[Effects of silver nanoparticles on pupation, eclosion, life span, apoptosis and protein expression in Drosophila melanogaster]. / 纳米银对果蝇化蛹、羽化、寿命及细胞凋亡和蛋白表达的影响.

Silver nanoparticle is widely used in the field of medicine because of its strong and effective antibacterial action. However, it has potential biological toxicity. In this study, the classical model organism, Drosophila melanogaster, was used to explore underlying mechanism for the toxic effects of silver nanoparticle. The pupation rate, eclosion time, eclosion rate and lifespan of Oregon R, w1118, and MTF mutants under different concentrations of silver nanoparticle were measured. The lacZ activity of rpr-lacZ strain was used to determine apoptosis of imaginal disc after treated with different concentrations of silver nanoparticle. The difference of intestinal protein expression in MTF mutants treated with different concentrations of silver nanoparticle was studied by SDS-PAGE. The amino acid sequence of differential proteins was further analyzed by mass spectrometry. The results showed that pupation rate and eclosion rate of MTF mutants significantly decreased when the concentration of silver nanoparticle increased to 200 µg·mL-1 and above. When the concentration of silver nanoparticle increased to 800 µg·mL-1, the rate of pupation and eclosion was significantly reduced, with the time of pupation and eclosion being not correlated to the concentration of silver nanoparticle. The concentrations of silver nanoparticle had no effect on the lifespan of Oregon R and w1118, while 200 µg·mL-1 silver nanoparticle significantly reduced the average lifespan of MTF mutant. Apoptosis increased with increasing concentration of silver nanoparticle. Results from SDS-PAGE and mass spectrometry analysis showed that the expression levels of proteins such as ATP kinase, heat shock protein and glucose metabolism related enzymes increased with increasing concentration of silver nanoparticle. Our results showed that high concentration of silver nanoparticle would reduce the survival rate of Drosophila, promote apoptosis and the expression of some proteins, which provided a theoretical basis for further understanding of the toxic mechanism of silver nanoparticle.

2-(2-Benzofuranyl)-2-imidazoline treatment within 5 hours after cerebral ischemia/reperfusion protects the brain.

We previously demonstrated that administering 2-(2-benzofuranyl)-2-imidazolin (2-BFI), an imidazoline I2 receptor agonist, immediately after ischemia onset can protect the brain from ischemic insult. However, immediate administration after stroke is difficult to realize in the clinic. Thus, the therapeutic time window of 2-BFI should be determined. Sprague-Dawley rats provided by Wenzhou Medical University in China received right middle cerebral artery occlusion for 120 minutes, and were treated with 2-BFI (3 mg/kg) through the caudal vein at 0, 1, 3, 5, 7, and 9 hours after reperfusion. Neurological function was assessed using the Longa's method. Infarct volume was measured by 2,3,5-triphenyltetrazolium chloride assay. Morphological changes in the cortical penumbra were observed by hematoxylin-eosin staining under transmission electron microscopy . The apoptosis levels in the ipsilateral cortex were examined with terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay. The protein expression of Bcl-2 and BAX was detected using immunohistochemistry. We found the following: Treatment with 2-BFI within 5 hours after reperfusion obviously improved neurological function. Administering 2-BFI within 9 hours after ischemia/reperfusion decreased infarct volume and alleviated apoptosis. 2-BFI administration at different time points after reperfusion alleviated the pathological damage of the ischemic penumbra and reduced the number of apoptotic neurons, but the protective effect was more obvious when administered within 5 hours. Administration of 2-BFI within 5 hours after reperfusion remarkably increased Bcl-2 expression and decreased BAX expression. To conclude, 2-BFI shows potent neuroprotective effects when administered within 5 hours after reperfusion, seemingly by up-regulating Bcl-2 and down-regulating BAX expression. The time window provided clinical potential for ischemic stroke by 2-BFI.

[The role of white gene and its mechanism during the formation of curly wing in Drosophila melanogaster.] / é»‘è ¹æžœè‡whiteåŸºå› åœ¨H2O2å½±å“ä¸‹å·ç¿ å½¢æˆä¸­çš„ä½œç”¨åŠæœºç†.

Curly wing is one of the most frequently used genetic markers in Drosophila melanogaster, but its molecular mechanism is remained unclear. Previous results have showed that physicochemical stimulation would affect the formation of the cruly wing. Our recent study found that H2O2 could not only affect the eclosion rate of D. melanogaster, but also induce the formation of curly wing. Here, we aimed to uncover the specific factors influencing the formation of curly wing in D. melanogaster via changing the concentration of H2O2 and the temperature as well as the time of H2O2 treatment. We measured the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-PX), in order to examine the effects of H2O2 on antioxidative capacity of D. melanogaster. The results showed that the eclosion rate of D. melanogaster was inversely correlated with the concentration of H2O2. The change of temperature, H2O2 concentration and the period of H2O2 treatment affected the degree of the curl and the proportion of the curly wing. The white mutant flies responded most significantly to these three conditions, the mini-white (white gene reverse mutation) flies could rescue the curly phenotype, and responded similarly to the wild type OR. H2O2 had effects on the formation of the curly wing which contained the Cy mutation, leading to increased rate of the curly wing. D. melanogaster treated with H2O2 would reduce the antioxidative capacity. Results from real-time quantitative PCR showed that H2O2 treatment resulted in a change in gene expression. The formation of curly wing was a complicated process, and H2O2 might act as a signaling molecule or indirectly affect certain factors in the formation of curly wing in D. melano-gaster. This process might share the same signaling pathway with the Cy mutant, or might be regulated by different modulating patterns.

[Biosynthesis and endocrine regulation of sex pheromones in moth].

The crucial importance of sex pheromones in driving mating behaviors in moths has been well demonstrated in the process of sexual communication between individuals that produce and recognize species specific pheromones. Sex-pheromone molecules from different moth species are chemically characteristic, showing different terminal functional groups, various carbon chain lengths, different position and configuration of double bond system. This review summarized information on the biosynthetic pathways and enzymes involved in producing pheromone molecules in different moths. Then we listed the components and their ratios in the sex pheromones of 15 moth species belonging to different subfamilies in Noctuidae. We also discussed the various viewpoints regarding how sex pheromones with specific ratios are produced. In the discussion we attempted to classify the pheromone molecules based on their producers, characteristics of their functional groups and carbon chain lengths. In particular, composition and ratio variations of pheromones in closely related species or within a species were compared, and the possible molecular mechanisms for these variations and their evolutionary significance were discussed. Finally, we reviewed the endocrine regulation and signal transduction pathways, in which the pheromone biosynthesis activating neuropeptide (PBAN) is involved. Comparing the biosynthetic pathways of sex pheromones among different species, this article aimed to reveal the common principles in pheromone biosynthesis among moth species and the characteristic features associated with the evolutionary course of individual species. Subsequently, some future research directions were proposed.

Transcriptome analysis of the Chinese white wax scale Ericerus pela with focus on genes involved in wax biosynthesis.

BACKGROUND: The Chinese white wax scale, Ericerus pela Chavannes is economically significant for its role in wax production. This insect has been bred in China for over a thousand years. The wax secreted by the male scale insect during the second-instar larval stage has been widespread used in wax candle production, wax printing, engraving, Chinese medicine, and more recently in the chemical, pharmaceutical, food, and cosmetics industries. However, little is known about the mechanisms responsible for white wax biosynthesis. The characterization of its larval transcriptome may promote better understanding of wax biosynthesis. METHODOLOGY/PRINCIPAL FINDINGS: In this study, characterization of the transcriptome of E. pela during peak wax secretion was performed using Illumina sequencing technology. Illumina sequencing produced 41,839 unigenes. These unigenes were annotated by blastx alignment against the NCBI Non-Redundant (NR), Swiss-Prot, KEGG, and COG databases. A total of 104 unigenes related to white wax biosynthesis were identified, and 15 of them were selected for quantitative real-time PCR analysis. We evaluated the variations in gene expression across different development stages, including egg, first/second instar larvae, male pupae, and male and female adults. Then we identified five genes involved in white wax biosynthesis. These genes were expressed most strongly during the second-instar larval stage of male E. pela. CONCLUSION/SIGNIFICANCE: The transcriptome analysis of E. pela during peak wax secretion provided an overview of gene expression information at the transcriptional level and a resource for gene mining. Five genes related to white wax biosynthesis were identified.

Molecular cloning and characterization of neutral ceramidase homologue from the red flour beetle, Tribolium castaneum.

Ceramidase plays an important role in regulating the metabolism of sphingolipids, such as ceramide, sphingosine (SPH), and sphingosine-1-phosphate (S1P), by controlling the hydrolysis of ceramide. Here we report the cloning and biochemical characterization of a neutral ceramidase from the red flour beetle Tribolium castaneum which is an important storage pest. The Tribolium castaneum neutral ceramidase (Tncer) is a protein of 696 amino acids. It shares a high degree of similarity in protein sequence to neutral ceramidases from various species. Tncer mRNA levels are higher in the adult stage than in pre-adult stages, and they are higher in the reproductive organs than in head, thorax, and midgut. The mature ovary has higher mRNA levels than the immature ovary. Tncer is localized to the plasma membrane. It uses various ceramides (D-erythro-C(6), C(12), C(16), C(18:1), and C(24:1)-ceramide) as substrates and has an abroad pH optimum for its in vitro activity. Tncer has an optimal temperature of 37 °C for its in vitro activity. Its activity is inhibited by Fe(2+). These results suggest that Tncer has distinct biochemical properties from neutral ceramidases from other species.

Role of Drosophila alkaline ceramidase (Dacer) in Drosophila development and longevity.

Ceramidases catalyze the hydrolysis of ceramides to generate sphingosine (SPH) and fatty acids, and ceramide metabolism is implicated in various biological responses in Drosophila melanogaster. Here we report the cloning, biochemical characterization, and functional analysis of a Drosophila alkaline ceramidase (Dacer). Dacer, a membrane-bound protein of 284 amino acids, shares homology with yeast and mammalian alkaline ceramidases. Overexpression of Dacer in High Five insect cells increases ceramidase activity in the alkaline pH range, indicating that Dacer is a bona fide alkaline ceramidase. Dacer mRNA is highly expressed in the midgut and at the pupal stage. An inactivation of Dacer by insertional mutagenesis increases the levels of ceramides in both Drosophila pupae and adult flies. Dacer inactivation increases Drosophila pre-adult development time, lifespan, and anti-oxidative stress capacity. Collectively, these results suggest that Dacer plays an important role in the Drosophila development and longevity by controlling the metabolism of ceramides.

A large scale screen for genes (3rd chromosome) related to Wingless signaling pathway.

A wing specific F1 genetic screen was carried out using the powerful Drosophila genetic system, combined with yeast FRT/FLP and GAL4/UAS system. Form the wing phenotypes and germline clone embryonic cuticle phenotypes observed in these mutant alleles, a number of mutant alleles of known or unknown genes were isolated. Among them, fifteen mutant alleles related to Wingless signal transduction were further isolated; the arm of these mutations located were determined, and their location in the chromosome were roughly mapped.