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1.
Zhonghua Gan Zang Bing Za Zhi ; 28(1): 47-52, 2020 Jan 20.
Artigo em Chinês | MEDLINE | ID: mdl-32023699

RESUMO

Objective: To establish and evaluate diagnostic efficacy and applicability of serum Golgi protein (GP) 73 based non-invasive diagnostic model with other conventional serological indicators for compensated stage hepatitis B cirrhosis. Methods: 666 cases with chronic hepatitis B (CHB) who had visited to the Fifth Medical Center of People's Liberation Army General Hospital from January 2010 to December 2017 were selected as the study subjects, and were classified according to compensated stage cirrhosis into clinical and pathological diagnosis group based on whether or not the liver histological examination was performed. A diagnostic model of compensated stage hepatitis B cirrhosis in the clinical diagnosis group was established. The current clinically used diagnostic model of liver cirrhosis, aspartate aminotransferase/platelet ratio index (APRI), fibrosis index (FIB)-4 and liver stiffness measurement (LSM) were compared. Eventually, the diagnostic model was verified step by step by pathological diagnosis group. Results: The area under the receiver operating characteristic curve (AUC) of GP73 and APRI, FIB-4, and LSM for cirrhosis patients in the clinical diagnosis group were 0.842, 0.857, 0.864, and 0.832, respectively. The diagnostic efficiency of the four indicators were of similar (P value > 0.05). A diagnostic model of compensated stage hepatitis B cirrhosis (GAPA) using logistic regression analysis was established: LogitP = 1/ [1 + exp (1.614-0.054 × GP73-0.045 × Age + 0.030 × PLT-0.015 × ALP)]. The AUC of the model was as high as 0.940 and the optimal cut-off value were 0.41. The corresponding diagnostic sensitivity and specificity were 0.92 and 0.82, respectively. The diagnostic efficiency was better than that of APRI, FIB-4, LSM and GP73 alone (P < 0.05). The AUC of GAPA was 0.877 in the pathological diagnosis group, which was similar to the diagnostic efficacy of LSM (0.891) and FIB-4 (0.847) (P > 0.1), but still superior to that of APRI (0.811) and GP73 alone (0.780) (P < 0.001). Conclusion: GAPA, a diagnostic model for compensated stage hepatitis B cirrhosis established in this study, has a good diagnostic efficacy in both the clinical and pathological diagnosis group, and has certain auxiliary diagnostic value in the areas where resources are relatively scarce or where LSM has not been developed.


Assuntos
Biomarcadores/metabolismo , Cirrose Hepática/diagnóstico , Fígado/metabolismo , Proteínas de Membrana/metabolismo , Aspartato Aminotransferases/metabolismo , Biópsia , Fibrose , Hepatite B , Humanos , Fígado/patologia , Proteínas de Membrana/sangue , Curva ROC , Índice de Gravidade de Doença
2.
J Cell Biol ; 219(2)2020 Feb 03.
Artigo em Inglês | MEDLINE | ID: mdl-31985747

RESUMO

IRE1ß is an ER stress sensor uniquely expressed in epithelial cells lining mucosal surfaces. Here, we show that intestinal epithelial cells expressing IRE1ß have an attenuated unfolded protein response to ER stress. When modeled in HEK293 cells and with purified protein, IRE1ß diminishes expression and inhibits signaling by the closely related stress sensor IRE1α. IRE1ß can assemble with and inhibit IRE1α to suppress stress-induced XBP1 splicing, a key mediator of the unfolded protein response. In comparison to IRE1α, IRE1ß has relatively weak XBP1 splicing activity, largely explained by a nonconserved amino acid in the kinase domain active site that impairs its phosphorylation and restricts oligomerization. This enables IRE1ß to act as a dominant-negative suppressor of IRE1α and affect how barrier epithelial cells manage the response to stress at the host-environment interface.


Assuntos
Retículo Endoplasmático/metabolismo , Endorribonucleases/metabolismo , Endorribonucleases/fisiologia , Proteínas de Membrana/fisiologia , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Serina-Treonina Quinases/fisiologia , Células CACO-2 , Endorribonucleases/genética , Células HEK293 , Humanos , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Modelos Moleculares , Proteínas Serina-Treonina Quinases/genética , Proteostase , Análise de Sequência de Proteína , Transdução de Sinais , Estresse Fisiológico , Resposta a Proteínas não Dobradas
3.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 51(1): 97-101, 2020 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-31950797

RESUMO

Objective: To analyse potential genetic cause of a family affected with hereditary elliptocytosis (HE). Methods: Peripheral blood samples from this HE family were collected. Targeted capture and high-throughput sequencing of 4 813 genetic disease-associated genes was performed in four members of the family. Possible causative genetic variation was obtained and further confirmed by Sanger sequencing. Fifty healthy control subjects were recruited for detection of the candidate variation. Results: High-throughput sequencing detected a nonsense mutation c.1215G>A(p.Trp405Ter)in exon 13 of the EPB41 gene in the proband and his mother presenting with moderate anemia. The pathogenicity of this loss-of-function mutation is very strong, because the G→A transition leads to introduce the premature stop codon instead of tryptophan codon at position 405, which producing a truncating protein with loss of important functional domains. This causative mutation is extremely rare in the population, and it has not yet been reported. The grandmother of the proband was heterozygous for the same mutation. Genotype-phenotype cosegregation was observed in this family. This mutation was not found in the 50 unrelated healthy controls. Conclusion: The c.1215G>A mutation of the EPB41 gene probably accounts for the disease in this HE family. This study reports a pathogenic EPB41 mutation in a Chinese HE family for the first time.


Assuntos
Proteínas do Citoesqueleto , Eliptocitose Hereditária , Proteínas de Membrana , Mutação , Proteínas do Citoesqueleto/genética , Eliptocitose Hereditária/genética , Heterozigoto , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Proteínas de Membrana/genética , Linhagem
4.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 37(1): 5-7, 2020 Jan 10.
Artigo em Chinês | MEDLINE | ID: mdl-31922585

RESUMO

OBJECTIVE: To analyze variants of PRRT2 gene in two children with paroxysmal kinesigenic dyskinesia. METHODS: Genomic DNA of the two children and their parents was extracted from peripheral venous blood samples. All exons and their flanking regions of the PRRT2 gene were subjected to PCR and Sanger sequencing. RESULTS: The two children were found to respectively harbor a c.282dupA and a c.715_716dupCC variant in exon 2 of the PRRT2 gene, which were both inherited from their mothers. Pooling together their frequencies in general population, genetic models, related literature and impact on protein function, the two novel variants were both predicted to be pathogenic. CONCLUSION: The c.282dupA and c.715_716dupCC variants probably underlie the disease in the two children.


Assuntos
Distonia , Proteínas de Membrana , Proteínas do Tecido Nervoso , Criança , Distonia/genética , Feminino , Humanos , Proteínas de Membrana/genética , Mutação , Proteínas do Tecido Nervoso/genética
5.
J Chem Theory Comput ; 16(1): 711-724, 2020 Jan 14.
Artigo em Inglês | MEDLINE | ID: mdl-31765139

RESUMO

Surfactant micelles are often utilized as membrane mimetics for structure determination and functional analysis of membrane proteins. The curved-surface effects of the micelle can perturb membrane protein structure. However, it is difficult to assess such effects and membrane mimetic artifacts by experimental and theoretical methods. Here, we propose an implicit micelle model (IMIC) to be used in molecular dynamics (MD) simulations of membrane proteins. IMIC is an extension of the IMM1 implicit membrane model and additionally introduces a superellipsoid approximation to represent the curved-surface effects. Most of the IMIC parameters are obtained from all-atom explicit solvent MD simulations of 12 membrane proteins in various micelles. The HIV envelope protein gp41, M13 major coat protein gp8, and amyloid precursor protein (APP) dimer are simulated via MD simulations with IMIC. These simulations clearly show how the micelle influences membrane protein structures compared to the bilayer environments. The MD simulations with IMIC provide reliable membrane protein structures in various micelle environments quickly with smaller computational cost than that for an explicit solvent/micelle model.


Assuntos
Proteínas de Membrana/química , Simulação de Dinâmica Molecular , Bicamadas Lipídicas/química , Micelas , Modelos Moleculares , Conformação Proteica , Termodinâmica , Água/química
6.
Fitoterapia ; 140: 104434, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31760067

RESUMO

Fritillaria cirrhosa bulbus is a Chinese folk herb famous for its antitussive, expectorant, anti-asthma and anti-inflammatory properties, and is widely used to treat respiratory diseases. However, the impacts of F. cirrhosa bulbus on oxidative stress are still unkown. In the present study, we investigated the potential effect and mechanism of six isosteroid alkaloids with different chemical structures from F. cirrhosa bulbus on protection against cigarette smoke-induced oxidative stress in RAW264.7 macrophages. The results showed that six isosteroid alkaloids reduced reactive oxygen species (ROS) production, elevated glutathione (GSH) level and promoted heme oxygenase (HO-1) expression, which is in association with induction of NF-E2-related factor 2 (Nrf2) nuclear translocation and up-regulation of Nrf2 expression. Among these alkaloids, verticinone, verticine, imperialine-3-ß-D-glucoside, delavine and peimisine exhibited more potent effect against CSE-induced oxidative stress than that of imperialine. These findings for the first time demonstrated that F. cirrhosa bulbus may play a protective role in cellular oxidative stress by activating Nrf2-mediated antioxidant pathway. Furthermore, the differences in antioxidant effects of these alkaloids were compared, as well as the corresponding structure-activity relationships were preliminarily elucidated. This suggested that F. cirrhosa bulbus might be a promising therapeutic treatment for the prevent of oxidative stress-related diseases.


Assuntos
Alcaloides/farmacologia , Fritillaria/química , Estresse Oxidativo/efeitos dos fármacos , Fumaça/efeitos adversos , Alcaloides/isolamento & purificação , Animais , Glutationa/metabolismo , Heme Oxigenase-1/metabolismo , Proteínas de Membrana/metabolismo , Camundongos , Estrutura Molecular , Fator 2 Relacionado a NF-E2/metabolismo , Compostos Fitoquímicos/isolamento & purificação , Compostos Fitoquímicos/farmacologia , Raízes de Plantas/química , Plantas Medicinais/química , Células RAW 264.7 , Espécies Reativas de Oxigênio/metabolismo , Relação Estrutura-Atividade , Produtos do Tabaco
7.
Gene ; 726: 144136, 2020 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-31629817

RESUMO

Chronic systolic heart failure (CSHF) was a complex syndrome. Recently, vagus nerve stimulation (VNS), a novel treatment method, has emerged for the treatment of CSHF. therefore the aim of this study was to explore the possible mechanism of VNS treatment alleviating CSHF in rats. Firstly, we found after VNS treatment for 72 h, the level of B-type natriuretic peptide in VNS group was lower than that in CSHF group. In addition, VNS treatment induced the elevated left ventricular ejection fraction level, reduced left ventricular end diastolic volume and left ventricular end systolic volume level in VNS group, suggesting a mitigation of CSHF by VNS. Then we found the level of miR-183-3p in CSHF group was much lower than that in VNS group by High-throughput sequencing. The further results indicated that Bcl-2 interacting protein 3 like (BNIP3L) was identified as the target gene of miR-183-3p, and the expression of BNIP3L was notably reduced in rats of VNS group compared with CSHF group. Moreover, the down-regulated expression of miR-183-3p increased BNIP3L-mediated autophagy in rats of CSHF group compared with VNS group. Further mechanism findings demonstrated that up-regulation of miR-183-3p reduced the expression of BNIP3L, while down-regulation of miR-183-3p facilitated the expression of BNIP3L in H9c2 cells. miR-183-3p could also regulate autophagy by targeting BNIP3L in vitro, which was manifested by overexpression of miR-183-3p to inhibit BNIP3L-mediated autophagy. Our data demonstrated that VNS treatment benefited CSHF via the up-regulation of miRNA-183-3p, which reduced the BNIP3L-mediated autophagy, providing a new therapeutic direction for CSHF.


Assuntos
Autofagia/genética , Insuficiência Cardíaca Sistólica/genética , Proteínas de Membrana/genética , MicroRNAs/genética , Proteínas Proto-Oncogênicas/genética , Regulação para Cima/genética , Animais , Regulação para Baixo/genética , Masculino , Ratos , Ratos Wistar , Volume Sistólico/genética , Ativação Transcricional/genética , Estimulação do Nervo Vago/métodos , Função Ventricular Esquerda/genética
8.
Oral Dis ; 26(1): 182-192, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31696592

RESUMO

This study aims to investigate the levels of SLIT3 in gingival crevicular fluid (GCF) of healthy and periodontal disease subjects, and their correlations to periodontal disease. A total of 45 periodontal patients and 45 periodontally healthy volunteers were enrolled. The clinical parameters, radiographic bone loss and the levels of SLIT3, receptor activator of NF-κB ligand (RANKL) and osteoprotegerin (OPG) in GCF were measured. The prevalences of Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia in subgingival plaque were also analyzed. The expression of SLIT3 and RANKL was detected in the periodontium of experimental periodontitis in rats and lipopolysaccharide (LPS)-induced mouse macrophage. The total amounts and concentrations of SLIT3 and RANKL were significantly higher in periodontitis than those in healthy, while the level of OPG was significantly lower (p < .05). Significant positive correlations were observed between the level of GCF SLIT3 and clinical attachment level and radiographic bone loss (p < .05). There existed a significant positive correlation between SLIT3 and RANKL (p < .05). Increased expression of SLIT3 and RANKL was observed in the periodontium of periodontal rats. SLIT3 expression was induced by LPS stimulation in macrophages. These results suggest that SLIT3 may act as a diagnostic indicator of periodontal disease and should be further investigated.


Assuntos
Líquido do Sulco Gengival/química , Proteínas de Membrana/metabolismo , Periodontite/metabolismo , Adulto , Animais , Placa Dentária/microbiologia , Feminino , Humanos , Masculino , Camundongos , Osteoprotegerina/metabolismo , Periodonto/metabolismo , Porphyromonas gingivalis/isolamento & purificação , Ligante RANK/metabolismo , Células RAW 264.7 , Ratos , Ratos Sprague-Dawley , Tannerella forsythia/isolamento & purificação , Treponema denticola/isolamento & purificação
9.
Toxicol Lett ; 319: 66-73, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31726083

RESUMO

Thallium ion (Tl+) and its neurotoxic products are widely known to cause severe neurological complications. However, the exact mechanism of action remains unknown, with limited therapeutic options available. This study aims to examine the toxic effects of Thallium (I) Nitrate (TlNO3) on primary hippocampal neurons of E17-E18 Wistar rat embryos, and the potential neuroprotective role of Nrf2- Keap1 signaling pathway against thallium-induced oxidative stress and mitochondrial dysfunction. TlNO3 induces a significant increase in reactive oxygen species levels and mitochondrial dysfunction in primary hippocampal neurons. Furthermore, the Nrf2-Keap1 signaling pathway played a protective role against TlNO3-induced hippocampal neuronal cytotoxicity. Moreover, mitochondrial fusion protein Mitofusin 2 (Mfn2) levels significantly decreased in hippocampal neurons when exposed to TlNO3, indicating that Mfn2 protein levels are linked to TlNO3-induced neurotoxicity. t-BHQ, a Nrf2 and phase II detoxification enzyme inducer, counteracted the oxidative damage in hippocampal neurons by activating the Nrf2-Keap1 signaling pathway after TlNO3 exposure; the activated Nrf2-Keap1 pathway could then maintain Mfn2 function by regulating Mfn2 protein expression. Thus, Nrf2-Keap1 pathway activation plays a protective role in Tl+-induced brain damage, and specific agonists have been identified to have great potential for treating thallium poisoning.


Assuntos
Hipocampo/efeitos dos fármacos , Proteína 1 Associada a ECH Semelhante a Kelch/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Fator 2 Relacionado a NF-E2/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Tálio/toxicidade , Animais , Encefalopatias/induzido quimicamente , Encefalopatias/patologia , Encefalopatias/prevenção & controle , Feminino , Hipocampo/citologia , Proteínas de Membrana/efeitos dos fármacos , Proteínas de Membrana/metabolismo , Proteínas Mitocondriais/efeitos dos fármacos , Proteínas Mitocondriais/metabolismo , Cultura Primária de Células , Ratos , Ratos Wistar , Espécies Reativas de Oxigênio/metabolismo
10.
Cell Mol Life Sci ; 77(2): 331-350, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31209506

RESUMO

Disintegrin and metalloproteinases (ADAMs) 10 and 17 can release the extracellular part of a variety of membrane-bound proteins via ectodomain shedding important for many biological functions. So far, substrate identification focused exclusively on membrane-anchored ADAM10 and ADAM17. However, besides known shedding of ADAM10, we identified ADAM8 as a protease capable of releasing the ADAM17 ectodomain. Therefore, we investigated whether the soluble ectodomains of ADAM10/17 (sADAM10/17) exhibit an altered substrate spectrum compared to their membrane-bound counterparts. A mass spectrometry-based N-terminomics approach identified 134 protein cleavage events in total and 45 common substrates for sADAM10/17 within the secretome of murine cardiomyocytes. Analysis of these cleavage sites confirmed previously identified amino acid preferences. Further in vitro studies verified fibronectin, cystatin C, sN-cadherin, PCPE-1 as well as sAPP as direct substrates of sADAM10 and/or sADAM17. Overall, we present the first degradome study for sADAM10/17, thereby introducing a new mode of proteolytic activity within the protease web.


Assuntos
Proteína ADAM10/metabolismo , Proteína ADAM17/metabolismo , Secretases da Proteína Precursora do Amiloide/metabolismo , Proteínas de Membrana/metabolismo , Metaloproteases/metabolismo , Aminoácidos/metabolismo , Animais , Linhagem Celular , Células HEK293 , Humanos , Camundongos , Miócitos Cardíacos/metabolismo
11.
Arch Virol ; 165(2): 355-366, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31845156

RESUMO

Picornaviruses infect a wide range of mammals including livestock such as cattle and swine. As with other picornavirus genera such as Aphthovirus, there is emerging evidence of a significant economic impact of livestock infections caused by members of the genera Enterovirus and Kobuvirus. While the human-infecting enteroviruses and kobuviruses have been intensively studied during the past decades in great detail, research on livestock-infecting viruses has been mostly limited to the genomic characterization of the viral strains identified worldwide. Here, we extend our previous studies of the structure and function of the complexes composed of the non-structural 3A proteins of human-infecting enteroviruses and kobuviruses and the host ACBD3 protein and present a structural and functional characterization of the complexes of the following livestock-infecting picornaviruses: bovine enteroviruses EV-E and EV-F, porcine enterovirus EV-G, and porcine kobuvirus AiV-C. We present a series of crystal structures of these complexes and demonstrate the role of these complexes in facilitation of viral replication.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Infecções por Enterovirus/metabolismo , Enterovirus Bovino/patogenicidade , Enterovirus Suínos/patogenicidade , Kobuvirus/patogenicidade , Proteínas de Membrana/metabolismo , Infecções por Picornaviridae/metabolismo , Animais , Bovinos , Linhagem Celular , Infecções por Enterovirus/veterinária , Infecções por Enterovirus/virologia , Enterovirus Suínos/genética , Células HEK293 , Humanos , Kobuvirus/genética , Infecções por Picornaviridae/veterinária , Infecções por Picornaviridae/virologia , Suínos , Proteínas Virais/genética , Proteínas Virais/metabolismo , Replicação Viral/genética
12.
Life Sci ; 242: 117186, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31862454

RESUMO

AIMS: This study was aimed to investigate the role of GSDME-mediated pyroptosis in cardiac injury induced by Doxorubicin (DOX), and to evaluate the role of BH3-only protein Bcl-2/adenovirus E1B 19-kDa-interacting protein 3 (Bnip3) in regulation of DOX-induced pyroptosis. MAIN METHODS: HL-1 cardiomyocytes and C57BL/6J mice were treated by DOX to establish DOX-induced cardiotoxicity in vitro and in vivo models, respectively. Cell transfection was applied to regulate the expression of caspase-3, GSDME and Bnip3. Western blot was used for measuring expression of protein level. LDH-cytotoxicity assay was used to detect the LDH release. The Flow cytometry analysis was used to detect the cell death. Echocardiography was used to determine the cardiac function. HE staining was used for observing pathological feature of heart tissues. KEY FINDINGS: Our results showed that GSDME-mediated pyroptosis was involved in DOX-induced cardiotoxicity in vivo. We showed that HL-1 cardiomyocytes exposed to DOX exhibited morphological features of pyroptosis in vitro. We also showed that DOX induced activation of caspase-3 and eventually triggered GSDME-dependent pyroptosis, which was reduced by the silence or inhibitor of caspase-3. We further showed that knockdown of GSDME inhibited DOX-induced cardiomyocyte pyroptosis in vitro. Finally, DOX increased the expression of Bnip3, whereas silencing of Bnip3 blunted cardiomyocyte pyroptosis induced by DOX, which was regulated through caspase-3 activation and GSDME cleavage. SIGNIFICANCE: Our findings revealed a novel pathway that cardiomyocyte pyroptosis is regulated through Bnip3-caspase-3-GSDME pathway following DOX treatment, suggesting that Bnip3-dependent pyroptosis may offer a novel therapeutic strategy to reduce cardiotoxicity induced by DOX.


Assuntos
Caspase 3/metabolismo , Doxorrubicina/farmacologia , Proteínas de Membrana/metabolismo , Proteínas Mitocondriais/metabolismo , Miócitos Cardíacos/efeitos dos fármacos , Piroptose/efeitos dos fármacos , Receptores Estrogênicos/metabolismo , Animais , Western Blotting , Ecocardiografia , Citometria de Fluxo , Masculino , Camundongos , Camundongos Endogâmicos C57BL
13.
Biosci Biotechnol Biochem ; 84(1): 154-158, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31794328

RESUMO

Malectin is a maltose-binding endoplasmic reticulum protein conserved in animals. In Arabidopsis thaliana, we identified four genes that encode malectin-like domain (MLD)- and leucine-rich repeat (LRR)-containing proteins (AtMLLRs): two were receptor-like proteins (AtMLLR1 and 2) and the other two were extracellular proteins (AtMLLR3 and 4). The promoter:G3GFP+promoter:GUS assay indicated the organ- and cell-specific expression of the AtMLLR2 and AtMLLR3 genes.Abbreviations: Cmr: chloramphenicol-resistance marker; G3GFP: G3 green fluorescent protein; GUS: ß-glucuronidase; KD: kinase domain; LRR: leucine-rich repeat; MLD: malectin-like domain; RLK: receptor-like kinase; SP: signal peptide; TMD: transmembrane domain; Tnos: nopaline synthase terminator.


Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Expressão Gênica , Lectinas/genética , Proteínas de Membrana/genética , Proteínas/genética , Retículo Endoplasmático/metabolismo , Glucuronidase/química , Proteínas de Fluorescência Verde/química , Leucina/genética , Microscopia de Fluorescência , Filogenia , Plantas Geneticamente Modificadas , Domínios Proteicos/genética , Coloração e Rotulagem
14.
Food Chem ; 307: 125565, 2020 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-31630022

RESUMO

The effectiveness of some non-specific proteases in reducing raw peanut allergenicity was investigated. Peanut kernels were treated by Alcalase, papain, Neutrase and bromelain, respectively. The residues of major peanut allergens Ara h 1, Ara h 2 and Ara h 6 were determined by sandwich ELISA and SDS-PAGE, and the allergenicities of treated peanuts were compared to that of untreated peanuts by western blot. All tested proteases were effective in reducing Ara h 1, but their effectiveness in hydrolyzing Ara h 2 and Ara h 6 varied greatly. The maximal reductions of extractable Ara h 1, Ara h 2 and Ara h 6 were 100%, 100% and 99.8%, respectively, achieved by Alcalase hydrolysis. Alcalase was more effective in overall allergenicity reduction; bromelain and Neutrase were the least effective in reducing Ara h 2 and Ara h 6, respectively. The hydrolysis of original allergens also produced some smaller peptides with strong IgE-binding.


Assuntos
Alérgenos/metabolismo , Arachis/química , Imunoglobulina E/imunologia , Hipersensibilidade a Amendoim/prevenção & controle , Peptídeo Hidrolases/metabolismo , Albuminas 2S de Plantas/análise , Albuminas 2S de Plantas/imunologia , Albuminas 2S de Plantas/metabolismo , Alérgenos/análise , Alérgenos/imunologia , Antígenos de Plantas/análise , Antígenos de Plantas/imunologia , Antígenos de Plantas/metabolismo , Western Blotting , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Humanos , Proteínas de Membrana/análise , Proteínas de Membrana/imunologia , Proteínas de Membrana/metabolismo , Proteínas de Plantas/análise , Proteínas de Plantas/imunologia , Proteínas de Plantas/metabolismo
15.
Anticancer Res ; 39(12): 6471-6478, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31810911

RESUMO

BACKGROUND/AIM: Basaloid squamous cell carcinoma of the oesophagus (BSCCE) has poorer prognosis than conventional oesophageal squamous cell carcinoma (ESCC). This study is the first report on highly expressed miRNAs in BSCCE and their target genes. MATERIALS AND METHODS: BSCCE and ESCC patients who underwent esophagectomy were selected for this study. Total RNA was extracted from formalin-fixed paraffin-embedded blocks to examine expression of miRNAs and target genes. miRNA mimic or inhibitor transfected cells were used in validation experiments. miRNA and mRNA quantification were performed by quantitative reverse transcription polymerase chain reaction (qRT-PCR). RESULTS: miRNA microarray analysis revealed four candidate miRNAs. Further investigations including cell line experiments demonstrated that miR-3687 was a candidate miRNA and progesterone receptor membrane component2 (PGRMC2) was its target gene. PGRMC2 was found to be related to cell proliferation and local progression. CONCLUSION: miR-3687 may be a candidate miRNA conferring BSCCE aggressiveness, and PGRMC2 is one of its target genes.


Assuntos
Carcinoma de Células Escamosas/cirurgia , Neoplasias Esofágicas/cirurgia , Proteínas de Membrana/genética , MicroRNAs/genética , Receptores de Progesterona/genética , Regulação para Cima , Idoso , Carcinoma de Células Escamosas/genética , Linhagem Celular Tumoral , Proliferação de Células , Progressão da Doença , Neoplasias Esofágicas/genética , Esofagectomia , Feminino , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Análise de Sequência com Séries de Oligonucleotídeos
16.
World J Microbiol Biotechnol ; 36(1): 4, 2019 Dec 12.
Artigo em Inglês | MEDLINE | ID: mdl-31832786

RESUMO

Colletotrichum gloeosporioides, one of the main agents of mango anthracnose, causes latent infections in unripe mango, and leads to huge economic losses during storage and transport. Dimethyl trisulfide (DMTS), one of the main volatile compounds produced by some microorganisms or plants, has shown antifungal activity against some phytopathogens in previous studies, but its effects on C. gloeosporioides and mechanisms of action have not been well characterized. In fumigation trials of conidia and mycelia of C. gloeosporioides for 2, 4, 6, 8, or 10 h, at a concentration of 100 µL/L of air space in vitro, DMTS caused serious damage to the integrity of plasma membranes, which significantly reduced the survival rate of spores, and resulted in abnormal hyphal morphology. Moreover, DMTS caused deterioration of subcellular structures of conidia and mycelia, such as cell walls, plasma membranes, Golgi bodies, and mitochondria, and contributed to leakage of protoplasm, thus promoting vacuole formation. In addition, to better understand the molecular mechanisms of the antifungal activity, the global gene expression profiles of isolate C. gloeosporioides TD3 treated in vitro with DMTS at a concentration of 100 µL/L of air for 0 h (Control), 1 h, or 3 h were investigated by RNA sequencing (RNA-seq), and over 62 Gb clean reads were generated from nine samples. Similar expressional patterns for nine differentially expressed genes (DEGs) in both RNA-seq and qRT-PCR assays showed the reliability of the RNA-seq data. In comparison to the non-treated control groups, we found DMTS suppressed expression of ß-1, 3-D-glucan, chitin, sterol biosynthesis-related genes, and membrane protein-related genes. These genes related to the formation of fungal cell walls and plasma membranes might be associated with the toxicity of DMTS against C. gloeosporioides. This is the first study demonstrating antifungal activity of DMTS against C. gloeosporioides on mango by direct damage of conidia and hyphae, thus providing a novel tool for postharvest control of mango anthracnose.


Assuntos
Antifúngicos/farmacologia , Colletotrichum/efeitos dos fármacos , Mangifera/microbiologia , Sulfetos/farmacologia , Quitina/metabolismo , Colletotrichum/isolamento & purificação , Contaminação de Alimentos , Microbiologia de Alimentos , Regulação Fúngica da Expressão Gênica , Hifas/efeitos dos fármacos , Proteínas de Membrana/metabolismo , Viabilidade Microbiana , Microscopia Eletrônica de Transmissão , Micélio/efeitos dos fármacos , Doenças das Plantas/microbiologia , RNA Fúngico/genética , RNA Fúngico/isolamento & purificação , Reprodutibilidade dos Testes , Alinhamento de Sequência , Análise de Sequência de RNA , Esporos Fúngicos/efeitos dos fármacos , Esporos Fúngicos/isolamento & purificação , Esteróis/metabolismo , beta-Glucanas/metabolismo
17.
BMC Complement Altern Med ; 19(1): 360, 2019 Dec 11.
Artigo em Inglês | MEDLINE | ID: mdl-31829159

RESUMO

BACKGROUND: Lingguizhugan decoction (LGZG), an ancient Chinese herbal formula, has been used to treat cardiovascular diseases in eastern Asia. We investigated whether LGZG has protective activity and the mechanism underlying its effect in an animal model of heart failure (HF). METHODS: A rat model of HF was established by administering eight intraperitoneal injections of doxorubicin (DOX) (cumulative dose of 16 mg/kg) over a 4-week period. Subsequently, LGZG at 5, 10, and 15 mL/kg/d was administered to the rats intragastrically once daily for 4 weeks. The body weight, heart weight index (HWI), heart weight/tibia length ratio (HW/TL), and serum BNP level were investigated to assess the effect of LGZG on HF. Echocardiography was performed to investigate cardiac function, and H&E staining to visualize myocardial morphology. Myocardial ultrastructure and T-tubule-sarcoplasmic reticulum (TT-SR) junctions were observed by transmission electron microscopy. The JP-2 protein level was determined by Western blotting. The mRNA level of CACNA1S and RyR2 and the microRNA-24 (miR-24) level were assayed by quantitative RT-PCR. RESULTS: Four weeks after DOX treatment, rats developed cardiac damage and exhibited a significantly increased BNP level compared with the control rats (169.6 ± 29.6 pg/mL versus 80.1 ± 9.8 pg/mL, P < 0.001). Conversely, LGZG, especially at the highest dose, markedly reduced the BNP level (93.8 ± 17.9 pg/mL, P < 0.001). Rats treated with DOX developed cardiac dysfunction, characterized by a strong decrease in left ventricular ejection fraction compared with the control (58.5 ± 8.7% versus 88.7 ± 4.0%; P < 0.001). Digoxin and LGZG improved cardiac dysfunction (79.6 ± 6.1%, 69.2 ± 2.5%, respectively) and preserved the left ventricular ejection fraction (77.9 ± 5.1, and 80.5 ± 4.9, respectively, P < 0.01). LGZG also improved the LVEDD, LVESD, and FS and eliminated ventricular hypertrophy, as indicated by decreased HWI and HW/TL ratio. LGZG attenuated morphological abnormalities and mitochondrial damage in the myocardium. In addition, a high dose of LGZG significantly downregulated the expression of miR-24 compared with that in DOX-treated rats (fold change 1.4 versus 3.4, P < 0.001), but upregulated the expression of JP-2 and antagonized DOX-induced T-tubule TT-SR microstructural remodeling. These activities improved periodic Ca2+ transients and cell contraction, which may underly the beneficial effect of LGZG on HF. CONCLUSIONS: LGZG exerted beneficial effects on DOX-induced HF in rats, which were mediated in part by improved TT-SR microstructural remodeling.


Assuntos
Cardiotônicos/farmacologia , Doxorrubicina/efeitos adversos , Insuficiência Cardíaca/induzido quimicamente , Extratos Vegetais/farmacologia , Retículo Sarcoplasmático/efeitos dos fármacos , Animais , Coração/efeitos dos fármacos , Masculino , Proteínas de Membrana/metabolismo , MicroRNAs , Proteínas Musculares/metabolismo , Miocárdio/química , Miocárdio/metabolismo , Ratos , Ratos Sprague-Dawley , Retículo Sarcoplasmático/ultraestrutura
18.
Biomed Khim ; 65(6): 513-519, 2019 Oct.
Artigo em Russo | MEDLINE | ID: mdl-31876522

RESUMO

The content of nuclear and membrane proteins of the placenta, as well as posttranslational modification of these proteins in physiological pregnancy and placental insufficiency (PI) were studied. Differential centrifugation, electrophoresis in polyacrylamide gel, spectrophotometric methods were used. It was found that with PN there is a decrease in the degree of production of the studied proteins of varying degrees relative to control parameters. For chromatin proteins, a more pronounced decrease in the content of non-histone proteins was found in comparison with histones. Among histone fractions, the maximum decrease was detected in the H2A fraction. The degree of change in the amount of membrane proteins depends on the detergent used. Changes in posttranslational protein modifications disorders are characterized by a decrease in the content of amine and amide (especially difficult to hydrolyze) groups and an increase in carbonyl derivatives of proteins. The revealed changes in the composition and structure of the nuclear and membrane proteins of the placenta, performing numerous regulatory functions, can be triggering links in the chain of molecular damage in the placenta at PI.


Assuntos
Núcleo Celular/química , Proteínas de Membrana/química , Placenta/química , Complicações na Gravidez , Processamento de Proteína Pós-Traducional , Eletroforese em Gel de Poliacrilamida , Feminino , Histonas/química , Humanos , Gravidez
19.
Nan Fang Yi Ke Da Xue Xue Bao ; 39(11): 1280-1286, 2019 Nov 30.
Artigo em Chinês | MEDLINE | ID: mdl-31852648

RESUMO

OBJECTIVE: To investigate the effect of Golgi phosphoprotein 3 (Golph3) on paclitaxel- induced apoptosis and autophagy in HeLa cells. METHODS: HeLa cells were transfected with a lentiviral vector expressing Golph3 or a small interfering RNA (siRNA) targeting Golph3 for up-regulation or down-regulation of Golph3 which was verified by Western blotting. The autophagic bodies in the cells were observed using transmission electron microscopy. The expression of autophagy markers p62 and LC3 were detected using Western blotting, and the cell apoptosis was examined by PI/Anexin V-FITC double staining and flow cytometry. The effects of blocking autophagy was evaluated by treatment of the cells with the autophagy inhibitor 3-MA. RESULTS: Transmission electron microscopy showed that the lentivirus-mediated overexpression of Golph3 significantly increased the number of autophagic bodies and interference of Golph3 expression significantly decreased autophagic bodies in HeLa cells. Western blotting showed that Golph3 overexpression caused an increased expression of LC3 and decreased the accumulation of p62 in the cells, and interference of Golph3 resulted in the reverse changes. The cell apoptosis induced by paclitaxel was significantly decreased in Golph3-overexpressing HeLa cells and increased in the cells with Golph3 knockdown (P>0.01). Treatment with 3-MA alone did not obviously affect HeLa cell apoptosis, but in cells with Golph3 knockdown, 3-MA significantly enhanced paclitaxel-induced apoptosis (P>0.01). CONCLUSIONS: Up-regulation of Golph3 promotes autophagy and inhibits paclitaxel-induced apoptosis, whereas suppression of Golph3 inhibits autophagy and enhances paclitaxel- induced apoptosis in HeLa cells.


Assuntos
Autofagia , Apoptose , Células HeLa , Humanos , Proteínas de Membrana , Paclitaxel , Fosfoproteínas
20.
Yi Chuan ; 41(12): 1110-1118, 2019 Dec 20.
Artigo em Chinês | MEDLINE | ID: mdl-31857282

RESUMO

Myogenesis is a complex physiological process that is mainly involved in the proliferation of myogenic stem cells to form myoblasts, which then differentiated and fused to form multinucleated myotubes. Many proteins have been found to be involved in myoblast fusion, but none of them are muscle-specific fusion proteins. In recent years, two muscle-specific transmembrane proteins, i.e. Myomaker and Myomerger, have been discovered and identified, which can coordinate and promote the fusion of myoblasts and thus participate in the process of myogenesis. In this review, we summarize the research progress of Myomaker and Myomerger in myogenesis, including their expression patterns and functional domains, as well as their participation in myoblast fusion mechanisms, aiming to provide relevant ideas for in-depth study of the myogenesis process and treatment of diseases related to myoblast fusion.


Assuntos
Proteínas de Membrana , Músculo Esquelético , Mioblastos , Animais , Diferenciação Celular , Fusão Celular , Humanos , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Desenvolvimento Muscular , Proteínas Musculares , Músculo Esquelético/citologia , Mioblastos/citologia
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