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1.
Arch Biochem Biophys ; 752: 109853, 2024 02.
Artigo em Inglês | MEDLINE | ID: mdl-38086523

RESUMO

Ventilator-induced lung injury (VILI) disturbs the disordered immune system and causes persistent inflammatory damage. 4-octyl itaconate (OI) is a synthetic cell-permeable itaconate derivative with antioxidant and anti-inflammatory effects. In this study, we assessed whether OI protects against VILI. OI was intraperitoneally injected for three days before mechanical ventilation (MV; 20 ml/kg at 70 breaths/min) for 2 h. Mouse lung vascular endothelial cells (MLVECs) were pretreated with OI (62.5, 125, and 250 µM) prior to cyclic stretch for 4 h. We found that OI attenuated VILI and inflammatory response. OI also increased superoxide dismutase, nuclear factor E2-related factor 2, and heme oxygenase-1 levels, and decreased reactive oxygen species and malondialdehyde levels. Furthermore, OI inhibited the expression of NLR family pyrin domain-containing 3 (NLRP3), caspase-1 p20, apoptosis-associated speck-like protein containing a CARD, and N-terminal fragment of gasdermin D. Therefore, OI attenuates VILI, potentially by suppressing oxidative stress and NLRP3 activation.


Assuntos
Proteína 3 que Contém Domínio de Pirina da Família NLR , Succinatos , Lesão Pulmonar Induzida por Ventilação Mecânica , Camundongos , Animais , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Células Endoteliais/metabolismo , Lesão Pulmonar Induzida por Ventilação Mecânica/tratamento farmacológico , Lesão Pulmonar Induzida por Ventilação Mecânica/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Pulmão/metabolismo
2.
Ecotoxicol Environ Saf ; 217: 112235, 2021 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-33873079

RESUMO

Ocean acidification (OA) has posed formidable threats to marine calcifiers. In response to elevated CO2 levels, marine calcifiers have developed multiple strategies to survive, such as taking advantage of apoptosis, but its regulation mechanism remains largely unknown. Here, we used the Pacific oyster Crassostrea gigas as model to understand the apoptotic responses and regulation mechanism at short- (7 d) to long-term (56 d) CO2 exposure (pH = 7.50). The apoptosis of hemocytes was significantly induced after short-term treatment (7-21 d) but was suppressed under long-term CO2 exposure (42-56 d). Similarly, caspase-3 and caspase-9 were also increased post short-term exposure and fell back to normal levels after long-term exposure. These data together indicated diverse regulation mechanisms of apoptosis through different exposure periods. Through analysis of the B-cell lymphoma 2 (Bcl-2) family mitochondrial apoptosis regulators, we showed that only CgBcl-XL's expression kept at high levels after 42- and 56-day CO2 exposure. CgBcl-XL shared sequence, and structural similarity with its mammalian counterpart, and knockdown of CgBcl-XL in hemocytes via RNA interference promoted apoptosis. The protein level of CgBcl-XL was significantly increased after long-term CO2 exposure (28-56 d), and its distribution in hemocytes became more concentrated and dense. Therefore, CgBcl-XL serves as an essential anti-apoptotic protein for tipping the balance of cell apoptosis, which may play a key role in survival under long-term CO2 exposure. These results reveal a potential adaptation strategy of oysters towards OA and the variable environment changes through the modulation of apoptosis.


Assuntos
Crassostrea/fisiologia , Aclimatação , Animais , Apoptose , Dióxido de Carbono/metabolismo , Dióxido de Carbono/fisiologia , Crassostrea/metabolismo , Hemócitos/metabolismo , Homeostase , Concentração de Íons de Hidrogênio , Mitocôndrias , Água do Mar/química
3.
Anal Bioanal Chem ; 412(25): 6927-6938, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32712814

RESUMO

Real-time fluorescence detection of nucleic acid exhibit excellent performance in analytical and diagnostic applications. However, the requirement of laboratory-based instrument and complex nucleic acid extraction greatly limits their application in point-of-care testing (POCT). Herein, a novel integrated silica membrane-based platform incorporating nucleic acid purification, amplification, and detection steps was developed. A universal and portable visualization platform was fabricated by incorporating denaturation bubble-mediated strand exchange amplification (SEA) reaction with silica membrane. The fluorescence signal of SYBR Green I with amplification products was visualized by the naked eye using a simple ultraviolet light on the silica membrane, and significant discrimination between the positive and negative samples could be easily and visually obtained. Besides, chitooligosaccharide-modified silica membrane allows the purification of nucleic acid in a totally aqueous system and enables in situ SEA. With the proposed integrated platform, 102-108 cfu/mL Vibrio parahaemolyticus could be successfully detected and excellent performance was also revealed for gram-positive pathogens. The detection limit of the method for artificially spiked oysters was 103 cfu/g and reached 100 cfu/g after 12 h enrichment. This proof-of-concept method could also be applied to a variety of nucleic acid amplification methods. We believe that the proposed silica membrane-based platform has great potential for the rapid and low-cost detection of nucleic acids especially in low-resource settings. Graphical abstract.


Assuntos
Microbiologia de Alimentos , Membranas Artificiais , Técnicas Microbiológicas/economia , Ácidos Nucleicos/isolamento & purificação , Dióxido de Silício/química , Animais , Custos e Análise de Custo , Limite de Detecção , Ostreidae/microbiologia , Testes Imediatos , Estudo de Prova de Conceito
4.
Fish Shellfish Immunol ; 88: 318-327, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30853654

RESUMO

A variety of combinations of leucine-rich repeat (LRR) and immunoglobulin-like (Ig) domains have been found and discovered in invertebrates and vertebrates, but the functions remain largely unexplored. In the present study, a novel LRR and Ig domain-containing protein (LRRIG), CgLRRIG-3, was identified and characterized from oyster Crassostrea gigas. It contained two typical LRR motifs, a LRRNT motif and an Ig domain and PSI-BALST and phylogeny analysis revealed that the sequence of CgLRRIG-3 was most related with leucine-rich repeat neuronal 1 proteins from vertebrate. Its mRNA transcripts were constitutively expressed in muscle, gill, hepatopancreas, mantle, gonad and hemocytes with the highest level in hepatopancreas. The mRNA expression level of CgLRRIG-3 in hemocytes could respond to the stimulations of variety PAMPs including lipopolysaccharide (LPS), peptidoglycan (PGN), glucan (GLU) and polyinosinic-polycytidylic acid (poly I:C). The recombinant proteins exhibited a wide PAMP binding repertoire to four typical PAMPs and could significantly induce the expression of CgTNF-1 and CgIL17-5 as well as increase phagocytosis in primary cultured oyster hemocytes. In hepatopancreas, CgLRRIG-3 was mainly distributed in the basolateral membrane of digestive tubule and the hemocoel sinusoid between the digestive tubules. And in hemocytes, the positive signal was mainly distributed in a special group of granulocytes. These results collectively indicated that CgLRRIG-3 could not only function as an immune effector.


Assuntos
Crassostrea/genética , Crassostrea/imunologia , Imunidade Inata , Receptores de Reconhecimento de Padrão/genética , Receptores de Reconhecimento de Padrão/imunologia , Sequência de Aminoácidos , Animais , Células Cultivadas , Hemócitos/metabolismo , Domínios de Imunoglobulina , Moléculas com Motivos Associados a Patógenos/farmacologia , Filogenia , Domínios Proteicos , Receptores de Reconhecimento de Padrão/química , Alinhamento de Sequência
5.
Heart Surg Forum ; 22(1): E019-E023, 2019 01 28.
Artigo em Inglês | MEDLINE | ID: mdl-30802192

RESUMO

BACKGROUND: To study the effect of miR-29b on myocardial infarction via Notch signaling pathway in rats. METHODS: The rat acute myocardial infarction (AMI) models were established and were divided into AMI group, sham group and normal group (N = 10 in each group). HE (Hemotoxylin and eosin) staining was used to detect whether the model was constructed successfully. MiR-29b mimics, inhibitors, mimics negative control (NC) were transfected into H9c2 (2-1) cells. Then, cells were divided into a mimics group, inhibitor group, NC group, and blank group. The relative expression levels of miR-29b, Notch1, DII4 and Hesl were detected by qRT-PCR. The expression of NICD1 was detected by Western blotting. RESULTS: The rat AMI model was successfully constructed. Compared with normal and sham groups, the miR-29b expression was down-regulated, while the expression of Notch1, DII4 and Hesl was increased, and the NICD1 protein expression was increased in the myocardial infarction area of the AMI group (P < .05). Compared with the NC and blank groups, the relative expression of Notch1, DII4, Hesl and NICD1 were upregulated in the mimics group (P < .05), whereas the expression of Notch1, DII4, Hesl and NICD1 in the inhibitor group was decreased (P < .05). CONCLUSION: MiR-29b inhibited myocardial fibrosis and cardiac hypertrophy by activating the Notch signaling pathway and protected myocardium against myocardial infarction.


Assuntos
Regulação da Expressão Gênica , MicroRNAs/genética , Infarto do Miocárdio/genética , RNA/genética , Receptor Notch1/genética , Remodelação Ventricular , Animais , Western Blotting , Modelos Animais de Doenças , Masculino , MicroRNAs/biossíntese , Infarto do Miocárdio/metabolismo , Infarto do Miocárdio/fisiopatologia , Ratos , Reação em Cadeia da Polimerase em Tempo Real , Receptor Notch1/metabolismo , Transdução de Sinais
6.
Anal Biochem ; 545: 38-42, 2018 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-29355484

RESUMO

A strand exchange amplification (SEA) method to detect foodborne pathogen Listeria monocytogenes was developed. SEA is a novel nucleic acid amplification method that only requires one pair of primers. The specie-specific primers were designed by targeting the 16S rRNA gene and the amplification reaction was performed as short as 60 min at 61 °C. Notably, SEA method could not only detect genomic DNA but also detect RNA by one step without requiring extra reverse transcription. The result could be visualized by naked eyes so that water bath pot would be the only equipment needed. Moreover, culture fluids and bacteria colony could be successfully detected without any pretreatment and the method displayed good specificity and strong anti-jamming capacity. These features greatly simplified the operating procedure and made SEA method be potential for developing point-of-care testing (POCT) devices to detect viable L. monocytogenes.


Assuntos
Listeria monocytogenes/isolamento & purificação , Técnicas de Amplificação de Ácido Nucleico/métodos , Microbiologia de Alimentos , Humanos , Listeria monocytogenes/genética , Testes Imediatos , RNA Ribossômico 16S/genética
7.
Fish Shellfish Immunol ; 77: 419-428, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29609030

RESUMO

Astakine is a cytokine-like factor containing a prokineticin domain, which directly participates in hematopoiesis and blood cell differentiation. In the present study, a novel Astakine gene was identified from Chinese mitten crab Eriocheir sinensis (designated as EsAst). The full-length cDNA of EsAst was of 1163 bp, consisting of a 5' untranslated region (UTR) of 120 bp, a 3' UTR of 656 bp, and an open reading frame (ORF) of 387 bp encoding a polypeptide of 128 amino acids. There were a signal peptide and a prokineticin domain with nine conserved cysteine residues in the deduced amino acid sequence of EsAst. EsAst shared higher similarity with Astakines from Penaeus monodon and Pacifastacus leniusculus, and it was closely clustered with the Astakine from shrimp P. monodon in the phylogenetic tree. The EsAst mRNA transcript was higher expressed in hemocytes and hepatopancreas. The relative expression level of EsAst in hemocytes was continuously increased from 1.5 to 48 h after Vibro anguillarum challenge compared that in the untreated control group. After Pichia pastoris GS115 challenge, the relative expression level of EsAst in hemocytes was also up-regulated. After rEsAst injection, ROS levels in HPT cells were also increased at 12 and 24 h, and the total hemocyte counts were also significantly increased at 6, 9, 12, and 24 h post rEsAst injection. The interference of EsAst expression with dsRNA injection could delay the recovery of hemocytes production post A. hydrophila stimulation. When mitochondrial complexes I was knock down by dsRNA, ROS levels were decreased and THCs were also decreased. Recovery of hemocyte production inducing by A. hydrophila stimulation and rEsAst injection were delayed with dsEsbc1 injection. When ROS levels were increased after RNAi of Lon protease, THCs were also increased. The expression levels of five genes (EsJNK, EsSTAT, EsPI3K, EsAKT1, EsP70S6K) involved in SAPK-JNK and mTOR signaling pathways were up-regulated at 12 and 24 h in rEsAst group and EsLon dsRNA group compared with that in EGFP dsRNA group, and were similar to the trend of ROS levels. These results collectively suggested that EsAst should be a novel Astakine to promote the production of hemocytes in a ROS-dependent way in E. sinensis.


Assuntos
Braquiúros/genética , Braquiúros/imunologia , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Fator de Crescimento do Endotélio Vascular Derivado de Glândula Endócrina/genética , Fator de Crescimento do Endotélio Vascular Derivado de Glândula Endócrina/imunologia , Aeromonas hydrophila/fisiologia , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/química , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/imunologia , Sequência de Bases , Perfilação da Expressão Gênica , Filogenia , Pichia/fisiologia , Distribuição Aleatória , Alinhamento de Sequência , Fator de Crescimento do Endotélio Vascular Derivado de Glândula Endócrina/química , Vibrio/fisiologia
8.
Fish Shellfish Immunol ; 74: 332-340, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29305333

RESUMO

Clip-domain serine proteinase is an important serine proteinase family involved in many biological processes, which is only found in invertebrates. In the present study, the full-length cDNA of a clip domain serine proteinase (designed as EsCDSP) gene was cloned from Chinese mitten crab Eriocheir sinensis using rapid amplification of cDNA ends (RACE) technique. It was of 1488 bp with an open reading frame (ORF) of 1134 bp encoding a polypeptide of 377 amino acids. There were a signal peptide, a clip domain, and a Tryp_SPc domain in the deduced amino acid sequence of EsCDSP. Highly conserved cysteine residues were identified in the clip domain and Tryp_SPc domain. EsCDSP shared similarities of 40%-61% with CDSPs from Penaeus monodon (ACP19562.1), Scylla paramamosain (CCW43200.1), Drosophila melanogaster (NP_649734.2) and Delia antiqua (AAW57295.1). It was clustered with other CDSPs from crabs in the phylogenetic tree. EsCDSP transcript was highly expressed in hemocytes and it could response to the stimulations of Vibro anguillarum and Pichia pastoris. rEsCDSP could activate proPO system and significantly increase the PO activity of HLS. In addition, rEsCDSP could bond to Aeromonas hydrophila, Vibro anguillarum and Vibro alginolyticus, and reduced the mortality rate causing by pathogen infection. All the results suggested that EsCDSP was an important immune response participator involved in activation of the proPO system of crab.


Assuntos
Braquiúros/genética , Braquiúros/imunologia , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Serina Proteases/genética , Serina Proteases/imunologia , Aeromonas hydrophila/fisiologia , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/química , Proteínas de Artrópodes/genética , Proteínas de Artrópodes/imunologia , Sequência de Bases , Perfilação da Expressão Gênica , Filogenia , Pichia/fisiologia , Alinhamento de Sequência , Serina Proteases/química , Vibrio/fisiologia
9.
Fish Shellfish Immunol ; 66: 140-147, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28476673

RESUMO

Ocean acidification (OA) has deleterious impacts on immune response and energy homeostasis status of Mollusca. In the present study, the apoptosis ratio of hemocytes and the adenosine triphosphate (ATP) allocation in gill tissues were determined after Pacific oysters Crassostrea gigas were exposed to elevated CO2 environment (pH = 7.50) for 16 days.The apoptosis ratio in CO2 exposure group (35.2%) was significantly higher (p < 0.05) than that in the control group, and the increased apoptosis ratio induced by elevated CO2 could be significantly inhibited (p < 0.05) by KH7, a specific inhibitor of a bicarbonate sensor soluble adenylyl cyclase (sAC). After CO2 exposure, sAC in oyster (CgsAC) was found to be clustered with mitochondria in the cytoplasm, and the pro-caspase-3 was cleaved into two small fragments. Moreover, the activities of caspase-3 and caspase-9 also increased post CO2 exposure and these increases could be inhibited by KH7. However, the activities of caspase-8 did not change significantly compared with that in the control group. After CO2 exposure, the ATP content in the gill increased significantly (p < 0.05) and such increase could also be inhibited by KH7. The ATP content in purified gill mitochondria decreased significantly (p < 0.05) after CO2 exposure, which was also inhibited by KH7. These results implied that the elevated CO2 could activate the mitochondria-CgsAC pathway of apoptosis and ATP metabolism in oyster, and this pathway played essential roles in maintaining the homeostasis and the balance of energy metabolism in response to OA.


Assuntos
Trifosfato de Adenosina/metabolismo , Adenilil Ciclases/metabolismo , Apoptose , Dióxido de Carbono/farmacologia , Crassostrea/fisiologia , Animais , Crassostrea/efeitos dos fármacos , Crassostrea/enzimologia , Mitocôndrias/metabolismo
10.
Fish Shellfish Immunol ; 60: 141-149, 2017 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-27871901

RESUMO

Translationally controlled tumor protein (TCTP) is initially described as a highly conserved protein implicated in cell growth, and it is subsequently confirmed to play important roles in mediating the innate immune response, especially the inflammatory. In the present study, the full-length cDNA sequence of a TCTP from Zhikong scallop Chlamys farreri (designed as CfTCTP) was cloned by rapid amplification of cDNA ends (RACE) technique based on the expression sequence tag (EST) analysis. It was of 1230 bp with an open reading frame (ORF) of 543 bp encoding a polypeptide of 180 amino acids. The deduced amino acid sequence contained a conserved TCTP signature sequence (from I47 to E58) and it shared 26.1%-48.9% similarities with previously identified TCTPs. CfTCTP was clustered with the TCTP from Argopectehs irradias in the phylogenetic tree and was designated into a single branch of mollusk with TCTP from Ruditapes philippinarum. The mRNA transcripts of CfTCTP were constitutively expressed in all the tested tissues, including haemocytes, muscle, mantle, gill, hepatopancreas, kidney and gonad, with the highest expression level in hepatopancreas. The mRNA expression level of CfTCTP in oocytes and fertilized eggs kept at a higher level, and was down-regulated from 2-cell embryos to the lowest level in gastrula. Then it was up-regulated in trochophore and dropped down in the late veliger larvae to the similar level as that in oocytes. After pathogen-associated molecular patterns (PAMPs) stimulation, the expression of CfTCTP mRNA in haemocytes was increased at 3 or 6 h, and fall down to the normal level at 24 h. The recombinant protein of CfTCTP could induce the release of histamine from BT-549 cells. All these results indicated that CfTCTP was a pro-inflammatory factor and it could be maternally transferred from female gonad to oocytes and offspring, and play pivotal role in the embryonic development and immune protection of scallops.


Assuntos
Biomarcadores Tumorais/genética , Imunidade Inata , Pectinidae/genética , Pectinidae/imunologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Biomarcadores Tumorais/química , Biomarcadores Tumorais/imunologia , DNA Complementar/genética , DNA Complementar/metabolismo , Embrião não Mamífero/imunologia , Imunomodulação , Moléculas com Motivos Associados a Patógenos/metabolismo , Pectinidae/classificação , Pectinidae/crescimento & desenvolvimento , Filogenia , Conformação Proteica , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Alinhamento de Sequência/veterinária , Proteína Tumoral 1 Controlada por Tradução
11.
Fish Shellfish Immunol ; 70: 308-318, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-28889011

RESUMO

Leucine-rich repeat (LRR) domain and immunoglobulin (Ig) domain are both competent immune recognition modules, and the immunological roles of LRR and Ig domain containing- proteins (LRRIGs) are speculated to be multifunctional and worth investigating. In the present study, two novel LRRIGs, CgLRRIG-1 and CgLRRIG-2, were identified and characterized from oyster Crassostrea gigas. Both of them contained an N-terminal LRR region, an Ig domain, a transmembrane region, and a C-terminal cytoplasmic tail. The mRNA transcripts of CgLRRIG-1 and CgLRRIG-2 were constitutively expressed in muscle, gill, hepatopancreas, mantle, gonad and hemocytes with the highest expression level in hepatopancreas. Their mRNA expression levels in hemocytes were significantly up-regulated after the stimulations with four PAMPs including peptidoglycan (PGN), lipopolysaccharide (LPS), glucan (GLU) and polyinosinic-polycytidylic acid (poly I:C) and one bacteria Vibrio anguillarum. The recombinant proteins, rCgLRRIG-1 and rCgLRRIG-2, could bind to PGN, LPS, GLU and poly I:C, and rCgLRRIG-2 exhibited higher binding affinity. Additionally, rCgLRRIG-1 and rCgLRRIG-2 could significantly induce the expression of CgTNF-1 and CgIL17-5 in cultured oyster hemocytes, and the activity of rCgLRRIG-2 was higher than that of rCgLRRIG-1. All these results indicated that CgLRRIG-1 and CgLRRIG-2 could function as immune effectors or pro-inflammatory factors as well as PRRs in oyster.


Assuntos
Crassostrea/genética , Crassostrea/imunologia , Citocinas/genética , Imunidade Inata , Receptores de Reconhecimento de Padrão/genética , Receptores de Reconhecimento de Padrão/imunologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Citocinas/imunologia , Expressão Gênica , Moléculas com Motivos Associados a Patógenos/farmacologia , Filogenia , Receptores de Reconhecimento de Padrão/química , Alinhamento de Sequência , Vibrio/fisiologia
12.
Fish Shellfish Immunol ; 61: 68-78, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-27940367

RESUMO

B-cell translocation gene 1 (BTG1) is a member of the anti-proliferative gene family, which plays important roles in regulation of cell cycle. In the present study, a B-cell translocation gene 1 molecule homologue (designed CgBTG1) are identified and characterized in oyster Crassostrea gigas. CgBTG1 contains a conserved BTG domain with Box A and Box B motifs, and it shares high similarities with both BTG1 and BTG2 proteins in vertebrates. CgBTG1 mRNA is predominantly expressed in hemocytes, and its expression level in hemocytes is significantly up-regulated at 6 h (5.40-fold, p < 0.01) post Vibrio splendidus stimulation. The apoptosis rate of oyster hemocytes is significantly decreased (p < 0.05) after CgBTG1 interfered by dsRNA (dsCgBTG1). This is indicated that CgBTG1 participated in the regulation of oyster hemocytes apoptosis. Furthermore, CgBTG1 could also induce the apoptosis of cancer cells (HeLa, A549 and BEL7402) in vitro. Compared with normal oysters, both vessel-like structures and muscle fibers in CgBTG1 interfered oysters are severely damaged after V. splendidus challenge in paraffin section, considering that CgBTG1 possessed an analogous feature of angiogenesis for maintenance of vessel-like structures in adductor muscle of oyster. The results suggests that CgBTG1 is a multi-functional molecule involved in the immune response of C. gigas against pathogen infection, which provides more clues for intensive studies of BTG family proteins in invertebrates.


Assuntos
Crassostrea/genética , Crassostrea/imunologia , Imunidade Inata , Proteínas de Neoplasias/genética , Vibrio/fisiologia , Animais , Linhagem Celular Tumoral , Clonagem Molecular , Crassostrea/metabolismo , DNA Complementar/genética , DNA Complementar/metabolismo , Células HEK293 , Hemócitos/imunologia , Hemócitos/metabolismo , Hemócitos/microbiologia , Humanos , Proteínas de Neoplasias/metabolismo , Especificidade de Órgãos , Fagocitose , Filogenia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Regulação para Cima
13.
Fish Shellfish Immunol ; 62: 341-348, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-28159695

RESUMO

Serotonin, also known as 5-hydroxytryptamine (5-HT), is a critical neurotransmitter in the neuroendocrine-immune regulatory network and involved in regulation of the stress response in vertebrates and invertebrates. In the present study, serotonin was found to be widely distributed in the tissues of Pacific oyster Crassostrea gigas, including haemolymph, gonad, visceral ganglion, mantle, gill, labial palps and hepatopancreas, and its concentration increased significantly in haemolymph and mantle after the oysters were exposed to air for 1 d. The apoptosis rate of haemocytes was significantly declined after the oysters received an injection of extra serotonin, while the activity of superoxide dismutase (SOD) in haemolymph increased significantly. After the stimulation of serotonin during air exposure, the apoptosis rate of oyster haemocytes and the concentration of H2O2 in haemolymph were significantly decreased, while the SOD activity was significantly elevated. Furthermore, the survival rate of oysters from 4th to 6th d after injection of serotonin was higher than that of FSSW group and air exposure group. The results clearly indicated that serotonin could modulate apoptotic effect and redox during air exposure to protect oysters from stress.


Assuntos
Ar , Crassostrea/fisiologia , Agonistas do Receptor de Serotonina/metabolismo , Serotonina/metabolismo , Animais , Apoptose , Crassostrea/enzimologia , Hemócitos/enzimologia , Hemócitos/imunologia , Hemócitos/fisiologia , Peróxido de Hidrogênio/metabolismo , Estresse Fisiológico , Superóxido Dismutase/metabolismo
14.
Fish Shellfish Immunol ; 57: 252-261, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-27542616

RESUMO

Receptor for activated C kinase 1 (RACK1) is a WD-domain repeating protein which involves in the mediation of various biological processes, including innate immune response. In the present study, a RACK1 (designed as EsRACK1) gene from Chinese mitten crab E. sinensis was cloned by rapid amplification of cDNA ends (RACE) technique. The full-length cDNA sequence of EsRACK1 was of 1117 bp with an open reading frame (ORF) of 957 bp encoding a polypeptide of 318 amino acids containing seven WD repeats. EsRACK1 shared 62%-99% similarities with previously identified RACK1s in amino acid sequence, and it was clustered with the RACK1 from Pacifastacus leniusculus in the phylogenetic tree. The mRNA transcripts of EsRACK1 were constitutively expressed in various tissues with the highest expression level in hepatopancreas. The expression of EsRACK1 mRNA in hemocytes were significantly up-regulated post the stimulations with Vibrio anguillarum and Pichia pastoris. After exposure to CdCl2 and pentachlorophenol, the transcripts of EsRACK1 in hemocytes were up-regulated at the late phase from 12 h. When EsRACK1 was knocked down by dsRNA based RNAi, the total hemocyte counts, new-born hemocytes and phosphorylation of JNK were all significantly decreased. In addition, EsRACK1 transcription and phosphorylation of JNK were both decreased in hematopoietic tissue post Aeromonas hydrophila challenge. All the results suggested that EsRACK1 was involved in the innate immune response of the crab and participated in the production of new-born hemocytes through activation of JNK.


Assuntos
Proteínas de Artrópodes/genética , Braquiúros/genética , Braquiúros/imunologia , Imunidade Inata , Receptores de Superfície Celular/genética , Poluentes Químicos da Água/toxicidade , Aeromonas hydrophila/fisiologia , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/química , Proteínas de Artrópodes/metabolismo , Sequência de Bases , Braquiúros/efeitos dos fármacos , Braquiúros/microbiologia , DNA Complementar/genética , DNA Complementar/metabolismo , Hematopoese/efeitos dos fármacos , Herbicidas/toxicidade , Metais Pesados/toxicidade , Filogenia , Pichia/fisiologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores de Quinase C Ativada , Receptores de Superfície Celular/química , Receptores de Superfície Celular/metabolismo , Alinhamento de Sequência , Distribuição Tecidual , Regulação para Cima , Vibrio/fisiologia
15.
J Immunoassay Immunochem ; 36(5): 517-31, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25611486

RESUMO

A simple, rapid sample extraction method for the determination of FQs was developed. Fishery samples were extracted with 2% of 5-sulfosalicylic acid dihydrate and the extracts were analyzed directly without any further purification or clean-up procedures. The FQs were determined with standards of 2% of 5-sulfosalicylic acid dihydrate in the concentration range of 0.1-25.6 µg L(-1), and the limit of detection (LOD) was 0.1 µg L(-1). The matrix interference originated from fishery samples was eliminated by 2% of 5-sulfosalicylic acid dihydrate and did not interact with horseradish peroxidase (HRP) labeled IgG in western blotting. No significant matrix interference was observed as samples extracted with 2% of 5-sulfosalicylic acid dihydrate. Recoveries of FQs in fishery muscle were between 72.37-94.35% in the concentrations range of 10-50 µg kg(-1).This extraction procedure was much rapider and simpler to conventional ELISA extraction procedure and could be used as a time-saving and cost-effective method for FQs monitoring in fishery samples.


Assuntos
Benzenossulfonatos/química , Ensaio de Imunoadsorção Enzimática/métodos , Pesqueiros , Fluoroquinolonas/análise , Salicilatos/química , Limite de Detecção , Músculos/química
16.
Anal Bioanal Chem ; 406(26): 6637-46, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25168115

RESUMO

A novel non-instrumental bioanalysis based on colloidal-gold immunochromatography in a modified glass capillary was developed and named capillary immunochromatographic assay (CICA). In this report, glass capillary was proposed as a support in immunochromatographic assay because of its excellent characteristics. Goat anti-rabbit IgG and parvalbumin (PV) were immobilized on the inner wall of the glass capillary as control zone and test zone, respectively. The CICA was constructed, and main variables for the performance were optimized. Using an important allergen of fish products (parvalbumin, PV) as the target, the analytical efficiency of the developed technique was investigated and the visual detection limit (VDL) and semi-quantitative limit of detection (LOD) were estimated to be 70 ng mL(-1) and 40 ng mL(-1), respectively. The coefficient of variation (CV) for the intra-assay and inter-assay was calculated for the PV concentration of 50 ng mL(-1), and the entire operation, including sample preparation, was consistently performed in 30 min. The developed technique was implemented and validated with different foodstuffs, including Scophthalmus maximus (Linnaeus), surimi products, and livestock, confirming sufficient accuracy and precision of results and verifying the method to be efficacious. These results enabled us to propose CICA as a new and promising technique for simple, rapid, and on-site screening of PV in biological samples.


Assuntos
Alérgenos/análise , Anticorpos Imobilizados/química , Cromatografia de Afinidade/instrumentação , Ouro/química , Nanopartículas Metálicas/química , Parvalbuminas/análise , Animais , Bovinos , Galinhas , Desenho de Equipamento , Peixes , Contaminação de Alimentos/análise , Cabras , Limite de Detecção , Carne/análise , Nanopartículas Metálicas/ultraestrutura , Coelhos , Suínos
18.
Mater Today Bio ; 25: 101008, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38495915

RESUMO

Massive blood loss due to injury is the leading cause of prehospital deaths in disasters and emergencies. Hemostatic materials are used to realize rapid hemostasis and protect patients from death. Researchers have designed and developed a variety of hemostatic materials. However, in addition to their hemostatic effect, hemostatic materials must be endowed with additional functions to meet the practical application requirements in different scenarios. Here, strategies for modifications of hemostatic materials for use in different application scenarios are listed: effective positioning at the site of deep and narrow wounds to stop bleeding, resistance to high blood pressure and wound movement to maintain wound formation, rapid and easy removal from the wound without affecting further treatment after hemostasis is completed, and continued function when retained in the wound as a dressing (such as antibacterial, antiadhesion, tissue repair, etc.). The problems encountered in the practical use of hemostatic materials and the strategies and progress of researchers will be further discussed in this review. We hope to provide valuable references for the design of more comprehensive and practical hemostatic materials.

19.
Sci Data ; 11(1): 963, 2024 Sep 04.
Artigo em Inglês | MEDLINE | ID: mdl-39232034

RESUMO

Dryocosmus kuriphilus, commonly known as the chestnut gall wasp, belongs to the family Cynipidae and is native to China. It is a highly invasive insect species causing serious damage to chestnut trees and has rapidly spread to various continents, including Europe, North America, and Oceania. The D. kuriphilus has become one of the important pests of chestnut plants in the world and is listed as a quarantine object by the European and Mediterranean Plant Protection Organization (EPPO). In this study, we used PacBio long reads, Illumina short reads, and Hi-C sequencing data to construct a chromosome-level assembly of the D. kuriphilus genome. The assembled genome includes 14,729 contigs with a total length of 2.28 Gb and a contig N50 of 0.8 Mb. With Hi-C technology, 2.17 Gb (95.02%) of contigs were anchored and oriented into the 10 pseudochromosomes with the scaffold N50 of 198.8 Mb and the scaffold N90 of 158.8 Mb. In total, 24,086 protein-coding genes were predicted in the assembled D. kuriphilus genome as the reference gene set. A total of 1.82 Gb repeats (occupying 79.7% of the genome), including 1.42 Gb of transposable elements and 0.40 Gb of tandem repeats, were identified in D. kuriphilus genome. In the evaluation of completeness, the BUSCO analysis determined a level of 98.1% completeness for the assembled genome sequences based on the Insecta database (OrthoDB version 10). The high-quality genome assembly of D. kuriphilus will not only provide a valuable reference for the study of its evolutionary history and genetic structure but also facilitate the research of host-pest interactions and invasiveness. Moreover, this genome assembly will promote in the development of effective management strategies to mitigate the economic and ecological impacts of this invasive pest on chestnut trees and ecosystems.


Assuntos
Vespas , Animais , Fagaceae/genética , Genoma de Inseto , Vespas/genética
20.
Int J Biol Macromol ; 276(Pt 2): 133914, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-39029842

RESUMO

Intelligent food packaging has recently gained significant attention due to the heightened consumer awareness regarding food quality. Although anthocyanins avoid safety issues, the instability and leakage of anthocyanins restrict their utilization in freshness indicator labels. In this study, we introduced an innovative metal-organic framework (UiO-66-NH2) synergistic pH-colorimetric label with fast ammonia-responsive, incorporating sodium alginate, red cabbage anthocyanin, and UiO-66-NH2. The cross-linked sodium alginate substrate enabled the label to possess superior insolubility. The microscopic morphology of the labels was intricately analyzed, while their sensitivity was rigorously tested utilizing ammonia as a representative gas. Due to the remarkable UV absorption capability of UiO-66-NH2 and various molecular interactions with anthocyanins, the label exhibited good UV absorption, enhanced stability, and optimized performance in reducing anthocyanin leakage, ensuring the stability and effectiveness of the labels in practical applications. The prepared label exhibited good specificity for volatile amines and ammonia gases, and robust anti-interference properties, enabling visualization and early detection of shrimp spoilage during storage at different temperatures. The strategy employed in this study presents promising new possibilities for developing intelligent packaging solutions for food products.


Assuntos
Alginatos , Antocianinas , Embalagem de Alimentos , Estruturas Metalorgânicas , Alginatos/química , Estruturas Metalorgânicas/química , Antocianinas/química , Embalagem de Alimentos/métodos , Concentração de Íons de Hidrogênio , Amônia/química , Amônia/análise , Qualidade dos Alimentos , Colorimetria/métodos
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