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1.
J Agric Food Chem ; 67(38): 10702-10712, 2019 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-31490688

RESUMO

Human milk oligosaccharides are complex carbohydrates with multibiofunctional health benefits to newborns. Human milk free oligosaccharides (HMOs) are well characterized. However, changes in the N/O-glycome during lactation are poorly reported. Herein, we qualitatively and quantitatively investigated N/O-glycome profiles and their alteration in human milk at different lactation stages. N-Glycans were mainly fucosylated and nonsialylated, nonfucosylated throughout lactation. O-Glycans mainly consisted of sialylated and nonsialylated, nonfucosylated in colostrum and transitional milk, and fucosylated and nonfucosylated, nonsialylated in mature milk. Fucosylated and sialylated N-glycans gradually decreased and increased, respectively, as lactation progressed; O-glycans showed the reverse. Interestingly, changes in HMO abundance decreased during lactation, complementing HMG N/O-glycome changes. In conclusion, temporal HMG glycosylation changes provide the groundwork for developing infant formula that is closer to breast milk at different lactation stages.


Assuntos
Glicoproteínas/química , Lactação , Leite Humano/química , Adulto , Colostro/química , Feminino , Glicoproteínas/metabolismo , Glicosilação , Humanos , Espectrometria de Massas , Leite Humano/metabolismo , Oligossacarídeos/química , Oligossacarídeos/metabolismo , Polissacarídeos/química , Polissacarídeos/metabolismo
2.
J Agric Food Chem ; 67(35): 9950-9957, 2019 Sep 04.
Artigo em Inglês | MEDLINE | ID: mdl-31403788

RESUMO

Protein glycosylation is a ubiquitous posttranslational modification that modulates protein properties, thereby influencing bioactivities within a system. Duck egg white (DEW) proteins exhibit diverse biological properties compared with their chicken egg white (CEW) counterparts, which might be related to glycosylation. N-Glycoproteome analysis of DEW was conducted, and a total of 231 N-glycosites from 68 N-glycoproteins were identified. Gene ontology analysis was used to elucidate the biofunctions of DEW N-glycoproteins and compare them with those of CEW, which showed that the differences mostly involved molecular functions and biological processes. The biological functions of DEW N-glycoproteins were illuminated through bioinformatics analysis and comparison with CEW orthologues, which showed different allergenicities and antibacterial abilities. These divergences might be initiated by specific alterations in glycosylation, which can enhance the proteolysis resistance and protein steric hindrance. These results provide new insights for discovering the effects of N-glycosylation on biofunctions during the divergence of homologous proteins.


Assuntos
Galinhas/genética , Patos/genética , Proteínas do Ovo/química , Glicoproteínas/química , Animais , Evolução Biológica , Galinhas/metabolismo , Patos/metabolismo , Proteínas do Ovo/genética , Proteínas do Ovo/metabolismo , Clara de Ovo/química , Glicoproteínas/genética , Glicoproteínas/metabolismo , Glicosilação , Proteômica
3.
World J Microbiol Biotechnol ; 35(8): 121, 2019 Jul 22.
Artigo em Inglês | MEDLINE | ID: mdl-31332590

RESUMO

The economics of bioflocculant production is coupled with the use of a low-cost substrate at appropriate culture conditions. The use of a waste substrate for this purpose offers an additional treatment measure to mitigate environmental pollution. We investigated the growth of Aspergillus flavus and its bioflocculant yield using chicken viscera hydrolysate as the sole media. The effects of culture conditions including time, pH, shaker speed, temperature and inoculum size on bioflocculant production were all investigated and optimised through response surface method based on the central component design (CCD) package of Design Expert. Next, the purified bioflocculant was physically and chemically characterised. Under optimised culture conditions (incubation time 72 h, pH 7, shaker speed 150 rpm, temperature 35 °C and inoculum 4%), 6.75 g/L yield of crude bioflocculant was recorded. The bioflocculant activity was mostly distributed in the cell-free supernatant with optimum efficiency of 91.8% at a dose of 4 mL/100 mL Kaolin suspension. The purified bioflocculant was a glycoprotein consisting of 23.46% protein and 74.5% sugar, including 46% neutral sugar and 2.01% uronic acid. The X-ray photoelectron spectroscopy fundamental analysis of the purified bioflocculant indicated that the mass proportion of C, O and N, were 63.46%, 27.87% and 8.86%, respectively. The bioflocculant is mainly composed of carbonyl, amino, hydroxyl, and amide functional groups. This study for the first time indicates a high potential of bioflocculant yield from chicken viscera at the appropriate culture conditions.


Assuntos
Aspergillus flavus/metabolismo , Meios de Cultura/química , Animais , Galinhas , Glicoproteínas/metabolismo , Concentração de Íons de Hidrogênio , Microscopia Eletrônica de Varredura , Espectroscopia Fotoeletrônica , Espectroscopia de Infravermelho com Transformada de Fourier , Temperatura Ambiente , Vísceras
4.
Nat Commun ; 10(1): 2498, 2019 06 07.
Artigo em Inglês | MEDLINE | ID: mdl-31175312

RESUMO

Allogeneic mesenchymal stem cells (MSCs) exhibit immunoregulatory function in human autoimmune diseases such as systemic lupus erythematosus (SLE), but the underlying mechanisms remain incompletely understood. Here we show that the number of peripheral tolerogenic CD1c+ dendritic cells (DCs) and the levels of serum FLT3L are significantly decreased in SLE patients especially with lupus nephritis, compared to healthy controls. Transplantation of allogeneic umbilical cord-derived MSCs (UC-MSCs) significantly up-regulates peripheral blood CD1c+DCs and serum FLT3L. Mechanistically, UC-MSCs express FLT3L that binds to FLT3 on CD1c+DCs to promote the proliferation and inhibit the apoptosis of tolerogenic CD1c+DCs. Conversely, reduction of FLT3L with small interfering RNA in MSCs abolishes the up-regulation of tolerogenic CD1c+DCs in lupus patients treated with MSCs. Interferon-γ induces FLT3L expression in UC-MSCs through JAK/STAT signaling pathway. Thus, allogeneic MSCs might suppress inflammation in lupus through up-regulating tolerogenic DCs.


Assuntos
Antígenos CD1/imunologia , Células Dendríticas/imunologia , Glicoproteínas/imunologia , Tolerância Imunológica/imunologia , Lúpus Eritematoso Sistêmico/terapia , Proteínas de Membrana/imunologia , Transplante de Células-Tronco Mesenquimais , Adulto , Antígenos CD1/metabolismo , Estudos de Casos e Controles , Células Dendríticas/metabolismo , Feminino , Glicoproteínas/metabolismo , Humanos , Interferon gama/farmacologia , Janus Quinases/efeitos dos fármacos , Janus Quinases/metabolismo , Lúpus Eritematoso Sistêmico/imunologia , Nefrite Lúpica/imunologia , Nefrite Lúpica/terapia , Masculino , Proteínas de Membrana/metabolismo , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/metabolismo , Pessoa de Meia-Idade , Fatores de Transcrição STAT/efeitos dos fármacos , Fatores de Transcrição STAT/metabolismo , Transdução de Sinais/efeitos dos fármacos , Transplante Homólogo , Adulto Jovem
5.
Medicine (Baltimore) ; 98(24): e16035, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31192960

RESUMO

BACKGROUND: The patient's pattern identification has been used for personalized medicine in traditional Korean medicine (TKM) and aims for patient-specific therapy by Korean medical doctors. The pattern identification in this trial will be diagnosed from body constitution questionnaire (BCQ) with a more objective diagnosis of it but this method still needs a more concrete scientific basis. Glycoproteins are well-known to be associated with diseases (especially cancers) so glycoproteomics can be applied to differentiate pattern identification types of lung cancer patients. Thus, for the first time proteomics approach will be applied to the pattern identification by comparing BCQ assessment in order to establish a scientific basis with clinical proteomics for precision medicine. METHODS: This observational trial will at first diagnose the pattern identification types of lung cancer patients with BCQ assessment and then elucidate their relationships with proteomics. Blood samples will be collected before surgery along with clinical information of participants. The patients' pattern identification in TKM will be diagnosed from BCQ assessment. Then, lung cancer patients will be divided and pooled into 3 lung cancer entire (LCE) groups according to their pattern identification types (Xu, Stasis, or Gentleness). Three lung cancer representative (LCR) groups will be selected and pooled from each LCE group by selecting those with the same control factors. The 3 LCE groups and the 3 LCR groups from lung cancer patients will be independently analyzed through the glycoproteomics approach based on the patients' pattern identification. Glycoproteins from the 6 groups will be identified through proteomics approach and then categorized for analysis. DISCUSSION: This study intends to diagnose pattern identification of patients in TKM with BCQ assessment and proteomics approach. The identification of the glycoproteins in each group will lead to the scientific foundation of personalized medicine in TKM according to patients' pattern identification for lung cancer therapy. We intend to(1) diagnose the pattern identification types of lung cancer patients with BCQ under the framework of TKM;(2) evaluate BCQ assessment with glycoproteomics approach for precision medicine. TRIAL REGISTRATION: ClinicalTrials.gov NCT03384680. Registered 27 December 2017. Retrospectively registered.


Assuntos
Constituição Corporal , Ensaios Clínicos como Assunto , Glicoproteínas/metabolismo , Neoplasias Pulmonares/terapia , Estudos Observacionais como Assunto , Medicina de Precisão , Biomarcadores Tumorais/metabolismo , Humanos , Neoplasias Pulmonares/diagnóstico , Medicina Tradicional Coreana , Seleção de Pacientes , Proteoma , Proteômica , Inquéritos e Questionários
6.
Aquat Toxicol ; 212: 54-69, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31075620

RESUMO

In this era of global climate change, ocean acidification is becoming a serious threat to the marine ecosystem. Despite this, it remains almost unknown how fish will respond to the co-occurrence of ocean acidification with other conventional environmental perturbations typically salinity fluctuation and high ammonia threat. Therefore, the present work evaluated the interactive effects of elevated pCO2, salinity reduction and high environmental ammonia (HEA) on the ecophysiological performance of European sea bass (Dicentrarchus labrax). Fish were progressively acclimated to seawater (32 ppt), to brackish water (10 ppt) and to hyposaline water (2.5 ppt). Following acclimation to different salinities for at least two weeks, fish were exposed to CO2-induced water acidification representing present-day (control pCO2, 400 µatm, LoCO2) and future (high pCO2, 1000 µatm, HiCO2) sea-surface CO2 level for 3, 7 and 21 days. At the end of each exposure period, fish were challenged with HEA for 6 h (1.18 mM representing 50% of 96 h LC50). Results show that, in response to the individual HiCO2 exposure, fish within each salinity compensated for blood acidosis. Fish subjected to HiCO2 were able to maintain ammonia excretion rate (Jamm) within control levels, suggesting that HiCO2 exposure alone had no impact on Jamm at any of the salinities. For 32 and 10 ppt fish, up-regulated expression of Na+/K+-ATPase was evident in all exposure groups (HEA, HiCO2 and HEA/HiCO2 co-exposed), whereas Na+/K+/2Cl- co-transporter was up-regulated mainly in HiCO2 group. Plasma glucose and lactate content were augmented in all exposure conditions for all salinity regimes. During HEA and HEA/HiCO2, Jamm was inhibited at different time points for all salinities, which resulted in a significant build-up of ammonia in plasma and muscle. Branchial expressions of Rhesus glycoproteins (Rhcg isoforms and Rhbg) were upregulated in response to HiCO2 as well as HEA at 10 ppt, with a more moderate response in 32 ppt groups. Overall, our findings denote that the adverse effect of single exposures of ocean acidification or HEA is exacerbated when present together, and suggests that fish are more vulnerable to these environmental threats at low salinities.


Assuntos
Equilíbrio Ácido-Base/efeitos dos fármacos , Ácidos/química , Amônia/toxicidade , Bass/fisiologia , Oceanos e Mares , Osmorregulação/efeitos dos fármacos , Salinidade , Amônia/sangue , Animais , Bass/sangue , Glicemia/metabolismo , Dióxido de Carbono/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Brânquias/efeitos dos fármacos , Brânquias/metabolismo , Glicoproteínas/metabolismo , Concentração de Íons de Hidrogênio , Íons/sangue , Ácido Láctico/sangue , Músculos/efeitos dos fármacos , Músculos/metabolismo , Análise de Componente Principal , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , ATPase Trocadora de Sódio-Potássio/metabolismo , Poluentes Químicos da Água/toxicidade
7.
Carbohydr Res ; 478: 33-45, 2019 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-31054381

RESUMO

Trypanosoma cruzi trans-sialidase (TcTS) is a cell surface protein that participates in the adhesion and invasion mechanisms of the parasite into the host cells, making it an attractive target for inhibitors design. In order to contribute to the knowledge of the interaction between TcTS and their acceptor substrates, we designed and synthesized a library of 20 benzyl lactosides substituted in C-6 of the glucose residue with a series of 1,2,3-triazole derivatives containing different aromatic substituents in the C-4 position. The library was prepared by alkyne-azide cycloaddition reaction catalyzed by Cu(I) ("click chemistry") between a benzyl ß-lactoside functionalized with an azide group in the C-6 position and a series of 2-propargyl phenyl ethers. Herein we analyzed the chromatographic behavior on high performance anion exchange chromatography (HPAEC) of the triazoyl-lactose derivatives and their activity as acceptors of TcTS and inhibitors of the sialylation of N-acetyllactosamine. The triazoyl derivatives were obtained with excellent yields and all of them behaved as moderate alternative substrates. The presence of bulky hydrophobic substituents dramatically increased the retention times in HPAEC but did not affect significantly their acceptor properties toward TcTS.


Assuntos
Amino Açúcares/antagonistas & inibidores , Glicoproteínas/metabolismo , Glicosídeos/farmacologia , Neuraminidase/metabolismo , Trypanosoma cruzi/enzimologia , Amino Açúcares/metabolismo , Configuração de Carboidratos , Glicoproteínas/química , Glicosídeos/síntese química , Glicosídeos/química , Interações Hidrofóbicas e Hidrofílicas , Neuraminidase/química , Especificidade por Substrato
8.
Mol Vis ; 25: 222-236, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31057322

RESUMO

Purpose: Glaucoma is characterized by optic nerve damage and retinal ganglion cell loss. The glycoprotein neuromedin B-associated (Gpnmb) gene is well-known to be involved in the glaucoma disease process. The purpose of this study is to identify a downstream gene through which Gpnmb affects the glaucoma phenotypes using a systems genetics approach. Methods: Retinal gene expression data for the BXD recombinant inbred (RI) strains (n=75) have previously been generated in our laboratory for a glaucoma study, and these data were used for genetic and bioinformatics analysis. Expression quantitative trait locus (eQTL) mapping and genetic correlation methods were used to identify a gene downstream of Gpnmb. Gene-set enrichment analysis was used to evaluate gene function and to construct coexpression networks. Results: The level of Gpnmb expression is associated with a highly statistically significant cis-eQTL. Stanniocalcin 1 (Stc1) has a significant trans-eQTL mapping to the Gpnmb locus. The expression of Gpnmb and Stc1 is highly correlated in the retina and other tissues, as well as with glaucoma-related phenotypes. Gene Ontology and pathway analysis showed that Stc1 and its covariates are highly associated with apoptosis, oxidative stress, and mitochondrial activity. A generated gene network indicated that Gpnmb and Stc1 are directly connected to and interact with other genes with similar biologic functions. Conclusions: These results suggest that Stc1 may be a downstream candidate of Gpnmb, and that both genes interact with other genes in a network to develop glaucoma pathogenesis through mechanisms such as apoptosis and oxidative stress.


Assuntos
Proteínas do Olho/genética , Redes Reguladoras de Genes , Glaucoma/genética , Glicoproteínas/genética , Glicoproteínas de Membrana/genética , Animais , Apoptose , Biologia Computacional/métodos , Modelos Animais de Doenças , Proteínas do Olho/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Ontologia Genética , Glaucoma/metabolismo , Glaucoma/patologia , Glicoproteínas/metabolismo , Humanos , Glicoproteínas de Membrana/metabolismo , Camundongos , Camundongos Transgênicos , Mitocôndrias/metabolismo , Mitocôndrias/patologia , Anotação de Sequência Molecular , Estresse Oxidativo , Fenótipo , Mapeamento de Interação de Proteínas , Locos de Características Quantitativas , Transdução de Sinais
9.
Life Sci ; 231: 116459, 2019 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-31075234

RESUMO

AIM: Head and neck squamous cell carcinoma (HNSCC) is one of the most prevalent types of cancer worldwide with unfavorable patient outcomes and relatively low survival rates. Long non-coding RNAs (lncRNAs) have been demonstrated to participate in the progression of HNSCC. The present study aimed to investigate the functional mechanism of lncRNA LINC00460 in HNSCC by mediating microRNA-206 (miR-206)/stanniocalcin-2 (STC2) axis. METHODS: The interactions among miR-206, LINC00460 and STC2 were identified, and the expression of LINC00460, miR-206 and STC2 in tissues and cells was determined. Gain- and loss-of function experiments were conducted to analyze effects of LINC00460, miR-206 and STC2 on the expression of apoptosis-related proteins, autophagy-related proteins, and the extents of AKT, ERK phosphorylation. Cell cycle distribution, apoptosis and the production of autophagosomes after transfection were evaluated to further explore the role of LINC00460/miR-206/STC2 axis in HNSCC. RESULTS: LINC00460 and STC2 were highly expressed while miR-206 was poorly expressed in HNSCC. Besides, miR-206 was found to bind to both LINC00460 and STC2. After the transfection of HNSCC cells with miR-206 mimic or si-LINC00460, the expression of STC2, AKT, ERK, as well as the extent of AKT, ERK phosphorylation all decreased, which facilitated the apoptosis and autophagy of HNSCC cells. CONCLUSION: Collectively, the apoptosis and autophagy of HNSCC can be facilitated by downregulating LINC00460, which highlights a novel target in the treatment of HNSCC.


Assuntos
Glicoproteínas/metabolismo , Neoplasias de Cabeça e Pescoço/genética , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , MicroRNAs/genética , RNA Longo não Codificante/genética , Carcinoma de Células Escamosas de Cabeça e Pescoço/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Apoptose/genética , Autofagia/fisiologia , Linfócitos T CD8-Positivos/metabolismo , Carcinogênese , Ciclo Celular/fisiologia , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Transformação Celular Neoplásica , Feminino , Glicoproteínas/genética , Neoplasias de Cabeça e Pescoço/metabolismo , Neoplasias de Cabeça e Pescoço/patologia , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/genética , Masculino , MicroRNAs/metabolismo , Pessoa de Meia-Idade , Transdução de Sinais , Carcinoma de Células Escamosas de Cabeça e Pescoço/metabolismo , Carcinoma de Células Escamosas de Cabeça e Pescoço/patologia , Ativação Transcricional , Regulação para Cima
10.
Invest Ophthalmol Vis Sci ; 60(6): 2034-2037, 2019 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-31067323

RESUMO

Antibodies are key reagents used in vision research, indeed across biomedical research, but they often do not reveal the whole story about a sample. It is important for researchers to be aware of aspects of antibodies that may affect or limit data interpretation. Federal agencies now require funded grants to demonstrate how they will authenticate reagents used. There is also a push for recombinant antibodies, enabled by phage display technology awarded the 2018 Nobel Prize in Chemistry, which allow for thorough validation and a fixed DNA sequence. Here, we discuss how issues surrounding antibodies are pertinent to detecting myocilin, a protein found in trabecular meshwork and associated with a portion of hereditary glaucoma. Confirmation of myocilin expression in tissues and cell culture has been adopted as validation standard in trabecular meshwork research; thus, a discussion of antibody characteristics and fidelity is critical. Further, based on our basic structural understanding of myocilin architecture and its biophysical aggregation properties, we provide a wish list for the characteristics of next-generation antibody reagents for vision researchers. In the long term, well-characterized antibodies targeting myocilin will enable new insights into its function and involvement in glaucoma pathogenesis.


Assuntos
Anticorpos/imunologia , Proteínas do Citoesqueleto/metabolismo , Proteínas do Olho/metabolismo , Glaucoma/imunologia , Glicoproteínas/metabolismo , Malha Trabecular/metabolismo , Proteínas do Citoesqueleto/imunologia , Proteínas do Olho/imunologia , Glaucoma/diagnóstico , Glaucoma/metabolismo , Glicoproteínas/imunologia , Humanos
11.
Nat Commun ; 10(1): 1813, 2019 04 18.
Artigo em Inglês | MEDLINE | ID: mdl-31000718

RESUMO

The asparagine (N)-linked Man9GlcNAc2 is required for glycoprotein folding and secretion. Understanding how its structure contributes to these functions has been stymied by our inability to produce this glycan as a homogenous structure of sufficient quantities for study. Here, we report the high yield chemoenzymatic synthesis of Man9GlcNAc2 and its biosynthetic intermediates by reconstituting the eukaryotic lipid-linked oligosaccharide (LLO) pathway. Endoplasmic reticulum mannosyltransferases (MTases) are expressed in E. coli and used for mannosylation of the dolichol mimic, phytanyl pyrophosphate GlcNAc2. These recombinant MTases recognize unique substrates and when combined, synthesize end products that precisely mimic those in vivo, demonstrating that ordered assembly of LLO is due to the strict enzyme substrate specificity. Indeed, non-physiological glycans are produced only when the luminal MTases are challenged with cytosolic substrates. Reconstitution of the LLO pathway to synthesize Man9GlcNAc2 in vitro provides an important tool for functional studies of the N-linked glycoprotein biosynthesis pathway.


Assuntos
Asparagina/metabolismo , Lipopolissacarídeos/biossíntese , Mananas/metabolismo , Manosiltransferases/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Asparagina/química , Retículo Endoplasmático/metabolismo , Glicoproteínas/metabolismo , Glicosilação , Mananas/química , Manosiltransferases/genética , Manosiltransferases/isolamento & purificação , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/isolamento & purificação
12.
MBio ; 10(2)2019 04 09.
Artigo em Inglês | MEDLINE | ID: mdl-30967458

RESUMO

Multidrug resistance (MDR) by bacterial pathogens constitutes a global health crisis, and resistance to treatment displayed by biofilm-associated infections (e.g., cystic fibrosis, surgical sites, and medical implants) only exacerbates a problem that is already difficult to overcome. Antimicrobial peptides (AMPs) are a promising class of therapeutics that may be useful in the battle against antibiotic resistance, although certain limitations have hindered their clinical development. The goal of this study was to examine the therapeutic potential of novel AMPs derived from the multifunctional respiratory host defense protein SPLUNC1. Using standard growth inhibition and antibiofilm assays, we demonstrated that a novel structurally optimized AMP, α4-short, was highly effective against the most common group of MDR bacteria while showing broad-spectrum bactericidal and antibiofilm activities. With negligible hemolysis and toxicity to white blood cells, the new peptide also demonstrated in vivo efficacy when delivered directly into the airway in a murine model of Pseudomonas aeruginosa-induced respiratory infection. The data warrant further exploration of SPLUNC1-derived AMPs with optimized structures to assess the potential application to difficult-to-cure biofilm-associated infections.IMPORTANCE The rise of superbugs underscores the urgent need for novel antimicrobial agents. Antimicrobial peptides (AMPs) have the ability to kill superbugs regardless of resistance to traditional antibiotics. However, AMPs often display a lack of efficacy in vivo. Sequence optimization and engineering are promising but may result in increased host toxicity. We report here the optimization of a novel AMP (α4-short) derived from the multifunctional respiratory protein SPLUNC1. The AMP α4-short demonstrated broad-spectrum activity against superbugs as well as in vivo efficacy in the P. aeruginosa pneumonia model. Further exploration for clinical development is warranted.


Assuntos
Anti-Infecciosos/uso terapêutico , Peptídeos Catiônicos Antimicrobianos/uso terapêutico , Produtos Biológicos/uso terapêutico , Glicoproteínas/metabolismo , Fosfoproteínas/metabolismo , Infecções por Pseudomonas/tratamento farmacológico , Infecções Respiratórias/tratamento farmacológico , Animais , Anti-Infecciosos/isolamento & purificação , Anti-Infecciosos/farmacologia , Peptídeos Catiônicos Antimicrobianos/isolamento & purificação , Peptídeos Catiônicos Antimicrobianos/farmacologia , Bactérias/efeitos dos fármacos , Produtos Biológicos/isolamento & purificação , Produtos Biológicos/farmacologia , Modelos Animais de Doenças , Camundongos , Testes de Sensibilidade Microbiana , Viabilidade Microbiana/efeitos dos fármacos , Resultado do Tratamento
13.
Oxid Med Cell Longev ; 2019: 2601394, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31001372

RESUMO

Oxidative stress plays an important role in various neurological disorders. Milk fat globule-epidermal growth factor-factor 8 (MFG-E8) is a regulatory protein for microglia. However, its involvement in microglial oxidative stress has not been established. In this study, we observed microglial oxidative stress in response to lipopolysaccharide (LPS) both in vitro and in vivo. LPS induced significant elevation of TNF-α, IL-6, MDA, and ROS and reduction of GSH and SOD in the mouse brains and primary microglia, which were reversed by MFG-E8 pretreatment. MFG-E8 induced the expression of Nrf-2 and HO-1 that was reduced by LPS incubation. Moreover, LPS-increased Keap-1 expression was reversed by MFG-E8. But the above tendencies were not seen when MFG-E8 was applied alone. The current study established the involvement of MFG-E8 in antioxidant effects during neuroinflammation. It may achieve the effects through the regulation of Keap-1/Nrf-2/HO-1 pathways.


Assuntos
Antígenos de Superfície/metabolismo , Glicolipídeos/metabolismo , Glicoproteínas/metabolismo , Lipopolissacarídeos/antagonistas & inibidores , Microglia/efeitos dos fármacos , Microglia/metabolismo , Proteínas do Leite/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/fisiologia , Animais , Inflamação/induzido quimicamente , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Lipopolissacarídeos/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Cultura Primária de Células
14.
Int J Mol Sci ; 20(8)2019 Apr 21.
Artigo em Inglês | MEDLINE | ID: mdl-31010077

RESUMO

Purple acid phosphatase (PAP) encoding genes are a multigene family. PAPs require iron (Fe) to exert their functions that are involved in diverse biological roles including Fe homeostasis. However, the possible roles of PAPs in response to excess Fe remain unknown. In this study, we attempted to understand the regulation of PAPs by excess Fe in tea plant (Camellia sinensis). A genome-wide investigation of PAP encoding genes identified 19 CsPAP members based on the conserved motifs. The phylogenetic analysis showed that PAPs could be clustered into four groups, of which group II contained two specific cysteine-containing motifs "GGECGV" and "YERTC". To explore the expression patterns of CsPAP genes in response to excessive Fe supply, RNA-sequencing (RNA-seq) analyses were performed to compare their transcript abundances between tea plants that are grown under normal and high iron conditions, respectively. 17 members were shown to be transcribed in both roots and leaves. When supplied with a high amount of iron, the expression levels of four genes were significantly changed. Of which, CsPAP15a, CsPAP23 and CsPAP27c were shown as downregulated, while the highly expressed CsPAP10a was upregulated. Moreover, CsPAP23 was found to be alternatively spliced, suggesting its post-transcriptional regulation. The present work implicates that some CsPAP genes could be associated with the responses of tea plants to the iron regime, which may offer a new direction towards a further understanding of iron homeostasis and provide the potential approaches for crop improvement in terms of iron biofortification.


Assuntos
Fosfatase Ácida/genética , Camellia sinensis/enzimologia , Glicoproteínas/genética , Ferro/metabolismo , Proteínas de Plantas/genética , Fosfatase Ácida/classificação , Fosfatase Ácida/metabolismo , Sequência de Aminoácidos , Camellia sinensis/genética , Genes de Plantas , Glicoproteínas/classificação , Glicoproteínas/metabolismo , Família Multigênica , Filogenia , Folhas de Planta/enzimologia , Folhas de Planta/genética , Proteínas de Plantas/classificação , Proteínas de Plantas/metabolismo , Raízes de Plantas/enzimologia , Raízes de Plantas/genética , Processamento de RNA , Alinhamento de Sequência , Transcriptoma
15.
Eur J Pharmacol ; 853: 289-298, 2019 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-30978318

RESUMO

C1q/TNF-related protein-9(CTRP9) is an adipose cytokine, a closest adiponectin paralog, which has anti-inflammatory, antioxidant, vasodilation and anti-atherosclerosis effects. In addition, it can increase insulin sensitivity, decrease blood glucose level and inhibit the apoptosis of endothelial cells. However, it remains unclear whether CTRP9 has beneficial effects on diabetic retinopathy (DR). An adenoviral vector expressing CTRP9 was intravenously injected into db/db mice, aged 12 weeks, at day 15 post injection, and the process was repeated. The transfection efficiency of CTRP9 was assessed by enzyme linked immunosorbent assay. We used RT-PCR, immunofluorescence, and Western blot to determine proinflammatory cytokines, adhesion molecules and tight-junction proteins. The breakdown of blood-retinal barrier (BRB) was evaluated using Evans blue and retinal staining. CTRP9 suppresses the expression of interleukin-1 beta, tumor necrosis factor-alpha, monocyte chemotactic protein-1 and adhesion molecules in the retina of db/db mice. CTRP9 can balance the expression of pigment epithelium-derived factor and vascular endothelial growth factor. CTRP9 can also inhibit the activation of nuclear factor Kappa B in the retina of db/db mouse. In addition, CTRP9 can prevent the breakdown of BRB and downregulation of tight-junction proteins in the retina of db/db mice. Evans blue assay revealed the breakdown of BRB and vascular leakage in the retinas of diabetic mice. CTRP9 can both qualitatively and quantitatively alleviate the vascular leakage in the early stage of diabetic retinas. CTRP9 can inhibit the inflammation of diabetic retinopathy and protect blood-retinal barrier via decreasing proinflammatory cytokines and preventing the downregulation of tight-junction proteins.


Assuntos
Adiponectina/metabolismo , Glicoproteínas/metabolismo , Retina/metabolismo , Animais , Glicemia/metabolismo , Peso Corporal/efeitos dos fármacos , Quimiocina CCL2/metabolismo , Inflamação/sangue , Inflamação/genética , Inflamação/metabolismo , Inflamação/patologia , Molécula 1 de Adesão Intercelular/metabolismo , Masculino , Camundongos , NF-kappa B/metabolismo , Fator de Crescimento Derivado de Plaquetas/metabolismo , Receptores de Adiponectina/genética , Transcrição Genética/efeitos dos fármacos , Molécula 1 de Adesão de Célula Vascular/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo
16.
BMC Plant Biol ; 19(1): 151, 2019 Apr 18.
Artigo em Inglês | MEDLINE | ID: mdl-30999851

RESUMO

BACKGROUND: Grafting is a technique widely used in horticulture. The processes involved in grafting are diverse, and the technique is commonly employed in studies focusing on the mechanisms that regulate cell differentiation or response of plants to abiotic stress. Information on the changes in the composition of the cell wall that occur during the grafting process is scarce. Therefore, this study was carried out for analyzing the composition of the cell wall using Arabidopsis hypocotyls as an example. During the study, the formation of a layer that covers the surface of the graft union was observed. So, this study also aimed to describe the histological and cellular changes that accompany autografting of Arabidopsis hypocotyls and to perform preliminary chemical and structural analyses of extracellular material that seals the graft union. RESULTS: During grafting, polyphenolic and lipid compounds were detected, along with extracellular deposition of carbohydrate/protein material. The spatiotemporal changes observed in the structure of the extracellular material included the formation of a fibrillar network, polymerization of the fibrillar network into a membranous layer, and the presence of bead-like structures on the surface of cells in established graft union. These bead-like structures appeared either "closed" or "open". Only three cell wall epitopes, namely: LM19 (un/low-methyl-esterified homogalacturonan), JIM11, and JIM20 (extensins), were detected abundantly on the cut surfaces that made the adhesion plane, as well as in the structure that covered the graft union and in the bead-like structures, during the subsequent stages of regeneration. CONCLUSIONS: To the best of our knowledge, this is the first report on the composition and structure of the extracellular material that gets deposited on the surface of graft union during Arabidopsis grafting. The results showed that unmethyl-esterified homogalacturonan and extensins are together involved in the adhesion of scion and stock, as well as taking part in sealing the graft union. The extracellular material is of importance not only due to the potential pectin-extensin interaction but also due to its origin. The findings presented here implicate a need for studies with biochemical approach for a detailed analysis of the composition and structure of the extracellular material.


Assuntos
Arabidopsis/fisiologia , Glicoproteínas/metabolismo , Pectinas/metabolismo , Proteínas de Plantas/metabolismo , Arabidopsis/anatomia & histologia , Arabidopsis/citologia , Arabidopsis/ultraestrutura , Parede Celular/metabolismo , Epitopos/metabolismo , Esterificação , Hipocótilo/citologia , Hipocótilo/fisiologia , Hipocótilo/ultraestrutura
17.
PLoS Pathog ; 15(3): e1007655, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30921434

RESUMO

Many persistent transmitted plant viruses, including rice stripe virus (RSV), cause serious damage to crop production worldwide. Although many reports have indicated that a successful insect-mediated virus transmission depends on a proper interaction between the virus and its insect vector, the mechanism(s) controlling this interaction remained poorly understood. In this study, we used RSV and its small brown planthopper (SBPH) vector as a working model to elucidate the molecular mechanisms underlying the entrance of RSV virions into SBPH midgut cells for virus circulative and propagative transmission. We have determined that this non-enveloped tenuivirus uses its non-structural glycoprotein NSvc2 as a helper component to overcome the midgut barrier(s) for RSV replication and transmission. In the absence of this glycoprotein, purified RSV virions were unable to enter SBPH midgut cells. In the RSV-infected cells, this glycoprotein was processed into two mature proteins: an amino-terminal protein (NSvc2-N) and a carboxyl-terminal protein (NSvc2-C). Both NSvc2-N and NSvc2-C interact with RSV virions. Our results showed that the NSvc2-N could bind directly to the surface of midgut lumen via its N-glycosylation sites. Upon recognition, the midgut cells underwent endocytosis followed by compartmentalization of RSV virions and NSvc2 into early and then late endosomes. The NSvc2-C triggered cell membrane fusion via its highly conserved fusion loop motifs under the acidic condition inside the late endosomes, leading to the release of RSV virions from endosomes into cytosol. In summary, our results showed for the first time that a rice tenuivirus utilized its glycoprotein NSvc2 as a helper component to ensure a proper interaction between its virions and SBPH midgut cells for its circulative and propagative transmission.


Assuntos
Glicoproteínas/fisiologia , Hemípteros/genética , Tenuivirus/metabolismo , Animais , Sistema Digestório/metabolismo , Sistema Digestório/virologia , Glicoproteínas/metabolismo , Insetos Vetores/metabolismo , Insetos Vetores/virologia , Insetos , Doenças das Plantas/virologia , Tenuivirus/patogenicidade , Vírion , Replicação Viral/fisiologia
18.
World J Gastroenterol ; 25(11): 1398-1408, 2019 Mar 21.
Artigo em Inglês | MEDLINE | ID: mdl-30918432

RESUMO

BACKGROUND: Liver cirrhosis is a major risk factor for hepatocellular carcinoma (HCC) development in chronic hepatitis B (CHB). Serum Mac-2 binding protein glycosylation isomer (M2BPGi) is a novel serological marker for fibrosis. The role of M2BPGi in prediction of HCC is unknown. AIM: To examine the role of serum M2BPGi in predicting HCC development in hepatitis B e antigen (HBeAg)-negative patients. METHODS: Treatment-naive CHB patients with documented spontaneous HBeAg seroconversion were recruited. Serum M2BPGi was measured at baseline (within 3 years from HBeAg seroconversion), at 5 years and 10 years after HBeAg seroconversion and expressed as cut-off index (COI). Multivariate cox regression was performed to identify predictors for HCC development. ROC analysis was used to determine the cut-off value of M2BPGi. RESULTS: Among 207 patients (57% male, median age at HBeAg seroconversion 40 years old) with median follow-up of 13.1 (11.8-15.5) years, the cumulative incidence of HCC at 15 years was 7%. Median M2BPGi levels were significantly higher in patients with HCC compared to those without HCC (baseline: 1.39 COI vs 0.38 COI, P < 0.001; 5-year: 1.45 COI vs 0.47 COI, P < 0.001; 10-year: 1.20 COI vs 0.55 COI, P = 0.001). Multivariate analysis revealed age at HBeAg seroconversion [odds ratio (OR) = 1.196, 95% confidence interval (CI): 1.034-1.382, P = 0.016] and baseline M2BPGi (OR = 4.666, 95%CI: 1.296-16.802, P = 0.018) were significant factors predictive of HCC. Using a cut-off value of 0.68 COI, baseline M2BPGi yielded AUROC of 0.883 with 91.7% sensitivity and 80.8% specificity. CONCLUSION: High serum M2BPGi within 3 years after HBeAg seroconversion was a strong predictor for subsequent HCC development in treatment-naive HBeAg-negative CHB patients.


Assuntos
Antígenos de Neoplasias/sangue , Biomarcadores Tumorais/sangue , Carcinoma Hepatocelular/diagnóstico , Proteínas de Transporte/sangue , Glicoproteínas/sangue , Hepatite B Crônica/sangue , Neoplasias Hepáticas/diagnóstico , Adulto , Antígenos de Neoplasias/metabolismo , Biomarcadores Tumorais/metabolismo , Carcinoma Hepatocelular/epidemiologia , Carcinoma Hepatocelular/patologia , Proteínas de Transporte/metabolismo , Feminino , Seguimentos , Glicoproteínas/metabolismo , Glicosilação , Antígenos E da Hepatite B/imunologia , Antígenos E da Hepatite B/isolamento & purificação , Vírus da Hepatite B/imunologia , Vírus da Hepatite B/isolamento & purificação , Hepatite B Crônica/imunologia , Hepatite B Crônica/patologia , Hepatite B Crônica/virologia , Humanos , Incidência , Neoplasias Hepáticas/epidemiologia , Neoplasias Hepáticas/patologia , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Isoformas de Proteínas/sangue , Isoformas de Proteínas/metabolismo , Curva ROC , Estudos Retrospectivos , Soroconversão
19.
Plant Mol Biol ; 100(1-2): 151-161, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30840202

RESUMO

KEY MESSAGE: Rice leucine-rich repeat extensin-like protein OsPEX1 mediates the intersection of lignin deposition and plant growth. Lignin, a major structural component of secondary cell wall, is essential for normal plant growth and development. However, the molecular and genetic regulation of lignin biosynthesis is not fully understood in rice. Here we report the identification and characterization of a rice semi-dominant dwarf mutant (pex1) with stiff culm. Molecular and genetic analyses revealed that the pex1 phenotype was caused by ectopic expression of a leucine-rich repeat extension-like gene, OsPEX1. Interestingly, the pex1 mutant showed significantly higher lignin content and increased expression levels of lignin-related genes compared with wild type plants. Conversely, OsPEX1-suppresssed transgenics displayed low lignin content and reduced transcriptional abundance of genes associated with lignin biosynthesis, indicating that the OsPEX1 mediates lignin biosynthesis and/or deposition in rice. When OsPEX1 was ectopically expressed in rice cultivars with tall stature that lacks the allele of semi-dwarf 1, well-known green revolution gene, the resulting transgenic plants displayed reduced height and enhanced lodging resistance. Our study uncovers a causative effect between the expression of OsPEX1 and lignin deposition. Lastly, we demonstrated that modulating OsPEX1 expression could provide a tool for improving rice lodging resistance.


Assuntos
Glicoproteínas/metabolismo , Lignina/biossíntese , Oryza/metabolismo , Desenvolvimento Vegetal , Proteínas de Plantas/metabolismo , Sequência de Bases , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Glicoproteínas/genética , Mutação/genética , Oryza/genética , Oryza/fisiologia , Fenótipo , Proteínas de Plantas/genética , Caules de Planta/metabolismo , Plantas Geneticamente Modificadas
20.
Methods Mol Biol ; 1944: 189-201, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30840244

RESUMO

Fibrosis is characterized by excessive deposition of collagen and additional extracellular matrix (ECM) components in response to chronic injuries. Liver fibrosis often results from chronic hepatitis C virus infection and alcohol abuse that can deteriorate to cirrhosis and liver failure. Current noninvasive diagnostic methods of liver fibrosis are limited in their ability to detect and differentiate between early and intermediate stages of fibrosis. New biomarkers of fibrosis that reflect ECM turnover are therefore badly needed. Procollagen C-proteinase enhancer 1 (PCPE-1), a connective tissue glycoprotein that functions as a positive regulator of C-terminal procollagen processing and subsequent collagen fibril assembly, is a promising candidate. Its tissue distribution and expression profile overlap those of collagen, and its expression in fibrosis is upregulated in parallel to the increase in collagen expression. The potential of PCPE-1 as a biomarker of liver fibrosis was recently established using a CCl4 mouse model of liver fibrosis by showing that the increase in collagen and PCPE-1 content in the fibrotic mouse liver was reflected by elevated plasma levels of PCPE-1. This was achieved using a newly developed highly sensitive, specific, accurate, and reproducible ELISA for mouse PCPE-1, which is based on commercially available antibodies and is offered as a new research tool in the field. A similar ELISA test was developed for human PCPE-1, and preliminary results with plasma from liver fibrosis patients revealed increased plasma concentrations of PCPE-1 in some patients. The protocols of both ELISA tests are outlined herein in great detail to permit their application by any laboratory with similar interests.


Assuntos
Ensaio de Imunoadsorção Enzimática/métodos , Proteínas da Matriz Extracelular/metabolismo , Fibroblastos/metabolismo , Glicoproteínas/metabolismo , Cirrose Hepática/diagnóstico , Cirrose Hepática/metabolismo , Animais , Biomarcadores/metabolismo , Células Cultivadas , Humanos , Camundongos
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