Key homeobox transcription factors regulate the development of the firefly's adult light organ and bioluminescence.

Adult fireflies exhibit unique flashing courtship signals, emitted by specialized light organs, which develop mostly independently from larval light organs during the pupal stage. The mechanisms of adult light organ development have not been thoroughly studied until now. Here we show that key homeobox transcription factors AlABD-B and AlUNC-4 regulate the development of adult light organs and bioluminescence in the firefly Aquatica leii. Interference with the expression of AlAbd-B and AlUnc-4 genes results in undeveloped or non-luminescent adult light organs. AlABD-B regulates AlUnc-4, and they interact with each other. AlABD-B and AlUNC-4 activate the expression of the luciferase gene AlLuc1 and some peroxins. Four peroxins are involved in the import of AlLUC1 into peroxisomes. Our study provides key insights into the development of adult light organs and flash signal control in fireflies.

An Overview of Aquatica Fu et al., a Phylogeny of Aquatic Fireflies Using Mitochondrial Genomes, a Description of Two New Species, and a New Record of Aquatic Fireflies in China (Coleoptera: Lampyridae: Luciolinae).

There are now seven species in the genus Aquatica Fu & Ballantyne, with all but one known from the characteristics of males, females, and larvae. Molecular information is combined with morphological taxonomy for the delimitation of both genus and species. The monophyly of the genus Aquatica is strongly supported in six trees, and its position as sister to the genus Nipponoluciola Ballantyne Kawashima Jusoh et Suzuki is stable across maximum likelihood and Bayesian inference results. Two new species of Aquatica Fu & Ballantyne, A. qingshen sp. nov. and A. xianning sp. nov., described from the features of males, females, and larvae, are closely related within a single clade within the genus Aquatica. Females have distinctively shaped median oviduct plates. A Chinese population identified as A. lateralis was found to be morphologically similar to the Japanese population, but genetic distances suggest that it is a distinctive species. No larvae are associated with this species. Definitions of the aquatic status of Luciolinae fireflies are expanded.

An Improved Chromosome-Level Genome Assembly of the Firefly Pyrocoelia pectoralis.

The endemic and endangered Chinese firefly Pyrocoelia pectoralis is a sexually dimorphic, nocturnal species. A previous attempt by this team to assemble a draft genome of P. pectoralis using PacBio and Illumina HiSeq X Ten platforms was limited in its usefulness by high redundancy and contamination. This prompted us to conduct an improved chromosome-level genome assembly of P. pectoralis. Ten chromosomes were further assembled based on Hi-C data to a 532.25 Mb final size with a 52.87 Mb scaffold N50. The total repeat lengths in the genome of P. pectoralis amount to 227.69 Mb; 42.78%. In total, 12,789 genes could be functionally annotated using at least one public database. Phylogenetic inference indicated that P. pectoralis and P. pyralis diverged ~51.41 million years ago. Gene family expansion and contraction analysis of 12 species were performed, and 546 expanded and 2660 contracted gene families were identified in P. pectoralis. We generated a high-quality draft of the P. pectoralis genome. This genome assembly should help promote research on the species' sexual dimorphism and its unique courtship behavior, which involves a combination of pheromonal and bioluminescent signals. It also can serve as a resource for accelerating genome-assisted improvements in the conservation of this species.

Sperm Competition and Paternity in the Endangered Firefly Pyrocoelia pectoralis (Coleoptera: Lampyridae: Lampyrinae).

The endangered terrestrial firefly Pyrocoelia pectoralis (Olivier) is endemic to China. Populations of P. pectoralis have decreased dramatically due to urbanization and pollution. Breeding and re-introduction to a suitable habitat may save the species from becoming extinct. Because of its polyandrous character, an investigation into the possibility of sperm competition and paternity outcomes from multiple matings was initiated to better understand its reproductive physiology. To achieve these goals, 13 SSR markers were developed. The results of paternity experiments indicate there is a significant difference between P3 and P1 or P2. The female reproductive system has three spermathecae which accept sperm from different matings, and no bursa or spermatophore-digesting organ is developed. Our research established that multiple inseminations with sperm from different males occur, leading to competition between ejaculates. The benefits of such competition include an increasing number of sperm in the ejaculates of competing males and the consequential increase in fertilized eggs (thus, fecundity), and thereby a higher chance of genetic diversity and fitness in the offspring of the firefly P. pectoralis.

LightFD: Real-Time Fault Diagnosis with Edge Intelligence for Power Transformers.

Power fault monitoring based on acoustic waves has gained a great deal of attention in industry. Existing methods for fault diagnosis typically collect sound signals on site and transmit them to a back-end server for analysis, which may fail to provide a real-time response due to transmission packet loss and latency. However, the limited computing power of edge devices and the existing methods for feature extraction pose a significant challenge to performing diagnosis on the edge. In this paper, we propose a fast Lightweight Fault Diagnosis method for power transformers, referred to as LightFD, which integrates several technical components. Firstly, before feature extraction, we design an asymmetric Hamming-cosine window function to reduce signal spectrum leakage and ensure data integrity. Secondly, we design a multidimensional spatio-temporal feature extraction method to extract acoustic features. Finally, we design a parallel dual-layer, dual-channel lightweight neural network to realize the classification of different fault types on edge devices with limited computing power. Extensive simulation and experimental results show that the diagnostic precision and recall of LightFD reach 94.64% and 95.33%, which represent an improvement of 4% and 1.6% over the traditional SVM method, respectively.

The complete mitochondrial genome of the firefly Curtos fulvocapitalis (Coleoptera: Lampyridae).

We report the complete mitochondrial genome of the firefly Curtos fulvocapitalis Jeng et Yang 1998. The circular genome is 16,398 bp and has a base composition of A (42.21%), C (11.22%), G (7.73%), and T (38.85%). Our sequence is similar to those of other metazoans, which contain 13 protein-coding genes. All 13 protein-coding genes were initiated by the ATN (ATT, ATA, and ATG) codon. Nine protein-coding genes stopped with TAA or TAG codons, and the other four genes had an incomplete termination codon, a single T. We sequenced the mitochondrial genome of fireflies to analyze phylogenetic relationships and determine the evolution of their flashing signals.

Selection and Validation of Suitable Reference Genes for RT-qPCR Analysis in the Rare Aquatic Firefly Aquatica leii (Coleoptera: Lampyridae).

Aquatica leii Fu and Ballantyne is a species of rare aquatic firefly and endemic in China. It is considered good material to study the molecular mechanism of sexual flash communication systems. To improve conservation and behavioral research strategies, large-scale genetic studies involving gene-expression analysis are required and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) is the most commonly used method. However, there have been very few reports on appropriate reference genes in any species of firefly. Here, we evaluated eight widely utilized reference genes including 18S, Actin, Reep5, Odc1, Tub, Gapdh, Ef1a and S27Ae for their expression stabilities in A. leii under three different conditions, i.e., life stage, tissue and dsRNA injection. Based on the gene stability ranking calculated by RefFinder, which integrates four algorithms (geNorm, delta Ct method, NormFinder, and BestKeeper), we recommend S27Ae and Reep5 as the most appropriate reference genes for molecular studies in different life stages; Ef1a and Odc1 for different tissues; Tub and Odc1 for RNAi studies. The most appropriate reference genes in all treatments are S27Ae and Tub. The results of this study will help improve accuracy and reliability to normalize RT-qPCR data in A. leii for further molecular analysis.

Reproductive Systems, Transfer and Digestion of Spermatophores in Two Asian Luciolinae Fireflies (Coleoptera: Lampyridae).

The internal reproductive anatomy of males and females of two Asian Luciolinae fireflies Emeia pseudosauteri (Geisthardt, 2004) and Abscondita chinensis (L., 1767) is described, and the time course for spermatophore transfer and digestion examined. E. pseudosauteri is sexually dimorphic, with a flightless female, and Abs. chinensis is sexually monomorphic, with the female flighted. Both are monandrous. Possible female accessory glands are described for the first time for both species. An overview of present knowledge of female reproductive anatomy in the Luciolinae reveals males of 18 species in 10 genera may produce spermatophores and permits speculation about spermatophore production in another 16 genera.

The genetic variations in the mitochondrial genomes of three Luciolinae fireflies.

This paper studied the complete mitochondrial genomes of three fireflies, Pygoluciola qingyu, Emeia pseudosauteri and Abscondita terminalis. We discussed the variations in the mitochondrial genomes of samples of each firefly from different populations. The mitochondrial genomes of Abs. terminalis and P. qingyu are very stable among their different populations. The mitochondrial genome of E. pseudosauteri shows some variations among the different populations, especially in the COI sequence.

The complete mitochondrial genome of the firefly, Diaphanes citrinus (Olivier), (coleoptera: Lampyridae).

We report the complete mitochondrial genome of firefly, Diaphanes citrinus (Olivier). The circular genome of 18,594 bp and has a base composition of A (41.26%), C (11.64%), G (10.18%), T (36.92%). Similar to other Metazoa, our sequence contains 13 protein-coding genes, 21 transfer RNA genes, two ribosomal RNA genes, and a non-coding AT-rich region. We sequenced the mitochondrial genome of fireflies to analyze phylogenetic relationships and deduce the evolution of their flashing signals.

The complete mitochondrial genome of the firefly, Luciola curtithorax (Coleoptera: Lampyridae).

We report the complete mitochondrial genome of firefly, Luciola curtithorax. The circular genome of 16,882 bp has a base composition of A (44.98%), C (11.84%), G (8.15%), and T (35.03%). Our sequence is similar to other Metazoa, which contains 13 protein-coding genes. All 13 protein-coding genes were initiated by the ATN (ATT, ATA, and ATG) codon. Ten protein-coding genes stopped with TAA or TAG codon and the other three genes have an incomplete termination codon, a single T--. We sequenced the mitochondrial genome of fireflies to analyze phylogenetic relationships and deduce the evolution of their flashing signals.

The complete mitochondrial genome of the firefly, Abscondita anceyi (Olivier) (Coleoptera: Lampyridae).

We report the complete mitochondrial genome of firefly, Abscondita anceyi (Olivier). The circular genome of 16,519 bp has a base composition of A (43.81%), C (11.80%), G (8.35%), and T (36.03%). Similar to other Metazoa, our sequence contains 13 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes, and a non-coding AT-rich region. We sequenced the mitochondrial genome of fireflies to analyse phylogenetic relationships and deduce the evolution of their flashing signals.

Long-read sequence assembly of the firefly Pyrocoelia pectoralis genome.

Background: Fireflies are a family of insects within the beetle order Coleoptera, or winged beetles, and they are one of the most well-known and loved insect species because of their bioluminescence. However, the firefly is in danger of extinction because of the massive destruction of its living environment. In order to improve the understanding of fireflies and protect them effectively, we sequenced the whole genome of the terrestrial firefly Pyrocoelia pectoralis. Findings: Here, we developed a highly reliable genome resource for the terrestrial firefly Pyrocoelia pectoralis (E. Oliv., 1883; Coleoptera: Lampyridae) using single molecule real time (SMRT) sequencing on the PacBio Sequel platform. In total, 57.8 Gb of long reads were generated and assembled into a 760.4-Mb genome, which is close to the estimated genome size and covered 98.7% complete and 0.7% partial insect Benchmarking Universal Single-Copy Orthologs. The k-mer analysis showed that this genome is highly heterozygous. However, our long-read assembly demonstrates continuousness with a contig N50 length of 3.04 Mb and the longest contig length of 13.69 Mb. Furthermore, 135 589 SSRs and 341 Mb of repeat sequences were detected. A total of 23 092 genes were predicted; 88.44% of genes were annotated with one or more related functions. Conclusions: We assembled a high-quality firefly genome, which will not only provide insights into the conservation and biodiversity of fireflies, but also provide a wealth of information to study the mechanisms of their sexual communication, bio-luminescence, and evolution.

The complete mitochondrial genome of the firefly, Pteroptyx maipo (Coleoptera: Lampyridae).

We report the complete mitochondrial genome of firefly, Pteroptyx maipo. The circular genome of 16,127 bp has a base composition of A (44.22%), C (11.60%), G (8.23%) and T (35.96%). Our sequence is similar to other Metazoa, which contains 13 protein-coding genes. All 13 protein-coding genes were initiated by the ATN (ATT, ATA and ATG) codon. Nine protein-coding genes stopped with TAA or TAG codon and the other four genes have an incomplete termination codon, a single T. We sequenced the mitochondrial genome of fireflies to analyze phylogenetic relationships and deduce the evolution of their flashing signals.

The complete mitochondrial genome of the firefly, Asymmetricata circumdata (Motschulsky) (Coleoptera: Lampyridae).

We report the complete mitochondrial genome of firefly, Asymmetricata circumdata (Motschulsky).The circular genome of 15,967 bp has a base composition of A (42.44%), C (12.83%), G (8.79%) and T (36.16%). Similar to other Metazoa, our sequence contains 13 protein-coding genes. All 13 protein-coding genes were initiated by the ATN (ATT, ATA and ATG) codon. Eight protein-coding genes stopped with TAA or TAG codon and the other 5 genes have an incomplete termination codon, a single T. We sequenced the mitochondrial genome of fireflies to analyze phylogenetic relationships and deduce the evolution of their flashing signals.

Design, expression and characterization of single chain Fv, Mms13 and the single chain Fv­mms13 fusion protein.

Single chain Fv (scFv) antibodies are attractive as tumor-targeting vehicles due to their smaller size compared with intact antibody molecules. Mms13 is a putative membrane anchor protein of magnetosome. The present study fused the scFV gene of type Ⅳ collagenase to mms13 using the splicing by overlap extension polymerase chain reaction technique. The genes of scFv, mms13 and the scFv-mms13 fusion gene were cloned into a pET30a(+) vector to construct pET30a(+)-scFv, pET30a(+)-mms13 and pET30a(+)-scFv-mms13 expression vectors. The three protein compositions were confirmed by DNA sequencing and western blot analysis, and their cellular locations were determined using SDS-PAGE. The results of enzyme-linked immunosorbent assays and immunofluorescence demonstrated that the ScFv and ScFv-mms13 fusion proteins bound to the type Ⅳ collagenase and the antigen-associated cancer cells SMMC-7721, MCF-7 and HepG2 cells, in a dose-dependent and saturable manner. Although the immunoreactivities of ScFv-mms13 to the type Ⅳ collagenase and associated tumor cells were marginally lower than the corresponding scFv (3G11), considerable binding ability to the antigen by ScFv-mms13 remained.

Purification and characterization of catalase from marine bacterium Acinetobacter sp. YS0810.

The catalase from marine bacterium Acinetobacter sp. YS0810 (YS0810CAT) was purified and characterized. Consecutive steps were used to achieve the purified enzyme as follows: ethanol precipitation, DEAE Sepharose ion exchange, Superdex 200 gel filtration, and Resource Q ion exchange. The active enzyme consisted of four identical subunits of 57.256 kDa. It showed a Soret peak at 405 nm, indicating the presence of iron protoporphyrin IX. The catalase was not apparently reduced by sodium dithionite but was inhibited by 3-amino-1,2,4-triazole, hydroxylamine hydrochloride, and sodium azide. Peroxidase-like activity was not found with the substrate o-phenylenediamine. So the catalase was determined to be a monofunctional catalase. N-terminal amino acid of the catalase analysis gave the sequence SQDPKKCPVTHLTTE, which showed high degree of homology with those of known catalases from bacteria. The analysis of amino acid sequence of the purified catalase by matrix-assisted laser desorption ionization time-of-flight mass spectrometry showed that it was a new catalase, in spite of its high homology with those of known catalases from other bacteria. The catalase showed high alkali stability and thermostability.

Studies on South-east Asian fireflies: Abscondita, a new genus with details of life history, flashing patterns and behaviour of Abs. chinensis (L.) and Abs. terminalis (Olivier) (Coleoptera: Lampyridae: Luciolinae).

Abscondita, a new genus of fireflies from South-east Asia, is described from males and females of Abs. anceyi (Olivier 1883), Abs. cerata (Olivier 1911), Abs. chinensis (L. 1767), Abs. perplexa (Walker 1858), Abs. promelaena (Walker 1858) and Abs. terminalis (Olivier 1883), all transferred from Luciola Laporte. Both L. dubia Olivier 1903 and L. dejeani Gemminger 1870 are synonymised with Luciola perplexa (Walker), and L. aegrota Olivier 1891 and L. melaspis Bourgeois 1909 with L. promelaena Walker. Females are characterised by their bursa plates. Larvae are associated and described for Abs. anceyi (Olivier), Abs. chinensis (L.) and Abs. terminalis (Olivier). Taxonomic issues regarding the identification of species with very similar colouration of pale dorsum and black tipped elytra are addressed and in some cases resolved. A neotype for Luciola chinensis (L.) is erected and Luciola praeusta (Kiesenwetter 1874) is synonymised with L. chinensis (L.). Descriptions of life histories, biology and flashing patterns of populations of Abs. chinensis and Abs. terminalis from central China are included. A bs. terminalis is the first Asian firefly known to possess multiple flash trains where males are documented to display with repeating flash trains.

Sexual dimorphism, mating systems, and nuptial gifts in two Asian fireflies (Coleoptera: Lampyridae).

Many insect taxa show conspicuous sexual dimorphism in which females lack functional wings and are therefore incapable of flight. In fireflies, this loss of female flight is correlated with male production of spermatophore nuptial gifts, with species with flightless females also lacking gift production. In this study we further examined the relationship between sexual dimorphism and nuptial gifts by examining the reproductive ecology of two Asian fireflies, the sexually monomorphic Aquatica ficta (Luciolinae) and dimorphic Pyrocoelia pectoralis (Lampyrinae). We describe the reproductive anatomy of P. pectoralis males and females, and the time course of A. ficta spermatophore transfer. Through a series of mating experiments, we determined the effect of the number of matings on female fecundity, egg hatching success and female lifespan. The spermatophore-producing A. ficta was found to be monandrous, and female lifespan doubled as a result of that single mating. P. pectoralis was found to lack spermatophores but females were polyandrous and fecundity and egg hatching success both increased with additional matings. These results contradict patterns known from North American firefly species and provide insight into the role of male-derived substances in mating systems.

[Treatment of 58 cases of penile squamous cell carcinoma].

OBJECTIVE: To search for rational and effective treatments for penile squamous cell carcinoma (PSCC). METHODS: We retrospectively analyzed the clinical data of 58 cases of pathologically confirmed PSCC, focusing on the treatment methods. RESULTS: Based on Jackson Staging, 25 of the 58 cases fell into stage I, 18 stage II, 11 stage III, and 4 stage IV. Fifty-three of the patients were treated by surgery, of whom 43 underwent limited resection of the tumor or partial amputation of the penis, and the other 10 received total penis amputation plus perineal urethrostomy and clearance of lymphoglandulae iliacae and inguinal lymph nodes, with the lymphoglandulae iliacae positive in 1 case and the inguinal lymph nodes positive in all. Thirty-seven cases received neoadjuvant hormonal therapy (thermotherapy plus chemotherapy) and combined postoperative chemotherapy, 12 postoperative chemotherapy only, and 4 merely surgery. Five of the total number underwent chemotherapy and/or radiotherapy without surgery. The 2-5 years follow-up of 48 patients found recurrence in 4 cases of partial penis amputation within 2 years, 4 deaths within 2 years, 7 deaths from 2 to 5 years. The 2- and 5-year survival rates were 91.7% and 77.1%, respectively. Ten of the cases were lost in follow-up. CONCLUSION: Surgery + neoadjuvant hormonal therapy + postoperative chemotherapy and/or radiotherapy is an effective method for PSCC, but whether it can reduce the recurrence of PSCC and improve the survival of the patients remains to be further studied.