Follicular fluid exosome-derived miR-339-5p enhances in vitro maturation of porcine oocytes via targeting SFPQ, a regulator of the ERK1/2 pathway.

In this study, we aimed to investigate cytoplasmic maturation and miRNA expression of mature oocytes cultured in porcine follicular fluid exosomes. We also examined the effect of miR-339-5p on oocyte maturation. Twenty eight differentially expressed miRNAs were detected using miRNA-seq. We then transfected cumulus oocyte complexes with miR-339-5p mimics and inhibitor during culture. The results showed that exosomes increased endoplasmic reticulum levels and the amount of lipid droplets, and decreased ROS levels, lipid droplet size, and percentage of oocytes with abnormal cortical granule distribution. Overexpressing miR-339-5p significantly decreased cumulus expansion genes, oocyte maturation-related genes, target gene proline/glutamine-rich splicing factor (SFPQ), ERK1/2 phosphorylation levels, oocyte maturation rate, blastocyst rate, and lipid droplet number, but increased lipid droplet size and the ratio of oocytes with abnormal cortical granule distribution. Inhibiting miR-339-5p reversed the decrease observed during overexpression. Mitochondrial membrane potential and ROS levels did not differ significantly between groups. In summary, exosomes promote oocyte cytoplasmic maturation and miR-339-5p regulating ERK1/2 activity through SFPQ expression, thereby elevating oocyte maturation and blastocyst formation rate in vitro.

Heat Shock Protein Genes Affect the Rapid Cold Hardening Ability of Two Invasive Tephritids.

Bactrocera dorsalis and Bactrocera correcta are two invasive species that can cause major economic damage to orchards and the fruit import and export industries. Their distribution is advancing northward due to climate change, which is threatening greater impacts on fruit production. This study tested the rapid cold-hardening ability of the two species and identified the temperature associated with the highest survival rate. Transcriptome data and survival data from the two Bactrocera species' larvae were obtained after rapid cold-hardening experiments. Based on the sequencing of transcripts, four Hsp genes were found to be affected: Hsp68 and Hsp70, which play more important roles in the rapid cold hardening of B. dorsalis, and Hsp23 and Hsp70, which play more important roles in the rapid cold hardening of B. correcta. This study explored the adaptability of the two species to cold, demonstrated the expression and function of four Hsps in response to rapid cold hardening, and explained the occurrence and expansion of these two species of tephritids, offering information for further studies.

Computational identification of long non-coding RNAs associated with graphene therapy in glioblastoma multiforme.

Glioblastoma multiforme represents the most prevalent primary malignant brain tumour, while long non-coding RNA assumes a pivotal role in the pathogenesis and progression of glioblastoma multiforme. Nonetheless, the successful delivery of long non-coding RNA-based therapeutics to the tumour site has encountered significant obstacles attributable to inadequate biocompatibility and inefficient drug delivery systems. In this context, the use of a biofunctional surface modification of graphene oxide has emerged as a promising strategy to surmount these challenges. By changing the surface of graphene oxide, enhanced biocompatibility can be achieved, facilitating efficient transport of long non-coding RNA-based therapeutics specifically to the tumour site. This innovative approach presents the opportunity to exploit the therapeutic potential inherent in long non-coding RNA biology for treating glioblastoma multiforme patients. This study aimed to extract relevant genes from The Cancer Genome Atlas database and associate them with long non-coding RNAs to identify graphene therapy-related long non-coding RNA. We conducted a series of analyses to achieve this goal, including univariate Cox regression, least absolute shrinkage and selection operator regression and multivariate Cox regression. The resulting graphene therapy-related long non-coding RNAs were utilized to develop a risk score model. Subsequently, we conducted Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses on the identified graphene therapy-related long non-coding RNAs. Additionally, we employed the risk model to construct the tumour microenvironment model and analyse drug sensitivity. To validate our findings, we referenced the IMvigor210 immunotherapy model. Finally, we investigated differences in the tumour stemness index. Through our investigation, we identified four promising graphene therapy-related long non-coding RNAs (AC011405.1, HOXC13-AS, LINC01127 and LINC01574) that could be utilized for treating glioblastoma multiforme patients. Furthermore, we identified 16 compounds that could be utilized in graphene therapy. Our study offers novel insights into the treatment of glioblastoma multiforme, and the identified graphene therapy-related long non-coding RNAs and compounds hold promise for further research in this field. Furthermore, additional biological experiments will be essential to validate the clinical significance of our model. These experiments can help confirm the potential therapeutic value and efficacy of the identified graphene therapy-related long non-coding RNAs and compounds in treating glioblastoma multiforme.

Molecular Engineering of Perylene Diimide Polymers with a Robust Built-in Electric Field for Enhanced Solar-Driven Water Splitting.

The built-in electric field of the polymer semiconductors could be regulated by the dipole moment of its building blocks, thereby promoting the separation of photogenerated carriers and achieving efficient solar-driven water splitting. Herein, three perylene diimide (PDI) polymers, namely oPDI, mPDI and pPDI, are synthesized with different phenylenediamine linkers. Notably, the energy level structure, light-harvesting efficiency, and photogenerated carrier separation and migration of polymers are regulated by the orientation of PDI unit. Among them, oPDI enables a large dipole moment and robust built-in electric field, resulting in enhanced solar-driven water splitting performance. Under simulated sunlight irradiation, oPDI exhibits the highest photocurrent of 115.1 µA cm-2 for photoelectrochemical oxygen evolution, which is 11.5 times that of mPDI, 26.8 times that of pPDI and 104.6 times that of its counterparts PDI monomer at the same conditions. This work provides a strategy for designing polymers by regulating the orientation of structural units to construct efficient solar energy conversion systems.

Comprehensive assessment of base excision repair (BER)-related lncRNAs as prognostic and functional biomarkers in lung adenocarcinoma: implications for personalized therapeutics and immunomodulation.

BACKGROUND: Lung adenocarcinoma (LUAD) is the most prevalent subtype of lung cancer, and comprehending its molecular mechanisms is pivotal for advancing treatment efficacy. This study aims to explore the prognostic and functional significance of base excision repair (BER)-related long non-coding RNAs (BERLncs) in LUAD. METHODS: A risk score model for BERLncs was developed using the least absolute shrinkage and selection operator regression and Cox regression analysis. Model validation and prognostic evaluation were performed using Kaplan-Meier and receiver-operating characteristic curve analyses. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses were conducted to elucidate the potential biological functions of BERLncs. Comparative analyses were carried out to investigate disparities in tumor mutation burden (TMB), immune infiltration, tumor immune dysfunction and exclusion (TIDE) score, chemosensitivity, and immune checkpoint gene expression between the two risk groups. RESULTS: A predictive risk score model comprising 19 BERLncs was successfully developed. Patients were divided into high-risk and low-risk groups according to the median risk score. The high-risk subgroup exhibited significantly inferior overall survival. Functional enrichment analysis revealed pathways associated with lung cancer development, notably the neuroactive ligand-receptor interaction pathway. High-risk patients demonstrated elevated TMB, diminished TIDE scores, and an immunosuppressive tumor microenvironment, while low-risk patients displayed potential benefits from immunotherapy. Additionally, the risk model identified potential anticancer agents. CONCLUSION: The risk score model based on BERLncs shows promise as a prognostic biomarker for LUAD patients, providing valuable insights for clinical decision-making, therapeutic strategies, and understanding of underlying biological mechanisms.

The Wnt pathway regulates wing morph determination in Acyrthosiphon pisum.

Wing dimorphism occurs in insects as a survival strategy to adapt to environmental changes. In response to environmental cues, mother aphids transmit signals to their offspring, and the offspring either emerge as winged adults or develop as wingless adults with degeneration of the wing primordia in the early instar stage. However, how the wing morph is determined in the early instar stage is still unclear. Here, we established a surgical sampling method to obtain precise wing primordium tissues for transcriptome analysis. We identified Wnt as a regulator of wing determination in the early second instar stage in the pea aphid. Inhibiting Wnt signaling via knockdown of Wnt2, Wnt11b, the Wnt receptor-encoding gene fz2 or the downstream targets vg and omb resulted in a decreased proportion of winged aphids. Activation of Wnt signaling via knockdown of miR-8, an inhibitor of the Wnt/Wg pathway, led to an increased proportion of winged aphids. Furthermore, the wing primordia of wingless nymphs underwent apoptosis in the early second instar, and cell death was activated by knockdown of fz2 under the wing-inducing condition. These results indicate that the developmental plasticity of aphid wings is modulated by the intrinsic Wnt pathway in response to environmental challenges.

Notch Signaling in Insect Development: A Simple Pathway with Diverse Functions.

Notch signaling is an evolutionarily conserved pathway which functions between adjacent cells to establish their distinct identities. Despite operating in a simple mechanism, Notch signaling plays remarkably diverse roles in development to regulate cell fate determination, organ growth and tissue patterning. While initially discovered and characterized in the model insect Drosophila melanogaster, recent studies across various insect species have revealed the broad involvement of Notch signaling in shaping insect tissues. This review focuses on providing a comprehensive picture regarding the roles of the Notch pathway in insect development. The roles of Notch in the formation and patterning of the insect embryo, wing, leg, ovary and several specific structures, as well as in physiological responses, are summarized. These results are discussed within the developmental context, aiming to deepen our understanding of the diversified functions of the Notch signaling pathway in different insect species.

Dual-duty NiCo2S4 nanosheet-based solar rechargeable batteries toward multi-scene solar energy conversion and storage.

Solar energy has the potential to be the next-generation power source if the intermittent nature can be overcome via rational energy storage engineering. The competitiveness of solar rechargeable batteries can be further enhanced if the demand for multiple energy storage scenarios can be met within one device. Moreover, active electrochemical materials with different energy storage types are the critical component of this energy storage system. In this work, dual-duty electrochemical functional materials were introduced to guide multi-scene solar energy storage device design and fabrication. Furthermore, dual-duty NiCo2S4 nanosheets were prepared and applied to solar rechargeable batteries. A photo-assisted aqueous polysulfide/iodide flow battery was designed and fabricated with a charging voltage as low as 0.05 V, showing the good electrocatalytic performance of NiCo2S4 nanosheets for aqueous redox couples. Moreover, the low charging voltage leads to 93.5% of input electric energy saving under one sun illumination (AM 1.5, 100 mW cm-2). On the other hand is the photo-assisted sodium-ion battery with a NiCo2S4 anode, showing a remarkably low charging voltage of 0.67 V and a high discharge medium voltage of 1.05 V. The battery can save about 67.6% of input electric energy under 1 sun illumination.

The mitochondrial ribosomal protein mRpL4 regulates Notch signaling.

Mitochondrial ribosomal proteins (MRPs) assemble as specialized ribosome to synthesize mtDNA-encoded proteins, which are essential for mitochondrial bioenergetic and metabolic processes. MRPs are required for fundamental cellular activities during animal development, but their roles beyond mitochondrial protein translation are poorly understood. Here, we report a conserved role of the mitochondrial ribosomal protein L4 (mRpL4) in Notch signaling. Genetic analyses demonstrate that mRpL4 is required in the Notch signal-receiving cells to permit target gene transcription during Drosophila wing development. We find that mRpL4 physically and genetically interacts with the WD40 repeat protein wap and activates the transcription of Notch signaling targets. We show that human mRpL4 is capable of replacing fly mRpL4 during wing development. Furthermore, knockout of mRpL4 in zebrafish leads to downregulated expression of Notch signaling components. Thus, we have discovered a previously unknown function of mRpL4 during animal development.

Insulin-like peptide 8 (Ilp8) regulates female fecundity in flies.

Introduction: Insulin-like peptides (Ilps) play crucial roles in nearly all life stages of insects. Ilp8 is involved in developmental stability, stress resistance and female fecundity in several insect species, but the underlying mechanisms are not fully understood. Here we report the functional characterization of Ilp8s in three fly species, including Bactrocera dorsalis, Drosophila mercatorum and Drosophila melanogaster. Methods: Phylogenetic analyses were performed to identify and characterize insect Ilp8s. The amino acid sequences of fly Ilp8s were aligned and the three-dimensional structures of fly Ilp8s were constructed and compared. The tissue specific expression pattern of fly Ilp8s were examined by qRT-PCR. In Bactrocera dorsalis and Drosophila mercatorum, dsRNAs were injected into virgin females to inhibit the expression of Ilp8 and the impacts on female fecundity were examined. In Drosophila melanogaster, the female fecundity of Ilp8 loss-of-function mutant was compared with wild type control flies. The mutant fruit fly strain was also used for sexual behavioral analysis and transcriptomic analysis. Results: Orthologs of Ilp8s are found in major groups of insects except for the lepidopterans and coleopterans, and Ilp8s are found to be well separated from other Ilps in three fly species. The key motif and the predicted three-dimensional structure of fly Ilp8s are well conserved. Ilp8 are specifically expressed in the ovary and are essential for female fecundity in three fly species. Behavior analysis demonstrates that Ilp8 mutation impairs female sexual attractiveness in fruit fly, which results in decreased mating success and is likely the cause of fecundity reduction. Further transcriptomic analysis indicates that Ilp8 might influence metabolism, immune activity, oocyte development as well as hormone homeostasis to collectively regulate female fecundity in the fruit fly. Discussion: Our findings support a universal role of insect Ilp8 in female fecundity, and also provide novel clues for understanding the modes of action of Ilp8.

Chronic exposure to the star polycation (SPc) nanocarrier in the larval stage adversely impairs life history traits in Drosophila melanogaster.

BACKGROUND: Nanomaterials are widely used as pesticide adjuvants to increase pesticide efficiency and minimize environmental pollution. But it is increasingly recognized that nanocarrier is a double-edged sword, as nanoparticles are emerging as new environmental pollutants. This study aimed to determine the biotoxicity of a widely applied star polycation (SPc) nanocarrier using Drosophila melanogaster, the fruit fly, as an in vivo model. RESULTS: The lethal concentration 50 (LC50) value of SPc was identified as 2.14 g/L toward third-instar larvae and 26.33 g/L for adults. Chronic exposure to a sub lethal concentration of SPc (1 g/L) in the larval stage showed long-lasting adverse effects on key life history traits. Exposure to SPc at larval stage adversely impacted the lifespan, fertility, climbing ability as well as stresses resistance of emerged adults. RNA-sequencing analysis found that SPc resulted in aberrant expression of genes involved in metabolism, innate immunity, stress response and hormone production in the larvae. Orally administrated SPc nanoparticles were mainly accumulated in intestine cells, while systemic responses were observed. CONCLUSIONS: These findings indicate that SPc nanoparticles are hazardous to fruit flies at multiple levels, which could help us to develop guidelines for further large-scale application.

Inhibition of 26S proteasome enhances AKAP3-mediated cAMP-PKA signaling during boar sperm capacitation.

This study investigates the effects of the ubiquitin-proteasome pathway (UPP) on porcine sperm capacitation and its interactions with the cAMP-PKA pathway. The semen of adult Landrace boars was divided into four groups: non-capacitated, capacitated, 10 µM/mL MG132, and 10 µM/mL DMSO groups. We characterized the parameters related to sperm dynamics using a computer-assisted sperm analysis system. The level of sperm protein tyrosine phosphorylation was detected using Western blotting, and the change of zinc ion signal was detected via flow cytometry. The relationship between A-kinase-anchor protein 3 (AKAP3), ubiquitin (Ub), and protein kinase A (PKA) was assessed by co-precipitation assays; to evaluate the interactions between the UPP and cAMP-PKA pathway, threonine, serine, and tyrosine phosphorylation were detected using Western blotting to evaluate the interaction between the UPP and cAMP-PKA pathway; Hoechst staining was used to detect the sperm-egg binding state and evaluate the effects of UPP inhibition. During capacitation, the levels of protein tyrosine, serine, and threonine phosphorylation and ubiquitination of porcine sperm increased, and sperm-egg binding was inhibited (P < 0.05). AKAP3 was degraded by UPP, and after inhibiting the 26 S proteasome, ubiquitinated AKAP3 accumulated in large quantities. Our findings indicate that, after the 26 S proteasome was inhibited, PKA was uncoupled from AKAP3 and degraded by UPP; the level of tyrosine phosphorylation induced by PKA-AKAP3 was reduced, the level of serine threonine phosphorylation increased, and the ubiquitination pathway interacted with the phosphorylation pathway and was involved in sperm capacitation.

Use of FLP/FRT System to Screen for Notch Signaling Regulators in the Drosophila Wing.

Mutations of genes encoding key components of the Notch signaling pathways often result in lethality at early developmental stages, making it difficult to decipher how they regulate the formation of specific cell types or organs. Mosaic analysis using the FLP/FRT system allows investigating the roles of essential genes during wing development in Drosophila melanogaster. This chapter describes the practical methods to isolate Notch signaling regulators by somatic mosaic screen. The fly stocks, cross schemes, and screen parameters are summarized. We also explain how to validate the roles of potential Notch signaling regulators.

Monochromatic light-enhanced photocatalytic CO2 reduction based on exciton properties of two-dimensional lead halide perovskites.

Converting CO2 into valuable solar fuels through photocatalysis has been considered a green and sustainable technology that is promising for alleviating global warming and providing energy in an environmentally friendly manner. However, traditional photocatalysts generally suffer from low surface-reactive reaction sites, inefficient light harvesting and rapid recombination of electron-hole pairs. Lead halide perovskite materials have been considered ideal semiconductor photocatalysts for photocatalytic CO2 reduction due to their tunable band gaps, strong light absorption, and low cost. Herein, a series of L2Csn-1PbnX3n+1 (L = ba, ha, oa; X = Cl, Br, I; n = 1, 2) 2D layered perovskites were synthesized by a facile solvothermal method. The effects of alkyl amine chain length, halogen atoms and inorganic layer number on their properties were studied. More importantly, these 2D materials were used as photocatalysts for CO2 reduction without any sacrificial agents. These 2D perovskites exhibited markedly increased performance in comparison with 3D bulk materials, benefitting from the larger surface-area-to-volume ratio and faster and more efficient exciton dissociation, which achieved the highest CO yield of 158.69 µmol g-1 h-1 and CH4 yield of 6.9 µmol g-1 h-1 through the design of the photocatalytic system. In addition, the influence of light source conditions on photocatalysis was studied systematically, including light source intensity and wavelength. The experimental results indicated that an appropriate solvent, high light intensity and monochromatic light source matching the wavelength of exciton absorption can effectively improve the photocatalytic efficiency.

JAK/STAT signaling regulates the Harmonia axyridis leg regeneration by coordinating cell proliferation.

Harmonia axyridis presents remarkable appendage regeneration capacity and can therefore be considered as an emerging regeneration research model. Amino acid sequences of the Janus kinase Hopscotch (Hahop) and the transcription factor STAT (HaStat), the main components of the JAK/STAT signaling pathway, conserved with their homologs in other models. The expression levels of these two genes were continuously up-regulated during the appendage regeneration process. To identify the functions of JAK/STAT signaling, we performed RNAi experiments of Hahop and HaStat in H. axyridis, and found regeneration defects following in HahopRNAi and HaStatRNAi treatments at different regeneration stages. Additionally, we confirmed that regeneration defects caused by the low-level of JAK/STAT activity were due to the inhibition of cell proliferation. The results of the current study suggest that JAK/STAT signaling regulates the entire regeneration process by coordinating cell proliferation of regenerating appendages.

Phenotypical and genetical characterization of the Mad1-2 allele during Drosophila wing development.

Growth and patterning of Drosophila wing depends upon the sequential organizing activities of Hedgehog (Hh) and Decapentaplegic (Dpp) signaling pathways. The Hh signaling directly activates the expression of dpp through the transcription factor cubitus interruptus (Ci). Dpp itself functions as a long-range morphogen to promote cell proliferation and differentiation through an essential transcription factor encoded by Mad. Here we report that the Mad1-2 allele exhibits phenotypes distinct from classical Dpp pathway mutants in the developing wing. The activity of Dpp signaling is attenuated in Mad1-2 mutant cells. However, activation of Dpp signaling is found in a subset of cells surrounding homozygous Mad1-2 clones when the clones are located at the anterior compartment of wing disc. Further analysis reveals that Mad1-2 mutant cells display high level of Hh signaling activity and accumulate significant amount of Ci. Unexpectedly, whole genome resequencing identifies multiple mutations in the 3'UTR region of Pka-C1 genomic loci in the Mad1-2 stock. We provide genetic and molecular evidence that the Pka-C1 mutations carried by Mad1-2 likely underlies the observed Hh signaling defects. Therefore, the contribution of Pka-C1 mutation should be taken in consideration when analyzing Mad1-2 phenotypes. The isolation of independent Mad and Pka-C1 alleles from the Mad1-2 stock further supports our conclusions.

The Ubiquitin Conjugating Enzyme UbcD1 is Required for Notch Signaling Activation During Drosophila Wing Development.

Notch signaling pathway plays crucial roles in animal development. Protein ubiquitination contributes to Notch signaling regulation by governing the stability and activity of major signaling components. Studies in Drosophila have identified multiple ubiquitin ligases and deubiquitinating enzymes that modify Notch ligand and receptor proteins. The fate of ubiquitinated substrates depend on topologies of the attached ubiquitin chains, which are determined by the ubiquitin conjugating enzymes (E2 enzymes). However, which E2 enzymes participate in Notch signal transduction remain elusive. Here, we report that the E2 enzyme UbcD1 is required for Notch signaling activation during Drosophila wing development. Mutations of UbcD1 lead to marginal nicks in the adult wing and reduction of Notch signaling targets expression in the wing imaginal disc. Genetic analysis reveal that UbcD1 functions in the signaling receiving cells prior to cleavage of the Notch protein. We provide further evidence suggesting that UbcD1 is likely involved in endocytic trafficking of Notch protein. Our results demonstrate that UbcD1 positively regulates Notch signaling and thus reveal a novel role of UbcD1 in development.

Substituent Modulation for Highly Bright 9-Borafluorene Derivatives with Carbazole Pendant.

A series of 3,6-di-tert-butyl carbazole-functionalized 9-borafluorene derivatives have been prepared with outstandingly strong photoluminescence with quantum yields up to ca. 100 and 94% for Mes*BF-pCz in solution and film, respectively. 1,3,5-Tris(trifluoromethyl)benzene (FMes)-substituted compounds exhibit enhanced Lewis acidity with coordination to weak nucleophiles like tetrahydrofuran, resulting in a long afterglow at low temperature. The large two-photon absorption cross-section of ca. 1103 GM for Mes*BF-pCz at 800 nm in CH2Cl2 indicated its potential application in bioimaging.

Characterization of a new mastermind allele identified from somatic mosaic screen.

The Notch signaling pathway is highly conserved and regulates various fundamental development events. Activation of Notch signaling relies on production of the Notch intracellular domain (NICD), which assembles a transcription factor complex to turn on down-stream targets expression. The mastermind (mam) gene encodes an essential co-activator that permits NICD activity in the cell nucleus. During a somatic mosaic screen in Drosophila, an uncharacterized gene l(2)S9998 is identified as a positive regulator of the Notch signaling pathway. Genetic analysis demonstrates that l(2)S9998 functions at the level of transcriptional activation of Notch targets in the signal receiving cells. Whole genome sequencing reveals that l(2)S9998 is a novel allele of the mam gene, which is further confirmed by complementation tests. Along with three molecularly defined transposon insertions isolated from the screen, four mutants of mam are shown to modulate Notch signaling during fly wing development. Our analysis provides additional genetic resources for understanding mam function and Notch signaling regulation.

The Ras/MAPK pathway is required for regenerative growth of wing discs in the black cutworm Agrotis ypsilon.

Regeneration is a common phenomenon in various organisms by which tissues restore the damaged or naturally detached parts. In insects, appendage regeneration takes place during the embryonic, larval and pupal stages for individual survival. The wing disc of black cutworm Agrotis ypsilon has the capacity of regeneration after ablation, but understanding of molecular mechanisms in wing disc regeneration is still limited. After ablation of partial or whole wing discs before the fifth instar larval stage, the adult wings appeared to be normal. In the last two larval stages, ablation of the left wing disc led to smaller corresponding adult wing. Cell proliferation was reduced in the ablated wing disc but was gradually recovered two days post ablation. Transcriptome analysis found that genes in the mitogen-activated protein kinase (MAPK) pathway were upregulated. Repression of gene expression in this pathway, including Ras oncogene at 64B (Ras64B), Downstream of raf1 (Dsor1), and cAMP-dependent protein kinase catalytic subunit 3 (Pka-C3) by RNA interference after ablation, led to diminishment of both adult wings, suggesting that the MAPK signaling is essential for wing growth. Additionally, cell proliferation was still decelerated by injecting Ras64B, Dsor, or Pka-C3 dsRNA two days after ablation, indicating that the MAPK signaling-regulated cell proliferation is essential for growth. These results provide molecular clues to the regulation of cell proliferation during regeneration in lepidopteran insects.