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1.
J Microencapsul ; 41(4): 296-311, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38709162

RESUMO

AIMS: To construct the microemulsion delivery system (ME) loading ATSO and NA and study their physicochemical characteristics to enhance their stability and water solubility. METHODS: By plotting ternary phase diagrams, the composition and proportions of the MEs were determined. The physicochemical characteristics and stability of MEs were evaluated by mean diameter, polydispersity index (PDI), pH, electrical conductivity, transmission electron microscopy (TEM), rheological behaviour measurement, and phase inversion temperature (PIT). RESULTS: The MEs was composed with EL-40 as a surfactant and specifically with the addition of ethanol as a cosurfactant in NA-loaded ME. The mean diameters of ATSO-loaded ME and NA-loaded ME were 39.65 ± 0.24 nm and 32.90 ± 2.65 nm, and PDI were 0.49 ± 0.01 and 0.28 ± 0.14, respectively. The TEM confirmed the spherical and smooth morphology of MEs. The rheological results indicated that MEs are dilatant fluids with the advantages of low viscosity, high fluidity, and tolerance to temperature fluctuations. The mean diameter and PDI of MEs showed no significant change after storage at 25 °C for 28 days and centrifugation. CONCLUSION: The prepared microemulsions could expand the application prospects of ATSO and NA products in cosmetics, medicine, foods and other fields.


Assuntos
Emulsões , Óleos de Plantas , Reologia , Emulsões/química , Óleos de Plantas/química , Acer/química , Ácidos Graxos/química , Sementes/química , Tensoativos/química , Estabilidade de Medicamentos , Viscosidade
2.
Arch Anim Nutr ; 78(1): 30-44, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38436931

RESUMO

Cassava protein (CP), barley protein (BP) and yellow pea protein (YPP) are important nutrient and integral constituent of staple in pet foods. It is known that the digestion of proteins directly influences their absorption and utilisation. In the present work, we performed in vitro simulated gastrointestinal digestion of three plant proteins as a staple for dog and cat food. The digestion rate of CP, BP and YPP in dog food was 56.33 ± 0.90%, 48.53 ± 0.91%, and 66.96 ± 0.37%, respectively, whereas the digestion rate of CP, BP, and YPP in cat food was 66.25 ± 0.72%, 43.42 ± 0.83%, and 58.05 ± 0.85%, respectively. Using SDS-polyacrylamide gel electrophoresis to determine the molecular weight (MW) of each protein and the products of their digestion, it was revealed that MW of digestion samples decreased, and MW during the small intestine phase was lower than that during the gastric phase. Peptide sequences of digested products were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS), and it was found that the total number of peptides in the small intestine digestion samples was higher than that in the gastric phase samples. The MW of peptides obtained from CP was within the range of 1000-1500 Da, while MW of peptides derived from BP and YPP was within the range of 400-2000 Da. In addition, free amino acids were mainly produced in the small intestine phase. Furthermore, the percentage of essential amino acids in the small intestine phase (63 ~ 82%) was higher than that in the gastric phase (37 ~ 63%). Taken together, these findings contribute to the current understanding of the utilisation of plant proteins in dog and cat foods and provide important insights into the selection and application of plant proteins as a staple in dog and cat foods.


Assuntos
Aminoácidos , Digestão , Peptídeos , Digestão/fisiologia , Aminoácidos/metabolismo , Aminoácidos/química , Animais , Peptídeos/metabolismo , Peptídeos/química , Ração Animal/análise , Proteínas de Plantas/metabolismo , Proteínas de Plantas/química , Hordeum/química , Hordeum/metabolismo , Manihot/química , Manihot/metabolismo , Pisum sativum/química , Pisum sativum/metabolismo , Cães , Proteínas de Ervilha/química , Proteínas de Ervilha/metabolismo , Gatos , Espectrometria de Massas em Tandem/veterinária , Trato Gastrointestinal/metabolismo , Trato Gastrointestinal/fisiologia , Trato Gastrointestinal/química
3.
Anal Chem ; 96(2): 676-684, 2024 01 16.
Artigo em Inglês | MEDLINE | ID: mdl-38173079

RESUMO

Identification of the phosphatidylserine (PS) discrepancies occurring on the cellular membrane during apoptotic processes is of the utmost importance. However, monitoring the quantity of PS molecules in real-time at a single-cell level currently remains a challenging task. Here, we demonstrate this objective by leveraging the specific binding and reversible interaction exhibited by the zinc(II) dipyridinamine complex (ZnDPA) with PS. Lipoic acid-functionalized ZnDPA (LP-ZnDPA) was subsequently immobilized onto the surface of an atomic force microscopy cantilever to form a force probe, ALP-ZnDPA, enabling a PS-specific dynamic imaging and detection mode. By utilizing this technique, we can not only create a heat map of the expression level of PS with submicron resolution but also quantify the number of molecules present on a single cell's surface with a detection limit of 1.86 × 104 molecules. The feasibility of the proposed method is demonstrated through the analysis of PS expression levels in different cancer cell lines and at various stages of paclitaxel-induced apoptosis. This study represents the first application of a force probe to quantify PS molecules on the surface of individual cells, providing insight into dynamic changes in PS content during apoptosis at the molecular level and introducing a novel dimension to current detection methodologies.


Assuntos
Fosfatidilserinas , Imagem Individual de Molécula , Fosfatidilserinas/química , Apoptose , Membrana Celular/metabolismo , Microscopia de Força Atômica/métodos , Análise Espectral
4.
J Sci Food Agric ; 104(2): 788-796, 2024 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-37669105

RESUMO

BACKGROUND: Calcium is important in the formation of bones and teeth, cell metabolism, and other physiological activities. In this work, casein phosphopeptide-calcium chelate (CPP-Ca) was synthesized and the optimal process parameters for the chelation reaction were obtained. The bioavailability of calcium in CPP-Ca was investigated by in vitro gastrointestinal simulated digestion. The existence of phytic acid and oxalic acid in the digestion system was evaluated to clarify the calcium holding ability of casein phosphopeptide (CPP). Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to identify oligopeptides from CPP-Ca. RESULTS: The optimal process parameters for the chelation reaction were: peptide concentration 7.76 mgmL-1 , pH 8.54, and reaction temperature 43.3 °C. The digestion in vitro results indicated that the calcium release rate of CPP-Ca in the stomach for 2 h reached 85%, and about 50% of the ionized calcium was re-chelated with CPP in the intestine. Phytic acid and oxalic acid could lead to a sharp decrease in soluble calcium but around 50% of the calcium was still retained in the form of chelates in the presence of CPP. The LC-MS/MS identified 19 casein-derived oligopeptides after digestion, and calcium modifications were found on eight peptides derived from ß-casein and αs2 -casein. CONCLUSIONS: This study clarified the excellent calcium holding capacity of CPP in the presence of phytic acid and oxalic acid. Liquid chromatography-tandem mass spectrometry also revealed peptide changes, and identified peptides that chelate with calcium. These findings provided significant insights that could be relevant to the further utilization and product development of peptide-calcium chelate in the food industry. © 2023 Society of Chemical Industry.


Assuntos
Cálcio , Fragmentos de Peptídeos , Cálcio/metabolismo , Fosfopeptídeos/química , Fosfopeptídeos/metabolismo , Caseínas/química , Cromatografia Líquida , Ácido Fítico , Espectrometria de Massas em Tandem , Cálcio da Dieta , Digestão , Oligopeptídeos , Ácido Oxálico
5.
Microbiol Spectr ; 11(4): e0059023, 2023 08 17.
Artigo em Inglês | MEDLINE | ID: mdl-37428087

RESUMO

Changes in diet and environment can lead to acute diarrhea in companion animals, but the composition and interactions of the gut microbiome during acute diarrhea remain unclear. In this multicenter case-control study, we investigated the relationship between intestinal flora and acute diarrhea in two breeds of cats. Acutely diarrheic American Shorthair (MD, n = 12) and British Shorthair (BD, n = 12) and healthy American Shorthair (MH, n = 12) and British Shorthair (BH, n = 12) cats were recruited. Gut microbial 16S rRNA sequencing, metagenomic sequencing, and untargeted metabolomic analysis were performed. We observed significant differences in beta-diversity (Adonis, P < 0.05) across breeds and disease state cohorts. Profound differences in gut microbial structure and function were found between the two cat breeds. In comparison to healthy British Shorthair cats, Prevotella, Providencia, and Sutterella were enriched while Blautia, Peptoclostridium, and Tyzzerella were reduced in American Shorthair cats. In the case-control cohort, cats with acute diarrhea exhibited an increased abundance of Bacteroidota, Prevotella, and Prevotella copri and a decreased abundance of Bacilli, Erysipelotrichales, and Erysipelatoclostridiaceae (both MD and BD cats, P < 0.05). Metabolomic analysis identified significant changes in the BD intestine, affecting 45 metabolic pathways. Moreover, using a random forest classifier, we successfully predicted the occurrence of acute diarrhea with an area under the curve of 0.95. Our findings indicate a distinct gut microbiome profile that is associated with the presence of acute diarrhea in cats. However, further investigations using larger cohorts of cats with diverse conditions are required to validate and extend these findings. IMPORTANCE Acute diarrhea is common in cats, and our understanding of the gut microbiome variations across breeds and disease states remains unclear. We investigated the gut microbiome of two cat breeds (British Shorthair and American Shorthair) with acute diarrhea. Our study revealed significant effects of breeds and disease states on the structure and function of the gut microbiota in cats. These findings emphasize the need to consider breed-related factors in animal nutrition and research models. Additionally, we observed an altered gut metabolome in cats with acute diarrhea, closely linked to changes in bacterial genera. We identified a panel of microbial biomarkers with high diagnostic accuracy for feline acute diarrhea. These findings provide novel insights into the diagnosis, classification, and treatment of feline gastrointestinal diseases.


Assuntos
Microbioma Gastrointestinal , Gatos , Animais , RNA Ribossômico 16S/genética , Estudos de Casos e Controles , Fezes/microbiologia , Diarreia/veterinária , Diarreia/microbiologia , Firmicutes/genética
6.
Arch Microbiol ; 205(3): 97, 2023 Feb 24.
Artigo em Inglês | MEDLINE | ID: mdl-36823480

RESUMO

For different breeds of dogs with acute diarrhea, the gut microbiota and metabolome profiles are unclear. This prospective observational study analyzed the gut microbiomes of poodles with acute diarrhea and Labrador retrievers with acute diarrhea based on 16S amplicon sequencing, with respective healthy dogs as controls. Fecal non-target metabolomics and metagenomics were performed on poodles with acute diarrhea. This study found that the diversity and structure of the microbial community differed significantly between the two breeds in cohorts of healthy dogs. Two breeds of dogs with acute diarrhea demonstrated different changes in microbial communities and metabolic functions. The metabolism of starch and sucrose was significantly decreased in dogs with acute diarrhea, which may be attributed to the reduced activity of dextran dextrinase. Non-targeted metabolomics identified 21 abnormal metabolic pathways exhibited by dogs with acute diarrhea, including starch, amino acid, bile acid metabolism, etc., and were closely related to specific intestinal flora. This study provided new insights into breed specificity and the development of dietary treatment strategy in canine gastrointestinal disease.


Assuntos
Microbioma Gastrointestinal , Microbiota , Cães , Animais , Metabolômica , Metaboloma , Amido/análise , Diarreia , Fezes , RNA Ribossômico 16S
7.
Vet Res Commun ; 47(2): 791-801, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-36456856

RESUMO

Exposure to environmental particulate matter (PM) causes lung damage in humans, but it is not sufficiently studied in companion animals. In this work, we found that organic extracts (OE) of PM induced oxidative stress and were more cytotoxic than water-soluble extracts (WE) of PM in canine (Canis familiaris) pulmonary alveolar type II epithelial (PAE) cells. Subsequently, we evaluated the alleviating effects of polyunsaturated fatty acid mixtures (eicosapentaenoic and docosahexaenoic acids), vitamin E, and lycopene on OE-exposed PAE cells. The results indicated that the three nutrients attenuated OE-induced oxidative stress. Compared with OE-exposed PAE cells, cells pretreated with the three nutrients exhibited a 1.7 ~ 2.2-fold reduction in reactive oxygen species, 15.58% ~ 19.96% increase in cell viability, 26.19% ~ 29.32% reduction in lactate dehydrogenase release and 33.87% ~ 40.10% reduction in intracellular malondialdehyde. Meanwhile, the activities of superoxide dismutase, catalase, and glutathione peroxidase increased by 35.22% ~ 47.70%, 45.36% ~ 64.13%, and 48.56% ~ 68.18%. Besides, microscopic observation revealed that nutrients improved cell morphology, as evidenced by reduced cell shrinkage and increased apposition. Finally, the mechanisms of OE toxicity and antioxidant nutrients were explored respectively. The results revealed that OE triggered cytotoxicity by directly disrupting antioxidant enzyme activity through activation of ROS, while nutrients inhibited OE-induced cellular oxidative stress via upregulation of the Nrf2/HO-1 signaling pathway. The present study may provide significant insights into the prevention of PM-induced lung diseases by antioxidant supplementation in animals.


Assuntos
Fator 2 Relacionado a NF-E2 , Material Particulado , Humanos , Cães , Animais , Material Particulado/toxicidade , Material Particulado/metabolismo , Licopeno/farmacologia , Licopeno/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Fator 2 Relacionado a NF-E2/farmacologia , Antioxidantes/farmacologia , Vitamina E/farmacologia , Pulmão , Estresse Oxidativo , Ácidos Graxos Insaturados/farmacologia , Espécies Reativas de Oxigênio/farmacologia
8.
Food Chem ; 401: 134218, 2023 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-36115235

RESUMO

In this work, CPP-Ca chelate was synthesized by chelating casein phosphopeptide (CPP) and calcium and characterized by Fourier transform infrared spectroscopy (FTIR), Scanning electron microscopy (SEM) Energy dispersive spectroscopy (EDS) and X-ray diffraction (XRD). The antioxidant activity and calcium holding capacity of CPP-Ca were evaluated and its secondary structure transition was monitored during gastrointestinal digestion by in situ Raman spectroscopy. The results demonstrated that calcium chelating rate reached 40 % and calcium ion was bound to CPP mainly through the interaction of carboxyl and amino groups. The result of calcium holding capacity confirmed the formation of calcium phosphate precipitates could be delayed by 10-15 min with increasing CPP concentration. In vitro simulated digestion revealed CPP-Ca exhibited excellent calcium solubility and its secondary structural changes occurred, especially α-helix and ß-sheet content. These findings provided significant insights into enhancing bioavailability of calcium supplements and developing of calcium functional foods for human and animals.


Assuntos
Caseínas , Fosfopeptídeos , Animais , Humanos , Caseínas/química , Cálcio/química , Antioxidantes , Cálcio da Dieta , Fosfatos de Cálcio , Digestão
9.
J Food Sci ; 87(9): 3925-3937, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-35904249

RESUMO

Acer truncatum seed oil (ATSO) contains abundant unsaturated fatty acids, with significant quantities of nervonic acid (NA, > 5%), which was authenticated as a new food resource in China. For the sake of minimizing animal consumption and the importance of NA for human health, extraction of NA from plants has become a research hotspot. In the present study, three extraction factors were determined to significantly influence the saponification reaction based on single-factor experiments: NaOH dosage, reaction time, and reaction temperature. These three factors were used to further optimize the saponification process through the response surface methodology, and the highest yield of mixed fatty acids was 83.12%. Moreover, the activation energy (40.8228 kJ/mol), the pre-exponential factor [2.568 × 106 m3 /(kmol·min)], and the kinetic equation [rA = kcA cB = 2.568 × 106 ·exp(- 4970 . 1 T ) $\frac{{{\rm{4970}}{\rm{.1}}}}{{\rm{T}}})$ cA cB ] of the ATSO saponification reaction were determined by combining the chemical reaction rate equation of the elementary reaction, the Arrhenius equation, and the NaOH concentration in the substrate. Finally, the mixed fatty acids of ATSO were crystallized by triple-stage low-temperature crystallization, and we achieved 25.05% purity for NA. This study provides a technological basis and strategy for specific fatty acid production from ASTO, as well as other vegetable oils important in the field of food and health supplement products. PRACTICAL APPLICATION: Nervonic acid (NA) is an essential component of neural cells and neural tissue, and it is vital for maintaining the normal work of nerve tissues in organisms and promotes neurodevelopment. NA has traditionally been mainly obtained from shark hunting, which is now restricted due to an international ban on shark fishing. The alternative way to produce NA cheaply and in large quantities is from plant sources. The techniques utilized in this study provide an effective method of NA separation from Acer truncatum seed oil for industrial production.


Assuntos
Acer , Acer/química , Cristalização , Ácidos Graxos/análise , Ácidos Graxos Monoinsaturados , Humanos , Cinética , Óleos de Plantas/química , Sementes/química , Hidróxido de Sódio , Tecnologia
10.
Heliyon ; 8(5): e09550, 2022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35663753

RESUMO

Ambient particulate matter (APM) is extremely harmful to life's health. In this study, we investigated cellular injury in cat (Felix catus) lung cells (FCA-L2) exposed to organic and water-soluble extracts from APM. As well, the protective effect of vitamin E (VE), lycopene and a mixture of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) (molar concentration ratio of 2:1) against this damage was evaluated. Organic and water-soluble extracts induced oxidative stress in FCA-L2 cells, as evidenced by excess reactive oxygen species production and mitochondrial damage, while treatment with VE, lycopene and EPA: DHA remarkably alleviated these indices. It was further found that treatment with EPA: DHA decreased lactate dehydrogenase and malondialdehyde, as well as increased activities of superoxide dismutase, glutathione peroxidase and catalase. Our study confirmed that nutrients mediates APM-induced oxidative stress via antioxidant proteins. Also, these findings could provide new insights into reducing APM-induced cytotoxicity by nutritional supplementation based on antioxidant compounds for animals.

11.
J Sci Food Agric ; 101(10): 4350-4360, 2021 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-33420734

RESUMO

BACKGROUND: Melt crystallization is typically recognized as a highly efficient and green method for oil fractionation. This work concentrated on novel layer melt crystallization for preparing desirable olein and stearin products from palm oil and the evaluation of fraction quality. Layer melt crystallization was performed at various temperatures and the effects on fractions were evaluated using iodine value (IV), solid fat content (SFC) and melting point. The lipid composition, thermal and crystallization properties, and phase behaviors of the final optimized fractions were determined using gas chromatography, high-performance liquid chromatography-atmospheric pressure chemical ionization mass spectrometry and differential scanning calorimetry. RESULTS: Increasing crystallization tube temperatures under the same jacket temperature increased the melting point and SFC, while decreasing the IV of the olein product. Opposite results were observed for the stearin product. Major fatty acids in fractions were determined as palmitic acid and oleic acid. 1,2-Dioleoyl-3-palmitoylglycerol and 1,3-dipalmitoyl-2-oleoylglycerol were identified as the main triacylglycerols in olein and stearin fractions, respectively. A critical effect of crystallization temperature on co-crystallization of oleins and stearins was revealed. A transition from plate-like crystal growth to spherulitic growth with spontaneous nucleation was indicated in palm oil and stearin fractions with increasing crystallization temperature. As for olein fractions, a temperature increase resulted in heterogeneous nucleation from instantaneous nucleation. CONCLUSIONS: Novel layer melt crystallization was successfully applied and optimized for fractionating palm oil. The composition and property changes of obtained fractions were analyzed and explained at both macroscopic and microscopic levels. © 2021 Society of Chemical Industry.


Assuntos
Lipídeos/química , Ácido Oleico/química , Óleo de Palmeira/química , Ácidos Esteáricos/química , Varredura Diferencial de Calorimetria , Cromatografia Líquida de Alta Pressão , Cristalização , Espectrometria de Massas , Temperatura
12.
J Sci Food Agric ; 101(2): 703-717, 2021 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-32710440

RESUMO

BACKGROUND: Ziziphus jujuba Miller cv. Dongzao is extremely susceptible to reddening, browning, nutritional loss, and perishability after harvest. In this study, we evaluated the mechanisms of calcium chloride and chitosan/nano-silica composite film treatments on the quality, especially in reddening, by physiological and metabolomic assays. RESULTS: The treatment delayed the decline of phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS), and chalcone isomerase (CHI) activities. Meanwhile, the treated groups retarded the increases in anthocyanin and quercetin contents by inhibiting the gene expressions of flavonol synthase (ZjFLS), dihydroflavonol 4-reductase (ZjDFR), and anthocyanidin synthase (ZjANS), while promoting leucoanthocyanidin reductase (ZjLAR) expression, which leads to retardation of fruit reddening. Anthocyanins were found to be responsible for post-harvest winter jujube reddening through principal component analysis. Results from the technique for order preference by similarity to an ideal solution indicated that the treated group delayed the decline of the quality of 'Dongzao' and extended its shelf life. CONCLUSION: The treatment induced the heightening of flavonoids metabolism. They enhanced the nutritional value and the ability to resist stress by delaying the decline of PAL, CHS, and CHI activities. Meanwhile, the treated groups retarded the increase in anthocyanin and quercetin contents by inhibiting the gene expressions of ZjFLS, ZjDFR, and ZjANS and promoting ZjLAR expression, which leads to retardation of fruit reddening. Anthocyanins are responsible for post-harvest winter jujube reddening. Coating treatment effectively delayed the decline of winter jujube quality. © 2020 Society of Chemical Industry.


Assuntos
Cloreto de Cálcio/farmacologia , Conservação de Alimentos/métodos , Frutas/química , Ziziphus/efeitos dos fármacos , Antocianinas/análise , Antocianinas/metabolismo , Conservação de Alimentos/instrumentação , Frutas/efeitos dos fármacos , Frutas/enzimologia , Frutas/genética , Regulação da Expressão Gênica de Plantas , Oxirredutases/genética , Oxirredutases/metabolismo , Oxigenases/genética , Oxigenases/metabolismo , Fenilalanina Amônia-Liase/genética , Fenilalanina Amônia-Liase/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Quercetina/análise , Quercetina/metabolismo , Ziziphus/química , Ziziphus/enzimologia , Ziziphus/genética
13.
J Agric Food Chem ; 68(33): 8996-9003, 2020 Aug 19.
Artigo em Inglês | MEDLINE | ID: mdl-32806119

RESUMO

In this work, a microemulsion emitting fluorescence was fabricated as a potential oral delivery system for bioactive compounds. In simulated oral administration, the microemulsion was characterized for its microstructure by 1hydrogen-nuclear magnetic resonance (1H-NMR). Results showed that microemulsions not only have good resistance to oral and gastric phases, but also lay a solid foundation for the release of bioactive compounds in the intestine. Fluorescence stability tests showed that microemulsions exhibit a remarkable fluorescence intensity in the digestive environment, indicating feasibility as a label-free delivery carrier. Moreover, in vitro release tests of bioactive compounds confirmed that an α-linolenic acid (ALA)-loaded microemulsion mainly released in the intestine, thereby achieving the aim of controlling the release of bioactive compounds. These results suggest that the synthesized fluorescent microemulsion, combining the favorable features of nontoxicity, antidigestive stability, remarkable fluorescence intensity, and controllable release, can be regarded as a promising label-free delivery carrier for oral administration.


Assuntos
Sistemas de Liberação de Medicamentos/instrumentação , Emulsões/química , Ácido alfa-Linolênico/química , Administração Oral , Emulsões/administração & dosagem , Fluorescência , Espectroscopia de Ressonância Magnética , Tamanho da Partícula , Ácido alfa-Linolênico/administração & dosagem
14.
J Agric Food Chem ; 67(41): 11518-11526, 2019 Oct 16.
Artigo em Inglês | MEDLINE | ID: mdl-31513385

RESUMO

In this work, the effects of α-linolenic acid (ALA) loaded in oil-in-water (O/W) and water-in-oil-in-water (W/O/W) microemulsions on cell viability, lactic dehydrogenase (LDH) viability, and reactive oxygen species (ROS) levels were examined using Cell Counting Kit-8 (CCK-8), an LDH assay kit, and a fluorescence microscope, respectively. The CCK-8 assay demonstrated that ALA inhibited MDA-MB-231 human breast cancer cell proliferation in a dose-dependent manner. Further, the results of LDH activity and ROS levels revealed that ALA-induced cancer cell damage was closely related to oxidative stress. Under the irradiation of ultraviolet light, the microemulsion without any added fluorescent dye would emit bright blue fluorescence, and the fluorescent images of the cells treated with ALA-loaded O/W and W/O/W microemulsions at different incubation times were taken, which exhibited long-term photostability and biocompatibility. In addition, the fluorescence mechanism of the microemulsion was explained by immobilizing surfactant molecules with aggregation-induced emission (AIE) properties at the water-oil interface through the microemulsion with a self-assembled structure. These findings showed the potential application of O/W and W/O/W microemulsions as the label-free delivery carriers in long-term imaging of living cells and real-time release monitoring of nutrients.


Assuntos
Proliferação de Células/efeitos dos fármacos , Ácido alfa-Linolênico/química , Ácido alfa-Linolênico/farmacologia , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Portadores de Fármacos/química , Emulsões/química , Emulsões/farmacologia , Fluorescência , Humanos , Óleos/química , Espécies Reativas de Oxigênio/metabolismo , Água/química
15.
J Agric Food Chem ; 67(15): 4273-4279, 2019 Apr 17.
Artigo em Inglês | MEDLINE | ID: mdl-30929442

RESUMO

In this work, the effects of curcumin on the viability, morphology, and nanomechanics of SH-SY5Y neuronal cells were investigated using a conventional cell viability test kit (CCK-8) and sophisticated AFM imaging and force measurement techniques. CCK-8 tests show that SH-SY5Y neuronal cells have a dose-response to curcumin in terms of viability that is dependent on the exposure durations. When exposed to a maximum dosage of 32 µg/mL used in the present study for 4 h, 24 h, and 48 h, the cell viability dropped to 73.4 ± 4.5%, 9.1 ± 3.2%, and 2.5 ± 1.2% of the control, correspondingly. AFM studies show that curcumin can induce the disappearance of synapses of the cells and the change of biomechanics. After exposure for 24 h at the concentration of 16 µg/mL, the viscous deformation of the cells decreased from 2.15 ± 0.02 to 1.58 ± 0.03 (×10-15 N·m), the elastic deformation increased from 1.26 ± 0.04 to 1.72 ± 0.07 (×10-15 N·m), and adhesion work decreased from 0.56 ± 0.07 to 0.39 ± 0.04 (×10-16 N·m). The morphological and mechanical changes obtained using AFM can be interpreted from optically observed cellular structural changes. The present study provides insights into the effects of curcumin on neuronal cells from both biological and biophysical aspects, which can help more comprehensively understand the interactions between curcumin and SH-SY5Y cells. The demonstrated techniques can be potentially used to assess the efficacy of bioactive constituents on cells or help screen drugs.


Assuntos
Curcumina/farmacologia , Neurônios/citologia , Neurônios/efeitos dos fármacos , Apoptose/efeitos dos fármacos , Fenômenos Biomecânicos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Humanos , Microscopia de Força Atômica , Neurônios/química
16.
Spectrochim Acta A Mol Biomol Spectrosc ; 214: 239-245, 2019 May 05.
Artigo em Inglês | MEDLINE | ID: mdl-30785043

RESUMO

The binding domains of lysozyme with ionic liquids (ILs, [C4mim]BF4, [C4mim]Cl, [C4mim]Br and [dmim]I) in aqueous solution was investigated by studying molecular interactions using spectroscopic techniques. Ultraviolet spectroscopy (UV) showed that the addition of ILs increased the absorption peak intensity of lysozyme at 210 nm by enhancing peptide bond valence electron transition. It is also found that a weak interaction between ILs and lysozyme chromophore groups was generated by analyzing the changes of absorption peak intensity near 280 nm. Fluorescence and Synchronous Fluorescence spectra results showed that four ILs had quenching effect on the fluorescent substances of lysozyme, and the quenching effect rose with increasing ILs concentration. Meanwhile, the interaction between lysozyme and ILs molecules is mainly based on Van der Waals force and two Tryptophan (Trp) residues (Trp62 or Trp108) at the active site of lysozyme molecules play a critical role in binding ILs to their own molecules.


Assuntos
Corantes Fluorescentes/química , Líquidos Iônicos/química , Muramidase/química , Sítios de Ligação , Estrutura Terciária de Proteína , Espectrometria de Fluorescência , Espectrofotometria Ultravioleta
17.
J Agric Food Chem ; 66(49): 13020-13030, 2018 Dec 12.
Artigo em Inglês | MEDLINE | ID: mdl-30507107

RESUMO

Our previous work has demonstrated that α-linolenic acid (ALA)-loaded oil-in-water (O/W) microemulsion could enhance ALA antioxidant capacity. Meanwhile, we also observed that synthesized microemulsion itself had fluorescence. In this work, we have prepared a multiple water-in-oil-in-water (W/O/W) microemulsion to further enhance ALA antioxidant capacity and activate this delivery carrier application potential with a free label. The compositions of primary water-in-oil (W/O) microemulsion were obtained using pseudo-ternary phase diagrams, and then W/O/W microemulsion was prepared adopting the "two-step heterotherm method". The conductivity of W/O/W microemulsion was measured to lie between 250.0 and 350.0 µs/cm. The spherical droplets with a mean particle diameter of 10.0-20.0 nm were confirmed by transmission electron microscopy and dynamic light scattering. Nuclear magnetic resonance confirmed that ALA diffused to the multiple water-oily interface simultaneously. In addition, the in vitro release and antioxidant capacity measurements of ALA-loaded W/O/W microemulsion concluded the sustained-release effect and excellent antioxidant capacity. The fluorescent intensity of W/O/W microemulsion was markedly increased in comparison to O/W microemulsion. The synthesized microemulsion could lead to important applications and have advantages of a label-free fluorescent carrier for optical imaging purposes.


Assuntos
Emulsões/química , Ácido alfa-Linolênico/administração & dosagem , Ácido alfa-Linolênico/química , Antioxidantes , Sistemas de Liberação de Medicamentos , Condutividade Elétrica , Fluorescência , Tamanho da Partícula , Pentanóis/química , Espectrometria de Fluorescência/métodos , Tensoativos , Água/química
18.
J Agric Food Chem ; 66(26): 6917-6925, 2018 Jul 05.
Artigo em Inglês | MEDLINE | ID: mdl-29932339

RESUMO

Various active ingredients play a crucial role in providing and supplementing the nutritional requirements of organisms. In this work, we attempted to chemically manipulate the interfacial microstructure of oil-water microemulsions (ME) with carbon dots (CDs), concentrating on substantially enhancing the antioxidant capacity of α-linolenic acid (ALA). To this end, CDs were synthesized and introduced into an ME. The molecular interaction of surfactant with CDs was investigated by Fourier-transform infrared spectroscopy (FTIR) and nuclear magnetic resonance (NMR). The microstructure of the ME was monitored by transmission electron microscopy (TEM) and cryo-electron microscopy (cryo-EM). The cryo-EM result showed the oil-water interface in the ME was better defined after the CDs were loaded, and 1H NMR proved the CDs were distributed mainly at the interface. On the basis of these results, interfacial models were proposed. Final evaluation results demonstrated the stabilizing effect and oxidation-inhibition ability of the ALA-loaded ME was substantially enhanced after the introduction of the CDs, indicating a "turn off" effect of the interface. Interestingly, CDs do not affect the in vitro release of ALA, indicating a "turn on" effect of the interface. This work provided a successful interface manipulation with a nanocarrier that can be used for a large diversity of food nutraceuticals.


Assuntos
Antioxidantes/química , Óleos/química , Ácido alfa-Linolênico/química , Carbono/química , Emulsões/química , Espectroscopia de Ressonância Magnética , Espectroscopia de Infravermelho com Transformada de Fourier , Água/química
19.
Food Chem ; 256: 311-318, 2018 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-29606454

RESUMO

α-Linolenic acid (ALA)-loaded microemulsion (ME) was prepared from isoamyl acetate, polyoxyethylene ether 35 (EL-35), ethanol and water. The dynamic phase behaviour was simulated using dissipative particle dynamics (DPD), which showed that spherical ME was formed at water/oil ratios of 1:9 and 9:1, while a lamellar structure with distinctive water-course and oil layer appeared at ratios of 3:7, 5:5, and 7:3. Phase stabilizing and anti-oxidation effect of environmental stresses on ALA-loaded microemulsion were investigated. Results showed that the ME region was large and had good environmental tolerance. Subsequently, the investigation of anti-oxidation stability revealed that more than 60% ALA of ALA-loaded ME could be protected from oxidation under environmental stresses. Furthermore, ALA-loaded ME was applied in aqueous-based foods. The transparency, precipitate, stratification and phase separation were used to evaluate influence of ME on product properties, confirming great feasibility and stability of ALA-loaded ME for practical applications.


Assuntos
Meio Ambiente , Modelos Teóricos , Óleos/química , Estresse Fisiológico , Água/química , Ácido alfa-Linolênico/química , Emulsões , Oxirredução
20.
ACS Appl Mater Interfaces ; 9(31): 25747-25754, 2017 Aug 09.
Artigo em Inglês | MEDLINE | ID: mdl-28703570

RESUMO

Nanoscale microemulsions have been utilized as delivery carriers for nutraceuticals and active biological drugs. Herein, we designed and synthesized a novel oil in water (O/W) fluorescent microemulsion based on isoamyl acetate, polyoxyethylene castor oil EL (CrEL), and water. The microemulsion emitted bright blue fluorescence, thus exhibiting its potential for active drug detection with label-free strategy. The microemulsion exhibited excitation-dependent emission and distinct red shift with longer excitation wavelengths. Lifetime and quantum yield of fluorescent microemulsion were 2.831 ns and 5.0%, respectively. An excellent fluorescent stability of the microemulsion was confirmed by altering pH, ionic strength, temperature, and time. Moreover, we proposed a probable mechanism of fluorochromic phenomenon, in connection with the aromatic ring structure of polyoxyethylene ether substituent in CrEL. Based on our findings, we concluded that this new fluorescent microemulsion is a promising drug carrier that can facilitate active drug detection with a label-free strategy. Although further research is required to understand the exact mechanism behind its fluorescence property, this work provided valuable guidance to develop new biosensors based on fluorescent microemulsion.


Assuntos
Corantes Fluorescentes/química , Portadores de Fármacos , Emulsões , Solubilidade , Tensoativos , Água
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