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1.
BMC Infect Dis ; 20(1): 507, 2020 Jul 13.
Artigo em Inglês | MEDLINE | ID: mdl-32660436

RESUMO

BACKGROUND: Group A streptococcus (GAS) is an important human pathogen responsible for a broad range of infections. Epidemiological surveillance has been crucial to detect changes in the geographical and temporal variation of the disease pattern. The objective of this study was to investigate the molecular epidemiological characteristics and antimicrobial resistance of GAS isolates from patients in Children's Hospital in Beijing. METHODS: From 2016 to 2017, pharyngeal swab samples were collected from the outpatients in Children's Hospital, Capital Institute of Pediatrics, who were diagnosed with scarlet fever. Antimicrobial susceptibility test was performed according to the distribution of conventional antibiotics and Clinical and Laboratory Standards Institute (CLSI) recommendations. The distribution of the macrolide-resistance genes (ermB, ermA, mefA), emm (M protein-coding gene) typing, and superantigens (SAg) gene profiling were examined by polymerase chain reaction (PCR). RESULTS: A total of 297 GAS isolates were collected. The susceptibility of the isolates to penicillin, ceftriaxone, and levofloxacin was 100%. The resistance rate to erythromycin and clindamycin was 98.3 and 96.6%, respectively. The dominant emm types were emm12 (65.32%), emm1 (27.61%), emm75 (2.69%), and emm89 (1.35%). Of the 297 isolates, 290 (97.64%) carried the ermB gene, and 5 (1.68%) carried the mefA gene, while none carried the ermA gene. The most common superantigen genes identified from GAS isolates were smeZ (96.97%), speC (92.59%), speG (91.58%), ssa (85.52%), speI (54.55%), speH (52.19%), and speA (34.34%). Isolates with the genotype emm1 possessed speA, speC, speG, speJ, speM, ssa, and smeZ, while emm12 possessed speC, speG, speH, speI, speM, ssa, and smeZ superantigens. CONCLUSIONS: The prevalent strain of GAS isolates in Beijing has a high resistance rate to macrolides; however, penicillin can still be the preferred antibiotic for treatment. Erythromycin resistance was predominantly mediated by ermB. The common emm types were emm12 and emm1. There was a correlation between emm and the superantigen gene. Thus, long-term monitoring and investigation of the emm types and superantigen genes of GAS prevalence are imperative.


Assuntos
Antibacterianos/uso terapêutico , Farmacorresistência Bacteriana/genética , Penicilinas/uso terapêutico , Escarlatina/tratamento farmacológico , Escarlatina/epidemiologia , Streptococcus pyogenes/imunologia , Adolescente , Antígenos de Bactérias/genética , Proteínas da Membrana Bacteriana Externa/genética , Proteínas de Bactérias/genética , Pequim/epidemiologia , Reanimação Cardiopulmonar , Proteínas de Transporte/genética , Criança , Pré-Escolar , Eritromicina/uso terapêutico , Feminino , Hospitais Pediátricos , Humanos , Lactente , Masculino , Testes de Sensibilidade Microbiana , Escarlatina/microbiologia , Streptococcus pyogenes/isolamento & purificação , Superantígenos/genética
2.
BMC Infect Dis ; 20(1): 480, 2020 Jul 06.
Artigo em Inglês | MEDLINE | ID: mdl-32631335

RESUMO

BACKGROUND: Influenza A virus (IAV) infection is a serious public health problem not only in South East Asia but also in European and African countries. Scientists are using network biology to dig deep into the essential host factors responsible for regulation of virus infections. Researchers can explore the virus invasion into the host cells by studying the virus-host relationship based on their protein-protein interaction network. METHODS: In this study, we present a comprehensive IAV-host protein-protein interaction network that is obtained based on the literature-curated protein interaction datasets and some important interaction databases. The network is constructed in Cytoscape and analyzed with its plugins including CytoHubba, CytoCluster, MCODE, ClusterViz and ClusterOne. In addition, Gene Ontology and KEGG enrichment analyses are performed on the highly IAV-associated human proteins. We also compare the current results with those from our previous study on Hepatitis C Virus (HCV)-host protein-protein interaction network in order to find out valuable information. RESULTS: We found out 1027 interactions among 829 proteins of which 14 are viral proteins and 815 belong to human proteins. The viral protein NS1 has the highest number of associations with human proteins followed by NP, PB2 and so on. Among human proteins, LNX2, MEOX2, TFCP2, PRKRA and DVL2 have the most interactions with viral proteins. Based on KEGG pathway enrichment analysis of the highly IAV-associated human proteins, we found out that they are enriched in the KEGG pathway of basal cell carcinoma. Similarly, the result of KEGG analysis of the common host factors involved in IAV and HCV infections shows that these factors are enriched in the infection pathways of Hepatitis B Virus (HBV), Viral Carcinoma, measles and certain other viruses. CONCLUSION: It is concluded that the list of proteins we identified might be used as potential drug targets for the drug design against the infectious diseases caused by Influenza A Virus and other viruses.


Assuntos
Interações Hospedeiro-Patógeno/genética , Vírus da Influenza A/metabolismo , Influenza Humana/metabolismo , Mapas de Interação de Proteínas/genética , Biologia de Sistemas/métodos , Proteínas de Transporte/genética , Proteínas de Ligação a DNA/genética , Hepacivirus/metabolismo , Hepatite C/metabolismo , Hepatite C/virologia , Humanos , Influenza Humana/virologia , Fatores de Transcrição/genética , Proteínas do Core Viral/genética , Proteínas não Estruturais Virais/genética , Replicação Viral
3.
J Biol Regul Homeost Agents ; 34(1): 69-82, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32392921

RESUMO

Baicalin has been used in China to treat inflammation-related diseases, such as inflammation-induced acute kidney injury (AKI). However, the specific mechanism of baicalin remains unclear. To observe the protective effects of baicalin on lipopolysaccharide (LPS)-induced inflammatory injury of renal tubular epithelial cells (HK-2 cells) and to explore its protective mechanism. LPS (1 mg/L) was used to induce an HK-2 cell inflammatory injury model in vitro. The cells were divided into seven groups: the normal control group, LPS-induced group, LPS plus 5 µmol/L baicalin treatment group, LPS plus 15 µmol/L baicalin treatment group, LPS plus 25 µmol/L baicalin treatment group, LPS plus 50 µmol/L baicalin treatment group, and LPS plus 75 µmol/L baicalin treatment group. 3-(4,5-dimethyl-2-thiazolyl)-2,5- diphenyl-2-H-tetrazolium bromide (MTT) assay was employed for detecting the relative survival rate of HK-2 cells. Enzyme-linked immunosorbent assay was used for detecting the levels of inflammatory factors, including interleukin-6 (IL-6), IL-1ß, and tumor necrosis factor-α (TNF-α). Moreover, the expression of inducible nitric oxide synthase (iNOS); cyclooxygenase-2 (COX-2); nuclear factor kB65 (NF-κB65); phosphorylated NF-κB inhibitory protein-α (p-IκB-α); NF-κB inhibitory protein (IκB); human thioredoxin interacting protein (TXNIP); and human NACHT, LRR, and PYD domain-containing protein 3 (NLRP3) were determined by Western blot analysis. The expression levels of NLRP3 and TXNIP mRNA and miR-223-3p were determined by RT-PCR. Results found that the relative survival rate of HK-2 cells treated with different baicalin concentrations was significantly increased (P<0.05) and the levels of the inflammatory factors IL-6, IL-1ß, and TNF-α were significantly decreased (P<0.05) compared with those of the LPS-induced group. The expression levels of the inflammatory proteins inducible nitric oxide synthase and cyclooxygenase-2 and the genes expressions of TXNIP and NLRP3 were significantly decreased in the cells (P<0.05), while the expression level of miR-223- 3p was significantly increased (P<0.05). These changes were induced in a dose-dependent manner. The results suggest that baicalin significantly inhibited the expression of inflammation-related proteins and alleviated LPS-induced inflammatory injury in HK-2 cells. The mechanism may be associated with the inhibition of activation of the TXNIP/NLRP3 inflammatory pathway, which might be mediated by increased expression of miR-223-3p. Thus, NLRP3 is a regulatory target of miR-223-3p.


Assuntos
Proteínas de Transporte/metabolismo , Células Epiteliais/efeitos dos fármacos , Flavonoides/farmacologia , MicroRNAs/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Transdução de Sinais , Células Cultivadas , Citocinas/metabolismo , Humanos , Inflamação , Túbulos Renais/citologia , Lipopolissacarídeos
4.
Am J Clin Nutr ; 111(6): 1150-1158, 2020 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-32393980

RESUMO

BACKGROUND: Saturated-fat intake and endotoxemia can impair cognition. However, their acute impact on cognitive performance is unknown. OBJECTIVE: This study assessed the impact of 2 high-fat meals and endotoxemia on attention. METHODS: In this double-blind, randomized crossover trial, 51 women (n = 32 breast cancer survivors, n = 19 noncancer controls; mean ± SD age: 53 ± 8 y) completed the Continuous Performance Test (CPT) and had their blood drawn to assess endotoxemia markers LPS binding protein (LBP), soluble CD14 (sCD14), and the LBP to sCD14 ratio 1 h prior to eating either a high-saturated-fat meal or a high-oleic-sunflower-oil meal. Women again completed the CPT 5 h postmeal. At 1 to 4 wk later, women completed the same protocol but consumed the other meal. RESULTS: In adjusted models, women had more difficulty distinguishing target stimuli from distractors after consuming the high-saturated-fat meal than they did after the oleic-sunflower-oil meal (B = 4.44, SE = 1.88, P = 0.02). Women with higher baseline LBP had less consistent response times (B = 0.002, SE = 0.0008, P = 0.04). Those with higher LBP and LBP:sCD14 were less able to sustain their attention throughout the entire CPT, as reflected by their progressively slower (B = 0.002, SE = 0.0006, P = 0.003; and B = 2.43, SE = 0.090, P = 0.008, respectively) and more erratic (B = 0.003, SE = 0.0008, P < 0.0001; and B = 3.29, SE = 1.17, P = 0.006, respectively) response times. Additionally, women with higher baseline LBP or sCD14 were less able to maintain or increase response speeds at higher interstimulus intervals (B = 0.002, SE = 0.0006, P = 0.02; and B = 0.006, SE = 0.003, P = 0.03, respectively), indicating greater difficulty adapting to changing task demands. Significant meal type by LBP and LBP:sCD14 interactions emerged (P < 0.05), such that high LBP and LBP:sCD14 erased between-meal cognitive differences, uniformly impairing performance. CONCLUSIONS: These results suggest that higher LBP, sCD14, and LBP:sCD14 and saturated-fat intake individually and jointly influence attention. Endotoxemia may override the relative cognitive benefit of healthier oil choices.This trial is registered at clinicaltrials.gov as NCT04247763.


Assuntos
Endotoxemia/psicologia , Proteínas da Fase Aguda , Adulto , Idoso , Atenção , Proteínas de Transporte/sangue , Cognição , Dieta Hiperlipídica/psicologia , Método Duplo-Cego , Endotoxemia/sangue , Endotoxemia/metabolismo , Feminino , Humanos , Receptores de Lipopolissacarídeos/sangue , Masculino , Refeições/psicologia , Glicoproteínas de Membrana/sangue , Pessoa de Meia-Idade , Período Pós-Prandial
5.
FEBS Lett ; 594(11): 1651-1660, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32449939

RESUMO

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has become a major global challenge. The virus infects host cells using its spike glycoprotein (S-protein) and has significantly higher infectivity and mortality rates among the aged population. Here, based on bioinformatic analysis, I provide evidence that some members of the upper respiratory tract (URT) commensal bacteria express viral S-protein -binding proteins. Based on this analysis and available data showing a decline in the population of these bacteria in the elderly, I propose that some URT commensal bacteria hamper SARS-CoV-2 infectivity and that a decline in the population of these bacteria contributes to the severity of infection. Further studies should provide a better understanding of the interaction of URT bacteria and SARS-CoV-2, which may lead to new therapeutic approaches.


Assuntos
Proteínas de Bactérias/metabolismo , Interações Microbianas , Sistema Respiratório/microbiologia , Glicoproteína da Espícula de Coronavírus/metabolismo , Envelhecimento , Betacoronavirus , Proteínas de Transporte/metabolismo , Biologia Computacional , Infecções por Coronavirus , Humanos , Modelos Moleculares , Orthomyxoviridae , Pandemias , Pneumonia Viral , Estrutura Secundária de Proteína , Proteobactérias/metabolismo
6.
PLoS One ; 15(4): e0232025, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32353019

RESUMO

The actin cytoskeleton plays a central role in establishing cell polarity and shape during embryonic morphogenesis. Daam1, a member of the Formin family of actin cytoskeleton regulators, is a Dvl2-binding protein that functions in the Wnt/Planar Cell Polarity (PCP) pathway. To examine the role of the Daam proteins in mammalian development, we generated Daam-deficient mice by gene targeting and found that Daam1, but not Daam2, is necessary for fetal survival. Embryonic development of Daam1 mutants was delayed most likely due to functional defects in the labyrinthine layer of the placenta. Examination of Daam2 and Daam1/2 double mutants revealed that Daam1 and Daam2 are functionally redundant during placental development. Of note, neural tube closure defects (NTD), which are observed in several mammalian PCP mutants, are not observed in Wnt5a or Daam1 single mutants, but arise in Daam1;Wnt5a double mutants. These findings demonstrate a unique function for Daam genes in placental development and are consistent with a role for Daam1 in the Wnt/PCP pathway in mammals.


Assuntos
Proteínas dos Microfilamentos/genética , Placentação/genética , Proteínas rho de Ligação ao GTP/genética , Citoesqueleto de Actina/genética , Citoesqueleto de Actina/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Animais , Proteínas de Transporte/metabolismo , Polaridade Celular , Citoesqueleto/metabolismo , Desenvolvimento Embrionário , Feminino , Forminas/genética , Forminas/metabolismo , Regulação da Expressão Gênica no Desenvolvimento/genética , Masculino , Camundongos/embriologia , Camundongos Endogâmicos C57BL , Camundongos Knockout , Proteínas dos Microfilamentos/metabolismo , Placenta/embriologia , Gravidez , Via de Sinalização Wnt , Proteínas rho de Ligação ao GTP/metabolismo
7.
Gene ; 753: 144802, 2020 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-32454178

RESUMO

Synchronous and timely regulation of multiple genes results in an effective defense response that decides the fate of the host when challenged with pathogens or unexpected changes in environmental conditions. One such gene, which is downregulated in response to multiple bacterial pathogens, is a putative nonspecific lipid transfer protein (nsLTP) of unknown function that we have named DISEASE RELATED NONSPECIFIC LIPID TRANSFER PROTEIN 1 (DRN1). We show that upon pathogen challenge, DRN1 is strongly downregulated, while a putative DRN1-targeting novel microRNA (miRNA) named DRN1 Regulating miRNA (DmiR) is reciprocally upregulated. Furthermore, we provide evidence that DRN1 is required for defense against bacterial and fungal pathogens as well as for normal seedling growth under salinity stress. Although nsLTP family members from different plant species are known to be a significant source of food allergens and are often associated with antimicrobial properties, our knowledge on the biological functions and regulation of this gene family is limited. Our current work not only sheds light on the mechanism of regulation but also helps in the functional characterization of DRN1, a putative nsLTP family member of hitherto unknown function.


Assuntos
Arabidopsis/genética , Proteínas de Transferência de Fosfolipídeos/genética , Estresse Salino/genética , Ácido Abscísico/metabolismo , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Resistência à Doença/genética , Secas , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Proteínas de Transferência de Fosfolipídeos/metabolismo , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Patologia Vegetal , Plantas Geneticamente Modificadas , Salinidade , Tolerância ao Sal/genética , Plântula/genética , Estresse Fisiológico/genética
8.
PLoS One ; 15(5): e0232427, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32369506

RESUMO

Hypertrophic cardiomyopathy (HCM) is the most frequent genetic cardiac disease with a prevalence of 1:500 to 1:200. While most patients show obstructive HCM and a relatively stable clinical phenotype (stage II), a small group of patients progresses to end-stage HCM (stage IV) within a relatively brief period. Previous research has shown sex-differences in stage II HCM with more diastolic dysfunction in female than in male patients. Moreover, female patients more often show progression to heart failure. Here we investigated if differences in functional and structural properties of the heart may underlie sex-differences in disease progression from stage II to stage IV HCM. Cardiac tissue from stage II and IV patients was obtained during myectomy (n = 54) and heart transplantation (n = 10), respectively. Isometric force was measured in membrane-permeabilized cardiomyocytes to define active and passive myofilament force development. Titin isoform composition was assessed using gel electrophoresis, and the amount of fibrosis and capillary density were determined with histology. In accordance with disease stage-dependent adverse cardiac remodeling end-stage patients showed a thinner interventricular septal wall and larger left ventricular and atrial diameters compared to stage II patients. Cardiomyocyte contractile properties and fibrosis were comparable between stage II and IV, while capillary density was significantly lower in stage IV compared to stage II. Women showed more adverse cellular remodeling compared to men at stage II, evident from more compliant titin, more fibrosis and lower capillary density. However, the disease stage-dependent reduction in capillary density was largest in men. In conclusion, the more severe cellular remodeling in female compared to male stage II patients suggests a more advanced disease stage at the time of myectomy in women. Changes in cardiomyocyte contractile properties do not explain the progression of stage II to stage IV, while reduced capillary density may underlie disease progression to end-stage heart failure.


Assuntos
Cardiomiopatia Hipertrófica/patologia , Remodelação Ventricular/fisiologia , Adolescente , Adulto , Idoso , Capilares/patologia , Miosinas Cardíacas/genética , Cardiomiopatia Hipertrófica/genética , Cardiomiopatia Hipertrófica/fisiopatologia , Proteínas de Transporte/genética , Estudos de Casos e Controles , Criança , Progressão da Doença , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Mutação , Miócitos Cardíacos/patologia , Miócitos Cardíacos/fisiologia , Cadeias Pesadas de Miosina/genética , Fatores de Risco , Caracteres Sexuais , Troponina T/genética , Remodelação Ventricular/genética , Adulto Jovem
9.
Yonsei Med J ; 61(6): 533-541, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32469177

RESUMO

PURPOSE: Ethanol elicits several inflammatory responses and affects the innate immune response. The aim of this study was to identify the mechanism by which ethanol affects uric acid-induced NLR family pyrin domain-containing 3 (NLRP3) inflammasome activation by regulation of aryl hydrocarbon receptor (AhR) and thioredoxin-interacting protein (TXNIP). MATERIALS AND METHODS: Human myeloid leukemia cells (U937 cells) were used to assess the role of ethanol in NLRP3 inflammasome activation induced by monosodium urate (MSU) crystals. Expression of target molecules, such as NLRP3 inflammasome components, AhR, and TXNIP, were measured using quantitative real-time PCR and Western blot analyses. The effect of ethanol-induced TXNIP on the NLRP3 inflammasome was assessed in human macrophages transfected with TXNIP siRNA. RESULTS: U937 cells treated with 100 mM ethanol for 24 h induced NLRP3 and interleukin (IL)-1ß expression. Ethanol increased reactive oxygen species generation in a time- and dose-dependent manner. AhR mRNA expression was downregulated in U937 cells treated with 100 mM ethanol, whereas CYP1A1 mRNA expression increased. Treatment with ethanol increased NLRP3 and IL-1ß mRNA and protein expression in U937 cells exposed to 1.0 mg/mL of MSU crystals for 24 h. TXNIP expression in U937 cells incubated with both 100 mM ethanol and 1.0 mg/mL of MSU crystals was significantly higher than in cells incubated with MSU crystals alone. Treatment with 100mM ethanol for 24 h downregulated NLRP3 and IL-1ß expression in MSU crystal-activated U937 cells transfected with TXNIP siRNA, compared to those with scramble siRNA. CONCLUSION: Ethanol stimulates uric acid-induced NLRP3 inflammasome activation through regression of AhR and upregulation of TXNIP.


Assuntos
Proteínas de Transporte/metabolismo , Etanol/toxicidade , Inflamassomos/metabolismo , Macrófagos/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Receptores de Hidrocarboneto Arílico/metabolismo , Ácido Úrico/toxicidade , Animais , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Lipopolissacarídeos/farmacologia , Macrófagos/efeitos dos fármacos , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Espécies Reativas de Oxigênio/metabolismo , Células U937
10.
J Environ Sci (China) ; 92: 129-140, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32430116

RESUMO

Simultaneous overabundance and scarcity of inorganic phosphate (Pi) is a critical issue driving the development of innovative water/wastewater treatment technologies that not only facilitate Pi removal to prevent eutrophication, but also recover Pi for agricultural reuse. Here, a cell-surface expressed high-affinity phosphate binding protein (PstS) system was developed, and its Pi capture and release potential was evaluated. E. coli was genetically modified to express PstS on its outer membrane using the ice nucleation protein (INP) as an anchoring motif. Verification of protein expression and localization were performed utilizing SDS-polyacrylamide gel electrophoresis (SDS-PAGE), western blot, and outer membrane separation analyses. Cell surface characterization was investigated through acid-base titration, X-ray photoelectron spectroscopy (XPS), and Fourier transform infrared spectroscopy (FTIR). These tests provided information on the macromolecular structure and composition of the bacteria surface as well as the proton-exchange properties of the surface functional groups (i.e., pKa values). Phosphate desorption and adsorption batch experiments were conducted to evaluate the effects of temperature, pH, and ionic strength on phosphate capture and release. The PstS surface-displayed cells demonstrated greater potential to release and capture phosphate compared to non-modified cells. Higher temperatures up to 40°C, basic pH conditions (pH = 10.5), and higher ionic strength up to 1.0 mol/L KCl promoted 20%-50% higher phosphate release.


Assuntos
Fosfatos , Fósforo , Adsorção , Proteínas de Transporte , Escherichia coli , Proteínas de Ligação a Fosfato
11.
Structure ; 28(5): 492-494, 2020 05 05.
Artigo em Inglês | MEDLINE | ID: mdl-32375057

RESUMO

In this issue of Structure, Rozbesky et al. (2020) report evidence for direct molecular interactions between Drosophila OTK with Sema1a and glycosaminoglycans, providing insights for OTK's mode of action in axon guidance and possibly in Wnt signaling.


Assuntos
Proteínas de Drosophila , Drosophila , Animais , Axônios , Proteínas de Transporte , Glicosaminoglicanos
12.
Zhong Nan Da Xue Xue Bao Yi Xue Ban ; 45(3): 305-313, 2020 Mar 28.
Artigo em Inglês, Chinês | MEDLINE | ID: mdl-32386023

RESUMO

OBJECTIVES: To investigate the role of pyruvate kinase isozyme type M2 (PKM2) in regulating the expression of matrix metalloproteinase-1 (MMP-1) in human umbilical vein endothelial cells (HUVECs) under high glucose exposure and the underlying mechanisms. METHODS: HUVECs were exposed to concentration gradient (5.5, 15.0, 30.0 mmol/L) of D-glucose for 72 h. Western blotting was used to detect the expression of PKM2, p-PKM2 Y105 as well as its upstream and downstream proteins. The formation of PKM2 tetramer/dimer was examined. Immunofluorescence was used to detect the translocation of PKM2. The specific siRNAs, TEPP-46, and rapamycin were used to analyze the regulatory relations among PKM2, mammalian target of rapamycin (mTOR) complex, and MMP-1. Co-immunoprecipitation was used to detect the interaction of mTOR complex and PKM2. RESULTS: High glucose exposure upregulated the level of p-PKM2 Y105, and promoted PKM2 dimer formation and nuclear translocation as well as the expression of MMP-1 and Rictor in HUVECs. Down-regulation of PKM2 expression or the treatment of TEPP-46 reversed the high glucose induced-upregulation of MMP-1. Inhibition of mTOR singnal pathway by rapacymin reversed the phosphorylation of PKM2 on tyrosine 105 and the expression of MMP-1 in high glucose-treated cells. Knockdown the expression of Rictor decreased the phosphorylation of PKM2 on tyrosine 105, while knockdown the expression of Raptor upregulated the phosphorylation of PKM2 on tyrosine 105 under high glucose condition. Co-immunoprecipitation indicated the direct interaction between Rictor and PKM2. CONCLUSIONS: The phosphorylation of PKM2 promotes high glucose-induced upregulation of MMP-1, and mTORC2 participates in the upregulation of PKM2 phosphorylation under high glucose condition.


Assuntos
Transdução de Sinais , Proteínas de Transporte , Glucose , Humanos , Metaloproteinase 1 da Matriz , Alvo Mecanístico do Complexo 2 de Rapamicina , Proteínas de Membrana , Fosforilação , Hormônios Tireóideos
13.
Gene ; 750: 144759, 2020 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-32423892

RESUMO

Zinc transporters play vital roles in regulating zinc content and localization by mobilizing zinc across cellular and intracellular membranes. Pacific oyster Crassostrea gigas is one of the most zinc-rich animals, which has been regarded as an excellent food for zinc supplement. But the information about zinc transporters and their involvements in zinc accumulation in oysters is still limited. In the present study, a total of 28 zinc transporter genes, including nine Zinc transporter genes (CgZnTs) and 19 Zrt/Irt-like protein genes (CgZIPs), were identified in C. gigas genome using a genome-wide search strategy. There were five ZIP10 homologs in C. gigas, which were much more than those in mammals, fish and other mollusks. Among oyster zinc transporters, immense variations were detected in their gene structure, protein length and physicochemical properties. Phylogenetic analysis showed that most of these transporters were distinctly clustered with their homologs from Homo sapiens, Danio rerio and other mollusks, and the most closely related transporters shared similar motif compositions. The highest zinc content was detected in the oyster mantle and gill, while the lowest level was found in the adductor muscle. The mRNA of all tested CgZnTs and CgZIPs were constitutively expressed in oyster tissues, and most of them were highly expressed in the gill or hepatopancreas. The analysis of RNA-seq data from gill and hepatopancreas showed that all the transporters exhibited divergent response patterns under zinc stress, except for CgZIP4 whose expression was almost undetectable in the two tissues. The results indicated that zinc transporters played important roles in the regulation of zinc homeostasis in C. gigas, which provided a solid foundation for further functional analysis of zinc transporters in oysters and other mollusks.


Assuntos
Proteínas de Transporte/metabolismo , Crassostrea/metabolismo , Zinco/metabolismo , Animais , Crassostrea/genética , Perfilação da Expressão Gênica/métodos , Genoma , Filogenia , Transcriptoma/genética
14.
Int J Food Microbiol ; 326: 108641, 2020 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-32371295

RESUMO

Thermotolerant Campylobacter is the leading bacterial cause of foodborne illness in humans worldwide. The objectives of this study were to estimate prevalence and to identify and characterize potential sources of thermotolerant Campylobacter contamination in broilers on farms and at the slaughterhouse; to evaluate the clonal relationship among thermotolerant Campylobacter isolates from different stages of the broiler meat supply chain, and to analyze the presence of virulence genes in different sources of thermotolerant Campylobacter. A total of 1210 samples were collected from three broiler meat supply chains in Santa Fe, Argentina. At the farms, the sampling collection included broilers one week prior to slaughter, wild-living birds, domestic dogs, wild rodents, farm workers' boots, litter, feed, drinking water, flies, and darkling beetles (Alphitobius diaperinus). At the slaughtering line, the samples taken were from the evisceration zone (broiler cecum, working surfaces, evisceration knives and workers' hands), from the chiller zone (surfaces and direct supply water) and from the packing zone (work surfaces, workers' hands and broiler carcasses). The samples taken along each supply chain were in the same batch. The isolates obtained were identified to the species level (C. jejuni and C. coli) by multiplex PCR and were analyzed using pulsed-field gel electrophoresis to compare different profiles according to the source. Finally, the presence of 11 virulence genes was examined (cadF, cdtA, cdtB, cdtC, ciaB, flaA, flhA, iam, wlaN, virB11, racR). From 254 isolates, 128 (50.4%) were Campylobacter jejuni and 126 (49.6%) Campylobacter coli. C. jejuni was the species most prevalent in farm and C. coli the species most prevalent at the slaughterhouse. We detected thermotolerant Campylobacter in samples of wild birds, darkling beetles, farm workers' boots, flies and litter. At the slaughterhouse, the prevalence varied along the process line. By analyzing PFGE results, C. jejuni showed 21 profiles with three predominant genotypes, while C. coli showed 14 profiles with four predominant genotypes. A high genotype diversity was found; however, relationships between isolates from different stages of the broiler meat chain, between broiler and potential sources of thermotolerant Campylobacter contamination and between strains in the farm and in the slaughterhouse were detected. Furthermore, there was evidence of cross-contamination at the slaughterhouse. FlaA, flhA genes were detected in all strains, and the third most prevalent virulence gene was cadF. Only those strains obtained from flies, wild-living birds and broiler carcass samples harbored 10 of 11 pathogenic genes. The prevalence of some pathogenic genes between C. jejuni and C. coli was different. This evidence should contribute the scientific basis to implement risk management measures in public health.


Assuntos
Campylobacter coli/genética , Campylobacter jejuni/genética , Doenças Transmitidas por Alimentos/microbiologia , Carne/microbiologia , Aves Domésticas/microbiologia , Matadouros/estatística & dados numéricos , Animais , Argentina , Proteínas da Membrana Bacteriana Externa/genética , Proteínas de Bactérias/genética , Infecções por Campylobacter/microbiologia , Campylobacter coli/isolamento & purificação , Campylobacter coli/patogenicidade , Campylobacter jejuni/isolamento & purificação , Campylobacter jejuni/patogenicidade , Proteínas de Transporte/genética , Ceco/microbiologia , Galinhas/microbiologia , Besouros/microbiologia , Dípteros/microbiologia , Cães , Água Potável/microbiologia , Eletroforese em Gel de Campo Pulsado , Flagelina/genética , Genótipo , Humanos , Indústria de Embalagem de Carne/estatística & dados numéricos , Proteínas de Membrana/genética , Prevalência , Roedores/microbiologia , Termotolerância , Virulência/genética
15.
Pestic Biochem Physiol ; 165: 104542, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32359552

RESUMO

The tea geometrid moth Ectropis obliqua Prout is one of the most serious moth pests in tea plants, and its sex pheromones have been identified as typical Type-II polyunsaturated hydrocarbons and epoxide derivatives. Therefore, the E. obliqua male olfactory system provides a good model to study the molecular basis of Type-II sex pheromone recognition as well as functional gene evolution towards structurally different types of moth sex pheromones. In this study, we identified the full-length sequence of a pheromone-binding protein, EoblPBP2 and revealed that it clustered together with the lepidopteran PBP2 subfamily, which binds Type I acetate pheromones. These findings suggest that the EoblPBP2 sequence and physiological function are conserved, although E. obliqua evolved Type II hydrocarbon and epoxide sex pheromones structurally different from Type I acetates. To examine this hypothesis, we studied the expression patterns and in vitro functions of EoblPBP2 in detail. Quantitative real-time PCR experiments showed that EoblPBP2 was predominantly expressed in male E. obliqua antennae. Fluorescence in situ hybridization further demonstrated that the EoblPBP2 gene was abundantly expressed in the pheromone-sensitive sensilla trichodea Str-I in male E. obliqua. The physiological function of recombinant EoblPBP2 was then examined using a competitive binding assay. The results showed that EoblPBP2 had high affinities for three E. obliqua Type II sex pheromone components and Type I acetate pheromones in comparison to some plant volatiles. These results indicate that PBP2 is involved in the detection of Type II pheromones in E. obliqua and it still retains high binding affinities to acetate pheromones and some green leaf ester volatiles.


Assuntos
Mariposas , Atrativos Sexuais , Animais , Proteínas de Transporte , Hibridização in Situ Fluorescente , Proteínas de Insetos , Masculino , Feromônios , Chá
16.
Cell Host Microbe ; 27(5): 687-688, 2020 05 13.
Artigo em Inglês | MEDLINE | ID: mdl-32407704

RESUMO

In this issue of Cell Host & Microbe, Yeung et al. (2020) and Lin et al. (2020) expose laboratory mice to a natural environment and use immune and microbiota characterization to show that fungi promote more human-like immunity. These studies will help develop animal models to more accurately resemble human immune responses.


Assuntos
Fungos , Microbiota , Animais , Proteínas Relacionadas à Autofagia , Proteínas de Transporte , Humanos , Imunidade , Camundongos , Modelos Animais , Proteína Adaptadora de Sinalização NOD2
17.
BMC Med Genet ; 21(1): 81, 2020 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-32295536

RESUMO

BACKGROUND: Prostate cancer is one of the five common cancers and has the second incidence rate and the third mortality rate in Iranian population. The purpose of this study was to evaluate the association of rs16901979, rs4242382 and rs1447295 on 8q24 locus, rs2735839 (KLK3 gene) and rs721048 (EHBP1 gene) with prostate adenocarcinoma through multi-stage approach to identify the polymorphisms associated with prostate cancer and use them as screening factors. Screening tests can identify people who may have a chance of developing the disease before detection and any symptoms. METHODS: The case-control study included 103 cases (prostate adenocarcinoma) and 100 controls (benign prostatic hyperplasia). Tetra-primer ARMS-PCR was used to genotyping of each participant. A Multi-stage approach was used for efficient genomic study. In this method, a smaller number of people can be used. Chi-squared, Fisher's exact test and logistic regression were used to investigate the SNPs associated with prostate cancer and Gleason score. RESULTS: In the first stage (59 men), the frequency of polymorphisms rs16901979, rs4242382, rs1447295, rs2735839 and rs721048 in the prostate adenocarcinoma group was evaluated compared to the control group (P-value < 0.3) in order to select meaningful polymorphisms. There was not any significant difference between genotype frequency rs16901979 (P = 0.671) and rs721048 (P = 0.474) in the case group compared to BPH. Therefore, these polymorphisms were eliminated, and in the second step (144 men), rs4242382, rs2735839 and rs1447295 were evaluated (P-value < 0.05). According to the total population (203 men), there was significant difference between genotype frequency rs4242382 (P = 0.001), rs2735839 (P = 0.000) and rs1447295 (P = 0.005) even after using Bonferroni correction (p = 0.016). The effect of these three polymorphisms on prostate cancer was not modified by age and PSA. There was a significant difference between the allelic frequency of A vs G (rs4242382, rs2735839) at all classes of Gleason score and A vs C (rs1447295) at Gleason score ≥ 8. CONCLUSIONS: The results of this study for rs2735839, rs4242382 and rs1447295 indicate the association of these polymorphisms with prostate adenocarcinoma predisposition in Iranian population. Exposure effect is homogeneous between different ages and PSA level categories. These three polymorphisms should be studied in a larger population to confirm these results.


Assuntos
Adenocarcinoma/genética , Proteínas de Transporte/genética , Calicreínas/genética , Antígeno Prostático Específico/genética , Neoplasias da Próstata/genética , Adenocarcinoma/patologia , Idoso , Frequência do Gene , Estudos de Associação Genética , Predisposição Genética para Doença , Genótipo , Humanos , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Gradação de Tumores , Polimorfismo de Nucleotídeo Único/genética , Próstata/patologia , Neoplasias da Próstata/patologia
18.
Science ; 368(6489)2020 04 24.
Artigo em Inglês | MEDLINE | ID: mdl-32327568

RESUMO

Misfolded luminal endoplasmic reticulum (ER) proteins undergo ER-associated degradation (ERAD-L): They are retrotranslocated into the cytosol, polyubiquitinated, and degraded by the proteasome. ERAD-L is mediated by the Hrd1 complex (composed of Hrd1, Hrd3, Der1, Usa1, and Yos9), but the mechanism of retrotranslocation remains mysterious. Here, we report a structure of the active Hrd1 complex, as determined by cryo-electron microscopy analysis of two subcomplexes. Hrd3 and Yos9 jointly create a luminal binding site that recognizes glycosylated substrates. Hrd1 and the rhomboid-like Der1 protein form two "half-channels" with cytosolic and luminal cavities, respectively, and lateral gates facing one another in a thinned membrane region. These structures, along with crosslinking and molecular dynamics simulation results, suggest how a polypeptide loop of an ERAD-L substrate moves through the ER membrane.


Assuntos
Proteínas de Transporte/química , Degradação Associada com o Retículo Endoplasmático , Glicoproteínas de Membrana/química , Proteínas de Membrana/química , Complexos Multiproteicos/química , Proteólise , Proteínas de Saccharomyces cerevisiae/química , Ubiquitina-Proteína Ligases/química , Proteínas de Transporte/metabolismo , Microscopia Crioeletrônica , Retículo Endoplasmático/metabolismo , Glicoproteínas de Membrana/metabolismo , Proteínas de Membrana/metabolismo , Simulação de Dinâmica Molecular , Complexos Multiproteicos/metabolismo , Domínios Proteicos , Dobramento de Proteína , Proteínas de Saccharomyces cerevisiae/metabolismo
19.
Proc Natl Acad Sci U S A ; 117(18): 9876-9883, 2020 05 05.
Artigo em Inglês | MEDLINE | ID: mdl-32303654

RESUMO

A massive intronic hexanucleotide repeat (GGGGCC) expansion in C9ORF72 is a genetic origin of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Recently, C9ORF72, together with SMCR8 and WDR41, has been shown to regulate autophagy and function as Rab GEF. However, the precise function of C9ORF72 remains unclear. Here, we report the cryogenic electron microscopy (cryo-EM) structure of the human C9ORF72-SMCR8-WDR41 complex at a resolution of 3.2 Å. The structure reveals the dimeric assembly of a heterotrimer of C9ORF72-SMCR8-WDR41. Notably, the C-terminal tail of C9ORF72 and the DENN domain of SMCR8 play critical roles in the dimerization of the two protomers of the C9ORF72-SMCR8-WDR41 complex. In the protomer, C9ORF72 and WDR41 are joined by SMCR8 without direct interaction. WDR41 binds to the DENN domain of SMCR8 by the C-terminal helix. Interestingly, the prominent structural feature of C9ORF72-SMCR8 resembles that of the FLNC-FNIP2 complex, the GTPase activating protein (GAP) of RagC/D. Structural comparison and sequence alignment revealed that Arg147 of SMCR8 is conserved and corresponds to the arginine finger of FLCN, and biochemical analysis indicated that the Arg147 of SMCR8 is critical to the stimulatory effect of the C9ORF72-SMCR8 complex on Rab8a and Rab11a. Our study not only illustrates the basis of C9ORF72-SMCR8-WDR41 complex assembly but also reveals the GAP activity of the C9ORF72-SMCR8 complex.


Assuntos
Proteínas Relacionadas à Autofagia/ultraestrutura , Proteína C9orf72/ultraestrutura , Proteínas de Transporte/ultraestrutura , Complexos Multiproteicos/ultraestrutura , Sequência de Aminoácidos/genética , Esclerose Amiotrófica Lateral/genética , Arginina/genética , Autofagia/genética , Proteínas Relacionadas à Autofagia/genética , Proteína C9orf72/genética , Proteínas de Transporte/genética , Microscopia Crioeletrônica , Filaminas/genética , Filaminas/ultraestrutura , Demência Frontotemporal/genética , Proteínas Ativadoras de GTPase/genética , Proteínas Ativadoras de GTPase/ultraestrutura , Predisposição Genética para Doença , Humanos , Complexos Multiproteicos/genética , Alinhamento de Sequência , Proteínas rab de Ligação ao GTP/genética
20.
Nature ; 580(7801): 93-99, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32238934

RESUMO

Prostate cancer is the second most common cancer in men worldwide1. Over the past decade, large-scale integrative genomics efforts have enhanced our understanding of this disease by characterizing its genetic and epigenetic landscape in thousands of patients2,3. However, most tumours profiled in these studies were obtained from patients from Western populations. Here we produced and analysed whole-genome, whole-transcriptome and DNA methylation data for 208 pairs of tumour tissue samples and matched healthy control tissue from Chinese patients with primary prostate cancer. Systematic comparison with published data from 2,554 prostate tumours revealed that the genomic alteration signatures in Chinese patients were markedly distinct from those of Western cohorts: specifically, 41% of tumours contained mutations in FOXA1 and 18% each had deletions in ZNF292 and CHD1. Alterations of the genome and epigenome were correlated and were predictive of disease phenotype and progression. Coding and noncoding mutations, as well as epimutations, converged on pathways that are important for prostate cancer, providing insights into this devastating disease. These discoveries underscore the importance of including population context in constructing comprehensive genomic maps for disease.


Assuntos
Grupo com Ancestrais do Continente Asiático/genética , Epigênese Genética , Epigenômica , Genoma Humano/genética , Genômica , Mutação , Neoplasias da Próstata/classificação , Neoplasias da Próstata/genética , Proteínas de Transporte/genética , Transformação Celular Neoplásica/genética , China , Estudos de Coortes , DNA Helicases/genética , Metilação de DNA , Proteínas de Ligação a DNA/genética , Regulação Neoplásica da Expressão Gênica , Fator 3-alfa Nuclear de Hepatócito/genética , Humanos , Masculino , Proteínas do Tecido Nervoso/genética , Neoplasias da Próstata/patologia , RNA-Seq , Transcriptoma/genética
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