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1.
Chemosphere ; 239: 124794, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31521929

RESUMO

Cadmium (Cd) pollution is considered one of the global environmental issues due to its adverse effects on plant and human health. With the rapid development of nanotechnology and the practical application of engineered nanoparticles (ENPs) in agriculture, the mechanisms underlying the interactions between NPs and heavy metal on their uptake, accumulation, and phytotoxicity in crops are still not fully understood. Therefore, the impact of TiO2 NPs (0, 100, 250 mg/L) and Cd (0, 50 µM) co-exposure on hydroponic maize (Zea mays L.) was determined under two exposure modes. Results showed that root co-exposure to TiO2 NPs and 100 mg/L Cd significantly enhanced Cd uptake and produced greater phytotoxicity in maize than foliar exposure to TiO2 NPs. Meanwhile, plant dry weight and chlorophyll content showed a reduction of 45.3% and 50.5%, respectively, when compared with single Cd treatment. In addition, the accumulation of Ti in shoots and roots increased by 1.61 and 4.29 times, respectively when root exposure to 250 mg/L TiO2 NPs. By contrast, foliar exposure of TiO2 NPs could markedly decrease shoot Cd contents from 15.2% to 17.8% and had a stronger influence on alleviating Cd-induced toxicity via increasing superoxide dismutase (SOD) and glutathione S-transferase (GST) activities and upregulating several metabolic pathways, including galactose metabolism and citrate cycle, alanine, aspartate and glutamate metabolism, as well as glycine, serine and threonine metabolism. This study provides a new strategy for the application of TiO2 NPs in crop safety production in Cd contaminated soils.


Assuntos
Cádmio/toxicidade , Poluentes do Solo/toxicidade , Titânio/farmacologia , Zea mays/efeitos dos fármacos , Aminoácidos/metabolismo , Cádmio/farmacocinética , Clorofila/metabolismo , Produtos Agrícolas/metabolismo , Galactose/metabolismo , Glutationa Transferase/metabolismo , Hidroponia , Nanopartículas Metálicas/administração & dosagem , Nanopartículas Metálicas/química , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/metabolismo , Proteínas de Plantas/metabolismo , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/metabolismo , Plântula/efeitos dos fármacos , Plântula/metabolismo , Poluentes do Solo/farmacocinética , Superóxido Dismutase/metabolismo , Titânio/administração & dosagem , Titânio/farmacocinética , Zea mays/metabolismo
2.
J Sci Food Agric ; 100(1): 92-101, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31435952

RESUMO

BACKGROUND: Oyster polypeptides have various biofunctions, such as anti-cancer and anti-oxidative stress, but whether it has the protective effects to primary ovarian failure (POF) remains poorly understand. To address this issue, daily gavage of oyster polypeptides was performed to investigate their protective effect, basing on d-galactose-induced POF model in C57BL/6 female mice. RESULTS: Oyster polypeptides restored the irregular estrous cycles and the abnormal serum follicle stimulating hormone (FSH), luteinizing hormone (LH) and progesterone (P) levels as well as the decreased mRNA expression level of Amh that were induced by d-galactose. The follicle development of POF mice was improved by increasing the primordial follicle ratio and decreasing the atretic follicle number after oral administration of oyster polypeptides. Moreover, in the oyster polypeptides treated mice, the total superoxide dismutase (T-SOD) activity was significantly increased, while the malondialdehyde levels were significantly decreased. The mRNA expression levels of stress-related genes (SOD2, SIRT1 and FOXO3a) were remarkably up-regulated after d-galactose induction, but the up-regulation was weakened or disappeared by the gavage of oyster polypeptides. In addition, oyster polypeptides treatment also reduced the apoptosis of the ovarian granulosa cells and down-regulated the mRNA expression levels of apoptosis-related genes (p53 and Bad but not Bcl-2). CONCLUSION: This study reveals that oyster polypeptides may protect ovary against d-galactose-induced POF by their anti-oxidative stress activity to rescue d-galactose-induced ovarian oxidative damage and therefore to prevent ovarian cells apoptosis, thereby tipping the abnormality trigged by POF to get close to the normal levels. © 2019 Society of Chemical Industry.


Assuntos
Ostreidae/química , Peptídeos/administração & dosagem , Insuficiência Ovariana Primária/tratamento farmacológico , Substâncias Protetoras/administração & dosagem , Animais , Apoptose/efeitos dos fármacos , Feminino , Galactose/efeitos adversos , Humanos , Hormônio Luteinizante/metabolismo , Malondialdeído/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Folículo Ovariano/efeitos dos fármacos , Folículo Ovariano/metabolismo , Ovário/efeitos dos fármacos , Ovário/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Insuficiência Ovariana Primária/induzido quimicamente , Insuficiência Ovariana Primária/genética , Insuficiência Ovariana Primária/metabolismo , Progesterona/metabolismo , Sirtuína 1/genética , Sirtuína 1/metabolismo , Superóxido Dismutase/genética , Superóxido Dismutase/metabolismo
3.
Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi ; 54(11): 843-849, 2019 Nov 07.
Artigo em Chinês | MEDLINE | ID: mdl-31795546

RESUMO

Objective: The aging model of guinea pigs induced by D-galactose was set up to investigate the changes of BK(Ca) expression and function on cochlear pericytes and their relationship with age-related hearing loss. Methods: Thirty healthy 8-week-old guinea pigs were randomly divided into three groups, with 10 in each group: D-galactose aging model group, subcutaneous injection of D-galactose (500 mg/kg) daily for 6 weeks; saline control group, the same amount of saline was injected into the neck of the aging model group for 6 weeks; the blank control group, no treatment was performed. The threshold of auditory brainstem response (ABR) was detected. The content of BK(Ca) in the perivascular cells of the guinea pig cochlear cells was detected by immunofluorescence technique. The changes of peripheral current density and BK(Ca) current were detected by patch clamp technique. The data were analyzed by GraphPad Prism software. Results: Compared with the saline group and the control group, the ABR threshold and the amplitude of the wave I were significantly decreased in the aging model group, and the difference was statistically significant (P<0.01). Compared with the control group, the expression of BK(Ca) in the vascular pericytes of guinea pigs in the aging model group was significantly reduced (1.00±0.08 vs 0.27±0.03,the difference was statistically significant P<0.01), and the cell current density and BK(Ca) net current value were also significantly reduced with statistically significant (P<0.01). Conclusions: D-galactose can successfully induce guinea pig aging model, in which BK(Ca) expression decreases and net current value decreases in pericytes of cochlear striavascularis, and changes in BK(Ca) expression and function may be related to age-related hearing loss.


Assuntos
Cóclea/metabolismo , Doenças Cocleares/metabolismo , Canais de Potássio Ativados por Cálcio de Condutância Alta/biossíntese , Pericitos/metabolismo , Presbiacusia/metabolismo , Animais , Cóclea/patologia , Cóclea/fisiopatologia , Doenças Cocleares/induzido quimicamente , Doenças Cocleares/patologia , Doenças Cocleares/fisiopatologia , Potenciais Evocados Auditivos do Tronco Encefálico , Galactose/administração & dosagem , Galactose/efeitos adversos , Cobaias , Modelos Animais , Presbiacusia/induzido quimicamente , Presbiacusia/patologia , Presbiacusia/fisiopatologia , Distribuição Aleatória
4.
J Agric Food Chem ; 67(46): 12863-12874, 2019 Nov 20.
Artigo em Inglês | MEDLINE | ID: mdl-31670949

RESUMO

A comprehensive quantitative characterization of Maillard reaction products was carried out for conventional (CON) and lactose-hydrolyzed (LH) ultrahigh temperature (UHT) milk during storage at 20, 30, and 40 °C for 1 year. The accumulation of 3-deoxyglucosone (3-DG) and 3-deoxygalactosone (3-DGal) in LH-UHT milk ranged from 20-fold (at 20 °C) to 44-fold (at 40 °C) higher than that in CON-UHT milk. High temperature storage (40 °C) significantly accelerated the accumulation of 3-DG, 3-DGal, and 5-hydroxymethyl furfural but not the majority of the analyzed advanced glycation endproducts (AGEs). The concentrations of major AGEs including N-ε-carboxymethyllysine (CML), N-ε-carboxyethyllysine (CEL), methylglyoxal-hydroimidazolone isomers (MG-H1/H3), glyoxal-hydroimidazolone isomers (G-H1/H3), and G-H2 detected in CON milk during storage were in the range 12-700, 1-14, 8-45, 4-13, and 1-30 µM, respectively, while they were 30-570, 2-88, 17-150, 9-20, and 5-34 µM, respectively, in LH milk. Pyrraline, S-(carboxymethyl)cysteine (CMC), and glyoxal-lysine dimer were detected in lower levels, while MG-H2, methylglyoxal-lysine dimer, argpyrimidine, glyoxal-lysine-amide, glycolic acid-lysine-amide, and pentosidine were not detected in any of the milk samples. This work demonstrates for the first time that five of the analyzed AGEs (CML, CEL, MG-H1/H3, G-H1/H3, and G-H2) could be selected as markers for evaluation of the extent of the Maillard reaction in UHT milk. These results contribute to a better understanding of how Maillard reactions progress during storage of UHT milk and can be used to develop strategies to inhibit Maillard reactions in LH milk.


Assuntos
Produtos Finais de Glicação Avançada/análise , Lactose/química , Leite/química , Animais , Bovinos , Desoxiglucose/análogos & derivados , Desoxiglucose/análise , Armazenamento de Alimentos , Galactose/análogos & derivados , Galactose/análise , Isomerismo , Lisina/análogos & derivados , Lisina/análise , Reação de Maillard , Aldeído Pirúvico/análise , Temperatura Ambiente
5.
BMC Complement Altern Med ; 19(1): 297, 2019 Nov 06.
Artigo em Inglês | MEDLINE | ID: mdl-31694618

RESUMO

BACKGROUND: Radix isatidis (Isatis indigotica Fort.) is an ancient medicinal herb, which has been applied to the prevention and treatment of influenza virus since ancient times. In recent years, the antioxidant activity of Radix isatidis has been widely concerned by researchers. Our previous studies have shown that Radix isatidis protein (RIP) has good antioxidant activity in vitro. In this study, the composition of the protein was characterized and its antioxidant activity in vivo was evaluated. METHODS: The model of oxidative damage in mice was established by subcutaneous injection of D-galactose for 7 weeks. Commercially available kits were used to determine the content of protein and several oxidation indexes in different tissues of mice. The tissue samples were stained with hematoxylin and eosin (H&E) and the pathological changes were observed by optical microscope. The molecular weight of RIP was analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The amino acid composition of RIP was determined by a non-derivative method developed by our research group. RESULTS: RIP significantly increased the activities of antioxidant enzymes such as SOD, CAT, GSH-Px and total antioxidant capability (TAOC) but decreased the MDA level in the serum, kidney and liver. H&E stained sections of liver and kidney revealed D-galactose could cause serious injury and RIP could substantially attenuate the injury. The analysis of SDS-PAGE showed that four bands with molecular weights of 19.2 kDa, 21.5 kDa, 24.8 kDa and 40.0 kDa were the main protein components of RIP. CONCLUSIONS: The results suggested that RIP had excellent antioxidant activity, which could be explored as a health-care product to retard aging and a good source of protein nutrition for human consumption.


Assuntos
Envelhecimento/efeitos dos fármacos , Antioxidantes/química , Medicamentos de Ervas Chinesas/química , Galactose/efeitos adversos , Proteínas de Plantas/administração & dosagem , Proteínas de Plantas/química , Envelhecimento/metabolismo , Animais , Antioxidantes/isolamento & purificação , Catalase/metabolismo , Humanos , Rim/efeitos dos fármacos , Rim/metabolismo , Fígado/efeitos dos fármacos , Fígado/metabolismo , Masculino , Malondialdeído/metabolismo , Camundongos , Camundongos Endogâmicos ICR , Peso Molecular , Estresse Oxidativo/efeitos dos fármacos , Proteínas de Plantas/isolamento & purificação , Raízes de Plantas/química , Superóxido Dismutase/metabolismo
6.
Nat Commun ; 10(1): 4548, 2019 10 07.
Artigo em Inglês | MEDLINE | ID: mdl-31591402

RESUMO

There are many industrially-relevant enzymes that while active, are severely limited by thermodynamic, kinetic, or stability issues (isomerases, lyases, transglycosidases). In this work, we study Lactobacillus sakei L-arabinose isomerase (LsLAI) for D-galactose to D-tagatose isomerization-that is limited by all three reaction parameters. The enzyme demonstrates low catalytic efficiency, low thermostability at temperatures > 40 °C, and equilibrium conversion < 50%. After exploring several strategies to overcome these limitations, we show that encapsulating LsLAI in gram-positive Lactobacillus plantarum that is chemically permeabilized enables reactions at high rates, high conversions, and elevated temperatures. In a batch process, this system enables ~ 50% conversion in 4 h starting with 300 mM galactose (an average productivity of 37 mM h-1), and 85% conversion in 48 h. We suggest that such an approach may be invaluable for other enzymatic processes that are similarly kinetically-, thermodynamically-, and/or stability-limited.


Assuntos
Aldose-Cetose Isomerases/metabolismo , Proteínas de Bactérias/metabolismo , Galactose/metabolismo , Hexoses/metabolismo , Lactobacillus plantarum/enzimologia , Aldose-Cetose Isomerases/genética , Proteínas de Bactérias/genética , Biocatálise , Estabilidade Enzimática , Concentração de Íons de Hidrogênio , Isomerismo , Cinética , Lactobacillus plantarum/genética , Lactobacillus plantarum/metabolismo , Temperatura Ambiente
7.
Eur J Histochem ; 63(3)2019 Sep 11.
Artigo em Inglês | MEDLINE | ID: mdl-31505925

RESUMO

In mammals, the alveolarization process develops predominantly after birth. Airway cells display a complex assemblage of glycans on their surface. These glycans, particularly terminal glycan extensions, are important effective carriers of information that change during the differentiation process. Nevertheless, few systematic data are reported about the cell surface sugar residue content during post-natal lung development. In the present work, we aimed to identify and semi-quantify N-acetylgalactosamine (GalNAc)/galactose (Gal) residues on the bronchioloalveolar cell surface in rat lung sections from 1-, 4-, 8- day old and adult animals and link these data with the lung glycocalyx composition. Horseradish peroxidase-conjugated lectin from Glycine max (soybean agglutinin, SBA) was used, and light microscopy methodologies were performed. SBA labelling intensity was studied before and after sialidase pre-treatment, at one-, four- and eight-day-old animals and adult animals. For semi-quantitative evaluation of SBA binding intensity, two investigators performed the analysis independently, blinded to the type of experiment. Reactivity of the lectin was assessed in bronchiolar and respiratory portion/alveolar epithelial cell surfaces. We evidenced a stronger positive reaction when lung sections were pre-treated with neuraminidase before incubation with the lectin in one- and four-day-old animals and adult animals. These results were not so manifest in eight-day-old animals. This binding pattern, generally points towards the presence of terminal but mainly sub-terminal GalNAc/Gal residues probably capped by sialic acids on the rat bronchiolar/respiratory tract epithelial cells. As this glycan extension is common in O- and N-glycans, our results suggest that these glycan classes can be present in bronchioloalveolar cells immediately after birth and exist during the postnatal period. The results observed in eight-day-old rat lung sections may be due to the dramatic lung morphologic changes and the possible underlying biological mechanisms that occur during this age-moment.


Assuntos
Acetilgalactosamina/metabolismo , Brônquios/citologia , Células Epiteliais/metabolismo , Galactose/metabolismo , Alvéolos Pulmonares/citologia , Animais , Brônquios/crescimento & desenvolvimento , Feminino , Histocitoquímica/métodos , Peroxidase do Rábano Silvestre/química , Neuraminidase/química , Lectinas de Plantas/química , Gravidez , Alvéolos Pulmonares/crescimento & desenvolvimento , Ratos Wistar , Proteínas de Soja/química
8.
J Agric Food Chem ; 67(40): 11158-11166, 2019 Oct 09.
Artigo em Inglês | MEDLINE | ID: mdl-31537069

RESUMO

There are ongoing interests in improving the galactooligosaccharide (GOS) synthesis efficiency of ß-galactosidase by protein engineering. In this study, an intelligent double-hydrophobic amino acid scanning strategy was proposed and employed to target nine residues forming the glycon-binding site (-1 subsite) of ß-galactosidase Bgal1-3. Two mutants C510V and H512I with significantly improved GOS synthesis efficiency were obtained. When 40% (w/v) lactose was used as a substrate, Bgal1-3 reached a maximum GOS yield of 45.3% at 16 h, while the mutants reached higher yields in a much shorter time (59.1% at 10 h for C510V, 51.5% at 2 h for H512I). When skim milk was treated with these enzymes, more GOS was produced (19.9 g/L for C510V, 12.7 g/L for H512I) than that for Bgal1-3 (10.3 g/L) at a lactose conversion of 90%. These results validated hydrophobicity scanning as an efficient method to engineer ß-galactosidases into promising catalysts for the preparation of GOS and GOS-enriched milk.


Assuntos
Bactérias/enzimologia , Proteínas de Bactérias/química , Galactose/química , Oligossacarídeos/química , beta-Galactosidase/química , Sequência de Aminoácidos , Bactérias/química , Bactérias/genética , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Biocatálise , Domínio Catalítico , Galactose/metabolismo , Interações Hidrofóbicas e Hidrofílicas , Lactose/química , Oligossacarídeos/metabolismo , Engenharia de Proteínas , Alinhamento de Sequência , beta-Galactosidase/genética , beta-Galactosidase/metabolismo
9.
Chem Commun (Camb) ; 55(81): 12204-12207, 2019 Oct 08.
Artigo em Inglês | MEDLINE | ID: mdl-31549691

RESUMO

Perfluorophenylboronic acid 1c catalyzes the direct stereoselective addition of alcohol nucleophiles to deactivated peracetylated d-galactal to give 2-deoxygalactosides in 55-88% yield with complete α-selectivity. The unprecedented results reported here also enable the synthesis of disaccharides containing the 2-deoxygalactose moiety directly from the deactivated peracetylated d-galactal. This convenient and metal-free glycosylation method works well with a wide range of alcohol nucleophiles as acceptors and tolerates a range of functional groups without the formation of the Ferrier byproduct and without the need for a large excess of nucleophiles or additives. The method is potentially useful for the synthesis of a variety of α-2-deoxygalactosides.


Assuntos
Ácidos Borônicos/química , Galactose/análogos & derivados , Galactosídeos/síntese química , Acetilação , Catálise , Dissacarídeos/química , Galactose/química , Glicosilação , Estrutura Molecular , Solventes/química , Estereoisomerismo , Temperatura Ambiente
10.
Plant Sci ; 287: 110179, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31481215

RESUMO

Stigmatic mucilage plays a crucial role in pollen-grain adhesion on the stigma in flowering plants. Little information is available regarding mucilage biosynthesis in orchid plants. In the present study, stigmatic mucilage is rich in galactose-containing polysaccharides, mainly consisted of galactose and arabinose in Dendrobium officinale. Thirteen galactosyltransferases involved in biosynthesis of the ß-1,3-galactose linkage polysaccharides, belonging to the CAZY GT31 family, were identified from D. officinale genome. A positive correlation between the mucilage content and the DoGALT2 expression at different stages was observed. DoGALT2 expressed overall sampled tissues with the highest in D. officinale stigmatic mucilage that contributes to pollen adhesion and elongation. DoGALT2 was targeted to Golgi, and had a GALT domain (PF01762) that was homologous to the characterized GALT2 in Arabidopsis. Compared to wild-type Arabidopsis, DoGALT2 overexpressing plants showed a higher content of galactose and galactose-containing alcohol-insoluble residues, and enhanced tolerance to abiotic stress. DoGALT2 complemented Arabidopsis GALT2 mutant (galt2-1), with an equivalent galactose with wild-type Arabidopsis but significantly higher than galt2-1. These findings provide evidence that DoGALT2 might be involved in regulating the biosynthesis of galactose-containing polysaccharides during D. officinale pollen development.


Assuntos
Dendrobium/enzimologia , Flores/enzimologia , Galactosiltransferases/metabolismo , Genes de Plantas/fisiologia , Mucilagem Vegetal/biossíntese , Proteínas de Plantas/metabolismo , Arabidopsis , Clonagem Molecular , Dendrobium/genética , Dendrobium/metabolismo , Flores/química , Flores/metabolismo , Galactose/metabolismo , Galactosiltransferases/genética , Mucilagem Vegetal/química , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Polissacarídeos/análise , Transcriptoma
11.
J Dairy Sci ; 102(11): 9740-9748, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31477300

RESUMO

ß-Galactosidase is one of the most important enzymes used in dairy industry. Here, a novel thermostable ß-galactosidase was cloned and overexpressed from Bacillus coagulans NL01 in Escherichia coli. The phylogenetic trees were constructed using neighbor-joining methods. Phylogeny and amino acid analysis indicated that this enzyme belonged to family 42 of glycoside hydrolases. The optimal pH and temperature were, respectively, 6.0 and 55 to 60°C. The purified enzyme had a 3.5-h half-life at 60°C. Enzyme activity was enhanced by Mn2+. Compared with other ß-galactosidases from glycoside hydrolase family 42, B. coagulans ß-galactosidase exhibited excellent hydrolysis activity. The Michaelis constant (Km) and maximum rate of enzymatic reaction (Vmax) values for p-nitrophenyl-ß-d-galactopyranoside and o-nitrophenyl-ß-d-galactopyranoside were 1.06 mM, 19,383.60 U/mg, and 2.73 mM, 5,978.00 U/mg, respectively. More importantly, the enzyme showed lactose hydrolysis ability superior to that of the commercial enzyme. The specific enzyme activity for lactose was 27.18 U/mg. A total of 104.02 g/L lactose in whey was completely hydrolyzed in 3 h with addition of 2.38 mg of pure enzyme per gram of lactose. In view of the high price of commercial ß-galactosidase, B. coagulans ß-galactosidase could be a promising prototype for development of commercial enzymes aimed at lactose treatment in the dairy industry.


Assuntos
Bacillus coagulans/enzimologia , Lactose/metabolismo , Soro do Leite/química , beta-Galactosidase/metabolismo , Sequência de Aminoácidos , Animais , Bacillus coagulans/classificação , Estabilidade Enzimática , Galactose/metabolismo , Temperatura Alta , Concentração de Íons de Hidrogênio , Hidrólise , Cinética , Filogenia , Alinhamento de Sequência , Especificidade por Substrato , Temperatura Ambiente , Soro do Leite/metabolismo , beta-Galactosidase/química
12.
Anal Chim Acta ; 1082: 37-48, 2019 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-31472711

RESUMO

Magnetic nanoparticles decorated with d-galactose and galactitol (Fe3O4@SiN-galactose and Fe3O4@SiN-galactitol) were synthesized and employed as sorbent in a magnetic solid phase extraction (MSPE) procedure prior the analysis of aminoglycosides (AGs) in honey samples by LC-MS/MS. AGs are broad spectrum antibiotics, characterized by aminosugars, widespread used in therapeutic and veterinary applications. AGs can be found in the environment and food of animal origin. Fe3O4@SiN-galactose and Fe3O4@SiN-galactitol were synthesized via copper catalyzed alkyne azide cycloaddition and the synthesis was efficiently followed by infrared spectroscopy. They were characterized by electron microscopy, thermogravimetric analysis and magnetization curves. The nature of the loading (acetonitrile:water, 50:50 v/v) and elution solution (formic acid 190 mM) were studied in order to optimize the MSPE. Quantitative difference between MSPE with Fe3O4@SiN-galactose and MSPE with Fe3O4@SiN-galactitol in terms of recovery was found. The final optimized method using Fe3O4@SiN-galactose and Fe3O4@SiN-galactitol was applied in the determination of AGs in honey. The MSPE performance of Fe3O4@SiN-galactitol was found to be superior to that of MSPE with Fe3O4@SiN-galactose. The limits of quantification were between 2 and 19 µg kg-1 for amikacin, dihydrostreptomycin, tobramicyn and gentamycin. A good correlation between predicted and nominal values of AGs in honey was found (trueness from 84% to 109%). This MSPE procedure not only requires a minimum amount of sorbent (1 mg) and sample (0.2 g), but it can also be accomplish in a rather short time.


Assuntos
Aminoglicosídeos/análise , Nanopartículas de Magnetita/química , Adsorção , Aminoglicosídeos/química , Cromatografia Líquida de Alta Pressão , Galactitol/química , Galactose/química , Mel/análise , Limite de Detecção , Extração em Fase Sólida/métodos , Espectrometria de Massas em Tandem
13.
Nat Commun ; 10(1): 3657, 2019 08 14.
Artigo em Inglês | MEDLINE | ID: mdl-31413260

RESUMO

We lack an understanding of how the full range of genetic variants that occur in individuals can interact. To address this shortcoming, here we combine diverse mutations between genes in a model regulatory network, the galactose (GAL) switch of budding yeast. The effects of thousands of pairs of mutations fall into a limited number of phenotypic classes. While these effects are mostly predictable using simple rules that capture the 'stereotypical' genetic interactions of the network, some double mutants have unexpected outcomes including constituting alternative functional switches. Each of these 'harmonious' genetic combinations exhibits altered dependency on other regulatory genes. These cases illustrate how both pairwise and higher epistasis determines gene essentiality and how combinations of mutations rewire regulatory networks. Together, our results provide an overview of how broad spectra of mutations interact, how these interactions can be predicted, and how diverse genetic solutions can achieve 'wild-type' phenotypic behavior.


Assuntos
Proteínas de Ligação a DNA/genética , Galactose/metabolismo , Proteínas Repressoras/genética , Proteínas de Saccharomyces cerevisiae/genética , Fatores de Transcrição/genética , Epistasia Genética , Galactoquinase/genética , Regulação Fúngica da Expressão Gênica , Redes Reguladoras de Genes , Proteínas de Transporte de Monossacarídeos/genética , Mutação , Saccharomyces cerevisiae/genética , Biologia de Sistemas , Transativadores/genética
14.
Medicine (Baltimore) ; 98(33): e16828, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31415402

RESUMO

RATIONALE: Congenital glucose-galactose malabsorption (CGGM) is a rare, autosomal recessive, hereditary disease that usuallypresents in newborns. CGGM manifests as severe diarrhea, hyperosmolar dehydration, and malnutrition. It does not respond to routine treatment and often is life-threatening. PATIENT CONCERNS: We described a Chinese infant girl with refractory diarrhea, who suffered from severe dehydration and malnutrition even if with fluid replacement therapy and fed with several special formulas. DIAGNOSES: The genetic analysis identified CGGM with SLC5A1 mutations. c.1436G > C (p.R479T) was a novel mutation. INTERVENTIONS: The patient was managed by free-glucose and galactose formula, and then special low-carbohydrate dietary therapy. OUTCOMES: The patient improved immediately after starting a free-glucose and galactose formula, and kept healthy with special low-carbohydrate diet. She had been followed up with nutritional management for 20 months. LESSONS: This report highlights the importance of differential diagnosis of congenital diarrhea and enteropathies. For CGGM, free-glucose and galactose milk powder was the most effective treatment. Low-carbohydrate diet gradually introduced was still a great challenge that requires continuing guidance from child nutritionists and dietitians. Long-term nutrition management was extremely important to ensure the normal growth and development of children.


Assuntos
Erros Inatos do Metabolismo dos Carboidratos/tratamento farmacológico , Dieta com Restrição de Carboidratos/métodos , Fórmulas Infantis , Síndromes de Malabsorção/tratamento farmacológico , Erros Inatos do Metabolismo dos Carboidratos/genética , China , Feminino , Galactose , Glucose , Humanos , Recém-Nascido , Síndromes de Malabsorção/genética , Mutação , Transportador 1 de Glucose-Sódio/genética
15.
Transplant Proc ; 51(6): 2043-2050, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31399182

RESUMO

BACKGROUND: Pigs are considered suitable animal donor models for xenotransplantation. For successful organ transplantation, immune rejection must be overcome. Xenotransplantation has recently been successfully performed using galactose-alpha1,3-galactose epitopes knockout (GalTKO) and a human membrane cofactor protein (hCD46) in a pig model. However, the growth and lifespan of the grafted organ have not been evaluated. Therefore, in the present study we evaluated aging and 84 senescence-related genes using the RT2 Profiler PCR array and whole blood samples from GalTKO/hCD46 Massachusetts General Hospital (MGH) pigs. METHODS: Experimental groups were double GalTKO/hCD46 (5-month-old), single GalTKO/hCD46 (2-year-old), and non-genetically modified (>3.5-year-old; control group within the same strain). Age-matched white hairless Yucatan (WHY) miniature pig groups were used as controls. RESULTS: Among the 19 senescence-related genes selected from the 84 genes for further evaluation, 13 were upregulated in the double GalTKO/hCD46 MGH pigs compared to control MGH pigs; however, in WHY pigs, only 4 genes were up- or down-regulated among the 19 genes. Moreover, in double GalTKO/hCD46 MGH and WHY pigs, the expression of the 19 genes changed only 1- to 2-fold, suggesting that there were no significant differences in senescence signals between the 2 pig lines. CONCLUSIONS: The present results indicate that the double GalTKO/hCD46 MGH pig might be a suitable model for human xenotransplantation studies. However, we used a limited number of experimental individuals, so further studies using larger experimental groups should be conducted to verify the present results.


Assuntos
Envelhecimento/genética , Animais Geneticamente Modificados , Galactose/deficiência , Proteína Cofatora de Membrana/genética , Transplante Heterólogo/métodos , Animais , Pré-Escolar , Modelos Animais de Doenças , Epitopos , Galactose/genética , Galactosiltransferases/genética , Técnicas de Inativação de Genes , Sobrevivência de Enxerto/imunologia , Xenoenxertos , Humanos , Pessoa de Meia-Idade , Suínos , Porco Miniatura
16.
J Agric Food Chem ; 67(35): 9820-9830, 2019 Sep 04.
Artigo em Inglês | MEDLINE | ID: mdl-31411471

RESUMO

Brain aging is commonly associated with neurodegenerative disorders, but the ameliorative effect of krill oil and the underlying mechanism remain unclear. In this study, the components of krill oil were measured, and the antiaging effects of krill oil were investigated in mice with d-galactose (d-gal)-induced brain aging via proteomics and gut microbiota analysis. Krill oil treatment decreased the expression of truncated dopamine- and cAMP-regulated phosphoproteins and proteins involved in the calcium signaling pathway. In addition, the concentrations of dopamine were increased in the serum (p < 0.05) and brain (p > 0.05) due to the enhanced expressions of tyrosine-3-monooxygenase and aromatic l-amino acid decarboxylase. Moreover, krill oil alleviated gut microbiota dysbiosis, decreased the abundance of bacteria that consume the precursor tyrosine, and increased the abundance of Lactobacillus spp. and short-chain fatty acid producers. This study revealed the beneficial effect of krill oil against d-gal-induced brain aging and clarified the underlying mechanism through proteomics and gut microbiota analysis.


Assuntos
Envelhecimento/efeitos dos fármacos , Encéfalo/fisiopatologia , Euphausiacea/química , Galactose/efeitos adversos , Microbioma Gastrointestinal/efeitos dos fármacos , Óleos/administração & dosagem , Envelhecimento/fisiologia , Animais , Bactérias/classificação , Bactérias/efeitos dos fármacos , Bactérias/genética , Bactérias/isolamento & purificação , Encéfalo/efeitos dos fármacos , Suplementos Nutricionais/análise , Humanos , Intestinos/efeitos dos fármacos , Intestinos/microbiologia , Masculino , Camundongos , Óleos/isolamento & purificação
17.
Int J Mol Sci ; 20(15)2019 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-31370181

RESUMO

The ability to control the glycosylation pattern of recombinant viral glycoproteins represents a major prerequisite before their use as vaccines. The aim of this study consisted of expressing the large soluble ectodomain of glycoprotein B (gB) from Human Cytomegalovirus (HMCV) in Nicotiana tabacum Bright Yellow-2 (BY-2) suspension cells and of comparing its glycosylation profile with that of gB produced in Chinese hamster ovary (CHO) cells. gB was secreted in the BY-2 culture medium at a concentration of 20 mg/L and directly purified by ammonium sulfate precipitation and size exclusion chromatography. We then measured the relative abundance of N-glycans present on 15 (BY-2) and 17 (CHO) out of the 18 N-sites by multienzymatic proteolysis and mass spectrometry. The glycosylation profile differed at each N-site, some sites being occupied exclusively by oligomannosidic type N-glycans and others by complex N-glycans processed in some cases with additional Lewis A structures (BY-2) or with beta-1,4-galactose and sialic acid (CHO). The profiles were strikingly comparable between BY-2- and CHO-produced gB. These results suggest a similar gB conformation when glycoproteins are expressed in plant cells as site accessibility influences the glycosylation profile at each site. These data thus strengthen the BY-2 suspension cultures as an alternative expression system.


Assuntos
Fragmentos de Peptídeos/química , Polissacarídeos/química , Proteínas do Envelope Viral/química , Sulfato de Amônio/química , Animais , Células CHO , Sequência de Carboidratos , Precipitação Química , Cromatografia em Gel/métodos , Cricetulus , Galactose/química , Expressão Gênica , Glicosilação , Humanos , Ácido N-Acetilneuramínico/química , Fragmentos de Peptídeos/isolamento & purificação , Células Vegetais/metabolismo , Polissacarídeos/isolamento & purificação , Proteólise , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Tabaco/citologia , Tabaco/metabolismo , Proteínas do Envelope Viral/genética , Proteínas do Envelope Viral/metabolismo
18.
J Agric Food Chem ; 67(36): 10059-10068, 2019 Sep 11.
Artigo em Inglês | MEDLINE | ID: mdl-31431007

RESUMO

Torularhodin is a natural product extracted from Sporidiobolus pararoseus and has a similar chemical structure to ß-carotene. The antioxidative effects of torularhodin were investigated using DPPH, ABTS, a cell oxidative damage model in vitro, and a d-galactose-induced liver-injured mouse model in vivo. Cell experiments demonstrated that torularhodin had a powerful effect on oxidative damage caused by H2O2 to AML12 cells. Torularhodin significantly reduced inflammatory cytokines and increased the activity of antioxidant enzymes both in mouse serum and the liver. The inhibition of d-galactose-induced oxidative damage in the liver was correlated with the torularhodin-mediated effects on improving the activity of Nrf2/HO-1, reducing the expression of Bax and NF-κB p65 by western blot analysis. RT-PCR results demonstrated torularhodin upregulated the antioxidative mRNA expression of Nrf2, NQO1, and HO-1 in the liver. In summary, torularhodin significantly scavenged free radicals and prevented oxidative damage in vitro and reduced d-galactose-induced liver oxidation via promotion of the Nrf2/HO-1 pathways in vivo.


Assuntos
Antioxidantes/administração & dosagem , Basidiomycota/química , Carotenoides/administração & dosagem , Doença Hepática Induzida por Substâncias e Drogas/tratamento farmacológico , Fator 2 Relacionado a NF-E2/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Animais , Doença Hepática Induzida por Substâncias e Drogas/etiologia , Doença Hepática Induzida por Substâncias e Drogas/genética , Doença Hepática Induzida por Substâncias e Drogas/metabolismo , Galactose/efeitos adversos , Heme Oxigenase-1/genética , Heme Oxigenase-1/metabolismo , Humanos , Fígado/efeitos dos fármacos , Fígado/metabolismo , Masculino , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Camundongos , Camundongos Endogâmicos ICR , NAD(P)H Desidrogenase (Quinona)/genética , NAD(P)H Desidrogenase (Quinona)/metabolismo , Fator 2 Relacionado a NF-E2/genética , NF-kappa B/genética , NF-kappa B/metabolismo , Transdução de Sinais/efeitos dos fármacos
19.
J Agric Food Chem ; 67(37): 10342-10351, 2019 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-31461273

RESUMO

Maltol, a maillard reaction product from ginseng (Panax ginseng C. A. Meyer), has been confirmed to inhibit oxidative stress in several animal models. Its beneficial effect on oxidative stress related brain aging is still unclear. In this study, the mouse model of d-galactose (d-Gal)-induced brain aging was employed to investigate the therapeutic effects and potential mechanisms of maltol. Maltol treatment significantly restored memory impairment in mice as determined by the Morris water maze tests. Long-term d-Gal treatment reduced expression of cholinergic regulators, i.e., the cholineacetyltransferase (ChAT) (0.456 ± 0.10 vs 0.211 ± 0.03 U/mg prot), the acetylcholinesterase (AChE) (36.4 ± 5.21 vs 66.5 ± 9.96 U/g). Maltol treatment prevented the reduction of ChAT and AChE in the hippocampus. Maltol decreased oxidative stress levels by reducing levels of reactive oxygen species (ROS) and malondialdehyde (MDA) production in the brain and by elevating antioxidative enzymes. Furthermore, maltol treatment minimized oxidative stress by increasing the phosphorylation levels of phosphatidylinositol-3-kinase (PI3K), protein kinase B (Akt), nuclear factor-erythroid 2-related factor 2 (Nrf2), and hemeoxygenase-1 (HO-1). The above results clearly indicate that supplementation of maltol diminishes d-Gal-induced behavioral dysfunction and neurological deficits via activation of the PI3K/Akt-mediated Nrf2/HO-1 signaling pathway in brain. Maltol might become a potential drug to slow the brain aging process and stimulate endogenous antioxidant defense capacity. This study provides the novel evidence that maltol may slow age-associated brain aging.


Assuntos
Envelhecimento/efeitos dos fármacos , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Galactose/efeitos adversos , Heme Oxigenase-1/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Extratos Vegetais/administração & dosagem , Pironas/administração & dosagem , Envelhecimento/metabolismo , Animais , Heme Oxigenase-1/genética , Humanos , Masculino , Malondialdeído/metabolismo , Camundongos , Camundongos Endogâmicos ICR , Fator 2 Relacionado a NF-E2/genética , Estresse Oxidativo/efeitos dos fármacos , Panax/química , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Espécies Reativas de Oxigênio/metabolismo
20.
Biomed Pharmacother ; 117: 109204, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31387177

RESUMO

We wished to investigate the role of a tilapia skin collagen polypeptide (TSCP; molecular weight <3 kDa) in alleviating liver and kidney injuries in aging mice induced by d-galactose (d-gal) and its underlying mechanism of action. First, we characterized TSCP. TSCP was passed through a 3-kDa ultrafiltration membrane, desalted in water by a solid-phase extraction column, purified further by reverse phase-high performance liquid chromatography, and analyzed by electrospray ionization mass spectrometry and tandem mass spectrometry. TSCP contained 17 types of amino acids (AAs) and 41 peptide chains of length 7 AAs to 22 AAs. The content of free AAs and total AAs of TSCP was 13.5% and 93.79%, respectively. Next, we undertook animal experiments. Mice were injected once-daily with D-gal (300 mg/kg body weight, s.c.) for 8 weeks, and TSCP was administered simultaneously once-daily by intragastric gavage. TSCP could visibly improve the decreased body weight, depressed appetite, and mental deterioration of mice triggered by d-gal. TSCP could also alleviate d-gal-induced damage to the liver and kidneys according to histopathology (especially high-dose TSCP). Consistent with these macroscopic and pathologic changes, TSCP could also prevent d-gal-induced increases in serum levels of alanine aminotransferase, aspartate transaminase, alkaline phosphatase, lipid peroxidation, creatinine and uric acid, as well as decreases in serum levels of immunoglobulin (Ig)G and IgM. Moreover, TSCP improved the activities of superoxide dismutase, catalase, and glutathione peroxidase, but also inhibited the increases in the levels of malondialdehyde and inducible nitric oxide synthase expression in the liver and kidneys of d-gal-treated mice. These results suggest that TSCP can alleviate the injuries to the liver and kidneys in aging mice induced by d-gal, and that its mechanism of action might be, at least partially, associated with attenuation of oxidative stress and enhancement of immune function.


Assuntos
Colágeno/farmacologia , Galactose/efeitos adversos , Rim/efeitos dos fármacos , Fígado/efeitos dos fármacos , Peptídeos/farmacologia , Substâncias Protetoras/farmacologia , Tilápia/metabolismo , Alanina Transaminase/metabolismo , Animais , Antioxidantes/metabolismo , Aspartato Aminotransferases/metabolismo , Catalase/metabolismo , Glutationa Peroxidase/metabolismo , Rim/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Fígado/metabolismo , Masculino , Malondialdeído/metabolismo , Camundongos , Estresse Oxidativo/efeitos dos fármacos , Superóxido Dismutase/metabolismo
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