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1.
Pestic Biochem Physiol ; 174: 104809, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33838710

RESUMO

Energy metabolism is important for the proliferation of microsporidia in infected host cells, but there is limited information on the host response. The energy metabolism response of silkworm (Bombyx mori) to microsporidia may help manage Nosema bombycis infections. We analyzed differentially expressed genes in the B.mori midgut transcriptome at two significant time points of microsporidia infection. A total of 1448 genes were up-regulated, while 315 genes were down-regulated. A high proportion of genes were involved in the phosphatidylinositol signaling system, protein processing in the endoplasmic reticulum, and glycerolipid metabolism at 48 h post infection (h p.i.), and a large number of genes were involved in the TCA cycle and protein processing at 120 h p.i. These results showed that the early stages of microsporidia infection affected the basic metabolism and biosynthesis processes of the silkworm. Knockout of Bm_nscaf2860_46 (Bombyx mori isocitrate dehydrogenase, BmIDH) and Bm_nscaf3027_062 (Bombyx mori hexokinase, BmHXK) reduced the production of ATP and inhibited microsporidia proliferation. Host fatty acid degradation, glycerol metabolism, glycolysis pathway, and TCA cycle response to microsporidia infection were also analyzed, and their importance to microsporidia proliferation was verified. These results increase our understanding of the molecular mechanisms involved in N. bombycis infection and provide new insights for research on microsporidia control. IMPORTANCE: Nosema bombycis can be vertically transmitted in silkworm eggs. The traditional prevention and control strategies for microsporidia are difficult and time-consuming, and this is a problem in silkworm culture. Research has mainly focused on host gene functions related to microsporidia infection and host immune responses after microsporidia infection. Little is known about the metabolic changes occurring in the host after infection. Understanding the metabolic changes in the silkworm host could aid in the recognition of host genes important for microsporidia infection and growth. We analyzed host metabolic changes and the main participating pathways at two time points after microsporidia infection and screened the microsporidia-dependent host energy metabolism genes BmIDH and BmHXK. The results revealed genes that are important for the proliferation of Nosema bombycis. These results illustrate how microsporidia hijack the host genome for their growth and reproduction.


Assuntos
Bombyx , Nosema , Animais , Bombyx/genética , Metabolismo Energético/genética , Perfilação da Expressão Gênica , Nosema/genética
2.
Pestic Biochem Physiol ; 174: 104824, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33838717

RESUMO

Acetamiprid is a new type of nicotinic insecticide that is widely used in pest control. Its environmental residues may cause silkworm cocooning disorder. In this study, silkworms that received continuous feeding of low concentration acetamiprid (0.15 mg/L) showed significantly decreased silk gland index and cocooning rate. Gene expression profiling of posterior silk glands (PSGs) revealed that the differentially expressed genes were significantly enriched in oxidative stress-related signal pathways with significant up-regulation. The contents of both H2O2 and MDA were increased, along with significantly elevated SOD and CAT activities, all of which reached maximal values at 48 h when H2O2 and MDA's contents were 10.46 and 7.98 nmol/mgprot, respectively, and SOD and CAT activities were 5.51 U/mgprot and 33.48 U/gprot, respectively. The transcription levels of antioxidant enzyme-related genes SOD, Mn-SOD, CuZn-SOD, CAT, TPX and GPX were all up-regulated, indicating that exposure to low concentration acetamiprid led to antioxidant response in silkworm PSG. The key genes in the FoxO/CncC/Keap1 signaling pathway that regulates antioxidant enzyme activity, FoxO, CncC, Keap1, NQO1, HO-1 and sMaf were all up-regulated during the whole process of treatment, with maximal values being reached at 72 h with 2.91, 1.46, 1.82, 2.52, 2.32 and 4.01 times of increases, respectively. These results demonstrate that exposure to low concentration acetamiprid causes oxidative stress in silkworm PSG, which may be the cause of cocooning disorder in silkworm. Our study provides a reference for the safety evaluation of environmental residues of acetamiprid on non-target insects.


Assuntos
Bombyx , Animais , Bombyx/genética , Bombyx/metabolismo , Crescimento e Desenvolvimento , Peróxido de Hidrogênio , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Proteína 1 Associada a ECH Semelhante a Kelch , Fator 2 Relacionado a NF-E2/metabolismo , Neonicotinoides , Estresse Oxidativo , Seda
3.
Int J Mol Sci ; 22(5)2021 Mar 08.
Artigo em Inglês | MEDLINE | ID: mdl-33800168

RESUMO

Transgenic silkworm expression systems have been applied for producing various recombinant proteins. Knocking out or downregulating an endogenous silk protein is considered a viable strategy for improving the ability of transgenic expression systems to produce exogenous proteins. Here, we report the expression of human epidermal growth factor (hEGF) in a P25 gene knockout silkworm. The hEGF gene regulated by the P25 gene promoter was integrated into a silkworm's genome. Five transgenic positive silkworm lineages were generated with different insertion sites on silkworm chromosomes and the ability to synthesize and secrete proteins into cocoons. Then, a cross-strategy was used to produce transgenic silkworms with a P25 gene knockout background. The results of the protein analysis showed that the loss of an endogenous P25 protein can increase the hEGF production to about 2.2-fold more than normal silkworms. Compared to those of transgenic silkworms with wild type (non-knockout) background, the morphology and secondary structure of cocoon silks were barely changed in transgenic silkworms with a P25 gene knockout background, indicating their similar physical properties of cocoon silks. In conclusion, P25 gene knockout silkworms may become an efficient bioreactor for the production of exogenous proteins and a promising tool for producing various protein-containing silk biomaterials.


Assuntos
Animais Geneticamente Modificados , Bombyx , Fator de Crescimento Epidérmico , Fibroínas/genética , Técnicas de Silenciamento de Genes , Animais , Animais Geneticamente Modificados/genética , Animais Geneticamente Modificados/metabolismo , Bombyx/genética , Bombyx/metabolismo , Fator de Crescimento Epidérmico/biossíntese , Fator de Crescimento Epidérmico/genética , Humanos , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo
4.
Arch Insect Biochem Physiol ; 106(3): e21768, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33644912

RESUMO

Bombyx mori Masculinizer protein (BmMasc) is essential for both masculinization and dosage compensation in B. mori. We previously identified a bipartite nuclear localization signal (NLS) of BmMasc and two essential residues (lysine at 274 [K274] and arginine at 275 [R275]) implicated in its function. Sequence comparison showed the presence of putative NLSs in lepidopteran Masc proteins, but their functional properties and critical residues are unknown. Here we characterized a putative NLS of Ostrinia furnacalis Masc (OfMasc) using B. mori ovary-derived BmN-4 cell line. Deletion and alanine scanning mutagenesis revealed that a putative NLS is required for nuclear localization of OfMasc. However, mutations at both K227 and R228, which correspond to K274 and R275 of BmMasc, respectively, do not greatly abolish the NLS activity. Additional mutagenesis analysis revealed that triple mutations at K227, R228, and K240 almost completely inhibited OfMasc nuclear localization. These results suggest that lepidopteran Masc proteins possess a common functional NLS, but the critical residues for its activity are different. Moreover, we examined the masculinizing activity of OfMasc derivatives and found that nuclear localization is not required for the masculinizing activity of OfMasc. The results from our studies indicate that lepidopteran Masc proteins function in the cytoplasm to drive masculinizing cascade.


Assuntos
Mariposas/genética , Sinais de Localização Nuclear , Animais , Bombyx/genética , Linhagem Celular , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Insetos , Sinais de Localização Nuclear/genética , Sinais de Localização Nuclear/metabolismo
5.
Arch Insect Biochem Physiol ; 106(4): e21783, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33719082

RESUMO

Vitamin C (VC) is an essential nutrient for many animals. However, whether insects, including Bombyx mori, can synthesize VC remains unclear. In this article, the optimized HPLC method was used to determine the content of l-ascorbic acid (AsA) in silkworm eggs, larvae and pupae, and the activity of l-gulono-1,4-lactone oxidase (GULO), a key enzyme in VC synthesis. The RNA interference method was used to determine the effect of the BmGulo-like gene on embryonic development and GULO activity in the pupal fat body. The AsA content increased significantly during E144 h-E168 h in the late embryonic stage and P48 h-P144 h in the middle-late pupal stage, in which exogenous VC was not ingested. Furthermore, the body AsA content in larvae fed VC-free feed also increased with larval stage. The GULO enzymatic activity was present in eggs and the fat bodies of larvae and pupae, even when the larvae were reared with fresh mulberry leaves. Moreover, the activity was higher in the later embryonic stages (E144 h-E168 h) and the early pupal stage (before P24 h). The GULO activity in the pupal fat body dramatically decreased when the screened BmGulo-like gene (BGIBMGA005735) was knocked down with small interfering RNA; in addition, the survival rate and hatching rate of eggs significantly decreased 21% and 44%, respectively, and embryonic development was delayed. Thus, Bombyx mori can synthesize AsA through the l-gulose pathway, albeit with low activity, and this synthesis ability varies with developmental stages.


Assuntos
Ácido Ascórbico/metabolismo , Bombyx/metabolismo , Animais , Bombyx/crescimento & desenvolvimento , Hexoses/metabolismo , Larva/crescimento & desenvolvimento , Larva/metabolismo , Pupa/crescimento & desenvolvimento , Pupa/metabolismo , Açúcares Ácidos/metabolismo
6.
J Insect Sci ; 21(2)2021 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-33764365

RESUMO

Lactic acid is widely used in the food, drugs, cosmetics, and other industries to maintain the microbial stability of low-pH products. However, it is unclear whether lactic acid can affect silkworm (Bombyx mori) growth and reproduction. This study investigated the effects of lactic acid on the growth and reproduction of the silkworm. We analyzed the growth, cocoon quality, and reproductive performance of fifth instar larvae fed on mulberry leaves saturated with different concentrations (0.01, 0.1, 1, and 10%) of lactic acid and the control. Results showed that 0.01, 0.1, and 1% lactic acid supplementation positively affects growth and female cocoon quality, with increased larval weight and female cocoon shell weight compared to the control group. In contrast, 10% lactic acid was toxic to the larvae and significantly decreased growth, leading to larval death. Our study provides a basic reference for the optimal amount of preservatives. In addition, this study can be a desirable intervention for sericulturists and can play an important role in getting high return from silkworm-rearing activities.


Assuntos
Bombyx/crescimento & desenvolvimento , Suplementos Nutricionais , Ácido Láctico , Animais , Peso Corporal , Fertilidade , Larva/crescimento & desenvolvimento , Reprodução
7.
Waste Manag ; 124: 72-81, 2021 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-33610113

RESUMO

Waste disposal and utilisation of its important components are pioneering goals for achieving sustainable development and a clean environment. Silkworm pupae (SWP) are considered a by-product of the sericulture industry and may contain a high concentration of selenium (Se) in some regions, making them a potentially hazardous waste posing health risks. This study examined six treatments of Se-rich SWP (0-100%) as a substrate for black soldier fly (BSF) larvae. Growth performance and protein content of BSF larvae were not affected by increasing SWP content. The total fat, mono-, and poly-unsaturated fatty acids in BSF pre-pupae increased with increasing SWP inclusion in the substrate, from P0 (pre-pupae fed control treatment) through P100 (pre-pupae fed 100% SWP treatment), by 18.83, 61.14, and 62.42%, respectively. The results of significance were: (1) BSF pre-pupae did not accumulate Se, maintaining the same amount of Se in their bodies (~0.18 mg/kg); (2) omega-3 fatty acids represented by linolenic acid in BSF increased by 1,223.35% from P0 to P100, with 70.65% recovered from the SWP; (3) valine percentage increased in BSF compared with the percentage of SWP by 25.30%; and (4) BSF larvae were observed reducing SWP waste by more than one-third. BSF larvae can reduce the waste from sericulture industry and exploit the beneficial properties and components of SWP, converting them into safe and highly nutritious products.


Assuntos
Bombyx , Dípteros , Ácidos Graxos Ômega-3 , Selênio , Animais , Larva , Valor Nutritivo
8.
Animal ; 15(2): 100112, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33573975

RESUMO

Silkworm (Bombyx mori L.) pupae are a by-product derived from silk production, which is often treated as waste and thus discarded: this can cause serious environmental problems and a loss of nutrients. Silkworm pupae are a rich source of protein and lipids, and the resulting protein meal can provide promising outcomes as livestock feed, notably for monogastric species. However, one possible issue that needs to be considered is the possible implication of the 1-Deoxynojirimycin (1-DNJ), a bio-compound of the silkworm that impairs glucose absorption, in poultry nutrition. Therefore, the present study evaluated the effect of the dietary inclusion of full-fat or defatted silkworm pupa meal (SWM) on the apparent digestibility of nutrients, feed choice and faecal microbiome in meat-producing quails. For the digestibility trial, a total of thirty-three 27-day-old Japanese quails (Coturnix coturnix japonica) were individually housed in digestibility cages and received three experimental diets: a control diet (control, commercial feed for fattening quails), and two other diets containing the 12.5% of either a full-fat SWM (SWM-FULL) or a defatted SWM (SWM-DEF). Subsequently, twenty-seven 33-day-old quails were simultaneously provided with Control, SWM-FULL and SWM-DEF diets for a 10-day feed choice trial. The results of the digestibility trial showed that the DM intake and excreta production were higher in both SWM groups than in the Control one (P < 0.001). The apparent digestibility of DM, organic matter, CP, ether extract, starch and energy was lower in both SWM groups than in the control group (P < 0.001), suggesting the possible implication of chitin and 1-DNJ. The feed choice test showed that quails preferred the Control diet (P < 0.001). From the microbiome analysis of the excreta, families such as Streptococcaceae (P < 0.05), Rikenellaceae and Eubacteriaceae (P < 0.01) and taxa at species level such as Lactobacillus delbrueckii (P < 0.05), Aneurinibacillus thermoaerophilus and Bacillus thermoamylovorans (P < 0.01) scored higher in SWM-FULL quails than in SWM-DEF and Control treatments. The present study demonstrated that a successful dietary inclusion of SWM for fattening quails needs to overcome the digestive criticalities caused by the of presence specific bio-compounds, namely chitin and 1-DNJ.


Assuntos
Ração Animal , Bombyx , Microbiota , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Animais , Bacillales , Bacillus , Coturnix , Dieta/veterinária , Digestão , Nutrientes , Pupa , Codorniz
9.
Arch Insect Biochem Physiol ; 106(3): 1-12, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33619747

RESUMO

Bombyx mori cypovirus (BmCPV) is one of the most important pathogens causing severe disease to silkworm. Emerging evidence indicates that long noncoding RNAs (lncRNAs) play importantly regulatory roles in virus infection and host immune response. To better understand the interaction between silkworm, Bombyx mori and BmCPV, we performed a comparative transcriptome analysis on lncRNAs and mRNAs between the virus-infected and noninfected silkworm larvae midgut at two time points postinoculation. A total of 16,753 genes and 1845 candidate lncRNAs were identified, among which 356 messenger RNA (mRNAs) and 41 lncRNAs were differentially expressed (DE). Target gene prediction revealed that most of DEmRNAs (123) were coexpressed with 28 DElncRNAs, suggesting that the expression of mRNA is mainly affected through trans- regulation by BmCPV-induced lncRNAs, and a regulatory network of DElncRNAs and DEmRNAs was then constructed. According to the network, many genes involved in apoptosis, autophagy, and antiviral response, such as ATG3, PDCD6, IBP2, and MFB1, could be targeted by different DElncRNAs, implying the essential roles of these genes and lncRNAs in BmCPV infection. In all, our studies revealed for the first time the alteration of lncRNA expression in BmCPV-infected larvae and its potential influence on BmCPV replication, providing a new perspective for host-cypovirus interaction studies.


Assuntos
Bombyx , RNA Longo não Codificante , Viroses , Animais , Bombyx/genética , Bombyx/imunologia , Bombyx/metabolismo , Bombyx/virologia , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Redes Reguladoras de Genes , Genes de Insetos , Interações entre Hospedeiro e Microrganismos , Imunidade , Larva/genética , Larva/imunologia , Larva/metabolismo , RNA Longo não Codificante/isolamento & purificação , RNA Longo não Codificante/metabolismo , RNA Mensageiro/metabolismo , Reoviridae , Viroses/imunologia , Viroses/metabolismo
10.
Nucleic Acids Res ; 49(4): 2306-2316, 2021 02 26.
Artigo em Inglês | MEDLINE | ID: mdl-33524157

RESUMO

Vast G-quadruplexes (GQs) are primarily folded by one, two, or four G-rich oligomers, rarely with an exception. Here, we present the first NMR solution structure of a trimolecular GQ (tri-GQ) that is solely assembled by the self-trimerization of d(GTTAGG), preferentially in Na+ solution tolerant to an equal amount of K+ cation. Eight guanines from three asymmetrically folded strands of d(GTTAGG) are organized into a two-tetrad core, which features a broken G-column and two width-irregular grooves. Fast strand exchanges on a timescale of second at 17°C spontaneously occur between folded tri-GQ and unfolded single-strand of d(GTTAGG) that both species coexist in dynamic equilibrium. Thus, this tri-GQ is not just simply a static assembly but rather a dynamic assembly. Moreover, another minor tetra-GQ that has putatively tetrameric (2+2) antiparallel topology becomes noticeable only at an extremely high strand concentration above 18 mM. The major tri-GQ and minor tetra-GQ are considered to be mutually related, and their reversible interconversion pathways are proposed accordingly. The sequence d(GTTAGG) could be regarded as either a reading frame shifted single repeat of human telomeric DNA or a 1.5 repeat of Bombyx mori telomeric DNA. Overall, our findings provide new insight into GQs and expect more functional applications.


Assuntos
DNA/química , Quadruplex G , Potássio/química , Sódio/química , Animais , Bombyx/genética , Temperatura Alta , Humanos , Modelos Moleculares , Ressonância Magnética Nuclear Biomolecular , Telômero/química
11.
ACS Appl Mater Interfaces ; 13(7): 8880-8888, 2021 Feb 24.
Artigo em Inglês | MEDLINE | ID: mdl-33576225

RESUMO

We report the unique actuation characteristics of moisture-driven, fully reversible soft biopolymer films fabricated from Bombyx mori silk. The instantaneous actuation is driven by the water vapor induced stress gradient generated across the thickness of the film, and it possesses subsecond response and actuation times. The excellent durability and consistent performance of the film without any noticeable fatigue are established by subjecting it to more than a thousand continuous actuation cycles. The weight-lifting capability of the film is fascinating, where a few tens of micrograms of water generate a colossal force required to lift hundreds of milligrams of weight. Several other potential uses of silk fibroin based soft actuators, such as an intelligent textile layer with the crescent-shaped windows that open on perspiring skin and an autonomous crawler, are also demonstrated. Interestingly, even moisture emanating from the human palm triggers the ultrafast actuation process. These silk films are fabricated using a simple facile solution-casting technique, which can be scaled up with relative ease.


Assuntos
Fibroínas/química , Seda/química , Animais , Bombyx , Tamanho da Partícula , Propriedades de Superfície , Volatilização
12.
J Insect Sci ; 21(1)2021 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-33560363

RESUMO

Insect innate immunity is initiated by the special recognition and binding of the foreign pathogens, which is accomplished by the pattern recognition receptors (PRRs). As an important type of PRRs, C-type lectins (CTLs) play various roles in insect innate immunity, including pathogen recognition, stimulation of prophenoloxidase, regulation of cellular immunity and so on. In this study, we have cloned the full-length cDNA of a CTL gene named CTL-S6 from the silkworm, Bombyx mori. The open reading frame (ORF) of B. mori CTL-S6 encodes 378 amino acids, which contain a secretion signal peptide. The mRNA of CTL-S6 exhibited the highest transcriptional level in the midgut. Its transcriptional level increased dramatically in fat body and hemocytes upon Escherichia coli or Micrococcus luteus challenge. Purified recombinant CTL-S6 could bind to bacterial cell wall components, including peptidoglycan (PGN, from Bacillus subtilis) and lipopolysaccharide (LPS, from E. coli 0111:B4), and recombinant CTL-S6 was involved in the encapsulation and melanization of hemocytes. Furthermore, the addition of recombinant CTL-S6 to the hemolymph of silkworm resulted in a significant increase in phenoloxidase activity. Overall, our results indicated that B. mori CTL-S6 may serve as a PRR for the recognition of foreign pathogens, prophenoloxidase pathway stimulation and involvement in the innate immunity.


Assuntos
Bombyx/genética , Bombyx/imunologia , Imunidade Inata , Proteínas de Insetos/genética , Lectinas Tipo C/genética , Sequência de Aminoácidos , Animais , Bombyx/crescimento & desenvolvimento , Escherichia coli/fisiologia , Corpo Adiposo/imunologia , Hemócitos/imunologia , Proteínas de Insetos/química , Proteínas de Insetos/metabolismo , Larva/genética , Larva/crescimento & desenvolvimento , Larva/imunologia , Lectinas Tipo C/química , Lectinas Tipo C/metabolismo , Micrococcus luteus/fisiologia , Filogenia , Alinhamento de Sequência
13.
Gene ; 777: 145450, 2021 Apr 20.
Artigo em Inglês | MEDLINE | ID: mdl-33516793

RESUMO

The Bombyx mori Shadow gene (BmShadow) belongs to the superfamily of cytochrome P450 genes. To elucidate the function of the BmShadow gene and its association with diapause, we employed the CRISPR/Cas9 system to knock out the BmShadow gene in the bivoltine strain Qiufeng. The mutant (BmShadow-/-) was obtained in G2, exhibiting a 42-base deletion corresponded exactly to the amino acids regions from positions 155 to 168. The larvae of BmShadow-/- cannot moult at the pre-moulting stage of the 2nd instar. When the BmShadow-/- larvae were fed with 20E analogue at the late stage of the 2nd instar, they were rescued and developed into the 3rd instar. Rescue experiments indicated that the 20E concentration of BmShadow-/- larvae was significantly lower than that in WT larvae, and the 20E concentration of BmShadow-/- larvae which fed 20E analogue was restored to normal levels. Interestingly, the BmShadow-/- larvae could not moult on the 1st instar when they hatched from eggs after being stored at 5 °C for 40 days or after hibernation, suggesting that the 20E transported from the mother was partially consumed in the diapause maintenance phase. Our study confirmed that BmShadow is involved in 20E synthesis and a 14-amino acids region from position 155 to 168 was essential for its function, also there appears to be no other compensation pathway in vivo, which offered an important potential target locus for the control of silkworm development and the biological control of agricultural and forestry pests.


Assuntos
Bombyx/genética , Sistema Enzimático do Citocromo P-450/genética , Muda/genética , Aminoácidos/metabolismo , Animais , Sistema Enzimático do Citocromo P-450/metabolismo , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Ecdisterona/genética , Ecdisterona/metabolismo , Proteínas de Insetos/genética , Larva/genética
14.
ACS Appl Mater Interfaces ; 13(2): 3024-3032, 2021 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-33404230

RESUMO

A wearable surface-enhanced Raman scattering (SERS) sensor has been developed as a patch type to utilize as a molecular sweat sensor. Here, the SERS patch sensor is designed to comprise a sweat-absorbing layer, which is an interface to the human skin, an SERS active layer, and a dermal protecting layer that prevents damage and contaminations. A silk fibroin protein film (SFF) is a basement layer that absorbs aqueous solutions and filtrates molecules larger than the nanopores created in the ß-sheet matrix of the SFF. On the SFF layer, a plasmonic silver nanowire (AgNW) layer is formed to enhance the Raman signal of the molecules that penetrated through the SERS patch in a label-free method. A transparent dermal protecting layer (DP) allows laser penetration to the AgNW layer enabling Raman measurement through the SERS patch without its detachment from the surface. The molecular detection capability and time-dependent absorption properties of the SERS patch are investigated, and then, the feasibility of its use as a wearable drug detection sweat sensor is demonstrated using 2-fluoro-methamphetamine (2-FMA) on the human cadaver skin. It is believed that the developed SERS patch can be utilized as various flexible and wearable biosensors for healthcare monitoring.


Assuntos
Técnicas Biossensoriais/instrumentação , Análise Espectral Raman/instrumentação , Suor/química , Dispositivos Eletrônicos Vestíveis , Animais , Bombyx/química , Estimulantes do Sistema Nervoso Central/análise , Monitoramento de Medicamentos/instrumentação , Fibroínas/química , Humanos , Metanfetamina/análogos & derivados , Metanfetamina/análise , Nanofios/química , Prata/química , Propriedades de Superfície
15.
Int J Biol Macromol ; 172: 124-132, 2021 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-33418047

RESUMO

The combination of biomimetic and 3D printing has created novel opportunities for the manufacture of 3D engineered materials. A sub-microscale E-Jet 3D printing method, inspired by the dehydration and protein enrichment process of silkworm, was developed to fabricate composite bone tissue scaffold with the characteristics of controllability, fast and inexpensive. By applying the resultant effects of thermal field and flow field to low viscous composite ink, the concentration gradient biopolymer ink was obtained near the needle tip, mimicking the advanced dehydration of natural spinning apparatus. After electrical shearing force were applied on concentration gradient ink, a stable and fine jet formed. Various printing modes (droplet, continuous fiber) and structure resolutions were achieved by adjusting local solvent evaporation. Thin film, high resolution 2D structures, high aspect ratio well-bonding 3D structures were fabricated. The printed result showed that a 100 µm-sized needle could be employed directly to print patterning down to 800 nm. The printed composite scaffold with controllability of fiber size and space has been proved the feasibility as a medium for bone tissue regeneration. It can be estimated that the novel biomimetic E-Jet 3D printing technique is a new and promising way for bone tissue repairing.


Assuntos
Bombyx/química , Regeneração Óssea/efeitos dos fármacos , Osso e Ossos/efeitos dos fármacos , Tecidos Suporte/química , Células 3T3 , Animais , Biopolímeros/química , Linhagem Celular , Hidrodinâmica , Camundongos , Impressão Tridimensional , Solventes/química , Engenharia Tecidual/métodos
16.
Ecotoxicol Environ Saf ; 208: 111582, 2021 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-33396105

RESUMO

In recent years, heavy metal pollution has caused immeasurable harm to the environment. As an emerging technology, phytoremediation technology has gained a place in the treatment of heavy metal pollution with its unique advantages. This study analyzes the toxic effects of mulberry (Morus alba) seeds, seedling growth and silkworm under heavy metal stress of lead (Pb) and cadmium (Cd), and explore the accumulation and migration of Pb and Cd in the soil-mulberry tree-silkworm system. The main results were as follows: (1) Seed germination and potted seedling experiments were conducted under heavy metal Pb and Cd stresses, and it was found that Pb and Cd had inhibitory effects on mulberry seed germination, growth and photosynthesis of mulberry seedlings, and as the concentration of heavy metals increased, the stronger the inhibitory effect. Moreover, Pb and Cd have a synergistic effect under compound stress. (2) The accumulation and transfer rules of Pb and Cd ions in mulberry were different. The content of Pb in mulberry was root > leaf > stem and the content of Cd was root > stem > leaf. The combined stress promoted the transfer of Pb and Cd from the underground part to the aerial portion of mulberry. (3) The silkworm feeds on mulberry leaves contaminated with heavy metals in this experiment and found that: with the increase of silkworm feeding, the heavy metal content in the silkworm body increased significantly, but the content remained in the silkworm body was less, most of it was excreted with silkworm excrement. Combined stress has no significant effect on the detoxification mechanism of silkworm. It is indispensable to think of the synergistic effect of heavy metals on plants germination when seeds are used for phytoremediation.


Assuntos
Bombyx/fisiologia , Cádmio/toxicidade , Cadeia Alimentar , Chumbo/toxicidade , Morus/fisiologia , Poluentes do Solo/toxicidade , Solo/química , Animais , Biodegradação Ambiental , Cádmio/análise , Cádmio/metabolismo , Frutas/química , Metais Pesados/análise , Fotossíntese , Folhas de Planta/química , Plântula/química , Poluentes do Solo/análise , Poluentes do Solo/metabolismo
17.
J Insect Sci ; 21(1)2021 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-33511414

RESUMO

Insect innate immunity is initiated by the special recognition and binding of the foreign pathogens, which is accomplished by the pattern recognition receptors (PRRs). As an important type of PRRs, C-type lectins (CTLs) play various roles in insect innate immunity, including pathogen recognition, stimulation of prophenoloxidase, regulation of cellular immunity and so on. In this study, we have cloned the full-length cDNA of a CTL gene named CTL-S6 from the silkworm, Bombyx mori. The open reading frame (ORF) of B. mori CTL-S6 encodes 378 amino acids, which contain a secretion signal peptide. The mRNA of CTL-S6 exhibited the highest transcriptional level in the midgut. Its transcriptional level increased dramatically in fat body and hemocytes upon Escherichia coli or Micrococcus luteus challenge. Purified recombinant CTL-S6 could bind to bacterial cell wall components, including peptidoglycan (PGN, from Bacillus subtilis) and lipopolysaccharide (LPS, from E. coli 0111:B4), and recombinant CTL-S6 was involved in the encapsulation and melanization of hemocytes. Furthermore, the addition of recombinant CTL-S6 to the hemolymph of silkworm resulted in a significant increase in phenoloxidase activity. Overall, our results indicated that B. mori CTL-S6 may serve as a PRR for the recognition of foreign pathogens, prophenoloxidase pathway stimulation and involvement in the innate immunity.


Assuntos
Escherichia coli/fisiologia , Imunidade Inata/genética , Proteínas de Insetos/genética , Lectinas Tipo C/genética , Micrococcus luteus/fisiologia , Receptores de Reconhecimento de Padrão/genética , Transcrição Genética , Sequência de Aminoácidos , Animais , Bombyx , Corpo Adiposo/imunologia , Perfilação da Expressão Gênica , Hemócitos/imunologia , Proteínas de Insetos/química , Proteínas de Insetos/metabolismo , Lectinas Tipo C/química , Lectinas Tipo C/metabolismo , Filogenia , Receptores de Reconhecimento de Padrão/química , Receptores de Reconhecimento de Padrão/metabolismo , Alinhamento de Sequência
18.
Arch Insect Biochem Physiol ; 106(2): e21762, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33415772

RESUMO

Bombyx mori nucleopolyhedrovirus (BmNPV) is one of primary silkworm pathogens and causes a serious damage of cocoon losses every year. Recent years, many works have been done to clarify the silkworm anti-BmNPV mechanism, and a significant progress has been made in screening and studying of genes and proteins related to BmNPV infection, but several of them lacked the proofs in vivo. In this study, to further validate the function of seven newly reported genes in vivo, including BmAtlatin-n, Bmferritin-heavy chain (BmFerHCH), Bmthymosin (BmTHY), Bmseroin1, Bmseroin2, Bmnuclear hormone receptors 96 (BmNHR96), and BmE3 ubiquitin-protein ligase SINA-like 10 (BmSINAL10), the response of them in the midgut, fat body, and hemolymph of differentially resistant strains (resistant strain YeA and susceptible strain YeB) at 48 h following BmNPV infection were analyzed. The results showed that the relative stable or upregulated expression level of BmAtlatin-n, BmTHY, Bmseroin1, and Bmseroin2 in YeA resistant strain following BmNPV infection further indicated their antiviral role in vivo, compared with susceptible YeB strain. Moreover, the significant downregulation of BmFerHCH, BmNHR96, and BmSINAL10 in both strains following BmNPV infection revealed their role in benefiting virus infection, as well as the upregulation of BmFerHCH in YeB midgut and BmSINAL10 in YeB hemolymph. These data could be used to complementary the proofs of the function of these genes in response to BmNPV infection.


Assuntos
Bombyx/genética , Bombyx/virologia , Genes de Insetos , Interações Hospedeiro-Patógeno , Nucleopoliedrovírus/fisiologia , Animais , Bombyx/crescimento & desenvolvimento , Bombyx/metabolismo , Corpo Adiposo/metabolismo , Trato Gastrointestinal/metabolismo , Hemolinfa/metabolismo , Larva/genética , Larva/crescimento & desenvolvimento , Larva/virologia
19.
Ecotoxicol Environ Saf ; 210: 111888, 2021 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-33421719

RESUMO

This study aimed to explore the toxicity of environmental residues of graphene oxide nanoparticles (GONPs) to reproduction of Lepidopteron insects using both ovary cell line (BmN) and individual female Bombyx mori as the research subjects. The results showed that GONPs dose dependently affect BmN cells. At higher concentrations (>25 mg/L), GONPs led to oxidative stress, ROS accumulation and DNA damage in BmN cells and significantly reduced their survival rate (p ≤ 0.05). Moreover, feeding female B. mori larvae with mulberry leaves treated with 25 mg/L GONPs significantly decreased their gonadosomatic index (GSI) by 40.84%, and increased oxidation levels and antioxidant enzyme activity in silkworm ovary tissues. Pathological analysis found that exposure to GONPs decreased the numbers of both oogonia and oocytes in ovarian tissues, increased the formation of peroxisome and vacuoles in follicle cells, reduced the transcription of genes (Vg, Ovo, Sxl-s, Sxl-l, and Otu) related to ovarian development in B. mori by 0.61, 0.65, 0.75, 0.72, and 0.42-fold, respectively, and lowered the amount of spawning by 52.25%. Overall, these results revealed that GONPs exposure is toxic to the reproduction of B. mori. The underlying mechanism is that oxidative stress due to GONPs causes oxidative damage to DNA, damages ovarian tissues, as well as hinders B. mori development and spawning. Thus, this study provides important experimental data for safety evaluation of reproductive toxicity due to GONPs exposure.


Assuntos
Bombyx/efeitos dos fármacos , Grafite/toxicidade , Nanopartículas/toxicidade , Animais , Bombyx/fisiologia , Linhagem Celular , Dano ao DNA , Feminino , Masculino , Oócitos/efeitos dos fármacos , Oócitos/metabolismo , Oócitos/patologia , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Reprodução/efeitos dos fármacos
20.
ACS Appl Mater Interfaces ; 13(4): 4861-4873, 2021 Feb 03.
Artigo em Inglês | MEDLINE | ID: mdl-33471499

RESUMO

A combination treatment strategy that relies on the synergetic effects of different therapeutic approaches has been considered to be an effective method for cancer therapy. Herein, a chemotherapeutic drug (doxorubicin, Dox) and a manganese ion (Mn2+) were co-loaded into regenerated silk fibroin-based nanoparticles (NPs), followed by the surface conjugation of phycocyanin (PC) to construct tumor microenvironment-activated nanococktails. The resultant PC-Mn@Dox-NPs showed increased drug release rates by responding to various stimulating factors (acidic pH, hydrogen peroxide (H2O2), and glutathione), revealing that they could efficiently release the payloads (Dox and Mn2+) in tumor cells. The released Dox could not only inhibit the growth of tumor cells but also generated a large amount of H2O2. The elevated H2O2 was decomposed into the highly harmful hydroxyl radicals and oxygen through an Mn2+-mediated Fenton-like reaction. Furthermore, the generated oxygen participated in photodynamic therapy (PDT) and produced abundant singlet oxygen. Our investigations demonstrate that these PC-Mn@Dox-NPs exhibit multiple bioresponsibilities and favorable biosafety. By integrating Dox-induced chemotherapy, Mn2+-mediated chemodynamic therapy, and PC-based PDT via cascade reactions, PC-Mn@Dox-NPs achieved enhanced in vitro and in vivo anticancer efficacies compared to all the mono- or dual-therapeutic approaches. These findings reveal that PC-Mn@Dox-NPs can be exploited as a promising nanococktail for cascade reaction-mediated synergistic cancer treatment.


Assuntos
Antibióticos Antineoplásicos/administração & dosagem , Doxorrubicina/administração & dosagem , Manganês/administração & dosagem , Neoplasias/tratamento farmacológico , Fármacos Fotossensibilizantes/administração & dosagem , Ficocianina/administração & dosagem , Animais , Antibióticos Antineoplásicos/farmacologia , Antibióticos Antineoplásicos/uso terapêutico , Bombyx/química , Cátions Bivalentes/administração & dosagem , Cátions Bivalentes/farmacologia , Cátions Bivalentes/uso terapêutico , Linhagem Celular Tumoral , Doxorrubicina/farmacologia , Doxorrubicina/uso terapêutico , Portadores de Fármacos/química , Fibroínas/química , Glutationa/metabolismo , Células Endoteliais da Veia Umbilical Humana , Humanos , Concentração de Íons de Hidrogênio , Manganês/farmacologia , Manganês/uso terapêutico , Camundongos , Nanopartículas/química , Neoplasias/metabolismo , Neoplasias/patologia , Fármacos Fotossensibilizantes/farmacologia , Fármacos Fotossensibilizantes/uso terapêutico , Ficocianina/farmacologia , Ficocianina/uso terapêutico , Microambiente Tumoral/efeitos dos fármacos
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