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1.
Front Plant Sci ; 15: 1431097, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38947949

RESUMO

Tender bamboo shoots undergo rapid senescence that influences their quality and commercial value after harvest. In this study, the tender sweet bamboo shoots ('Wensun') were packed by a passive modified atmosphere packaging (PMAP) to inhibit the senescence process, taking polyethylene package as control. The increase in CO2 and the decrease in O2 gas concentrations in the headspace atmosphere of the packages were remarkably modified by PMAP treatments. The modified gas atmosphere packaging inhibited the changes in firmness, as well as the content of cellulose, total pectin, and lignin in the cell walls of bamboo shoots. The enzymatic activities of cellulase, pectinase, and polygalacturonase that act on cell wall polysaccharides, and phenylalanine ammonia lyase, cinnamyl alcohol dehydrogenase, peroxidase, and laccase regulating the lignin biosynthesis were modified by PMAP treatment different from control during storage. The expression levels of the lignin biosynthesis genes PePAL3/4, PeCAD, Pe4CL5, PeC4H, PeCCOAOMT, PeCOMT, cellulose synthase PeCESA1, and related transcription factors PeSND2, PeKNAT7, PeMYB20, PeMYB63, and PeMYB85 were clearly regulated. These results suggest that PMAP efficiently retards the changes in lignin and cell wall polysaccharides, thus delaying the senescence of tender sweet bamboo shoots during storage.

2.
Food Chem X ; 21: 101129, 2024 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-38298353

RESUMO

Fresh puffer fish (Takifugu obscurus) are susceptible to microbial contamination and have a very short shelf-life of chilled storage. Hence, this study aimed to evaluate the effects of plasma-activated lactic acid (PALA) on microbiota composition and quality attributes of puffer fish fillets during chilled storage. The results showed that PALA treatment effectively reduced the growth of bacteria and attenuated changes in physicochemical indicators (total volatile basic nitrogen, pH value, K value, and biogenic amines) of puffer fish fillets. Additionally, insignificant changes were observed in lipid oxidation during the first 8 days (p > 0.05). Illumina-MiSeq high-throughput sequencing revealed that PALA effectively inhibited the growth of Pseudomonas in puffer fish fillets and maintained the diverse characteristics of the microbial community. In combination with sensory analysis, PALA extended the shelf life of puffer fish fillets for 4 days, suggesting that PALA could be considered a potential fish fillet preservation method.

3.
J Environ Manage ; 328: 116945, 2023 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-36512947

RESUMO

The contamination of organoarsenic is becoming increasingly prominent while SR-AOPs were confirmed to be valid for their remediation. This study has found that the novel metal/carbon catalyst (Fe/C-Mn) prepared by solid waste with hierarchical pores could simultaneously degrade roxarsone (ROX) and remove As(V). A total of 95.6% of ROX (20 mg/L) could be removed at the concentration of 1.0 g/L of catalyst and 0.4 g/L of oxidant in the Fe/C-Mn/PMS system within 90 min. The scavenging experiment and electrochemical test revealed that both single-electron and two-electron pathways contributed to the ROX decomposition. Spectroscopic analysis suggested the ROX has been successfully mineralized while As(V) was fixed with the surface Fe and Mn. Density functional theory (DFT) calculation and chromatographic analysis indicated that the As7, N8, O9 and O10 sites of ROX molecule were vulnerable to being attacked by nucleophilic, electrophilic and radical, resulting in the formation of several intermediates such as phenolic compounds. Additionally, the low metal leaching concentration during recycling and high anti-interference ability in various water matrices manifested the practicability of Fe/C-Mn/PMS system.


Assuntos
Roxarsona , Roxarsona/química , Roxarsona/metabolismo , Manganês , Carvão Vegetal , Metais , Eletrólitos
4.
J Colloid Interface Sci ; 630(Pt B): 714-726, 2023 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-36347098

RESUMO

In this study, a novel carbon-wrapped-iron hierarchical porous catalyst (Fe/C-Mn800) was prepared from electrolytic manganese residue (EMR) and sewage sludge (SS), which showed outstanding degradation ability toward benzohydroxamic acid (BHA, nearly 90 % was removed within 60 min) with low metal leaching rate. Mechanism exploration found transition metal ions (Fe and Mn) can serve as electron acceptors and facilitate the generation of persistent free radicals (PFRs). These transition metal ions and PFRs mainly participated in the single-electron pathway via activating PMS to generate a large amount of reactive oxygen species (ROS). While the electron negative graphitic N and CO groups not only improve the electronegatively of catalyst, but also acted as the electron sacrificers to favor the electron transfer and directly oxidized the absorbed BHA through the ternary activated outer-sphere complexes. Eley-Rideal (E-R) and Langmuir-Hinshelwood (L-H) analysis further demonstrated the crucial role of pre-adsorption during the degradation process. This work provided a deep insight into the degradation mechanism of metal/carbon composite and promising opportunity widened the horizon of the high-value utilization of EMR and SS.


Assuntos
Peróxidos , Resíduos Sólidos , Carbono/química , Radicais Livres , Metais , Peróxidos/química , Porosidade , Esgotos/química
5.
Sci Rep ; 12(1): 10893, 2022 06 28.
Artigo em Inglês | MEDLINE | ID: mdl-35764785

RESUMO

Fresh mulberry leaf vegetable is nutritive and becoming popular. However, available preservation technologies are deficient. In present work, the effects of two kinds of modified atmosphere packaging on postharvest quality of fresh mulberry leaf vegetable stored at 4 °C were evaluated. The respiration rate of samples in the modified polyethylene packages (MP20) was 12.88-22.65% lower than that in normal polyethylene packaging (CK). The content of total soluble solids, soluble protein, and total polyphenol in MP20 was less changed than that in CK, and the vitamin C retention was higher as well. Moreover, the lignin content in MP20 was lower than that in CK during storage (19.79% vs 13.38% at day 8), and that was significantly positively related to the polyphenol oxidase and peroxidase activities inhibition. Taken together, a packaging with moderate gas permeability (MP20) is suitable for nutrition maintenance and lignification inhibition of fresh mulberry leaf vegetable during cold storage.


Assuntos
Morus , Verduras , Atmosfera , Folhas de Planta , Polietileno
6.
ACS Nano ; 15(9): 14253-14262, 2021 09 28.
Artigo em Inglês | MEDLINE | ID: mdl-34409836

RESUMO

As a powerful signal amplification tool, the DNA walker has been widely applied to detect rare microRNA (miRNA) in vivo. Despite the significant advances, a near-infrared (NIR) light controllable DNA walker for signal amplification powered by an endogenous initiator has not been realized, which is crucial for spatiotemporal imaging of miRNA in living cells with high sensitivity. Herein, we constructed a NIR-photoactivatable DNA walker system, which was powered by endogenous adenosine triphosphate (ATP) for in situ miRNA imaging with spatial and temporal resolution. The system was very stable with an extremely low fluorescent background for the bioimaging in living cells. We employed upconversion nanoparticles (UCNPs) as the carriers of the DNA probe and transducers of converting NIR to UV light. Coupled with the DNA walker fueled by intracellular ATP, a smart system based on the NIR light initiated DNA walker was successfully developed for precise spatiotemporal control in living cells. Triggered by NIR light, the DNA walker could autonomously and progressively travel along the track with the assistance of intracellular ATP. The system has been successfully applied for in situ miRNA imaging in different cell lines with highly spatial and temporal resolution. This strategy can expand NIR photocontrol the DNA walker for precise imaging in a biological system.


Assuntos
Trifosfato de Adenosina , MicroRNAs , DNA/genética , MicroRNAs/genética
7.
J Food Sci ; 86(9): 3884-3895, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34333772

RESUMO

Chinese kale is one of the most popular vegetables in southern China and Asia, but it has a short shelf-life. The effect of high oxygen atmospheric packaging (HOAP) treatment on the respiration rate as well as chlorophyll content and the expression of their metabolism-related genes of the soluble proteins in Chinese kale during storage were assessed. The results showed that Chinese kale subjected to HOAP treatment showed stimulated respiration rate and regulated expression of chlorophyll metabolism-related genes, such as BrChlases, BrPPH (pheophytin pheophorbide hydrolase), BrPAO (pheidea oxygenase gene), BrRCCR (red chlorophyll catabolite reductase), and BrSAG12 (senescence-associated gene), compared to the Chinese kale in the control. The activities of chlorophyll enzymes, that is, Chlase and Mg-dechelatase, were also influenced by HOAP treatment during storage. Furthermore, the total content of soluble proteins was stimulated to accumulate, and the intensity of protein bands, detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis profiling, increased in HOAP-treated samples. Based on the current results, as well as the results of our previous study regarding HOAP treatment of other vegetables, we speculate that HOAP may function by regulating the respiration rate and the accumulation of functional proteins, especially chlorophyll catabolic and antioxidant enzymes, to maintain the freshness of Chinese kale during storage. PRACTICAL APPLICATION: HOAP treatment could be a potential method for delaying quality changes and extending the shelf-life of Chinese kale after harvest.


Assuntos
Brassica , Embalagem de Alimentos , Oxigênio , Brassica/química , Brassica/efeitos dos fármacos , Brassica/metabolismo , China , Clorofila/análise , Embalagem de Alimentos/métodos , Embalagem de Alimentos/normas , Oxigênio/farmacologia
8.
ACS Sens ; 6(5): 1925-1932, 2021 05 28.
Artigo em Inglês | MEDLINE | ID: mdl-33881313

RESUMO

The efficient capture and sensitive detection of circulating tumor cells (CTCs) play a vital role in cancer diagnosis and prognosis. However, CTCs in the peripheral blood are very rare and heterogeneous, which make them difficult to isolate and detect. Herein, a novel colorimetric nanobioplatform was successfully developed for the highly efficient capture and highly sensitive detection of heterogeneous CTCs, which consisted of two parts: the multivalent aptamer-modified gold nanoparticles as the capture unit and two kinds of aptamer-functionalized pH-sensitive allochroic dyes (thymolphthalein and curcumin) @ molybdenum disulfide nanoflakes (MoS2 NFs) acting as the visual simultaneous detection of heterogeneous CTCs. Using MCF-7 and HeLa cells as the CTC models, the capture unit can effectively isolate the CTCs due to the multivalent probe with improved affinity. The two allochroic dyes can display obvious color changes under alkaline conditions (pH 12.5) in the presence of MCF-7 and HeLa cells, which provided a rapid and sensitive strategy for visualizing simultaneous detection of heterogeneous CTCs as low as 5 cells mL-1. This nanoplatform possessed a high sensitivity toward CTC detection owing to high dye loading capacity of MoS2 NFs and allochroic dyes with excellent pH sensitivity. It can successfully distinguish and quantitatively detect the targeted heterogeneous CTCs from numerous interfering cells in diluted whole blood. It can also be used to detect CTCs from lysed blood samples from cancer patients, indicating promising application for cancer diagnosis.


Assuntos
Nanopartículas Metálicas , Células Neoplásicas Circulantes , Colorimetria , Corantes , Ouro , Células HeLa , Humanos , Concentração de Íons de Hidrogênio
9.
Anal Chem ; 92(4): 3111-3117, 2020 02 18.
Artigo em Inglês | MEDLINE | ID: mdl-31968939

RESUMO

The integrative bioplatform for capture, detection and release of circulating tumor cells (CTCs) is of great significance in clinical diagnosis and biomedical research. To fulfill this demand, we introduced a near-infrared (NIR) light-switched bioplatform for efficient isolation and downstream analysis of CTCs. The platform was created by first modifying the PEG-MoS2 nanoflakes (NFs)@gelatin nanocomposite on the ITO surface, and then introducing the MUC1 aptamer as a specific recognition element via coupling reaction between aptamer and gelatin to achieve the specific capture for CTCs. Subsequently, the captured cells are released under a NIR light irradiation (808 nm) by using MoS2 NFs as the NIR-regulated control element. Significantly, this platform could capture and release of CTCs with an excellent capture/release efficiency of 89.5% and 92.5%, respectively. Furthermore, the electrochemical bioplatform exhibited a wide linear range for the detection of CTCs from 50 to 1 × 106 cells mL-1 with a detection limit of 15 cells mL-1. After 5 days of reculture, the released cells still maintain good cell shape and proliferation capacity. Moreover, the bioplatfrom is a simple, versatile, and universal system for the recognition, capture, release, and detection of different types of CTCs. Therefore, this bioplatform shows potential applications on the early diagnosis of cancers.


Assuntos
Separação Celular/métodos , Dissulfetos/química , Gelatina/química , Raios Infravermelhos , Molibdênio/química , Nanoestruturas/química , Células Neoplásicas Circulantes/patologia , Eletroquímica , Células HeLa , Humanos , Células MCF-7 , Polietilenoglicóis/química
10.
Insect Sci ; 27(4): 675-686, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30912872

RESUMO

During insect larval-pupal metamorphosis, proteins in the hemolymph are absorbed by the fat body for the maintenance of intracellular homeostasis; however, the type of proteins and how these proteins are internalized into the fat body are unclear. In Bombyx mori, the developmental profiles of total proteins in the hemolymph and fat body showed that hemolymph-decreased protein bands (55-100 kDa) were in accordance with those protein bands that increased in the fat body. Inhibition of clathrin-dependent endocytosis predominantly blocked the transportation of 55-100 kDa proteins from the hemolymph into the fat body, which was further verified by RNA interference treatment of Bmclathrin. Six hexamerins were shown to comprise ∼90% of the total identified proteins in both the hemolymph and fat body by mass spectrum (MS) analysis. In addition, hemolymph-specific proteins were mainly involved in material transportation, while fat body-specific proteins particularly participated in metabolism. In this paper, four hexamerins were found for the first time, and potential proteins absorbed by the fat body from the hemolymph through clathrin-dependent endocytosis were identified. This study sheds light on the protein absorption mechanism during insect metamorphosis.


Assuntos
Bombyx/fisiologia , Clatrina/metabolismo , Endocitose , Corpo Adiposo/fisiologia , Hemolinfa/fisiologia , Proteínas de Insetos/metabolismo , Absorção Fisiológica , Animais , Bombyx/crescimento & desenvolvimento , Larva/crescimento & desenvolvimento , Larva/fisiologia
11.
Carbohydr Polym ; 225: 115255, 2019 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-31521314

RESUMO

Selectivity of deproteinization and demineralization using natural deep eutectic solvents for preparation of insect chitin (Hermetia illucens) was investigated. The relationships between the pH and pKa values of NADESs and demineralization, deproteinization and crystallinity indexes of the obtained chitin were discussed, respectively. It was found that the acidic NADESs consist of basic hydrogen bond acceptor (HBA) and acidic hydrogen bond donor (HBD), or the alkali NADESs composed of acid HBA and alkali HBD were better for chitin products. After reused for three times, demineralization and deproteinization abilities of NADESs were not significantly decreased. Mechanistic analysis indicated that H+ released from HBA or HBD was the main reason for demineralization, whereas intermolecular and intramolecular hydrogen bond in NADESs facilitates the removal of protein. Overall, this report provides a preliminary reference for design of the sustainable NADESs for preparation of chitin from natural resources.


Assuntos
Quitina/química , Quitina/isolamento & purificação , Dípteros/metabolismo , Solventes/química , Animais , Ligação de Hidrogênio
12.
Anal Chem ; 91(11): 7181-7188, 2019 06 04.
Artigo em Inglês | MEDLINE | ID: mdl-31067856

RESUMO

A luminescence resonance energy transfer (LRET) system was successfully developed using near-infrared (NIR) Ag2S nanodots (NDs) as the energy acceptors and upconversion nanoparticles (UCNPs) as the energy donors. The system possessing the properties of NIR excitation (980 nm) and NIR emission (795 nm) was used for the ratiometric detection and bioimaging of pH in tumor cells and zebrafish. Glutathione and mercaptopropionic acid (MPA) co-modified Ag2S NDs (GM-Ag2S NDs) were prepared by ligand exchange with an excellent pH-responsive property over a pH range of 4.0 to 9.0. The NIR GM-Ag2S NDs were covalently grafted with silica coated UCNPs, and an efficient LRET platform was developed via modulation of the thickness of the silica coating. Due to the LRET process between UCNPs and GM-Ag2S NDs, a ratiometric luminescence nanoprobe with the properties of NIR excitation-NIR emission was constructed for pH biosensing and bioimaging. On the basis of high contrast bioimaging, the nanoplatform can distinguish between tumor and normal tissue in the zebrafish model.


Assuntos
Transferência Ressonante de Energia de Fluorescência , Nanopartículas/química , Imagem Óptica , Compostos de Prata/química , Ácido 3-Mercaptopropiônico/química , Animais , Glutationa/química , Células HeLa , Humanos , Concentração de Íons de Hidrogênio , Raios Infravermelhos , Peixe-Zebra
13.
Biosens Bioelectron ; 137: 148-153, 2019 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-31100597

RESUMO

Herein, a ratiometric fluorescent method was developed for alkaline phosphatase (ALP) detection based on near-infrared (NIR) Ag2S quantum dots (QDs) and calcein through the competitive approach. The system based on Ag2S QDs and calcein shows green (maximum emission at 512 nm from calcein) and near infrared (NIR) fluorescence (maximum 798 nm from Ag2S QDs) under the same excitation wavelength (468 nm). In the presence of Ce3+, the fluorescence intensity of calcein is decreased due to static quenching, while the fluorescence intensity of Ag2S QDs is enhanced through aggregation induced emission (AIE). The p-nitrophenyl phosphate is hydrolyzed by ALP, and the yield phosphate ions bind with Ce3+ with higher affinity than these of Ag2S QDs and calcein. Therefore, the green fluorescence from calcein is recovered while NIR fluorescence from Ag2S QDs is decreased. On the basis of these findings, a ratiometric fluorescence assay was developed for the measurement of ALP activity. The ratio of fluorescence intensity at 512 and 798 nm (F512/F798) was well associated with the ALP concentration ranging from 2 to 100 mU/mL with the detection limit of 1.28 mU/mL. The method was successfully applied for detecting ALP in human serum with an acceptable recovery and bioimaging intracellular ALP with good performance. In addition, the approach was also employed for the screening ALP inhibitor.


Assuntos
Fosfatase Alcalina/isolamento & purificação , Técnicas Biossensoriais , Pontos Quânticos/química , Fosfatase Alcalina/química , Fluoresceínas/química , Fluorescência , Humanos , Limite de Detecção , Compostos de Prata/química
14.
Dev Comp Immunol ; 95: 59-67, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30708025

RESUMO

Peptidoglycan recognition proteins (PGRPs) are members of an important class of pattern recognition receptors in insects that can specifically recognize peptidoglycan (PGN) in bacterial cell walls and participate in immune regulation and bacterial clearance. Although the role of PGRPs in regulating the innate immune response in Drosophila melanogaster has been studied, little is known regarding PGRPs in Lepidoptera species. In this study, five short (S)-type Bombyx mori PGRPs (BmPGRPs) were cloned, expressed, and evaluated for their function in innate immunity. B. mori larvae that were injected with the gram-positive bacterium Bacillus megaterium or the gram-negative bacterium Escherichia coli exhibited a rapid and significant upregulation in S-type BmPGRP expression. The results showed that the five evaluated BmPGRPs have significant agglutination activity toward E. coli and B. megaterium and more notable amidase activity toward meso-diaminopimelic acid peptidoglycan (DAP-PGN). Furthermore, only in the presence of BmPGRP-S5 did B. mori larval hemocytes exhibit significant phagocytosis against E. coli and B. megaterium.


Assuntos
Bombyx/imunologia , Proteínas de Transporte/imunologia , Imunidade Inata , Proteínas de Insetos/imunologia , Animais , Bacillus megaterium/imunologia , Bombyx/microbiologia , Proteínas de Transporte/isolamento & purificação , Proteínas de Transporte/metabolismo , Linhagem Celular , Drosophila melanogaster , Escherichia coli/imunologia , Hemócitos/imunologia , Hemócitos/metabolismo , Proteínas de Insetos/isolamento & purificação , Proteínas de Insetos/metabolismo , Larva/citologia , Larva/imunologia , Larva/metabolismo , Fagocitose/imunologia , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismo , Regulação para Cima
15.
Dev Comp Immunol ; 88: 94-103, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-30009928

RESUMO

Lipopolysaccharide (LPS) is a common component of the outermost cell wall in Gram-negative bacteria. In mammals, LPS serves as an endotoxin that can be recognized by a receptor complex of TLR4 (Toll-like receptor 4) and MD-2 (myeloid differentiation-2) and subsequently induce a strong immune response to signal the release of tumor necrosis factor (TNF). In Drosophila melanogaster, no receptors for LPS have been identified, and LPS cannot activate immune responses. Here, we report a protein, BmEsr16, which contains an ML (MD-2-related lipid-recognition) domain, may function as an LPS receptor in the silkworm Bombyx mori. We showed that antibacterial activity in the hemolymph of B. mori larvae was induced by Escherichia coli, peptidoglycan (PGN) and LPS and that the expression of antimicrobial peptide genes was also induced by LPS. Furthermore, both the expression of BmEsr16 mRNA in the fat body and the expression of BmEsr16 protein in the hemolymph were induced by LPS. Recombinant BmEsr16 bound to LPS and lipid A, as well as to PGN, lipoteichoic acid, but not to laminarin or mannan. More importantly, LPS-induced immune responses in the hemolymph of B. mori larvae were blocked when the endogenous BmEsr16 protein was neutralized by polyclonal antibody specific to BmEsr16. Our results suggest that BmEsr16 may function as a key accessory protein for LPS signaling in B. mori.


Assuntos
Bombyx/imunologia , Imunidade Inata , Proteínas de Insetos/imunologia , Receptores de Lipopolissacarídeos/imunologia , Lipopolissacarídeos/imunologia , Animais , Escherichia coli/imunologia , Proteínas de Escherichia coli/imunologia , Proteínas de Escherichia coli/metabolismo , Hemolinfa/imunologia , Proteínas de Insetos/química , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Larva/imunologia , Receptores de Lipopolissacarídeos/química , Receptores de Lipopolissacarídeos/genética , Receptores de Lipopolissacarídeos/metabolismo , Lipopolissacarídeos/química , Lipopolissacarídeos/metabolismo , Simulação de Acoplamento Molecular , Peptidoglicano/química , Peptidoglicano/imunologia , Domínios Proteicos/imunologia , RNA Mensageiro/metabolismo , Alinhamento de Sequência , Transdução de Sinais/imunologia
16.
Parasitol Res ; 116(10): 2741-2746, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28815299

RESUMO

A new microsporidium was isolated from Chilo suppressalis (Walker) (Lepidoptera: Pyralidae), one of the most important rice pests in China. The morphology and molecular systematics of this novel microsporidium were described in this study. The spores were long oval and measured 3.17 × 1.64 µm on fresh smears. Ultrastructure of the spores was characteristic for the genus Nosema: a diplokaryon, 10-12 polar filament coils of the same type, and posterior vacuole. Small subunit rRNA gene sequence data and phylogenetic analysis further confirmed that the microsporidian species from C. suppressalis belong to the true Nosema sub-group of the genus Nosema. Besides, the microsporidium Nosema sp. CS could cause systemic infection of Bombyx mori and infect silkworms through vertical transmission. Therefore, mulberry field pest control should be carefully monitored, and sanitation of mulberry leaves is essential to control the pebrine disease in sericulture.


Assuntos
Lepidópteros/microbiologia , Nosema/classificação , Nosema/crescimento & desenvolvimento , Oryza/parasitologia , Filogenia , Animais , Bombyx/microbiologia , China , Nosema/genética , Nosema/isolamento & purificação
17.
Int J Mol Sci ; 18(2)2017 Feb 10.
Artigo em Inglês | MEDLINE | ID: mdl-28208575

RESUMO

The silkworm Bombyx mori is an economically important species. White muscardine caused by Beauveria bassiana is the main fungal disease in sericulture, and understanding the silkworm responses to B. bassiana infection is of particular interest. Herein, we investigated the molecular mechanisms underlying these responses in two silkworm strains Haoyue (HY, sensitive to B. bassiana) and Kang 8 (K8, resistant to B. bassiana) using an RNA-seq approach. For each strain, three biological replicates for immersion treatment, two replicates for injection treatment and three untreated controls were collected to generate 16 libraries for sequencing. Differentially expressed genes (DEGs) between treated samples and untreated controls, and between the two silkworm strains, were identified. DEGs and the enriched Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways of the two strains exhibited an obvious difference. Several genes encoding cuticle proteins, serine proteinase inhibitors (SPI) and antimicrobial peptides (AMP) and the drug metabolism pathway involved in toxin detoxification were considered to be related to the resistance of K8 to B. bassiana. These results revealed insight into the resistance and susceptibility of two silkworm strains against B. bassiana infection and provided a roadmap for silkworm molecular breeding to enhance its resistance to B. bassiana.


Assuntos
Doenças dos Animais/genética , Doenças dos Animais/microbiologia , Beauveria , Bombyx/genética , Bombyx/microbiologia , Resistência à Doença , Predisposição Genética para Doença , Análise de Sequência de RNA , Animais , Bombyx/classificação , Biologia Computacional/métodos , Resistência à Doença/genética , Perfilação da Expressão Gênica , Sequenciamento de Nucleotídeos em Larga Escala , Especificidade da Espécie , Transcriptoma
18.
PLoS One ; 9(9): e105637, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25255321

RESUMO

Albendazole is a broad-spectrum parasiticide with high effectiveness and low host toxicity. No method is currently available for measuring albendazole and its metabolites in silkworm hemolymph. This study describes a rapid, selective, sensitive, synchronous and reliable detection method for albendazole and its metabolites in silkworm hemolymph using ultrafast liquid chromatography tandem triple quadrupole mass spectrometry (UFLC-MS/MS). The method is liquid-liquid extraction followed by UFLC separation and quantification in an MS/MS system with positive electrospray ionization in multiple reaction monitoring mode. Precursor-to-product ion transitions were monitored at 266.100 to 234.100 for albendazole (ABZ), 282.200 to 208.100 for albendazole sulfoxide (ABZSO), 298.200 to 159.100 for albendazole sulfone (ABZSO2) and 240.200 to 133.100 for albendazole amino sulfone (ABZSO2-NH2). Calibration curves had good linearities with R2 of 0.9905-0.9972. Limits of quantitation (LOQs) were 1.32 ng/mL for ABZ, 16.67 ng/mL for ABZSO, 0.76 ng/mL for ABZSO2 and 5.94 ng/mL for ABZSO2-NH2. Recoveries were 93.12%-103.83% for ABZ, 66.51%-108.51% for ABZSO, 96.85%-105.6% for ABZSO2 and 96.46%-106.14% for ABZSO2-NH2, (RSDs <8%). Accuracy, precision and stability tests showed acceptable variation in quality control (QC) samples. This analytical method successfully determined albendazole and its metabolites in silkworm hemolymph in a pharmacokinetic study. The results of single-dose treatment suggested that the concentrations of ABZ, ABZSO and ABZSO2 increased and then fell, while ABZSO2-NH2 level was low without obvious change. Different trends were observed for multi-dose treatment, with concentrations of ABZSO and ABZSO2 rising over time.


Assuntos
Albendazol/metabolismo , Antiparasitários/metabolismo , Bombyx/metabolismo , Cromatografia Líquida de Alta Pressão/métodos , Hemolinfa/metabolismo , Espectrometria de Massas em Tandem/métodos , Albendazol/análogos & derivados , Albendazol/isolamento & purificação , Albendazol/farmacocinética , Métodos Analíticos de Preparação de Amostras , Animais , Antiparasitários/isolamento & purificação , Antiparasitários/farmacocinética , Estabilidade de Medicamentos , Estudos de Viabilidade , Congelamento , Limite de Detecção , Modelos Lineares , Extração Líquido-Líquido , Fatores de Tempo
19.
Arch Insect Biochem Physiol ; 86(1): 33-45, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24719308

RESUMO

Destruxin A (DA), a cyclodepsipeptidic secondary metabolite of the entomopathogenic fungus, Metarhizium anisopliae, is an important anti-immunity agent against insect hemocytes. To understand the mechanism of the molecular responses to DA, fifth-instar larvae of the silkworm, Bombyx mori, were injected with 2 µg of DA. The proteomics of hemocytes were then investigated using two-dimensional electrophoresis and mass spectrometry, and validated qPCR. As a result, a total of 47 differently expressed protein spots were detected and 22 proteins in 26 spots were identified. There are eight immunity-related proteins, including three downregulated proteins (antitrypsin isoform 3, p50 protein, and calreticulin precursor) and five upregulated proteins (C-type lectin 10 precursor, serine proteinase-like protein, paralytic peptide, PPO-1, and PPO-2). Four resistance- and/or stress-related proteins (arginine kinase, carboxylesterase clade H, member 1, aminoacylase, and thiol peroxiredoxin) were upregulated. Ten proteins with other or unknown functions were also recorded. Five selected proteins were verified with qPCR. These results provide new insights into the molecular mechanism of host immune response to DA challenge.


Assuntos
Bombyx/efeitos dos fármacos , Bombyx/imunologia , Depsipeptídeos/toxicidade , Hemócitos/efeitos dos fármacos , Hemócitos/imunologia , Micotoxinas/toxicidade , Animais , Proteínas de Insetos , Larva , Proteoma
20.
PLoS One ; 6(3): e18109, 2011 Mar 29.
Artigo em Inglês | MEDLINE | ID: mdl-21479226

RESUMO

Antimicrobial peptides are small-molecule proteins that are usually encoded by multiple-gene families. They play crucial roles in the innate immune response, but reports on the functional divergence of antimicrobial peptide gene families are rare. In this study, 14 paralogs of antimicrobial peptides belonging to cecropin, moricin and gloverin families were recombinantly expressed in pET expression systems. By antimicrobial activity tests, peptides representing paralogs in the same family of cecropin and moricin families, displayed remarkable differences against 10 tested bacteria. The evolutionary rates were relatively fast in the two families, which presented obvious functional divergence among paralogs of each family. Four peptides of gloverin family had similar antimicrobial spectrum and activity against tested bacteria. The gloverin family showed similar antimicrobial function and slow evolutionary rates. By induced transcriptional activity, genes encoding active antimicrobial peptides were upregulated at obviously different levels when silkworm pupae were infected by three types of microbes. Association analysis of antimicrobial activities and induced transcriptional activities indicated that the antimicrobial activities might be positively correlated with induced transcriptional activities in the cecropin and moricin families. These results suggest that representative BmcecB6, BmcecD and Bmmor as the major effector genes have broad antimicrobial spectrum, strong antimicrobial activity and high microbe-induced expression among each family and maybe play crucial roles in eliminating microbial infection.


Assuntos
Peptídeos Catiônicos Antimicrobianos/genética , Bombyx/genética , Perfilação da Expressão Gênica , Proteínas Recombinantes/genética , Homologia de Sequência de Aminoácidos , Substituição de Aminoácidos/genética , Animais , Peptídeos Catiônicos Antimicrobianos/metabolismo , Peptídeos Catiônicos Antimicrobianos/farmacologia , Sequência de Bases , Bombyx/efeitos dos fármacos , Bombyx/microbiologia , Eletroforese em Gel de Poliacrilamida , Regulação da Expressão Gênica/efeitos dos fármacos , Variação Genética/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Família Multigênica/genética , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacologia , Transcrição Gênica/efeitos dos fármacos , Ultrafiltração
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