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1.
Sci Rep ; 10(1): 7222, 2020 Apr 24.
Artigo em Inglês | MEDLINE | ID: mdl-32332824

RESUMO

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

2.
Sci Rep ; 10(1): 7221, 2020 Apr 24.
Artigo em Inglês | MEDLINE | ID: mdl-32332832

RESUMO

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

3.
Eur J Cardiothorac Surg ; 56(6): 1090-1096, 2019 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-31329842

RESUMO

OBJECTIVES: To study the perioperative outcomes and long-term survival rates in patients undergoing thoracic endovascular aortic repair (TEVAR) for uncomplicated type B dissection. METHODS: A total of 751 patients with uncomplicated type B dissection who underwent TEVAR at our centre between May 2001 and December 2013 were retrospectively reviewed. The mean age of all patients (619 males and 132 females) was 52.8 ± 10.9 years. The follow-up period ranged from 1 to 170 months (median 70 months). RESULTS: Five patients died during the perioperative period (mortality rate 0.7%). Four patients (0.5%) developed retrograde type A dissection. Two patients (0.3%) developed paraplegia and 1 patient developed incomplete paralysis (0.1%). There were no postoperative cerebral infarctions. The 5- and 10-year survival rates were 96.5% [95% confidence interval (CI) 95.0-98.0%] and 83.0% (95% CI 77.9-88.4%), respectively. The 5- and 10-year reintervention rates were 4.6% (95% CI 3.0-6.2%) and 7.9% (95% CI 5.3-10.5%), respectively. CONCLUSIONS: Although the application of TEVAR for patients with uncomplicated dissection is still under debate, many patients who have undergone TEVAR have benefitted substantially from the treatment. Our data showed that TEVAR had low mortality and complication rates both in the short- and long-term follow-up periods. TEVAR may be considered as a first choice for patients with uncomplicated type B dissection.


Assuntos
Aneurisma da Aorta Torácica/cirurgia , Dissecção Aórtica/cirurgia , Implante de Prótese Vascular/métodos , Procedimentos Endovasculares/métodos , Adulto , Aorta/cirurgia , Implante de Prótese Vascular/efeitos adversos , Implante de Prótese Vascular/mortalidade , Procedimentos Endovasculares/efeitos adversos , Procedimentos Endovasculares/mortalidade , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias/etiologia , Reoperação , Estudos Retrospectivos , Resultado do Tratamento
4.
Int J Biol Macromol ; 113: 961-970, 2018 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-29462677

RESUMO

The complete mitochondrial genome (mitogenome) of Biston marginata (Lepidoptera: Geometridae) was determined and annotated. The circular genome is 15,470bp long and it contains the entire set of 37 genes usually present in lepidopteran mitogenomes. The nucleotide composition of the genome is highly A+T biased, accounting for 81.20%, with a slightly positive AT skewness (0.028), indicating the occurrence of more As than Ts, as found in other Geometridae species. Except for cox1 gene starts with non-canonical initial codon CGA, all protein-coding genes start with ATN codon. Three of the 13 PCGs (protein coding gene) had an incomplete termination codon, T or TA, while the others terminated with TAA. All tRNA genes are predicted to fold into typical clover-leaf secondary structure, except for the trnS1 (AGN), in which the dihydrouridine (DHU) arm could not form a stable stem-loop structure. The A+T-rich region of 343bp is comprised of non-repetitive sequences, but have several distinctive features, including the motif "ATAGA" followed by a 19bp poly-T stretch, a microsatellite-like (TA)7 element next to the ATTTA motif. The phylogenetic analyses support the view that the B. marginata is closely related to the Biston pantrinaria, and confirm that Biston marginata belongs to the family Geometridae.


Assuntos
Genoma Mitocondrial/genética , Lepidópteros/classificação , Lepidópteros/genética , Filogenia , Animais , Códon/genética , DNA Intergênico/genética , Sequência Rica em GC , Fases de Leitura Aberta/genética , RNA Ribossômico/genética , RNA de Transferência/genética
5.
Dev Comp Immunol ; 78: 114-123, 2018 01.
Artigo em Inglês | MEDLINE | ID: mdl-28958702

RESUMO

Cathepsins are a group of protease, located in lysosome and play a vital role in physiological process. Here, we reported cathepsin L-like protease (Ap-cathL), which contained an open reading frame of 1155 bp and encoding 385 amino acid residues protein. The I29 inhibitor domain and peptidase C1A (clan CA of cysteine proteases, papain family C1 subfamily) putative conserved domains were detected in Ap-cathL. Quantitative real-time PCR (qRT-PCR) analysis revealed that Ap-cathL highly expressed in the fat body and midgut. The high expression during the molting stage, pupal stage and following 20E (20-hydroxyecdysone) treatment indicated that it maybe involved in the process of molting and metamorphosis. In addition, depletion of Ap-cathL influenced the expression of apoptosis pathway related genes. The protease inhibitor and RNA interference experiments showed that Ap-cathL was involved in the fat body dissociation of A. pernyi. These results suggest that Ap-cathL may involve in the process of metamorphosis and fat body dissociation of A. pernyi.


Assuntos
Catepsina L/metabolismo , Corpo Adiposo/fisiologia , Proteínas de Insetos/metabolismo , Metamorfose Biológica/genética , Muda/genética , Mariposas/fisiologia , Peptídeo Hidrolases/metabolismo , Animais , Apoptose/genética , Catepsina L/genética , Células Cultivadas , Clonagem Molecular , Ecdisterona/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Insetos/genética , Peptídeo Hidrolases/genética , RNA Interferente Pequeno/genética
6.
Dev Comp Immunol ; 81: 187-192, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29225004

RESUMO

The SOCS (Suppressor of cytokine signaling) family members are a potential negative regulator of cytokine signaling pathway and play a key role to maintain immunological functions in animals. SOCS-6 is an important member of the SOCS family, however the functions of this gene have rarely been explored among eukaryotes. Herein, we cloned and expressed SOCS-6 gene from Bombyx mori (Dazao) (BmSOCS-6), and anti-rabbit antibodies were prepared using purified recombinant BmSOCS-6 protein. Under normal physiological conditions, the BmSOCS-6 expression was observed at varied levels in six tissues, with most greatly expressed in fat body and hemocytes. After immune challenge with viral, fungal and bacterial pathogens, the BmSOCS-6 showed distinctly varied expression patterns in tissue, time and microbe dependent manner. By contrast, recombinant BmSOCS-6 protein strongly enhanced the expression of epidermal growth factor receptor (EGFR) pathway related genes, while the depletion of BmSOCS-6 by double stranded RNA suppressed their production. Altogether we concluded that BmSOCS-6 may improve the efficiency of EGFR signaling pathway in B. mori (Dazao).


Assuntos
Bombyx/imunologia , Receptores ErbB/metabolismo , Corpo Adiposo/fisiologia , Hemócitos/fisiologia , Infecções/imunologia , Proteínas de Insetos/genética , Proteínas Supressoras da Sinalização de Citocina/genética , Animais , Clonagem Molecular , Citocinas/metabolismo , Regulação da Expressão Gênica , Humanos , Imunidade Inata , Proteínas de Insetos/metabolismo , Filogenia , RNA Interferente Pequeno/genética , Proteínas Recombinantes/genética , Transdução de Sinais , Proteínas Supressoras da Sinalização de Citocina/metabolismo
7.
J Insect Physiol ; 103: 47-56, 2017 11.
Artigo em Inglês | MEDLINE | ID: mdl-29032156

RESUMO

Suppressors of cytokine signaling (SOCS) are a potent negative regulator of diverse cytokine-related responses to maintain various physiological processes in animals. Here, we obtained the SOCS2-12 gene sequence of Bombyx mori (Dazao) (BmSOCS2-12) from the National Center for Biotechnology Information (NCBI) to study its expression profile in different tissues, as well as in the immune tissues following larval exposure to pathogens. Further, we investigated the role of BmSOCS2-12 in producing antimicrobial peptides (AMPs) and as a regulator of ecdysteroid signaling transduction. The quantitative real-time PCR analysis revealed unequal transcript levels of BmSOCS2-12 in the different tissues, however the gene's expression was highest in those of fat body and hemocyte. The challenge with pathogens significantly upregulated the transcript level of BmSOCS2-12 in both fat body and hemocyte when compared with the control. By contrast, recombinant BmSOCS2-12 protein injections strongly suppressed the expression of AMPs, while the knockdown of BmSOCS2-12 by double-stranded RNA enhanced their production. Administration of 20-hydroxyecdysone significantly downregulated the BmSOCS2-12 expression in fat body, and the depletion of BmSOCS2-12 enhanced the transcript levels of 20-hydroxyecdysone-responsive genes at 48 h. Altogether, BmSOCS2-12 may have multiple functional roles in the physiology of B. mori (Dazao), since it negatively regulates the expression of AMPs and ecdysteroid signaling transduction.


Assuntos
Peptídeos Catiônicos Antimicrobianos/metabolismo , Bombyx/metabolismo , Proteínas Supressoras da Sinalização de Citocina/metabolismo , Sequência de Aminoácidos , Animais , Bombyx/genética , Bombyx/imunologia , Ecdisterona/metabolismo , Expressão Gênica , Regulação da Expressão Gênica , Filogenia , Proteínas Supressoras da Sinalização de Citocina/química
8.
J Invertebr Pathol ; 150: 6-14, 2017 11.
Artigo em Inglês | MEDLINE | ID: mdl-28859880

RESUMO

Cathepsins are key members of mammalian papain-like cysteine proteases that play an important role in the immune response. In this study, a fragment of cDNA encoding cathepsin O proteinase (ApCathepsin O) was cloned from Antheraea pernyi. It contains an open reading frame of 1170bp and encodes a protein with 390 amino acid residues, including a conserved I29 inhibitor domain and a peptidase C1A (clan CA of cysteine proteases, papain family C1 subfamily) domain. Comparison with other previously reported cathepsin O proteins showed identity ranging from 45% to 79%. Quantitative real-time PCR (qRT-PCR) and Western blot analysis revealed that ApCathepsin O was highly expressed in the fat body; furthermore, the high expression during the pupal stage indicated that it might be involved during metamorphosis. After exposure to four different heat-killed pathogens (Escherichia coli, Beauveria bassiana, Micrococcus luteus, and A. pernyi nucleopolyhedrovirus), the expression levels of ApCathepsin O mRNA significantly increased and showed variable expression patterns. This indicates that ApCathepsin O is potentially involved in the innate immune system of A. pernyi. Interestingly, ApCathepsin O expression was upregulated after 20-hydroxyecdysone (20E) injection, which suggested that it might be regulated by 20E. In conclusion, ApCathepsin O is a protease that may play an important role in the innate immune response and metamorphosis of A. pernyi.


Assuntos
Catepsinas/metabolismo , Imunidade Inata/fisiologia , Proteínas de Insetos/metabolismo , Metamorfose Biológica/fisiologia , Mariposas/metabolismo , Animais , Catepsinas/genética , Clonagem Molecular , Proteínas de Insetos/genética , Mariposas/genética
9.
Zootaxa ; 4254(5): 501-519, 2017 Apr 20.
Artigo em Inglês | MEDLINE | ID: mdl-28609941

RESUMO

In this study, a complete mitochondrial genome (mitogenome) sequence of Abraxas suspecta (Lepidoptera: Geometridae) is isolated and characterized. The complete DNA is 15,547 bp length and contains 2 ribosomal RNA genes, 23 putative transfer RNA (tRNA) genes including an extra tRNAAsn (AUU), 13 protein-coding genes and an adenine (A) + thymine (T)-rich region. The nucleotide composition and gene organization are identical to those of other lepidopteran, except for the presence of an extra copy of trnN (AUU). Of the 38 genes, twenty-five genes (9 PCGs and 16 tRNAs) are encoded by heavy strand (H-strand), while thirteen are encoded by light strand (L-strand). Among the 13 PCGs, 12 PCGs employ ATN as initiation codon, while cytochrome c oxidase subunit 1 (cox1) utilizes CGA as initiation codon. Four of the 13 PCGs have the incomplete termination codon T, while the remainder terminated with the canonical stop codon. All tRNA genes are folded into the typical clover-leaf structure of mitochondrial tRNAs, except for the tRNASer (AGN) gene, in which the DHU arm fails to form a stable stem-loop structure. The A+T-rich region is 532 bp long, and contains some conserved regions, including 'ATAGA' motif followed by a 17bp poly-T stretch, a microsatellite-like element (AT)8(AAT)3 and also a poly-A element. A short Phylogenetic analysis based on 13 PCGs using maximum likelihood (ML) and Bayesian inference (BI) revealed that A. suspecta resides in the Geometridae family. We present the method and approach to use moths as model organisms for further genetic and evolutionary biology studies.


Assuntos
Genoma Mitocondrial , Mariposas , Animais , Sequência de Bases , Teorema de Bayes , Lepidópteros , Filogenia , RNA de Transferência , Análise de Sequência de DNA
10.
PLoS One ; 12(6): e0178773, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28598968

RESUMO

In the present study, the complete sequence of the mitochondrial genome (mitogenome) of Daphnis nerii (Lepidoptera: Sphingidae) is described. The mitogenome (15,247 bp) of D.nerii encodes13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), two ribosomal RNA genes (rRNAs) and an adenine (A) + thymine (T)-rich region. Its gene complement and order is similar to that of other sequenced lepidopterans. The 12 PCGs initiated by ATN codons except for cytochrome c oxidase subunit 1 (cox1) gene that is seemingly initiated by the CGA codon as documented in other insect mitogenomes. Four of the 13 PCGs have the incomplete termination codon T, while the remainder terminated with the canonical stop codon. This mitogenome has six major intergenic spacers, with the exception of A+T-rich region, spanning at least 10 bp. The A+T-rich region is 351 bp long, and contains some conserved regions, including 'ATAGA' motif followed by a 17 bp poly-T stretch, a microsatellite-like element (AT)9 and also a poly-A element. Phylogenetic analyses based on 13 PCGs using maximum likelihood (ML) and Bayesian inference (BI) revealed that D. nerii resides in the Sphingidae family.


Assuntos
Genoma Mitocondrial , Mariposas/classificação , Mariposas/genética , Filogenia , Animais , Composição de Bases , Códon , Genes Mitocondriais , Genômica/métodos , Sequenciamento de Nucleotídeos em Larga Escala , Fases de Leitura Aberta , RNA Ribossômico/genética , RNA de Transferência/genética
11.
Dev Comp Immunol ; 76: 45-55, 2017 11.
Artigo em Inglês | MEDLINE | ID: mdl-28545959

RESUMO

Genes encoding proteins of serpins superfamily are widely distributed in invertebrates. In insects, serpins play important roles in regulating immune responses and other physiological processes. Here, we report the cloning and characterization of cDNA of Apserpin-14 from Chinese oak silkworm (Antheraea pernyi). The Apserpin-14 gene contains 1206 bp open reading frame, encoding a predicted 401 amino acid residue protein. We expressed the recombinant Apserpin-14 protein in Escherichia coli and then purified protein was used to prepare rabbit anti-Apserpin-14 polyclonal antibodies. Quantitative real-time PCR analysis revealed that mRNA level of Apserpin-14 was highest in the fat body, whereas, among developmental stages the 5th instar and pupal stage showed greatest expression. Furthermore, Escherichia coli, Beauveria bassiana, Micrococcus luteus and nuclear polyhedrosis virus challenge enhanced Apserpin-14 transcript in both the fat body and hemocyte. Recombinant Apserpin-14 added to hemolymph inhibited spontaneous melanization and suppressed prophenoloxidase activation stimulated by M. luteus, but did not affect phenoloxidase (PO) activity. Injection of recombinant Apserpin-14 protein into A. pernyi larvae significantly reduced the transcript levels of antimicrobial peptides in the fat body, while its depletion by double stranded RNA enhanced their expression. We concluded that Apserpin-14 likely involved in regulation of proPO activation and production of antimicrobial peptides, implying its important role in the innate immune system of A. pernyi.


Assuntos
Bombyx/metabolismo , Catecol Oxidase/metabolismo , Precursores Enzimáticos/metabolismo , Proteínas de Insetos/metabolismo , Peptídeos/metabolismo , Serpinas/metabolismo , Sequência de Aminoácidos , Animais , DNA Complementar/metabolismo , Corpo Adiposo/metabolismo , Hemócitos/metabolismo , Hemolinfa/metabolismo , Mariposas/metabolismo , Proteínas Recombinantes/metabolismo , Homologia de Sequência
12.
Genome ; 60(2): 128-138, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-28084809

RESUMO

In the present study, we sequenced the complete mitochondrial genome (mitogenome) of Agrius convolvuli (Lepidoptera: Sphingidae) and compared it with previously sequenced mitogenomes of lepidopteran species. The mitogenome was a circular molecule, 15 349 base pairs (bp) long, containing 37 genes. The order and orientation of genes in the A. convolvuli mitogenome were similar to those in sequenced mitogenomes of other lepidopterans. All 13 protein-coding genes (PCGs) were initiated by ATN codons, except for the cytochrome c oxidase subunit 1 (cox1) gene, which seemed to be initiated by the codon CGA, as observed in other lepidopterans. Three of the 13 PCGs had the incomplete termination codon T, while the remainder terminated with TAA. Additionally, the codon distributions of the 13 PCGs revealed that Asn, Ile, Leu2, Lys, Phe, and Tyr were the most frequently used codon families. All transfer RNAs were folded into the expected cloverleaf structure except for tRNASer(AGN), which lacked a stable dihydrouridine arm. The length of the adenine (A) + thymine (T)-rich region was 331 bp. This region included the motif ATAGA followed by a 19-bp poly-T stretch and a microsatellite-like (TA)8 element next to the motif ATTTA. Phylogenetic analyses (maximum likelihood and Bayesian methods) showed that A. convolvuli belongs to the family Sphingidae.


Assuntos
Genoma Mitocondrial , Ipomoea batatas/parasitologia , Lepidópteros/genética , Animais , Composição de Bases , Biologia Computacional/métodos , DNA Intergênico , Ordem dos Genes , Sequenciamento de Nucleotídeos em Larga Escala , Lepidópteros/classificação , Anotação de Sequência Molecular , Fases de Leitura Aberta , Filogenia
13.
Artigo em Inglês | MEDLINE | ID: mdl-28008655

RESUMO

In present study, a Cecropin-like peptide from Antheraea pernyi (ApCec) was cloned and characterized. The full-length ApCec cDNA encoded a protein with 64 amino acids including a putative 22-amino-acid signal peptide, a 4-amino-acid propeptide, and a 38-amino-acid mature peptide. ApCec gene was highly expressed in Malpighian tubules of A. pernyi after induction for 24 h by Escherichia coli in PBS. Pro-ApCec (including propeptide and mature peptide) and M-ApCec (just mature peptide) were synthesized chemically and analyzed by HPLC and mass spectroscopy. The antibacterial activity of M-ApCec is more potent than pro-ApCec against E. coli K12 or B. subtilus in both minimum inhibitory concentration and inhibition zone assays. Hemolytic assay results showed M-ApCec possessed a low cytotoxicity to mammalian cells. The secondary structure of M-ApCec forms α-helical structure, shown by circular dichroism spectroscopy. Transmission electron microscopy analysis suggested that M-ApCec killed bacteria by disrupting bacterial cell membrane integrity. Our results indicate ApCec may play an important role in defending from pathogenic bacteria in A. pernyi, and it may be as a potential candidate for applications in antibacterial drug development and agriculture.


Assuntos
Antibacterianos/farmacologia , Cecropinas/genética , Cecropinas/farmacologia , Proteínas de Insetos/genética , Mariposas/genética , Sequência de Aminoácidos , Animais , Bacillus subtilis/efeitos dos fármacos , Cecropinas/química , Cecropinas/metabolismo , Clonagem Molecular , DNA Complementar/genética , DNA Complementar/metabolismo , Escherichia coli/fisiologia , Escherichia coli K12/efeitos dos fármacos , Regulação da Expressão Gênica , Proteínas de Insetos/química , Proteínas de Insetos/metabolismo , Larva/genética , Larva/crescimento & desenvolvimento , Larva/metabolismo , Mariposas/crescimento & desenvolvimento , Mariposas/metabolismo , Filogenia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Alinhamento de Sequência
14.
Sci Rep ; 6: 39153, 2016 12 15.
Artigo em Inglês | MEDLINE | ID: mdl-27974854

RESUMO

The complete mitochondrial genome (mitogenome) of Leucoma salicis (Lepidoptera: Lymantriidae) was sequenced and annotated. It is a circular molecule of 15,334 bp, containing the 37 genes usually present in insect mitogenomes. All protein-coding genes (PCGs) are initiated by ATN codons, other than cox1, which is initiated by CGA. Three of the 13 PCGs had an incomplete termination codon, T or TA, while the others terminated with TAA. The relative synonymous codon usage of the 13 protein-coding genes (PCGs) was consistent with those of published lepidopteran sequences. All tRNA genes had typical clover-leaf secondary structures, except for the tRNASer (AGN), in which the dihydrouridine (DHU) arm could not form a stable stem-loop structure. The A + T-rich region of 325 bp had several distinctive features, including the motif 'ATAGA' followed by an 18 bp poly-T stretch, a microsatellite-like (AT)7 element, and an 11-bp poly-A present immediately upstream of tRNAMet. Relationships among 32 insect species were determined using Maximum Likelihood (ML), Neighbor Joining (NJ) and Bayesian Inference (BI) phylogenetic methods. These analyses confirm that L. salicis belongs to the Lymantriidae; and that Lymantriidae is a member of Noctuoidea, and is a sister taxon to Erebidae, Nolidae and Noctuidae, most closely related to Erebidae.


Assuntos
Genoma Mitocondrial , Lepidópteros/genética , Mariposas/genética , Animais , Sequência de Bases , Teorema de Bayes , Códon de Terminação , DNA Mitocondrial/química , DNA Mitocondrial/isolamento & purificação , DNA Mitocondrial/metabolismo , Lepidópteros/classificação , Funções Verossimilhança , Mariposas/classificação , Conformação de Ácido Nucleico , Fases de Leitura Aberta/genética , Filogenia , RNA Ribossômico/classificação , RNA Ribossômico/genética , RNA de Transferência/química , RNA de Transferência/classificação , RNA de Transferência/genética , Análise de Sequência de DNA
15.
Protein Pept Lett ; 23(10): 878-883, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27586183

RESUMO

The receptor for activated C kinase (RACK) is an important scaffold protein with regulatory functions in cells. However, its role in the immune response of Antheraea pernyi to pathogen challenge remains unclear. To investigate the biological functions of RACK in the wild silkworm A. pernyi, cloning was performed and the expression patterns of the RACK gene were analyzed. Sequence analysis revealed that the RACK gene was 1120 bp containing a 960-bp open reading frame. The deduced RACK protein sequence reveals the higher identity with its homologs from other insects. SDS-PAGE and western blot analysis demonstrated successful expression of a 36-kDa recombinant RACK protein in Escherichia coli. The titer of a rabbit-raised antibody against recombinant RACK protein was about 1: 20000, determined by ELISA. Real-time PCR analysis showed that RACK expression was higher in fat bodies than in other examined A. pernyi tissues. The expression of RACK mRNA in fat bodies of fifth larvae of A. pernyi was obviously induced after nucleopolyhedrovirus, E. coli or Beauveria bassiana challenge. However, the expression patterns of RACK were different in response to these pathogens. Our data suggest that RACK may play a role in the innate immune responses of A. pernyi.


Assuntos
Mariposas/enzimologia , Receptores de Superfície Celular/química , Sequência de Aminoácidos , Animais , Clonagem Molecular , Eletroforese em Gel de Poliacrilamida , Receptores de Quinase C Ativada , Receptores de Superfície Celular/genética , Homologia de Sequência de Aminoácidos , Seda
16.
Fish Shellfish Immunol ; 56: 162-168, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-27417230

RESUMO

Cathepsin B is a lysosomal cysteine protease of the papain-like enzyme family with multiple biological functions. In the present study, a cathepsin B gene (named PcCTSB) was cloned and characterized from the red crayfish, Procambarus clarkii. The cDNA fragments of PcCTSB was 990 bp in length. It encoded a putative protein of 329 amino acid residues with predicted molecular weight of 36.4 kDa and isoelectric point of 7.020. Sequence alignment revealed that PcCTSB protein is 53.6%-80.4% identical with those from other 10 species. The predicted tertiary structure of PcCTSB protein was highly similar to that of animals. The results of the phylogenetic analysis indicated that the PcCTSB protein could be clustered with the Eriocheir sinensis cathepsin B protein. The recombinant protein of PcCTSB was expressed successfully in Escherichia coli cells. The mRNA expressions of PcCTSB were detected in all tested tissues, particularly high in the hepatopancreas. After lipopolysaccharide (LPS) challenge, the expression levels of PcCTSB were up-regulated significantly at different time points compared with control. Our results suggested that the PcCTSB might play an important role in defending against the pathogenes infection.


Assuntos
Proteínas de Artrópodes/genética , Astacoidea/genética , Astacoidea/imunologia , Catepsina B/genética , Regulação da Expressão Gênica/imunologia , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/química , Proteínas de Artrópodes/metabolismo , Astacoidea/classificação , Astacoidea/metabolismo , Sequência de Bases , Catepsina B/química , Catepsina B/metabolismo , Clonagem Molecular , DNA Complementar/genética , DNA Complementar/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Lipopolissacarídeos/farmacologia , Filogenia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Alinhamento de Sequência
17.
J Invertebr Pathol ; 138: 10-7, 2016 07.
Artigo em Inglês | MEDLINE | ID: mdl-27261060

RESUMO

Yippee was first identified as a protein that physically interacts with the Hemolin protein of Hyalophora cecropia. In this study, we identified a gene with a 366bp open reading frame (ORF) that encodes a 121 amino acid protein containing a conserved Yippee domain. We named this gene Ap-Yippee (Yippee gene from Antheraea pernyi), and investigated the role of the protein in the host immune response. A recombinant Ap-Yippee protein was expressed in Escherichia coli cells, and polyclonal antibodies were produced against the recombinant protein. Real-time PCR and a Western blot analysis revealed that Ap-Yippee is expressed in the hemocytes, Malpighian tubules, midgut, silk gland, epidermis, and fat bodies of A. pernyi, with the highest expression level observed in Malpighian tubules. The fifth instar larvae of A. pernyi were challenged by injecting them with nucleopolyhedrovirus (AP-NPV), the Gram-negative bacterium E. coli, the Gram-positive bacterium Micrococcus luteus, or the entomopathogenic fungus, Beauveria bassiana. These challenges with diverse pathogens resulted in differential expression patterns of the protein. A knockdown of the Ap-Yippee gene by small interfering RNA (siRNA) transfection had a significant influence on the expression of the hemolin in the pupae which was confirmed by qRT-PCR and Western blot. Furthermore, a possible protein-protein interaction between Ap-Yippee and Hemolin was explored by Far-Western blotting. Therefore, our data suggest that the Ap-Yippee protein is involved in a pathway that regulates the immune response of insects.


Assuntos
Imunidade Inata/imunologia , Proteínas de Insetos/imunologia , Mariposas/genética , Mariposas/imunologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Western Blotting , Proteínas de Transporte/biossíntese , Proteínas de Transporte/imunologia , Imunoglobulinas/imunologia , Imunoglobulinas/metabolismo , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Alinhamento de Sequência , Transcriptoma
18.
BMC Cardiovasc Disord ; 16: 119, 2016 05 31.
Artigo em Inglês | MEDLINE | ID: mdl-27246834

RESUMO

BACKGROUND: This study evaluated the protective effect of Echinatin against myocardial ischemia/reperfusion (I/R) injury in rats. METHODS: The effect of Echinatin on cardiac function in rats subjected to I/R was demonstrated through improved Langendorff retrograde perfusion technology. Adult Sprague-Dawley rats were randomly divided into five groups, and myocardial infarct size was macroscopically estimated through 2,3,5-triphenyltetrazolium chloride staining. The coronary effluent was analyzed for the release of lactate dehydrogenase (LDH) and creatine kinase (CK) to assess the degree of cardiac injury. The concentrations of malondialdehyde (MDA), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) were determined along with superoxide dismutase (SOD) activity using ELISA. Finally, cardiomyocyte apoptosis analysis was conducted with POD, an in situ cell death detection kit. RESULTS: Echinatin (0.5 and 2.5 µg/mL) pretreatment enhanced the maximum up/down rate of the left ventricular pressure (±dp/dtmax), improved the heart rate, increased the left ventricular developed pressure (LVDP), enhanced the coronary flow, and reduced the CK and LDH levels in the coronary flow of the treated group compared with the I/R group. Echinatin limited the contents of CK and LDH, improved the LVDP, reduced the contents of MDA, IL-6, and TNF-α, and increased the SOD activity. The infarct size and cell apoptosis in the hearts of the rats in the Echinatin-treated group were smaller and lower, respectively, than those in the hearts of the rats in the I/R control group. CONCLUSION: Echinatin exerts a protective effect against I/R-induced myocardial injury on hearts. This effect may be attributed to the antioxidant and anti-inflammatory activities of this compound.


Assuntos
Cardiotônicos/farmacologia , Chalconas/farmacologia , Traumatismo por Reperfusão Miocárdica/prevenção & controle , Função Ventricular Esquerda/efeitos dos fármacos , Animais , Anti-Inflamatórios/farmacologia , Antioxidantes/farmacologia , Apoptose/efeitos dos fármacos , Circulação Coronária/efeitos dos fármacos , Creatina Quinase/metabolismo , Citoproteção , Relação Dose-Resposta a Droga , Frequência Cardíaca/efeitos dos fármacos , Mediadores da Inflamação/metabolismo , Interleucina-6/metabolismo , Preparação de Coração Isolado , L-Lactato Desidrogenase/metabolismo , Malondialdeído/metabolismo , Traumatismo por Reperfusão Miocárdica/metabolismo , Traumatismo por Reperfusão Miocárdica/patologia , Traumatismo por Reperfusão Miocárdica/fisiopatologia , Miocárdio/metabolismo , Miocárdio/patologia , Estresse Oxidativo/efeitos dos fármacos , Ratos Sprague-Dawley , Superóxido Dismutase/metabolismo , Fatores de Tempo , Fator de Necrose Tumoral alfa/metabolismo , Pressão Ventricular/efeitos dos fármacos
19.
Sci Rep ; 6: 26387, 2016 05 25.
Artigo em Inglês | MEDLINE | ID: mdl-27222440

RESUMO

In this study, we sequenced the complete mitochondrial genome of Eligma narcissus and compared it with 18 other lepidopteran species. The mitochondrial genome (mitogenome) was a circular molecule of 15,376 bp containing 13 protein-coding genes (PCGs), 22 transfer RNA (tRNA) genes, two ribosomal RNA (rRNA) genes and an adenine (A) + thymine (T) - rich region. The positive AT skew (0.007) indicated the occurrence of more As than Ts. The arrangement of 13 PCGs was similar to that of other sequenced lepidopterans. All PCGs were initiated by ATN codons, except for the cytochrome c oxidase subunit 1 (cox1) gene, which was initiated by the CGA sequence, as observed in other lepidopterans. The results of the codon usage analysis indicated that Asn, Ile, Leu, Tyr and Phe were the five most frequent amino acids. All tRNA genes were shown to be folded into the expected typical cloverleaf structure observed for mitochondrial tRNA genes. Phylogenetic relationships were analyzed based on the nucleotide sequences of 13 PCGs from other insect mitogenomes, which confirmed that E. narcissus is a member of the Noctuidae superfamily.


Assuntos
Genoma Mitocondrial , Mariposas/genética , Sequência Rica em At , Animais , Sequência de Bases , Códon , Sequência Conservada , Proteínas de Insetos/genética , Sequências Repetidas Invertidas , Proteínas Mitocondriais/genética , Anotação de Sequência Molecular , Filogenia , RNA Ribossômico/genética , RNA de Transferência/genética , Sequenciamento Completo do Genoma
20.
Artigo em Inglês | MEDLINE | ID: mdl-25187437

RESUMO

The complete mitochondrial genome (mitogenome) of Plutella xylostella (Lepidoptera: Plutellidae) was determined (GenBank accession No. KM023645). The length of this mitogenome is 16,014 bp with 13 protein-coding genes (PCGs), 2 rRNA genes, 22 tRNA genes and an A + T-rich region. It presents the typical gene organization and order for completely sequenced lepidopteran mitogenomes. The nucleotide composition of the genome is highly A + T biased, accounting for 81.48%, with a slightly positive AT skewness (0.005). All PCGs are initiated by typical ATN codons, except for the gene cox1, which uses CGA as its start codon. Some PCGs harbor TA (nad5) or incomplete termination codon T (cox1, cox2, nad2 and nad4), while others use TAA as their termination codons. The A + T-rich region is located between rrnS and trnM with a length of 888 bp.


Assuntos
DNA Mitocondrial/química , Genoma Mitocondrial , Mariposas/genética , Animais , Sequência de Bases , Dados de Sequência Molecular , Conformação de Ácido Nucleico , Motivos de Nucleotídeos , Análise de Sequência de DNA
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