respectively. The PF for royal jelly formation was 0.04 when acetamiprid was detected in beebread alone, and it was 0.12 when acetamiprid was only detected in honey. Finally, the predicted exposure concentration of acetamiprid in royal jelly was calculated to be 2.05 µg/kg using the PF without significant difference with the 90th percentile value (3.64 µg/kg) in the actual sample. However, the value was 16.62 µg/kg without considering the PF. This study establishes a methodology for the correct evaluation of the risk to bee larva of acetamiprid residues in bee pollen and honey sac contents and the residual levels in royal jelly.

A Blockchain-Based Authentication and Authorization Scheme for Distributed Mobile Cloud Computing Services.

Authentication and authorization constitute the essential security component, access control, for preventing unauthorized access to cloud services in mobile cloud computing (MCC) environments. Traditional centralized access control models relying on third party trust face a critical challenge due to a high trust cost and single point of failure. Blockchain can achieve the distributed trust for access control designs in a mutual untrustworthy scenario, but it also leads to expensive storage overhead. Considering the above issues, this work constructed an authentication and authorization scheme based on blockchain that can provide a dynamic update of access permissions by utilizing the smart contract. Compared with the conventional authentication scheme, the proposed scheme integrates an extra authorization function without additional computation and communication costs in the authentication phase. To improve the storage efficiency and system scalability, only one transaction is required to be stored in blockchain to record a user's access privileges on different service providers (SPs). In addition, mobile users in the proposed scheme are able to register with an arbitrary SP once and then utilize the same credential to access different SPs with different access levels. The security analysis indicates that the proposed scheme is secure under the random oracle model. The performance analysis clearly shows that the proposed scheme possesses superior computation and communication efficiencies and requires a low blockchain storage capacity for accomplishing user registration and updates.

Apilactobacillus kunkeei Alleviated Toxicity of Acetamiprid in Honeybee.

Nowadays, colony collapse disorder extensively affects honeybees. Insecticides, including acetamiprid, are considered as critical factors. As prevalent probiotics, we speculated that supplementation with lactic acid bacteria (LAB) could alleviate acetamiprid-induced health injuries in honeybees. Apilactobacillus kunkeei was isolated from beebread; it significantly increased the survival of honeybees under acetamiprid exportation (from 84% to 92%). Based on 16S rRNA pyrosequencing, information on the intestinal bacteria of honeybees was acquired. The results showed that supplementation with A. kunkeei significantly increased survival and decreased pollen consumption by honeybees under acetamiprid exportation. Under acetamiprid exportation, some opportunistic and pathogenic bacteria invaded the intestinal regions. Subsequently, the community richness and diversity of symbiotic microbiota were decreased. The community structure of intestinal bacteria was changed and differentiated. However, with the supplementation of A. kunkeei, the community richness and community diversity of symbiotic microbiota showed an upward trend, and the community structure was stabilized. Our results showed that A. kunkeei alleviated acetamiprid-induced symbiotic microbiota dysregulation and mortality in honeybees. This demonstrates the importance of symbiotic microbiota in honeybees and supports the application of Apilactobacillus kunkeei as probiotics in beekeeping.

Toxic effects of detected pyrethroid pesticides on honeybee (Apis mellifera ligustica Spin and Apis cerana cerana Fabricius).

To obtain the presence of environmental contaminants in honeybee and compare the toxicity of the detected pesticides to Apis mellifera ligustica Spin and Apis cerana cerana Fabricius. In this work, 214 honeybee samples were collected to simultaneous monitoring 66 pesticides between 2016 and 2017 in China. A modified QuEChERS extraction method coupled with multi-residue analytical methods by Ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) and Gas chromatography-mass spectrum (GC-MS). Among, four pyrethroid pesticides were selected to test and compare the acute oral toxicities of two honeybees. And the survival risk of beta-cypermethrin was analyzed to them. Using this method, 21 compounds were detected, including 3 neonicotinoids, 5 pyrethroids, 5 organophosphorus and 8 others. Importantly, detected frequencies of pyrethroid pesticides were accounted for 53.3%. Among, acute toxicity values (LD50) of four pyrethroid pesticides to the A.m. ligustica were higher than of that the A.c. cerana. When they were exposed to the same concentration of beta-cypermethrin (0.2906 mg/L), the survival rate of the A.m. ligustica (40.0%) was higher than the A.c. cerana (18.9%). Our work is valuable to analyze multiple pesticide residues of honeybees and evaluate the survival risk of two honeybee species, which also provides a basis for the risk assessment.

Proteome analysis reveals the molecular basis of honeybee brain and midgut response to sulfoxaflor.

Sulfoxaflor is a widely used pesticide in agriculture. However, the molecular effects of sublethal sulfoxaflor on honeybees (Apis mellifera L.) remain elusive. Here, the effects of a sublethal dose of sulfoxaflor (0.05 µg/bee) on the brain and midgut proteome response of the honeybee were investigated. Exposure to sublethal sulfoxaflor doses did not cause significant honeybee death, but it induced significant alterations in the brain and midgut proteomes. After sulfoxaflor challenge, 135 and 28 proteins were differentially regulated in the brain and midgut, respectively. The up-regulated proteins were mainly implicated in energy metabolism, neurotransmitter transport and drug metabolism processes, and included in particular enzymes of the citrate cycle and cellular respiration process, such as ATP citrate synthase, malate dehydrogenase, cytochrome b-c1 complex subunits, and NADH dehydrogenase. These findings suggest that honeybees enhance energy metabolism in the midgut and brain to resist sulfoxaflor challenge. Notably, treatment with sulfoxaflor resulted in a 6.8 times increase in expression levels of the major royal jelly protein 1 (MRJP1) in the brain, and knockdown of MRJP1 mRNA expression using RNA interference significantly decreased the survival rate, indicating that MRJP1 may play an important role in sulfoxaflor tolerance. Our data reveals that sulfoxaflor influences multiple processes related to both metabolism and the nervous system, and provides novel insights into the molecular basis of the honeybee brain and midgut response to sublethal dose of sulfoxaflor.

Community composition, bacterial symbionts, antibacterial and antioxidant activities of honeybee-associated fungi.

BACKGROUND: Fungi associated with insects represent one potentially rich source for the discovery of novel metabolites. However, a comprehensive understanding of the fungal communities of Apis mellifera ligustica remains elusive. RESULTS: Here, we investigated the phylogenetic diversity and community composition of honeybee-associated fungi using combination of culture-dependent and culture-independent approaches. A total of forty-five fungi were isolated and purified from the Apis mellifera ligustica, royal jelly, and honeycomb, which belonged to four classes and eleven different genera. Furthermore, 28 bacterial 16S rRNA gene sequences were obtained by PCR from the fungal metagenome. High-throughput sequencing analyses revealed that the fungal communities were more diverse, a total of 62 fungal genera were detected in the honeybee gut by culture-independent method, whereas only 4 genera were isolated by culture-dependent method. Similarly, 247 fungal genera were detected in the honeycomb, whereas only 4 genera were isolated. In addition, we assessed the antibacterial and antioxidant activities of fungal isolates. Most fungal crude extracts obtained from the cultivation supernatant exhibited antioxidant activities. Only two fungal crude extracts displayed moderate activity against Escherichia coli and Staphylococcus aureus. Chemical analysis of Chaetomium subaffine MFFC22 led to the discovery of three known compounds, including cochliodinol (1), emodin (2), chrysophanol (3). Among them, cochliodinol (1) showed intense DPPH radical scavenging activity with the 50% inhibitory concentration (IC50) of 3.06 µg/mL, which was comparable to that of the positive ascorbic acid (IC50 = 2.25 µg/mL). Compound 2 displayed weak inhibitory activities against Micrococcus tetragenus and S. aureus. CONCLUSIONS: This research provided a fundamental clue for the complex interactions among honeybees, fungi, bacterial symbionts, and the effects on the honeybee. Furthermore, the diversity of honeybee-associated fungi had great potential in finding the resource of new species and antioxidants.

The effects of beta-cypermethrin, chlorbenzuron, chlorothalonil, and pendimethalin on Apis mellifera ligustica and Apis cerana cerana larvae reared in vitro.

BACKGROUND: Declines in bee populations and diversity have drawn international attention. The long-term use of chemical pesticides has affected bee behavior and physiology. This study aimed to investigate the effects of chronic exposure to four commonly used chemical pesticides (beta-cypermethrin, chlorbenzuron, chlorothalonil and pendimethalin) on the growth of Apis mellifera ligustica and Apis cerana cerana larvae reared in vitro. RESULTS: Pesticide type and concentration were the main factors affecting honeybee fitness. Beta-cypermethrin and chlorbenzuron had chronic toxic effects on bee larvae. They reduced the fitness of A. m. ligustica and A. c. cerana even at low doses of 323.5 ng g-1 for beta-cypermethrin and 62.6 ng g-1 for chlorbenzuron in bee bread. The effects were positively associated with the dietary amounts of pesticides. By contrast, chlorothalonil and pendimethalin exposure did not affect bee larvae despite changes in enzyme activities. Caution is still needed with chlorothalonil, which led to a decrease in harvest adult bee numbers at a high dose (6937.2 ng g-1 ). Furthermore, a difference in pesticide resistance was observed, suggesting that A. m. ligustica may tolerate toxic effects better than A. c. cerana. CONCLUSION: This study sheds new light on chronic toxicity in bee larvae exposed to residues in bee bread. The results could guide the scientific and rational use of chemical pesticides to reduce the potential risks to A. m. ligustica and A. c. cerana. © 2021 Society of Chemical Industry.

Diversity and antibacterial potential of the Actinobacteria associated with Apis mellifera ligustica.

Insect-associated Actinobacteria are a potentially rich source of novel natural products with antibacterial activity. Here, the community composition of Actinobacteria associated with Apis mellifera ligustica was investigated by integrated culture-dependent and independent methods. A total of 61 strains of Streptomyces genera were isolated from the honeycomb, larva, and different anatomical parts of the honeybee's body using the culture-dependent method. Amplicon sequencing analyses revealed that the actinobacterial communities were dominated by the family of Bifidobacteriaceae and Microbacteriaceae in the honeybee gut, and Nocardiaceae and Pseudonocardiaceae in the honeycomb, whereas only Streptomyces genera were isolated by the culture-dependent method. Culture-independent analyses showed more diverse actinobacterial communities than those of culture-dependent methods. The antibacterial bioassay showed that most crude extracts of representative isolates exhibited antibacterial activities. Among them, the crude extract of Streptomyces sp. FCF01 showed the best antibacterial activities against Staphylococcus aureus, Micrococcus tetragenus, and Pseudomonas syringae pv. actinidiae (Psa) with the disc diameter of inhibition zone diameter (IZD) of 23.00, 15.00, and 13.33 mm, respectively. Chemical analysis of Streptomyces sp. FCF01 led to the isolation of three secondary metabolites, including mayamycin (1), mayamycin B (2), and N-(2-Hydroxyphenyl) acetamide (3). Among them, compound 1 displayed strong antibacterial activity against S. aureus, M. tetragenus, and Psa with minimum inhibitory concentrations (MIC) values of 6.25, 12.5, and 6.25 µg/ml, respectively. In addition, two novel derivative compounds 1a and 1b were synthesized by acetylation of compound 1. Both compounds 1a and 1b displayed similar antibacterial activities with those of metabolite 1. These results indicated that Streptomyces species associated with honeybees had great potential in finding antibiotics.

Analysis of honey bee exposure to multiple pesticide residues in the hive environment.

Since the loss of honeybees in hives could have a greater impact on colony health than those of their foraging bees, it is imperative to know beehives' pesticide exposure via oral ingestion of contaminated in-hive matrices. Here, a 4-year monitoring survey of 64 pesticide residues in pollen, nectar and related beehive matrices (beebread and honey) from China's main honey producing areas was carried out using a modified version of the QuEChERS multi-residue method. The results showed that 93.6% of pollen, 81.5% of nectar, 96.6% of beebread, and 49.3% of honey containing at least one target pesticide were detected either at or above the method detection limits (MDLs), respectively, with up to 19 pesticides found per sample. Carbendazim was the most frequently detected pesticide (present in >85% of the samples), and pyrethroids were also abundant (median concentration = 134.3-279.0 µg/kg). The transfer of pesticides from the environment into the beehive was shown, but the pesticide transference ratio may be affected by complex factors. Although the overall risk to colony health from pesticides appears to be at an acceptable level, the hazard quotient/hazard index (HQ/HI) value revealed that pyrethroids were clearly the most influential contributor, accounting for up to 45% of HI. Collectively, these empirical findings provide further insights into the extent of contamination caused by agricultural pesticide use on honeybee colonies.

Overwintering honeybees maintained dynamic and stable intestinal bacteria.

Honeybee is an important pollinator for maintaining ecological balance. However, scientist found the bizarre mass death of bees in winter. Meanwhile, some reported that the differences composed of intestinal bacteria between healthy honeybees and CCD honeybees. It is essential that explored dynamic changes to the intestinal bacteria in overwintering honeybees. We collected bee samples before overwintering, during prophase of overwintering, metaphase of overwintering, anaphase of overwintering, telophase of overwintering, and after overwintering. By using high-throughput sequencing targeting the V3-V4 regions of the 16S rDNA, the abundance of the intestinal bacteria were analyzed in overwintering honeybees. A total of 1,373,886 high-quality sequences were acquired and Proteobacteria (85.69%), Firmicutes (10.40%), Actinobacteria (3.66%), and Cyanobacteria (1.87%) were identified as major components of the intestinal bacteria. All core honeybee intestinal bacteria genera, such as Gilliamella, Bartonella, Snodgrassella, Lactobacillus, Frischella, Commensalibacter, and Bifidobacterium were detected. The abundance of Actinobacteria, Bartonella, and Bifidobacterium increased initially and then decreased in winter honeybees. There were no significant differences in the richness and evenness of the microbiota in overwintering honeybees; however, there was a statistically significant difference in the beta diversity of the intestinal bacteria after overwintering compared with that in other groups. Our results suggested that honeybees maintained their intestinal ecosystem balance, and increased the abundance of gut probiotics in response to environmental and nutrition pressures in winter.

Age and Behavior-Dependent Differential miRNAs Expression in the Hypopharyngeal Glands of Honeybees (Apis mellifera L.).

This study aims to investigate the expression differences of miRNAs in the hypopharyngeal glands (HPGs) of honeybees at three developmental stages and to explore their regulation functions in the HPGs development. Small RNA sequencing was employed to analyze the miRNA profiles of HPGs in newly-emerged bees (NEB), nurse bees (NB), and forager bees (FB). Results showed that a total of 153 known miRNAs were found in the three stages, and ame-miR-276-3p, ame-miR-375-3p, ame-miR-14-3p, ame-miR-275-3p, and ame-miR-3477-5p were the top five most abundant ones. Furthermore, the expression of 11 miRNAs, 17 miRNAs, and 18 miRNAs were significantly different in NB vs. FB comparison, NB vs. NEB comparison, and in FB vs. NEB comparison, respectively, of which ame-miR-184-3p and ame-miR-252a-5p were downregulated in NB compared with that in both the FB and NEB, while ame-miR-11-3p, ame-miR-281-3p, and ame-miR-31a-5p had lower expression levels in FB compared with that in both the NB and NEB. Bioinformatic analysis showed that the potential target genes of the differentially expressed miRNAs (DEMs) were mainly enriched in several key signaling pathways, including mTOR signaling pathway, MAPK signaling pathway-fly, FoxO signaling pathway, Hippo signaling pathway-fly. Overall, our study characterized the miRNA profiles in the HPGs of honeybees at three different developmental stages and provided a basis for further study of the roles of miRNAs in HPGs development.

Tissue Architecture Influences the Biological Effectiveness of Boron Neutron Capture Therapy in In Vitro/In Silico Three-Dimensional Self-Assembly Cell Models of Pancreatic Cancers.

Pancreatic cancer is a leading cause of cancer death, and boron neutron capture therapy (BNCT) is one of the promising radiotherapy techniques for patients with pancreatic cancer. In this study, we evaluated the biological effectiveness of BNCT at multicellular levels using in vitro and in silico models. To recapture the phenotypic characteristic of pancreatic tumors, we developed a cell self-assembly approach with human pancreatic cancer cells Panc-1 and BxPC-3 cocultured with MRC-5 fibroblasts. On substrate with physiological stiffness, tumor cells self-assembled into 3D spheroids, and the cocultured fibroblasts further facilitated the assembly process, which recapture the influence of tumor stroma. Interestingly, after 1.2 MW neutron irradiation, lower survival rates and higher apoptosis (increasing by 4-fold for Panc-1 and 1.5-fold for BxPC-3) were observed in 3D spheroids, instead of in 2D monolayers. The unexpected low tolerance of 3D spheroids to BNCT highlights the unique characteristics of BNCT over conventional radiotherapy. The uptake of boron-containing compound boronophenylalanine (BPA) and the alteration of E-cadherin can partially contribute to the observed susceptibility. In addition to biological effects, the probability of induced α-particle exposure correlated to the multicellular organization was speculated to affect the cellular responses to BNCT. A Monte Carlo (MC) simulation was also established to further interpret the observed survival. Intracellular boron distribution in the multicellular structure and related treatment resistance were reconstructed in silico. Simulation results demonstrated that the physical architecture is one of the essential factors for biological effectiveness in BNCT, which supports our in vitro findings. In summary, we developed in vitro and in silico self-assembly 3D models to evaluate the effectiveness of BNCT on pancreatic tumors. Considering the easy-access of this 3D cell-assembly platform, this study may not only contribute to the current understanding of BNCT but is also expected to be applied to evaluate the BNCT efficacy for individualized treatment plans in the future.

Identification of Olfactory Genes From the Greater Wax Moth by Antennal Transcriptome Analysis.

The olfactory system is used by insects to find hosts, mates, and oviposition sites. Insects have different types of olfactory proteins, including odorant-binding proteins (OBPs), chemosensory proteins (CSPs), odorant receptors (ORs), ionotropic receptors (IRs), and sensory neuron membrane proteins (SNMPs) to perceive chemical cues from the environment. The greater wax moth, Galleria mellonella, is an important lepidopteran pest of apiculture. However, the molecular mechanism underlying odorant perception in this species is unclear. In this study, we performed transcriptome sequencing of G. mellonella antennae to identify genes involved in olfaction. A total of 42,544 unigenes were obtained by assembling the transcriptome. Functional classification of these unigenes was determined by searching against the Gene Ontology (GO), eukaryotic orthologous groups (KOG), and the Kyoto Encyclopedia of Genes and Genomes (KEGG) databases. We identified a total of 102 olfactory-related genes: 21 OBPs, 18 CSPs, 43 ORs, 18 IRs, and 2 SNMPs. Results from BLASTX best hit and phylogenetic analyses showed that most of the genes had a close relationship with orthologs from other Lepidoptera species. A large number of OBPs and CSPs were tandemly arrayed in the genomic scaffolds and formed gene clusters. Reverse transcription-quantitative PCR results showed that GmelOBP19 and GmelOR47 are mainly expressed in male antennae. This work provides a transcriptome resource for olfactory genes in G. mellonella, and the findings pave the way for studying the function of these genes.

IL-33 Alleviated Brain Damage via Anti-apoptosis, Endoplasmic Reticulum Stress, and Inflammation After Epilepsy.

Interleukin (IL)-33 belongs to a novel chromatin-associated cytokine newly recognized by the IL-1 family, and its specific receptor is the orphan IL-1 receptor (ST2). Cumulative evidence suggests that IL-33 plays a crucial effect on the pathological changes and pathogenesis of central nervous system (CNS) diseases and injuries, such as recurrent neonatal seizures (RNS). However, the specific roles of IL-33 and its related molecular mechanisms in RNS remain confused. In the present study, we investigated the protein expression changes and co-localized cell types of IL-33 or ST2, as well as the effect of IL-33 on RNS-induced neurobehavioral defects, weight loss, and apoptosis. Moreover, an inhibitor of IL-33, anti-IL-33 was performed to further exploited underlying mechanisms. We found that administration of IL-33 up-regulated the expression levels of IL-33 and ST2, and increased the number of its co-localization with Olig-2-positive oligodendrocytes and NeuN-positive neurons at 72 h post-RNS. Noteworthily, RNS-induced neurobehavioral deficits, bodyweight loss, and spatial learning and memory impairment, as well as cell apoptosis, were reversed by IL-33 pretreatment. Additionally, the increase in IL-1ß and TNF-α levels, up-regulation of ER stress, as well as a decrease in anti-apoptotic protein Bcl-2 and an increase in pro-apoptotic protein CC-3 induced by RNS are prevented by administration of IL-33. Moreover, IL-33 in combination with Anti-IL-33 significantly inverted the effects of IL-33 or Anti-IL-33 alone on apoptosis, ER stress, and inflammation. Collectively, these data suggest that IL-33 attenuates RNS-induced neurobehavioral disorders, bodyweight loss, and spatial learning and memory deficits, at least in part through mechanisms involved in inhibition of apoptosis, ER stress, and neuro-inflammation.

Annexin A1-derived peptide Ac2-26 facilitates wound healing in diabetic mice.

Impaired wound healing is a common complication of diabetes. In diabetic wounds, macrophages present dysfunctional efferocytosis and abnormal phenotypes, which could result in excessive neutrophil accumulation and prolonged inflammation, thereby eventually hindering wound repair. ANXA1 N-terminal peptide Ac2-26 exhibits a high potential in mitigating inflammation and improving repair; however, its efficacy in diabetic wound repair remains unclear. In this study, a cutaneous excisional wound model was built in genetically diabetic mice. Ac2-26 or a vehicle solution was employed locally in wound sites. Subsequently, wound zones were measured and sampled at different time intervals post-wounding. Using hematoxylin-eosin and Masson's trichrome staining, we observed the histopathological variations and collagen deposition in wound samples. Based on immunohistochemistry and immunofluorescence, the numbers of neutrophils, macrophages, and CD206-positive macrophages in the wound samples were determined. Cytokine expression in wound samples was studied by immunoblot assay. Results showed that Ac2-26 treatment could facilitate diabetic wound closure, down-regulate the number of neutrophils, and improve angiogenesis and collagen deposition. In addition, Ac2-26 application expedited macrophage recruitment and up-regulated the percentage of macrophages expressing CD206, which is a marker for M2 macrophages. Moreover, Ac2-26 inhibited the expressions of TNF-α and IL-6 and up-regulated the expressions of IL-10, TGF-ß, and VEGFA during diabetic wound healing. Hence, based on the aforementioned findings, Ac2-26 application in diabetic wounds could exert anti-inflammatory and pro-repair effects by reducing neutrophil accumulation and facilitating M2 macrophage development.