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1.
Eur J Nucl Med Mol Imaging ; 51(7): 2012-2022, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38326656

RESUMO

PURPOSE: Fibroblast-activated protein (FAP) is highly expressed in cancer-associated fibroblasts (CAFs) of many solid cancers, but low or absent in normal tissues. Our study aimed to develop a novel FAP-specific tracer, namely [18F]FAP-2286, and evaluated its performance in comparison with well-established agents such as [18F]FAPI-42 and [68Ga]Ga-FAP-2286 in preclinical research, as well as 2-[18F]FDG in pilot clinical study. METHODS: [18F]FAP-2286 was manually synthesized in accordance with Good Manufacturing Practice (GMP). Subsequent investigations encompassed cell uptake, competitive binding affinity, internalization and efflux assays using HT-1080hFAP cell lines. PET imaging and biodistribution studies were conducted in HEK-293ThFAP, A549hFAP, HT-1080hFAP tumor-bearing mice as well as HEK-293T, A549 and HT-1080 control groups. Furthermore, clinical evaluation of [18F]FAP-2286 was performed in fifteen patients with various cancers compared to 2-[18F]FDG PET. RESULTS: The radiolabeling yield of [18F]FAP-2286 was 30.53 ± 5.20%, with a radiochemical purity exceeding 97%. In cell assays, [18F]FAP-2286 showed specific uptake, high internalization fraction and low cellular efflux. Rapid tumor uptake and satisfactory tumor retention was observed on micro-PET imaging and cancer patients. Meanwhile, the clinical research demonstrated that [18F]FAP-2286 may represent an alternative for low glucose-metabolism malignant tumors PET imaging such as gastric cancers. CONCLUSION: [18F]FAP-2286 showed superior imaging quality including rapid and high target uptake and satisfactory retention in both tumor-bearing mice and cancer patients. It may emerge as a promising candidate for early or delayed phase imaging and 2-[18F]FDG non-avid cancers PET scan.


Assuntos
Fluordesoxiglucose F18 , Neoplasias , Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada , Humanos , Animais , Camundongos , Fluordesoxiglucose F18/farmacocinética , Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada/métodos , Neoplasias/diagnóstico por imagem , Neoplasias/metabolismo , Masculino , Feminino , Linhagem Celular Tumoral , Células HEK293 , Distribuição Tecidual , Pessoa de Meia-Idade , Compostos Radiofarmacêuticos/farmacocinética , Idoso , Proteínas de Membrana , Endopeptidases
2.
BMC Infect Dis ; 24(1): 1149, 2024 Oct 13.
Artigo em Inglês | MEDLINE | ID: mdl-39396949

RESUMO

BACKGROUND: The best antiviral treatment for chronic hepatitis B (CHB) poses a complex challenge. The treatment effect of the combination of nucleoside analogues (NAs) and pegylated interferon alpha (PegIFN) was still in debate. METHODS: We studied patients treated with NAs and PegIFN-2b at our institution from November 2019 to January 2022. Logistic regression identified independent factors influencing clinical cure. The predictive accuracy of the formula was assessed using the Receiver operating characteristic (ROC) curve at different time points (before therapy, 12 weeks, and 24 weeks into treatment). RESULTS: A total of 120 patients were enrolled in the final analysis. Among the cohort of patients under study, 71 (59.1%) patients had clinical cure while 49 (40.9%) patients did not. Hepatitis B surface antigen (HBsAg) at baseline and age were the powerful variables predicting the clearance of HBsAg. The area under the ROC (AUC) was 0.907 for pre-treatment predictive model, 0.958 for 12-week predictive model and 0.747 for 24-week predictive model. CONCLUSION: This study provided predictive formulas for clinical cure, offering valuable insights for CHB treatment. PegIFN and NAs exhibited efficacy. Future research that explores additional factors, such as HBV genotype, in a larger cohort study is needed.


Assuntos
Antivirais , Hepatite B Crônica , Interferon-alfa , Polietilenoglicóis , Proteínas Recombinantes , Humanos , Hepatite B Crônica/tratamento farmacológico , Masculino , Interferon-alfa/uso terapêutico , Antivirais/uso terapêutico , Feminino , Adulto , Pessoa de Meia-Idade , Polietilenoglicóis/uso terapêutico , Proteínas Recombinantes/uso terapêutico , Resultado do Tratamento , Nucleosídeos/uso terapêutico , Antígenos de Superfície da Hepatite B/sangue , Quimioterapia Combinada , Curva ROC , Vírus da Hepatite B/efeitos dos fármacos , Vírus da Hepatite B/genética , Estudos Retrospectivos , Interferon alfa-2
3.
Cell Physiol Biochem ; 51(5): 2111-2122, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30522100

RESUMO

BACKGROUND/AIMS: Sestrin 2 is associated with the pathophysiology of several diseases. The aim of this study was to investigate the effects and potential mechanisms of Sestrin 2 in rat hepatic stellate cells (HSCs) during liver fibrogenesis. METHODS: In this study, Sestrin 2 protein expression was detected in rat HSC-T6 cells challenged with transforming growth factor-ß (TGF-ß) and in mice treated with carbon tetrachloride (CCl4), a well-known model of hepatic fibrosis. Next, HSC-T6 cells and fibrotic mice were transfected with lentivirus. The mRNA expression levels of markers of liver fibrosis [alpha-smooth muscle actin (α-SMA) and collagen 1A1 (Col1A1)] were analyzed by quantitative reverse transcription-polymerase chain reaction (RT-PCR). Cell death and proliferation were evaluated by the MTT assay, and biochemical markers of liver damage in serum [alanine transaminase (ALT) and aspartate transaminase (AST)] were also measured using a biochemical analyzer. Histopathological examination was used to evaluate the degree of liver fibrosis, and protein expression [phospho-adenosine monophosphate-activated protein kinase (p-AMPK), AMPK, phospho-mammalian target of rapamycin (p-mTOR), and mTOR] was determined by western blotting. RESULTS: We found that Sestrin 2 was elevated in both the HSC-T6 cell and hepatic fibrosis models. In vitro, overexpression of Sestrin 2 attenuated the mRNA levels of α-SMA and Col1A1, suppressed α-SMA protein expression, and modulated HSC-T6 cell proliferation. In vivo, overexpression of Sestrin 2 reduced the ALT and AST levels as well as the α-SMA and Col1A1 protein expression in the CCl4 model of liver fibrosis. Moreover, the degree of liver fibrosis was ameliorated. Interestingly, overexpression of Sestrin 2 increased p-AMPK but decreased p-mTOR protein expression. CONCLUSION: Our findings indicate that Sestrin 2 may attenuate the activation of HSCs and ameliorate liver fibrosis, most likely via upregulation of AMPK phosphorylation and suppression of the mTOR signaling pathway.


Assuntos
Proteínas Quinases Ativadas por AMP/metabolismo , Células Estreladas do Fígado/patologia , Cirrose Hepática/patologia , Proteínas Nucleares/metabolismo , Transdução de Sinais , Serina-Treonina Quinases TOR/metabolismo , Animais , Linhagem Celular , Células Estreladas do Fígado/metabolismo , Fígado/metabolismo , Fígado/patologia , Cirrose Hepática/metabolismo , Masculino , Camundongos Endogâmicos C57BL , Peroxidases , Fosforilação , Ratos
4.
Microb Cell Fact ; 14: 56, 2015 Apr 16.
Artigo em Inglês | MEDLINE | ID: mdl-25880855

RESUMO

BACKGROUND: Measurement of mitochondrial ATP synthesis is a critical way to compare cellular energetic performance. However, fractionation of mitochondria requires large amounts of cells, lengthy purification procedures, and an extreme caution to avoid damaging intact mitochondria, making it the highest barrier to high-throughput studies of mitochondrial function. To evaluate 45 genes involved in oxidative phosphorylation in Saccharomyces cerevisiae, we aimed to develop a simple and rapid method to measure mitochondrial ATP synthesis. RESULTS: To obtain functional mitochondria, S. cerevisiae cells were lysed with zymolyase followed by two-step, low- then high-speed centrifugation. Using a firefly luciferin-luciferase assay, the ATP synthetic activity of the mitochondria was determined. Decreasing the ATP synthesis in the presence of mitochondrial inhibitors confirmed functionality of the isolated crude mitochondria. Deletion of genes encoding mitochondrial ATP synthesis-related protein showed their dependency on the oxidative phosphorylation in S. cerevisiae. CONCLUSIONS: Compared with conventional procedures, this measurement method for S. cerevisiae Mitochondrial ATP Synthetic activity in High-throughput (MASH method) is simple and requires a small amount of cells, making it suitable for high-throughput analyses. To our knowledge, this is the first report on a rapid purification process for yeast mitochondria suitable for high-throughput screening.


Assuntos
Trifosfato de Adenosina/metabolismo , Ensaios de Triagem em Larga Escala/métodos , Mitocôndrias/metabolismo , Fosforilação Oxidativa
5.
J Sci Food Agric ; 95(14): 2874-9, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25428886

RESUMO

BACKGROUND: Currently there is much interest in the application of physical modification techniques such as heat-moisture treatment (HMT). The effects of HMT on normal and waxy rice starches, subject to different levels of moisture content and duration of heating, were investigated. RESULTS: Water solubility index (determined at 90 °C) decreased after HMT for normal and waxy rice starches, while swelling power (determined at 90 °C) showed inconsistent results (decrease for normal type, increase for waxy type) after HMT. Values in pasting parameters of normal and waxy rice starch increased initially, but the extent of increase slowed down with moisture content and length of treatment increasing. HMT decreased gelatinization temperatures with 4 h and 8 h treatment, but when length of treatment was prolonged to 16 h gelatinization temperature increased. Degree of crystallinity decreased for all treatments, and decreased much more at higher levels of moisture content. CONCLUSION: Variations in levels of moisture content and duration of heating had significant effects on physicochemical and crystalline properties to different extents.


Assuntos
Grão Comestível/química , Temperatura Alta , Oryza/química , Amido/química , Água , Cristalização , Humanos , Solubilidade , Viscosidade
6.
EJNMMI Res ; 14(1): 87, 2024 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-39352615

RESUMO

BACKGROUND: Fibroblast activation protein (FAP) has gained attention as a promising molecular target with potential utility for cancer diagnosis and therapy. [68Ga]Ga-labeled FAP-targeting peptides have been successfully applied to positron emission tomography (PET) imaging of various tumor types. To meet the applicable demand for peptide-based FAP tracers with high patient throughput, we herein report the radiosynthesis, preclinical evaluation, and the first-in-human imaging of a novel [18F]F-labeled FAP-targeting peptide. RESULTS: [18F]AlF-FAP-NUR was automatedly prepared within 45 min with a non-decay corrected radiochemical yield of 18.73 ± 4.25% (n = 3). Compared to [68Ga]Ga-FAP-2286, the [18F]F-labeled peptide demonstrated more rapid, higher levels of cellular uptake and internalization, and lower levels of cellular efflux in HT1080-FAP cells. Micro-PET imaging and biodistribution studies conducted on xenograft mice models revealed a similar distribution pattern between the two tracers. However, [18F]AlF-FAP-NUR demonstrated significantly higher tumor-specific uptake resulting in improved Tumor-Background Ratios (TBRs). In the patients, a significant accumulation of [18F]AlF-FAP-NUR was found in the primary tumor. High uptake of the tracer within the bladder indicated that its major route of excretion was through urine. CONCLUSIONS: Based on the physical imaging properties and longer half-life of [18F]F, [18F]AlF-FAP-NUR exhibited promising characteristics such as enhanced tumor-specific accumulation and elevated TBRs, which made it a viable candidate for further clinical investigation. TRIAL REGISTRATION: www.Chictr.org.cn , ChiCTR2300076976 Retrospectively registered 25 October 2023. at, URL: https://www.chictr.org.cn/showproj.html?proj=206753 .

7.
Front Microbiol ; 15: 1442604, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39171262

RESUMO

This study investigated the impact of seasonal environmental factors on microorganisms and flavor compounds during acetic acid fermentation (AAF) of Zhenjiang aromatic vinegar (ZAV). Environmental factors were monitored throughout the fermentation process, which spanned multiple seasons. Methods such as headspace solid phase microextraction gas chromatography-mass spectrometry (HS-SPME-GC-MS), high performance liquid chromatography (HPLC), and high-throughput sequencing were employed to examine how these environmental factors influenced the flavor profile and microbial community of ZAV. The findings suggested that ZAV brewed in autumn had the strongest flavor and sweetness. The key microorganisms responsible for the flavor of ZAV included Lactobacillus acetotolerans, Lactobacillus plantarum, Lactobacillus reuteri, Lactobacillus fermentum, Acetobacter pasteurianus. Moreover, correlation analysis showed that room temperature had a significant impact on the composition of the microbial community, along with other key seasonal environmental factors like total acid, pH, reducing sugar, and humidity. These results provide a theoretical foundation for regulating core microorganisms and environmental factors during fermentation, enhancing ZAV quality.

8.
Front Microbiol ; 15: 1428095, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39109213

RESUMO

Introduction: Baloxavir Marboxil is a per oral small-molecule antiviral for the treatment of influenza. While the efficacy and safety of Baloxavir Marboxil have been thoroughly characterized across an extensive clinical trial, studies on the effectiveness of Baloxavir Marboxil in a real-world setting are still scarce. Methods: We conducted an ambispective, observational, multi-center study that enrolled uncomplicated in-fluenza outpatients treated with Baloxavir Marboxil or Oseltamivir in East China. The primary endpoint was time from treatment to alleviation of all influenza symptoms (TTAIS). The secondary endpoints included time from treatment to alleviation of fever (TTAF) and household transmission during the duration of influenza. Results: A total of 509 patients were enrolled. The median TTAIS in the Baloxavir Marboxil group and the Oseltamivir group was 28.0 h (IQR, 20.0 to 50.0) and 48.0 h (IQR, 30.0 to 67.0), respectively. The median TTAF in the Baloxavir Marboxil group and the Oseltamivir group was 18 h (IQR, 10.0-24.0) and 30.0 h (IQR, 19.0-48.0). In the COX multivariable analysis, Baloxavir Marboxil reduced the duration of influenza symptoms (HR = 1.36 [95%CI:1.12-1.64], p = 0.002) and the duration of fever (HR = 1.93 [95%CI:1.48-2.52], p < 0.001) compared to Oseltamivir. When antiviral drugs were given within 12-48 h after symptom onset, the Baloxavir Marboxil group had a significantly shorter TTAIS compared to the Oseltamivir group. There was no significant difference in the rate of adverse events between the two group (p = 0.555). Discussion: Baloxavir Marboxil was superior to Oseltamivir in alleviating influenza symptoms in outpatients with uncomplicated influenza. Our findings suggested that compared to Oseltamivir, Baloxavir Marboxil might be more appropriate for patients with influenza 12- 48 h after symptom onset.

9.
Metab Eng ; 20: 84-91, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-24055789

RESUMO

The heat treatment of recombinant mesophiles having heterologous thermotolerant enzymes results in the one-step preparation of highly selective biocatalytic modules. The assembly of these modules enables us to readily construct an artificial metabolic pathway in vitro. In this work, we constructed a non-natural, cofactor-balanced, and oxygen-insensitive pathway for n-butanol production using 16 thermotolerant enzymes. The whole pathway was divided into 7 parts, in each of which NAD(H)-dependent enzymes were assigned to be the last step, and the fluxes through each part were spectrophotometrically determined. This real-time monitoring technique enabled the experimental optimization of enzyme level to achieve a desired production rate. Through the optimized pathway, n-butanol could be produced from glucose with a molar yield of 82% at a rate of 8.2 µmol l(-1) min(-1). Our approach would be widely applicable to the rational optimization of artificial metabolic pathways as well as to the in vitro production of value-added biomolecules.


Assuntos
1-Butanol/metabolismo , Acetobacter/enzimologia , Proteínas de Bactérias/química , Proteínas de Bactérias/metabolismo , Glucose/metabolismo , Thermus thermophilus/enzimologia , 1-Butanol/química , Sistema Livre de Células/química , Sistema Livre de Células/enzimologia
10.
Front Microbiol ; 14: 1126238, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36970705

RESUMO

Inoculation strategy is a significant determinant of the flavor quality of Zhenjiang aromatic vinegar. Herein, the comparative analyses of the effects of various inoculation strategies on the physicochemical properties, microbial community structure, and flavoring characteristics of Zhenjiang aromatic vinegar were performed. The results showed that the contents of total acid (6.91 g/100 g), organic acid (2099.63 ± 4.13 mg/100 g) and amino acid (3666.18 ± 14.40 mg/100 g) in the direct inoculation strategy were higher than those in the traditional inoculation strategy (6.21 ± 0.02 g/100 g, 1939.66 ± 4.16 mg/100 g and 3301.46 ± 13.41 mg/100 g). At the same time, it can effectively promote the production of acetoin. The diversity of strains under the traditional inoculation strategy was higher than that under the direct inoculation strategy, and the relative abundance of major microbial genera in the fermentation process was lower than that under the direct inoculation strategy. In addition, for two different inoculation strategies, pH was proved to be an important environmental factor affecting the microbial community structure during acetic acid fermentation. The correlation between main microbial species, organic acids, non-volatile acids, and volatile flavor compounds is more consistent. Therefore, this study may help to develop direct injection composite microbial inoculants to replace traditional starter cultures in future research.

11.
Microb Cell Fact ; 11: 120, 2012 Sep 06.
Artigo em Inglês | MEDLINE | ID: mdl-22950411

RESUMO

BACKGROUND: The integration of biotechnology into chemical manufacturing has been recognized as a key technology to build a sustainable society. However, the practical applications of biocatalytic chemical conversions are often restricted due to their complexities involving the unpredictability of product yield and the troublesome controls in fermentation processes. One of the possible strategies to overcome these limitations is to eliminate the use of living microorganisms and to use only enzymes involved in the metabolic pathway. Use of recombinant mesophiles producing thermophilic enzymes at high temperature results in denaturation of indigenous proteins and elimination of undesired side reactions; consequently, highly selective and stable biocatalytic modules can be readily prepared. By rationally combining those modules together, artificial synthetic pathways specialized for chemical manufacturing could be designed and constructed. RESULTS: A chimeric Embden-Meyerhof (EM) pathway with balanced consumption and regeneration of ATP and ADP was constructed by using nine recombinant E. coli strains overproducing either one of the seven glycolytic enzymes of Thermus thermophilus, the cofactor-independent phosphoglycerate mutase of Pyrococcus horikoshii, or the non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase of Thermococcus kodakarensis. By coupling this pathway with the Thermus malate/lactate dehydrogenase, a stoichiometric amount of lactate was produced from glucose with an overall ATP turnover number of 31. CONCLUSIONS: In this study, a novel and simple technology for flexible design of a bespoke metabolic pathway was developed. The concept has been testified via a non-ATP-forming chimeric EM pathway. We designated this technology as "synthetic metabolic engineering". Our technology is, in principle, applicable to all thermophilic enzymes as long as they can be functionally expressed in the host, and thus would be potentially applicable to the biocatalytic manufacture of any chemicals or materials on demand.


Assuntos
Engenharia Metabólica , Trifosfato de Adenosina/metabolismo , Escherichia coli/metabolismo , Glucose/metabolismo , Gliceraldeído-3-Fosfato Desidrogenases/genética , Gliceraldeído-3-Fosfato Desidrogenases/metabolismo , Concentração de Íons de Hidrogênio , Ácido Láctico/metabolismo , Malato Desidrogenase/genética , Malato Desidrogenase/metabolismo , Fosfoglicerato Mutase/genética , Fosfoglicerato Mutase/metabolismo , Pyrococcus/enzimologia , Temperatura , Thermococcus/enzimologia , Thermus thermophilus/enzimologia
12.
Acta Biomater ; 125: 29-40, 2021 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-33582362

RESUMO

Bacterial infections are serious threats to public health due to lack of advanced techniques to rapidly and accurately diagnose these infections in clinics. Although bacterial infections can be treated with broad-spectrum antibiotics based on empirical judgment, the emergence of antimicrobial resistance has attracted global attention due to long-term misuse and abuse of antibiotics by humans in recent decades. Therefore, it is imperative to selectively discriminate and precisely eliminate pathogenic bacteria. Herein, in addition to the conventional methods for bacterial identification, we comprehensively reviewed the recently developed theranostic platforms for specific discrimination and selective killing of bacteria according to their different interactions with the target bacteria, such as electrostatic and hydrophobic interactions, molecular recognition, microenvironment response, metabolic labeling, bacteriophage targeting, and others. These theranostic agents not only benefit from improved therapeutic efficiency but also present limited susceptibility to induce bacterial resistance. The strategies summarized in this review will open up new avenues in developing effective antimicrobial materials to accurately diagnose and treat bacterial infections in the post-antibiotic era. STATEMENT OF SIGNIFICANCE: Bacterial infections are difficult to be rapidly and accurately diagnosed, and are generally treated with broad-spectrum antibiotics, which leads to the development of drug resistance. By integrating imaging modalities and therapeutic methods in a single treatment, various theranostic agents have been developed to address the abovementioned issues. Therefore, the emerging theranostic platforms for selective identification and elimination of bacteria based on the distinct interactions of the theranostic agents with the target bacteria are summarized in this review. We believe that the information provided in this review will guide researchers in designing advanced antibacterial theranostics for practical applications in the post-antibiotic era.


Assuntos
Infecções Bacterianas , Medicina de Precisão , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Bactérias , Infecções Bacterianas/diagnóstico , Infecções Bacterianas/tratamento farmacológico , Humanos
13.
J Biomed Mater Res A ; 109(10): 1979-1989, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-33822474

RESUMO

Native decellularized extracellular matrix provides an adequate platform for tissues and organs and promotes the development of organogenesis and tissue remodeling. However, thrombosis poses a great challenge that hinders the transplantation for a substantial organ in vivo. Therefore, anticoagulation and re-reendothelialization of organ biological scaffolds are the primary concerns to be addressed before orthotopic transplantation. Herein, a heparinized decellularized kidney scaffold (HEP-DKSs) was prepared using end-point attachment technology, followed by binding the vascular endothelial growth factor (VEGF) to greatly improve the hemocompatibility and angiogenesis of DKSs. Based on the anticoagulant, co-culture of human umbilical vein endothelial cells, and subcapsular transplantation of kidney experiments, HEP-VEGF-DKSs are shown to reduce platelet adhesion, which is crucial for subsequent vascularization and slow release of heparin and VEGF, suggesting its ability of improve neovascularization. Taken together, these data indicated an optimal anticoagulation function of HEP-VEGF-DKSs and the potential of vascularization for regeneration of whole decellularized kidney.


Assuntos
Heparina/farmacologia , Rim/citologia , Neovascularização Fisiológica , Alicerces Teciduais/química , Animais , Anticoagulantes/farmacologia , Técnicas de Cocultura , Liberação Controlada de Fármacos , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Humanos , Rim/efeitos dos fármacos , Neovascularização Fisiológica/efeitos dos fármacos , Ratos Sprague-Dawley , Resistência à Tração , Fator A de Crescimento do Endotélio Vascular/metabolismo
14.
Food Chem ; 276: 754-760, 2019 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-30409658

RESUMO

Starch granule-associated proteins (SGAPs) including granule-surface proteins and granule-channel proteins in waxy, low- and high-amylose rice starch were extracted and identified. The in vitro digestibility of starch was investigated before and after the extraction of granule-channel proteins or total SGAPs. The results showed that 10 types of major differentially expressed proteins (DEPs) including 14-3-3-like protein and ribosomal protein were found among starches. In addition, the lack of only granule-channel proteins or total SGAPs led to significant and different changes in the levels of rapidly digestible starch, slowly digestible starch and resistant starch. Possible mechanisms are related to the accessibility of amylase into starch granules and structural properties of SGAPs. This study provides more information about DEPs in rice starch with different amylose content and supports further study on the relationship between SGAPs and in vitro starch digestibility.


Assuntos
Oryza/química , Proteínas de Plantas/isolamento & purificação , Amido/química , Amilose/análise , Amilose/isolamento & purificação , Fracionamento Químico/métodos , Digestão , Proteínas de Plantas/análise , Proteínas de Plantas/farmacocinética , Proteínas de Vegetais Comestíveis/análise , Proteínas de Vegetais Comestíveis/isolamento & purificação , Proteínas de Vegetais Comestíveis/farmacocinética , Amido/ultraestrutura
15.
Colloids Surf B Biointerfaces ; 172: 298-307, 2018 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-30173097

RESUMO

Functionalized-nanoparticles have been developed as novel therapeutic delivery platform for simultaneous drug loading and therapy over the past decade. Rationally-designed biocompatible nanosystem simultaneously with multistimuli-responsive property and synergistic therapeutic potential are highly desirable for modern biological applications. Herein, Cu2Se nanoparticles (Cu2SeNPs) with suitable size have been functionalized by bull serum albumin (BSA) through a simply, facile and controllable method. As a result, Cu2SeNPs modified by BSA (BSA-Cu2SeNPs) showed excellent biocompatibility and stability. The strong absorbance of BSA-Cu2SeNPs at near infrared region imparts them with high photothermal efficiency. Then loading doxorubicin (DOX, anticancer drug) on the surface of BSA-Cu2SeNPs, and consequently, a novel multifunctional nanosystem of BSA-Cu2SeNPs-DOX is designed. The BSA-Cu2SeNPs can achieve high DOX loading capacity (approximately 157 µg DOX per mg of Cu2Se). Furthermore, a rational and precise release of DOX from the BSA-Cu2SeNPs-DOX could be easily realized under the stimulates of the pH and temperature, which remarkably improved antitumor efficacy of combined chemotherapy and photothermal therapy triggered by 808 nm NIR laser. Thus, the BSA-Cu2SeNPs-DOX could serve as an ideal nanoplatform for cancer diagnosis and treatment in future. The results of cell experiments show that the BSA-Cu2SeNPs-DOX exhibited favorable selective cellular uptake cells. Under the NIR laser irradiation, BSA-Cu2SeNPs-DOX could induce the excessive expression of ROS, eventually leading to the death of U251 cells. Both in vitro and in vivo experiments indicate that the nanosystem of BSA-Cu2SeNPs-DOX showed excellent synergistic therapeutic effect and multistimuli-responsive drug vehicle, which will exert huge potential for future clinical application.


Assuntos
Doxorrubicina/farmacologia , Portadores de Fármacos/química , Hipertermia Induzida , Nanopartículas/química , Fototerapia , Soroalbumina Bovina/química , Antineoplásicos/farmacologia , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Doxorrubicina/química , Sinergismo Farmacológico , Endocitose/efeitos dos fármacos , Humanos , Nanopartículas/ultraestrutura , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais/efeitos dos fármacos , Espectroscopia de Infravermelho com Transformada de Fourier , Temperatura
16.
Mol Med Rep ; 15(4): 2308-2312, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28260040

RESUMO

Endothelial dysfunction and impaired endothelial regenerative capacity are key contributors to the high incidence of cardiovascular disease in patients with chronic kidney disease (CKD). Uremic toxins are associated with this pathogenesis. Previous studies have revealed that a uremic toxin, para­cresol (p­cresol), exerts an antiproliferation effect on human endothelial progenitor cells (EPCs), but the mechanism remains unclear. In the present study, reactive oxygen species (ROS) were confirmed to function as signaling molecules that regulate growth factor­dependent EPC proliferation. EPCs were treated with p­cresol for 72 h, using a concentration range typically found in CKD patients. ROS production was analyzed by fluorescence microscopy and flow cytometry, and protein expression levels of nicotinamide adenine dinucleotide phosphate oxidase, a major source of ROS, were analyzed by western blot analysis. mRNA expression levels of antioxidant genes were assessed by reverse transcription­quantitative polymerase chain reaction analysis. The results revealed that p­cresol partially inhibits ROS production, and this effect may be associated with a significant reduction in cytochrome b­245 alpha and beta chain expression in EPCs. An increase of glutathione peroxidase 4 mRNA expression was also detected. In conclusion, the present study revealed that the antiproliferation effect of p­cresol on EPCs might act via its antioxidant activity. The results of the present study may facilitate understanding of uremic toxin toxicity on the cardiovascular system.


Assuntos
Proliferação de Células , Cresóis/metabolismo , Células Progenitoras Endoteliais/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Uremia/metabolismo , Catalase/genética , Linhagem Celular , Células Progenitoras Endoteliais/citologia , Células Progenitoras Endoteliais/patologia , Regulação da Expressão Gênica , Glutationa Peroxidase/genética , Humanos , Glicoproteínas de Membrana/metabolismo , NADPH Oxidase 2 , NADPH Oxidases/metabolismo , Fosfolipídeo Hidroperóxido Glutationa Peroxidase , Superóxido Dismutase/genética , Superóxido Dismutase-1/genética , Uremia/genética , Uremia/patologia
17.
Int J Biol Macromol ; 84: 428-33, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26718868

RESUMO

Changes in the physicochemical properties and starch digestibility of white salted noodles (WSN) at different cooking stage were investigated. The noodles were dried in fresh air and then cooked for 2-12 min by boiling in distilled water to determine the properties of cooking quality, textural properties and optical characteristic. For starch digestibility, dry noodles were milled and sieved into various particle size classes ranging from 0.5 mm to 5.0 mm, and hydrolyzed by porcine pancreatic α-amylase. The optimal cooking time of WSN determined by squeezing between glasses was 6 min. The results showed that the kinetics of solvation of starch and protein molecules were responsible for changes of the physicochemical properties of WSN during cooking. The susceptibility of starch to α-amylase was influenced by the cooking time, particle size and enzyme treatment. The greater value of rapidly digestible starch (RDS) and lower value of slowly digestible starch (SDS) and resistant starch (RS) were reached at the optimal cooking stage ranging between 63.14-71.97%, 2.47-10.74% and 23.94-26.88%, respectively, indicating the susceptibility on hydrolysis by enzyme was important in defining the cooked stage. The study suggested that cooking quality and digestibility were not correlated but the texture greatly controls the digestibility of the noodles.


Assuntos
Culinária , Farinha/análise , Amido/química , Fatores de Tempo , Triticum/química
18.
Int J Biol Macromol ; 93(Pt A): 34-40, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27565294

RESUMO

Impact of heat-moisture treatment (HMT) on nutritional properties of normal maize starch (NMS) under various reaction conditions was investigated. NMS was adjusted to moisture levels of 20%, 25%, 30% and 35% and heated at 80, 100, 120 and 140°C for 4, 8, 12 and 16h. Response surface methodology (RSM) based on Box-Behnken design (BBD) was employed to obtain the optimal combination of moisture level (X1: 20-30%), length of heating (X2: 4-12h), and temperature (X3: 100-140°C). The optimum reaction condition decreased rapidly digestible starch (RDS) from 87.10% to 82.21%, when NMS was subjected to HMT at 23.6% moisture content and heated at 114.8°C for 9.04h. The ANOVA measurement and confirmation experiments were performed to verify the predictive value and the RSM model, indicating that temperature was the main factor to determine the digestion rate of HMT NMS. The results suggested that RDS was not correlated to heating length but positively correlated to temperature and moisture content. Reaction condition had no correlations with slowly digestible starch (SDS) and resistant starch (RS). This study could provide more information for producing low-glycemic index products.


Assuntos
Amido/química , Zea mays/química , Carboidratos da Dieta , Índice Glicêmico , Temperatura Alta , Humanos , Água/química
19.
Food Chem ; 173: 1125-32, 2015 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-25466134

RESUMO

Changes in the properties of normal maize starch (NMS) and waxy maize starch (WMS) after heat-moisture treatment (HMT) under various reaction conditions were investigated. NMS and WMS were adjusted to moisture levels of 20%, 25% and 30% and heated at 100 °C for 2, 4, 8 and 16 h. The results showed that moisture content was the most important factor in determining pasting properties for NMS, whereas the heating length was more important for WMS. Swelling power decreased in NMS but increased in WMS, and while the solubility index decreased for both samples, the changes were largely determined by moisture content. The gelatinisation temperatures of both samples increased with increasing moisture content but remained unchanged with increasing heating length. The Fourier transform infrared (FT-IR) absorbance ratio was affected to different extents by the moisture levels but remained constant with increasing the heating length. The X-ray intensities increased but relative crystallinity decreased to a greater extent with increasing moisture content. This study showed that the levels of moisture content and length of heating had significant impacts on the structural and physicochemical properties of normal and waxy maize starches but to different extents.


Assuntos
Amilopectina/química , Amilose/química , Temperatura Alta , Zea mays/química , Fenômenos Químicos , Manipulação de Alimentos , Solubilidade , Espectroscopia de Infravermelho com Transformada de Fourier , Água/química , Difração de Raios X
20.
Biotechnol Biofuels ; 8: 48, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25802553

RESUMO

BACKGROUND: Marine microalgae are among the most promising lipid sources for biodiesel production because they can be grown on nonarable land without the use of potable water. Marine microalgae also harvest solar energy efficiently with a high growth rate, converting CO2 into lipids stored in the cells. Both light intensity and nitrogen availability strongly affect the growth, lipid accumulation, and fatty acid composition of oleaginous microalgae. However, very few studies have systematically examined how to optimize lipid productivity by adjusting irradiance intensity, and the metabolic dynamics that may lead to improved lipid accumulation in microalgae have not been elucidated. Little is known about the mechanism of lipid synthesis regulation in microalgae. Moreover, few studies have assessed the potential of using marine microalgae as oil producers. RESULTS: In this work, a newly isolated marine microalga, Chlamydomonas sp. JSC4, was selected as a potential lipid producer, and the effect of photobioreactor operations on cell growth and lipid production was investigated. The combined effects of light intensity and nitrogen depletion stresses on growth and lipid accumulation were further explored in an effort to markedly improve lipid production and quality. The optimal lipid productivity and content attained were 312 mg L(-1) d(-1) and 43.1% per unit dry cell weight, respectively. This lipid productivity is the highest ever reported for marine microalgae. Metabolic intermediates were profiled over time to observe transient changes during lipid accumulation triggered by combined stresses. Finally, metabolite turnover was also assessed using an in vivo (13)C-labeling technique to directly measure the flow of carbon during lipid biosynthesis under stress associated with light intensity and nitrogen deficiency. CONCLUSIONS: This work demonstrates the synergistic integration of cultivation and dynamic metabolic profiling technologies to develop a simple and effective strategy for enhancing oil production in a marine microalga. The knowledge obtained from this study could be useful in assessing the feasibility of marine microalgae biodiesel production and for understanding the links between dynamic metabolic profiles and lipid biosynthesis during the course of microalgal cultivation.

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