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1.
Gene ; 806: 145922, 2022 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-34454032

RESUMO

Gastric cancer (GC)-derived cell lines were generally used in basic cancer research and drug screening. However, it is always concerned about the difference between cultured cells and primary tumor by oncologists. To address this question, we compared differentially expressed genes (DEGs) in primary cancers, healthy tissues, and cell lines both in vitro and in silico. Seven reported genes with decreased expression in GCs by DNA methylation were analyzed in our cohort studies and experimentally validation. Selected datasets from TCGA (The Cancer Genome Atlas), CCLE (The Broad Institute Cancer Cell Line Encyclopedia), and GTEx (The Genotype-Tissue Expression project) were used to represent GCs, GC-derived cell lines, and healthy tissues respectively in the in silico analysis. Thirty gastric tissues together with six cell lines were used for validations. Unexpectedly, we experimentally found that reported cancer-related downregulated genes were only found in cancer cell lines but not in biopsies. The unchanged gene expressions in primary GCs were generally consistent with our cohort study, using information from cancerous (TCGA) and healthy tissues (GETx). Substantial differences were also found between DEGs of cancer tissues (TGCA)/ healthy tissues (GTEx) pair and cell lines (CCLE)/ healthy tissues (GTEx) pair, which confirmed the significant differences between primary cancer and cancer cell lines. Moreover, elevated expression of YWHAQ (14-3-3 δ) and THBS1 were observed in the GC biopsies, which might be potential biomarkers for GC diagnosis, considering the increased YWHAQ and THBS1 associated with poor survival rates in gastric cancer patients. In sum, it is suggested that cautions should be taken when using GC cell lines to study genes that show great differences between cell lines and tissues.


Assuntos
Proteínas 14-3-3/genética , Regulação Neoplásica da Expressão Gênica , Proteínas de Neoplasias/genética , Neoplasias Gástricas/genética , Trombospondinas/genética , Proteínas 14-3-3/metabolismo , Idoso , Atlas como Assunto , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Linhagem Celular Tumoral , Metilação de DNA , Conjuntos de Dados como Assunto , Epigênese Genética , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Modelos Biológicos , Proteínas de Neoplasias/metabolismo , Cultura Primária de Células , Prognóstico , Neoplasias Gástricas/diagnóstico , Neoplasias Gástricas/mortalidade , Neoplasias Gástricas/patologia , Análise de Sobrevida , Trombospondinas/metabolismo , Células Tumorais Cultivadas
2.
Gene ; 806: 145920, 2022 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-34455026

RESUMO

Depression is deemed a mood disorder characterized by a high rate of relapse. Therefore, overcoming of the recurrent depression is globally expecting. Kososan, a traditional Japanese herbal medicine, has been clinically used for mild depressive mood, and our previous studies have shown some evidence for its antidepressive-like efficacy in experimental animal models of depression. However, it remains unclear whether kososan has beneficial effects on recurrent depression. Here, we examined its effect using a mouse model of modified repeated social defeat stress (SDS) paradigm. Male BALB/c mice were exposed to a 5-min SDS from unfamiliar aggressive CD-1 mice for 5 days. Kososan extract (1.0 kg/kg/day) or an antidepressant milnacipran (60 mg/kg/day) was administered orally for 26 days (days 7-32) to depression-like mice with social avoidant behaviors on day 6. Single 5 min of SDS was subjected to mice recovered from the social avoidance on day 31, and then the recurrence of depression-like behaviors was evaluated on day 32. Hippocampal gene expression patterns were also assayed by DNA microarray analysis. Water- or milnacipran-administered mice resulted in a recurrence of depression-like behaviors by re-exposure of single SDS, whereas kososan-administered mice did not recur depression-like behaviors. Distinct gene expression patterns were also found for treating kososan and milnacipran. Collectively, this finding suggests that kososan exerts a preventive effect on recurrent depression-like behaviors in mice. Pretreatment of kososan is more useful for recurrent depression than that of milnacipran.


Assuntos
Antidepressivos/farmacologia , Depressão/prevenção & controle , Medicamentos de Ervas Chinesas/farmacologia , Proteínas do Tecido Nervoso/genética , Derrota Social , Estresse Psicológico/tratamento farmacológico , Administração Oral , Animais , Depressão/genética , Depressão/fisiopatologia , Depressão/psicologia , Modelos Animais de Doenças , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Ontologia Genética , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Hipocampo/fisiopatologia , Japão , Masculino , Medicina Kampo/métodos , Camundongos , Camundongos Endogâmicos BALB C , Milnaciprano/farmacologia , Anotação de Sequência Molecular , Proteínas do Tecido Nervoso/classificação , Proteínas do Tecido Nervoso/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos , Recidiva , Estresse Psicológico/genética , Estresse Psicológico/fisiopatologia
3.
Gene ; 806: 145928, 2022 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-34455027

RESUMO

Cytochrome P450 Family 19 (CYP19) is a crucial enzyme to catalyze the conversion of androgens to estrogens. However, the regulatory mechanism of goose CYP19 gene remains poorly understood. The present study attempted to obtain the full-length coding sequence (CDS) and 5'-flanking sequence of CYP19 gene, to investigate its expression and distribution profiles in different sized follicles, and to analyze the transcriptional regulatory mechanism of CYP19 gene in goose. Results showed that its CDS consisted of 1512 nucleotides and the encoded amino acid sequence contained a classical P450 structural domain. Homology analysis showed that there were high homologies of nucleotide and amino acid sequences between goose and other avian species. Its promoter sequence spanned from -1925 bp to the transcription start site (ATG) and several transcriptional factors were predicted in this region. Further analysis from luciferase assay showed that the luciferase activity was the highest spanning from -118 to -1 bp by constructing deletion promoter reporter vector. In addition, result from quantitative real-time polymerase chain reaction indicated that the mRNA level of CYP19 gene were highly expressed in theca layer of the fifth largest follicle, and the cellular location was in the theca externa cells by immunohistochemistry. Taken together, it could be concluded that the transcription activity of CYP19 gene was activated by transcriptional factors in its proximal region of promoter to promote the synthesis of estrogens, regulating the selection of pre-hierarchical into hierarchical follicle in goose.


Assuntos
Proteínas Aviárias/genética , Família 19 do Citocromo P450/genética , Gansos/genética , Regulação Enzimológica da Expressão Gênica , RNA Mensageiro/genética , Transcrição Genética , Sequência de Aminoácidos , Animais , Proteínas Aviárias/metabolismo , Família 19 do Citocromo P450/metabolismo , Feminino , Gansos/classificação , Regulação da Expressão Gênica no Desenvolvimento , Folículo Ovariano/citologia , Folículo Ovariano/crescimento & desenvolvimento , Folículo Ovariano/metabolismo , Filogenia , Regiões Promotoras Genéticas , RNA Mensageiro/metabolismo , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Sítio de Iniciação de Transcrição
4.
Gene ; 806: 145929, 2022 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-34461150

RESUMO

The body color of Neocaridina denticulate sinensis is a compelling phenotypic trait, in which a cascade of carotenoid metabolic processes plays an important role. The study was conducted to compare the transcriptome of cephalothoraxes among three pigmentation phenotypes (red, blue, and chocolate) of N. denticulate sinensis. The purpose of this study was to explore the candidate genes associated with different colors of N. denticulate sinensis. Nine cDNA libraries in three groups were constructed from the cephalothoraxes of shrimps. After assembly, 75022 unigenes were obtained in total with an average length of 1026 bp and N50 length of 1876 bp. There were 45977, 25284, 23605, 21913 unigenes annotated in the Nr, Swissprot, KOG, and KEGG databases, respectively. Differential expression analysis revealed that there were 829, 554, and 3194 differentially expressed genes (DEGs) in RD vs BL, RD vs CH, and BL vs CH, respectively. These DEGs may play roles in the absorption, transport, and metabolism of carotenoids. We also emphasized that electron transfer across the inner mitochondrial membrane (IMM) was a key process in pigment metabolism. In addition, a total of 6328 simple sequence repeats (SSRs) were also detected in N. denticulate sinensis. The results laid a solid foundation for further research on the molecular mechanism of integument pigmentation in the crustacean and contributed to developing more attractive aquatic animals.


Assuntos
Proteínas de Artrópodes/genética , Carotenoides/metabolismo , Decápodes/genética , Pigmentação/genética , Transcriptoma , Animais , Organismos Aquáticos , Proteínas de Artrópodes/classificação , Proteínas de Artrópodes/metabolismo , Transporte Biológico , Cor , Bases de Dados Genéticas , Decápodes/anatomia & histologia , Decápodes/metabolismo , Água Doce , Regulação da Expressão Gênica , Biblioteca Gênica , Ontologia Genética , Repetições de Microssatélites , Mitocôndrias/metabolismo , Membranas Mitocondriais/metabolismo , Anotação de Sequência Molecular , Característica Quantitativa Herdável
5.
Gene ; 806: 145935, 2022 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-34478821

RESUMO

Soluble molecules of programmed death ligand 1 (sPD-L1) are known to modulate T-cell depletion, an important mechanism of hepatitis B virus (HBV) persistence and liver disease progression. In addition, PD-L1 polymorphisms in the 3'-UTR can influence PD-L1 expression and have been associated with cancer risk, although not definitively. The purpose of this study was to investigate the association of PD-L1 polymorphisms and circulating levels of sPD-L1 in HBV infection and live disease progression. In this study, five hundred fifty-one HBV infected patients of the three clinically well-defined subgroups chronic hepatitis B (CHB, n = 186), liver cirrhosis (LC, n = 142) and hepatocellular carcinoma (HCC, n = 223) and 240 healthy individuals (HC) were enrolled. PD-L1 polymorphisms (rs2297136 and rs4143815) were genotyped by in-house validated ARMS assays. Logistic regression models were applied in order to determine the association of PD-L1 polymorphisms with HBV infection as well as with progression of related liver diseases. Plasma sPD-L1 levels were quantified by ELISA assays. The PD-L1 rs2297136 AA genotype was associated with HBV infection susceptibility (HBV vs. HC: OR = 1.6; 95%CI = 1.1-2.3; p = 0.0087) and disease progression (LC vs. CHB: OR = 1.8; 95%CI = 1.1-2.9; p = 0.018). Whereas, the rs2297136 GG genotype was a protective factor for HCC development. Plasma sPD-L1 levels were significantly high in HBV patients (p < 0.0001) and higher in the LC followed by CHB and HCC groups. High sPD-L1 levels correlated with increased liver enzymes and with advanced liver disease progression (Child-pugh C > B > A, p < 0.0001) and BCLC classification (BCLC D > C > B > A, p = 0.031). We could, for the first time, conclude that PD-L1 rs2297136 polymorphism and plasma sPD-L1 protein levels associate with HBV infection and HBV-related liver disease progression.


Assuntos
Antígeno B7-H1/genética , Carcinoma Hepatocelular/genética , Vírus da Hepatite B/patogenicidade , Hepatite B Crônica/genética , Cirrose Hepática/genética , Neoplasias Hepáticas/genética , Polimorfismo Genético , Regiões 3' não Traduzidas , Adulto , Idoso , Antígeno B7-H1/sangue , Biomarcadores/sangue , Carcinoma Hepatocelular/diagnóstico , Carcinoma Hepatocelular/etiologia , Carcinoma Hepatocelular/virologia , Estudos de Casos e Controles , Progressão da Doença , Feminino , Expressão Gênica , Predisposição Genética para Doença , Vírus da Hepatite B/crescimento & desenvolvimento , Hepatite B Crônica/complicações , Hepatite B Crônica/diagnóstico , Hepatite B Crônica/virologia , Humanos , Fígado/metabolismo , Fígado/patologia , Fígado/virologia , Cirrose Hepática/diagnóstico , Cirrose Hepática/etiologia , Cirrose Hepática/virologia , Neoplasias Hepáticas/diagnóstico , Neoplasias Hepáticas/etiologia , Neoplasias Hepáticas/virologia , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes
6.
Food Chem ; 366: 130532, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34274702

RESUMO

One major pepsinogen, PG-I, and two minor pepsinogens, PG-II and PG-III were purified from lizardfish stomach by ammonium sulfate precipitation and two chromatographic columns. The three purified PGs migrated as single bands in native-PAGE gels with molecular weights (MW) ranging from 36 to 38 kDa. Each PG was converted to pepsin (P) at pH 2.0, and the MW were determined as 32 kDa (for P-I), 31 kDa (for P-II) and 30 kDa (for P-III). The optimum pH and temperature of pepsins were 2.0-3.5, and 40-50 °C. All 3 pepsins were strongly inhibited by pepstatin A. Divalent cations slightly stimulated the pepsin activities, but ATP had no effect on the pepsins. Purified pepsins were effective in the hydrolysis of various proteins. Km and kcat of the three pepsins for hemoglobin hydrolysis were 107.64-276.61 µM and 18.30-32.68 s-1, respectively. The new pepsins have potential for use in protein food procession and modification.


Assuntos
Pepsina A , Pepsinogênios , Sequência de Aminoácidos , Animais , Peixes/metabolismo , Pepsina A/metabolismo , Pepsinogênios/metabolismo , Estômago
7.
Food Chem ; 366: 130553, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34284194

RESUMO

A method for real-time monitoring of lipid digestion based on photonic crystals formed from silica was developed. As an effective "net", the highly ordered silica colloidal crystal (SCC) film provides structural support for lipid payload. This method based on optical interferometric film kinetics was used to record the whole kinetics progress of olive oil hydrolysis by lipase in real time and calculate the kinetic Michaelis constant. The kinetic parameters were compared with the results determined by the titration method. The effects of bile salt content on lipase and olive oil layer were studied. This method provides a potential evaluation system for real-time digestion and degradation of edible oil in the food field. It also provides a basis for further real-time evaluation of lipid bioavailability in food systems by real-time recording the release and degradation of lipids in the food nano-matrix.


Assuntos
Lipídeos , Lipólise , Digestão , Interferometria , Lipase/metabolismo
8.
Food Chem ; 366: 130569, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34298394

RESUMO

This study aimed to investigate the effects of fermented soymilk (FSM) with Lactobacillus brevis CICC 23,474 and L. brevis CICC 23,470 on the structural changes and allergenicity of major allergenic proteins in soymilk (SM). Spectroscopy and liquid chromatograph-tandem mass spectrometer (LC-MS/MS) were used to characterize changes in protein spatial structure and epitopes. The antigenicity and potential allergenicity were evaluated by immunoblotting, enzyme-linked immunosorbent assay (ELISA) and KU812 cell degranulation assay. Results suggested that the advanced structure of proteins was destroyed. Antigenicity was also significantly reduced, and five human IgE-binding linear epitopes (i.e., E5-E33, R27-S41, D414-A437, G253-I265 and V449-S471) were destroyed by fermentation. Furthermore, after in vitro simulated gastrointestinal digestion, FSM showed lower IgG/IgE-binding capacity and weaker degranulation ability of KU812 cells. All these findings demonstrated that fermentation with Lactobacillus can destroy the conformational and linear epitopes of proteins and reduce the potential allergenicity of SM.


Assuntos
Lactobacillus , Leite de Soja , Alérgenos/metabolismo , Cromatografia Líquida , Fermentação , Humanos , Lactobacillus/metabolismo , Espectrometria de Massas em Tandem
9.
Food Chem ; 366: 130523, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34303206

RESUMO

Our study offers a novel sn-1,3 specific lipase MAJ1 from marine member Janibacter sp. strain HTCC2649 for preparing long-medium-long (LML) type structured triacylglycerols (TAGs). Firstly, the resin ECR1030 was selected as a suitable support for the immobilization of lipase MAJ1. An efficient synthesis of LML-type structured TAGs by the immobilized lipase MAJ1-catalyzed interesterification of methyl palmitate and tricaprylin was studied in a solvent-free system. The reaction conditions, including substrate molar ratio, temperature and enzyme loading, were optimized. Under the optimum conditions (immobilized lipase MAJ1 of 45 U/g, substrate molar ratio of 4:1, temperature of 35 °C, reaction time of 24 h), the structured TAGs with double long chains (DLCST) were obtained in a yield of 44.3 mol%. Secondly, multi-dimensional mass spectrometry-based shotgun lipidomics (MDMS-SL) was employed to quantify each TAG positional isomer in DLCST. The content of 1,3-dipalmitoyl-2-capryloyl-sn-glycerol in DLCST was 97.6% determined by the MDMS-SL technology.


Assuntos
Enzimas Imobilizadas , Lipase , Catálise , Esterificação , Lipase/metabolismo , Solventes , Temperatura , Triglicerídeos
10.
Food Chem ; 366: 130616, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34311240

RESUMO

The lipase from Bacillus licheniformis NCU CS-5 was immobilized onto ß-cyclodextrin (CD) grafted and aminopropyl-functionalized chitosan-coated Fe3O4 magnetic nanocomposites (Fe3O4-CTS-APTES-GA-ß-CD). Fourier transform infrared spectroscopy, thermogravimetry analysis, X-ray diffraction, scanning electron microscopy and transmission electron microscopy showed that not only the functionalized magnetic nanoparticles were synthesized but also the immobilized lipase was successfully produced. The immobilized lipase exhibited higher optimal pH value (10.5) and temperature (60℃) than the free lipase. The pH and thermal stabilities of the immobilized lipase were improved significantly compared to the free lipase. The immobilized lipase remained more than 80% of the relative activity at temperature of 60 ℃ and pH 12.0. The immobilized lipase also remained over 80% of its relative activity after 28 days of storage and 15 cycles of application. The application of the immobilized lipase in esterification of isoamyl acetate and pentyl valerate showed that maximum esterification efficiency was achieved in n-hexane having 68.0% and 89.2% respectively. Therefore, these results indicated that the Fe3O4-CTS-APTES-GA-ß-CD nanoparticles are novel carriers for immobilizing enzyme, and the immobilized lipase can be used as an innovative green approach to the synthesis of fruity flavor esters in food industry.


Assuntos
Quitosana , Ciclodextrinas , Nanopartículas de Magnetita , Nanocompostos , Estabilidade Enzimática , Enzimas Imobilizadas/metabolismo , Esterificação , Ésteres , Concentração de Íons de Hidrogênio , Lipase/metabolismo , Fenômenos Magnéticos
11.
Food Chem ; 366: 130636, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34314929

RESUMO

The study aimed to determine the efficiency of advanced chelate compounds-based trace minerals (OTM) in laying hens. Laying hens (240, 32 weeks old) were assigned to one of the following five groups: NOTM (no added trace minerals), CONTM (standard mineral salts), and three experimental groups in which chelates were used to replace 33, 66, and 100% of mineral salts (OTM33, OTM66, and OTM100, respectively). Each treatment had six replicates with eight hens per replicate. After 18 weeks, performance and physicochemical properties of eggs in all experimental groups was better than those in the NOTM group. Among the treatments, OTM66 and OTM100 produced the best results in terms of laying performance, yolk PUFA/SFA ratio, Zn and Se contents, and malondialdehyde concentration in both serum and yolk. In conclusion, up to 66% OTM supplementation was beneficial for performance, lipid and mineral composition of yolk, and oxidative status.


Assuntos
Galinhas , Ácidos Graxos , Ração Animal/análise , Animais , Dieta , Suplementos Nutricionais , Gema de Ovo/metabolismo , Ácidos Graxos/metabolismo , Feminino , Minerais/metabolismo , Estresse Oxidativo
12.
Food Chem ; 366: 130650, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34330030

RESUMO

Mixed lauric acid esters (MLE) with antibacterial and antioxidative activities were produced through lipase-catalyzed two-step esterification in solvent-free system without purification. In the first reaction, erythorbyl laurate was synthesized for 72 h. Successive reaction for 6 h at molar ratio of 1.0 (lauric acid to glycerol) produced MLE containing erythorbyl laurate and glyceryl laurate with small amounts of residual substrates, by converting 99.52% of lauric acid. MLE addition (0.5-2.0%, w/w) to Tween 20-stabilized emulsions decreased droplet size, polydispersity index, and zeta-potential, possibly enhancing the emulsion stability. In the emulsions, MLE at 0.5 and 2.0% (w/w) caused 4.4-4.6 and 5.9-6.1 log reductions of Gram-positive (Staphylococcus aureus, Listeria monocytogenes) and Gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa), respectively, within 12 h. Lipid hydroperoxide concentrations decreased to 50.8-98.3% in the presence of 0.5-2.0% (w/w) MLE. These findings support a novel approach without needing purification to produce multi-functional food additives for emulsion foods.


Assuntos
Ésteres , Lipase , Antibacterianos/farmacologia , Antioxidantes , Catálise , Esterificação , Ácidos Láuricos , Lipase/metabolismo , Solventes
13.
Food Chem ; 366: 130699, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34348221

RESUMO

In this work, magnetic chitosan (MCH) beads were synthesized by phase-inversion method, and grafted with polydopamine (PDA) and then used for direct immobilization of Candida rugosa lipase by Schiff base reaction. The amount of immobilized enzyme and the retained activity were found to be 47.3 mg/g and 72.8%, respectively, at pH 7.0, and at 25 °C. The apparent Km (9.7 mmol/L), and Vmax (384 U/mg) values of the immobilized lipase were significantly changed compared to the free lipase. The MCH@PDA-lipase was better thermal and storage stability at different temperatures than those of the free lipase. In hexane medium, the esterification reaction results showed that the maximum conversions of isoamylalcohol and isopentyl alcohol to isoamyl acetate and isopentyl acetate using the MCH@PDA-lipase were found to be 98.4 ± 1.3% and 73.7 ± 0.7%, respectively. These results showed that the MCH@PDA-lipase can be used as an operative immobilized enzyme system for many biotechnological applications.


Assuntos
Quitosana , Lipase , Estabilidade Enzimática , Enzimas Imobilizadas/metabolismo , Ésteres , Concentração de Íons de Hidrogênio , Lipase/metabolismo , Fenômenos Magnéticos , Saccharomycetales
14.
Food Chem ; 367: 130701, 2022 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-34388635

RESUMO

Carotenoids are colored compounds with important physiological functions. The Haida golden scallop, which has an orange adductor muscle, is a carotenoid-enriched variety of scallop Mizuhopecten yessoensis, an important aquaculture shellfish. In this study, we investigated the tissue distribution of the carotenoids, pectenolone and pectenoxanthin, in both Haida golden scallop and normal Yesso scallop. Both carotenoids were detected in all the sampled tissues of the two scallops, except in the adductor muscle of normal scallop. There were significantly more carotenoids in Haida golden scallop than in normal scallop, in the tissues of the mantle, female gonad, kidney, and adductor muscle. Increased carotenoid concentrations were detected in Haida golden scallop adductor muscle during the spring spawning season, indicating the effects of reproduction on muscle carotenoids accumulation. This study was the first systematic investigation of carotenoid distribution in Yesso scallop tissues and will benefit future research on carotenoid accumulation and function in scallops.


Assuntos
Citrus sinensis , Pectinidae , Animais , Carotenoides/metabolismo , Citrus sinensis/metabolismo , Músculo Esquelético/metabolismo , Pectinidae/metabolismo , Estações do Ano , Distribuição Tecidual
15.
Food Chem ; 367: 130767, 2022 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-34391996

RESUMO

This study aimed to investigate the effect of caspase-3 inhibitor in mitochondrial apoptosis activation on structure protein degradation during postmortem storage. Mitochondrial dysfunction, apoptotic factors, structure protein degradation and the myofibrillar rupture index between the control and caspase-3 inhibitor groups were determined. The results show caspase-3 inhibitor repressed the mitochondrial membrane permeability and mitochondrial swelling, as well as increased mitochondrial membrane potential, causing a decrease in the release of cytochrome c from mitochondria to cytoplasm and caspase-9/3 activities (P < 0.05). Subsequently, small myofibrillar proteins (desmin and troponin-T) were susceptible to degradation, initiating texture deterioration. By contrast, giant structure proteins (titin and nebulin) were degraded during later postmortem storage, predominantly contributing to fish softening. The results further suggest that caspase-3 is involved in degradation of structure proteins during postmortem through mitochondrial apoptosis pathways.


Assuntos
Esocidae , Mitocôndrias , Animais , Apoptose , Caspase 3/genética , Caspase 3/metabolismo , Caspase 9/metabolismo , Esocidae/metabolismo , Mitocôndrias/metabolismo , Membranas Mitocondriais/metabolismo
16.
Food Chem ; 366: 130422, 2022 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-34392082

RESUMO

Tea cream, produced by interactions among tea ingredients, is undesirable in tea beverage industry. The interaction between bovine serum albumin (BSA) and theaflavin-3,3'-digallate (TFDG, an important component in tea cream and functional substance of black tea) was investigated by fluorescence spectroscopy, ultraviolet-visible (UV-vis) absorption spectroscopy, synchronous fluorescence spectroscopy, fourier-transform infrared (FT-IR) spectroscopy, and molecular docking technique. Multi-spectroscopic experiments demonstrated that TFDG interacted with BSA via static quenching, and the microenvironment around BSA became more hydrophobicity. FT-IR showed that the α-helix of BSA was increased when binding with TFDG. Thermodynamic parameters and molecular docking demonstrated that hydrophobic interactions and hydrogen bonds dominated the interaction between TFDG and BSA. The mechanism proposed in this research could further develop some nanoparticles to excellent biochemical properties while reducing the formation of tea cream, and explore the potential of BSA as transport carrier for TFDG.


Assuntos
Soroalbumina Bovina , Biflavonoides , Sítios de Ligação , Catequina/análogos & derivados , Dicroísmo Circular , Simulação de Acoplamento Molecular , Ligação Proteica , Soroalbumina Bovina/metabolismo , Espectrometria de Fluorescência , Espectrofotometria Ultravioleta , Espectroscopia de Infravermelho com Transformada de Fourier , Termodinâmica
17.
J Cell Sci ; 135(5)2022 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-33912961

RESUMO

Septins colocalize with membrane sterol-rich regions and facilitate recruitment of cell wall synthases during wall remodeling. We show that null mutants missing an Aspergillus nidulans core septin present in hexamers and octamers (ΔaspAcdc11, ΔaspBcdc3 or ΔaspCcdc12) are sensitive to multiple cell wall-disturbing agents that activate the cell wall integrity MAPK pathway. The null mutant missing the octamer-exclusive core septin (ΔaspDcdc10) showed similar sensitivity, but only to a single cell wall-disturbing agent and the null mutant missing the noncore septin (ΔaspE) showed only very mild sensitivity to a different single agent. Core septin mutants showed changes in wall polysaccharide composition and chitin synthase localization. Mutants missing any of the five septins resisted ergosterol-disrupting agents. Hexamer mutants showed increased sensitivity to sphingolipid-disrupting agents. Core septins mislocalized after treatment with sphingolipid-disrupting agents, but not after ergosterol-disrupting agents. Our data suggest that the core septins are involved in cell wall integrity signaling, that all five septins are involved in monitoring ergosterol metabolism, that the hexamer septins are required for sphingolipid metabolism and that septins require sphingolipids to coordinate the cell wall integrity response.


Assuntos
Aspergillus nidulans , Septinas , Aspergillus nidulans/metabolismo , Parede Celular/metabolismo , Metabolismo dos Lipídeos , Septinas/genética , Septinas/metabolismo , Esfingolipídeos/metabolismo
18.
Food Chem ; 367: 130655, 2022 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-34371277

RESUMO

Interactions between the dimeric form of ß-lactoglobulin and vanillic acid were investigated at pH 7.2, using a variety of spectroscopic techniques and molecular dynamics (MD) simulations. FTIR and CD studies showed alterations in the secondary structure of the protein upon its interaction with the ligand. Fluorescence measurements indicated that the dimeric complex with the phenolic acid produced a large dissociation constant (KD) compared to the monomeric counterpart at acidic pH (part A of this series). Stoichiometry of 1:1 was identified for the ß-lactoglobulin-vanillic acid complex by Job plot analysis at neutral pH suggesting two ligand molecules can participate in binding with the dimer. Molecular docking and MD simulations suggested that the top-ranked binding sites of the ligand were located at the entrance of each ß-barrel structure of the dimer. These simulations also allowed identification of the contribution of water molecules, in the form of protein-water-ligand bridging interactions, to the complexes.


Assuntos
Lactoglobulinas , Simulação de Dinâmica Molecular , Sítios de Ligação , Concentração de Íons de Hidrogênio , Lactoglobulinas/metabolismo , Simulação de Acoplamento Molecular , Ligação Proteica , Ácido Vanílico
19.
J Cell Sci ; 135(5)2022 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-34156466

RESUMO

Store-operated Ca2+ entry is a central component of intracellular Ca2+ signaling pathways. The Ca2+ release-activated channel (CRAC) mediates store-operated Ca2+ entry in many different cell types. The CRAC channel is composed of the plasma membrane (PM)-localized Orai1 channel and endoplasmic reticulum (ER)-localized STIM1 Ca2+ sensor. Upon ER Ca2+ store depletion, Orai1 and STIM1 form complexes at ER-PM junctions, leading to the formation of activated CRAC channels. Although the importance of CRAC channels is well described, the underlying mechanisms that regulate the recruitment of Orai1 to ER-PM junctions are not fully understood. Here, we describe the rapid and transient S-acylation of Orai1. Using biochemical approaches, we show that Orai1 is rapidly S-acylated at cysteine 143 upon ER Ca2+ store depletion. Importantly, S-acylation of cysteine 143 is required for Orai1-mediated Ca2+ entry and recruitment to STIM1 puncta. We conclude that store depletion-induced S-acylation of Orai1 is necessary for recruitment to ER-PM junctions, subsequent binding to STIM1 and channel activation.


Assuntos
Canais de Cálcio , Cálcio , Acilação , Cálcio/metabolismo , Canais de Cálcio/genética , Canais de Cálcio/metabolismo , Sinalização do Cálcio , Membrana Celular/metabolismo , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Proteína ORAI1/genética , Proteína ORAI1/metabolismo , Molécula 1 de Interação Estromal/genética , Molécula 1 de Interação Estromal/metabolismo
20.
BMC Gastroenterol ; 21(1): 359, 2021 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-34600475

RESUMO

BACKGROUND: Activation of Adenosine 5'-monophosphate-activated protein kinase/Sirtuin1 (AMPK/SIRT1) exerts an effect in alleviating obesity and gut damage. Sodium nitroprusside (SNP), a nitric oxide (NO) donor, has been reported to activate AMPK. This study was to investigate the effect of SNP on HFD induced gut dysfunction and the mechanism. METHODS: SNP was applied on lipopolysaccharide (LPS) stimulated Caco-2 cell monolayers which mimicked intestinal epithelial barrier dysfunction and HFD-fed mice which were complicated by gut dysfunction. Then AMPKα/SIRT1 pathway and gut barrier indicators were investigated. RESULTS: SNP rescued the loss of tight junction proteins ZO-1 and occludin, the inhibition of AMPKα/SIRT1 in LPS stimulated Caco-2 cell monolayers, and the effects were not shown when AMPKa1 was knocked-down by siRNA. SNP also alleviated HFD induced obesity and gut dysfunction in mice, as indicated by the decreasing of intestinal permeability, the increasing expression of ZO-1 and occludin, the decreasing levels of pro-inflammatory cytokine IL-6, and the repairing of gut microbiota dysbiosis. These effects were complicated by the increased colonic NO content and the activated AMPKα/SIRT1 signaling. CONCLUSIONS: The results may imply that SNP, as a NO donor, alleviates HFD induced gut dysfunction probably by activating the AMPKα/SIRT1 signaling pathway.


Assuntos
Proteínas Quinases Ativadas por AMP , Sirtuína 1 , Proteínas Quinases Ativadas por AMP/metabolismo , Animais , Células CACO-2 , Dieta Hiperlipídica , Humanos , Camundongos , Nitroprussiato/farmacologia , Transdução de Sinais , Sirtuína 1/genética , Sirtuína 1/metabolismo
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