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1.
Food Chem ; 337: 127761, 2021 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-32777565

RESUMO

Amino and thiolated aptamers are the main aptamers used to construct label-free electrochemical impedimetric aptasensors. In this study, the modification performance and electrochemical properties of amino aptamers and thiolated aptamers were studied in the construction of label-free impedimetric sensors. The results showed that the initial modification density of amino aptamers was higher than that of thiol aptamers. Aptamers can recognize and bind OTA to generate electrical signals. The higher the density of aptamer modification was, the better the electric signals were. If only considering the initial modification density, amino aptamers were more suitable for the preparation of aptasensors than thiolated aptamers. However, the modification density of the amino aptamer decreased with the prolonged immersion time in 1 mM HCl solution, which suggests that the stability of this sensor was poor. However, the thiolated aptamer maintained relatively constant density and could be reused. Thus, the thiolated aptasensor had a wide range and good reproducibility and stability for the determination of ochratoxin A (OTA). In addition, this study proved that gold nanoparticles play an important role in signal amplification by increasing the effective gold surface to fix more aptamers in the process of sensor preparation.

2.
Pest Manag Sci ; 2020 Oct 23.
Artigo em Inglês | MEDLINE | ID: mdl-33098218

RESUMO

BACKGROUND: The sheath blight, caused by Rhizoctonia solani, can be effectively controlled by the application of the succinate dehydrogenase inhibitor (SDHI) thifluzamide. Although the resistant risk of thifluzamide in R. solani had been reported, but the thifluzamide-resistance mechanism and the evolution of thifluzamide-resistance in R. solani have not been investigated in detail. RESULTS: No differences were found between the sequences of the SDHA, SDHC and SDHD protein among the thifluzamide-sensitive isolates and the thifluzamide-resistant mutants, but a single point mutation H249Y was found in SDHB protein. Two different types of thifluzamide-resistant R. solani mutants were characterized: homokaryotic type, carrying only the resistance allele, and heterokaryotic type, retaining the wild-type allele in addition to the resistance allele. The resistance level differed according to nuclear composition at position of codon 249 in sdhB gene. Molecular docking results suggested that the point mutation (H249Y) might significantly altered the affinity of thifluzamide and SDHB protein. Heterokaryotic mutants were able to evolve into a homokaryon when repeatedly cultured on agar media or rice plants in the presence of thifluzamide, but thifluzamide treatment had no effect on the genotypes of the homokaryotic mutants or the sensitive isolates. CONCLUSION: This study showed that H249Y in SDHB protein could cause thifluzamide-resistance in R. solani. Fungicide application could promote heterokaryotic mutants to evolve into a homokaryon. This article is protected by copyright. All rights reserved.

3.
PLoS One ; 15(8): e0236898, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32785280

RESUMO

The development of fungal fruiting bodies from a hyphal thallus is inducible under low temperature (cold stress). The molecular mechanism has been subject to surprisingly few studies. Analysis of gene expression level has become an important means to study gene function and its regulation mechanism. But identification of reference genes (RGs) stability under cold stress have not been reported in famous medicinal mushroom-forming fungi Cordyceps militaris. Herein, 12 candidate RGs had been systematically validated under cold stress in C. militaris. Three different algorithms, geNorm, NormFinder and BestKeeper were applied to evaluate the expression stability of the RGs. Our results showed that UBC and UBQ were the most stable RGs for cold treatments in short and long periods, respectively. 2 RGs (UBC and PP2A) and 3 RGs (UBQ, TUB and CYP) were the suitable RGs for cold treatments in short and long periods, respectively. Moreover, target genes, two-component-system histidine kinase genes, were selected to validate the most and least stable RGs under cold treatment, which indicated that use of unstable expressed genes as RGs leads to biased results. Our results provide a good starting point for accurate reverse transcriptase quantitative polymerase chain reaction normalization by using UBC and UBQ in C. militaris under cold stress and better support for understanding the mechanism of response to cold stress and fruiting body formation in C. militaris and other mushroom-forming fungi in future research.


Assuntos
Resposta ao Choque Frio/genética , Cordyceps/genética , Cordyceps/fisiologia , Perfilação da Expressão Gênica/normas , Histidina Quinase/genética , Cordyceps/enzimologia , Padrões de Referência
4.
Toxins (Basel) ; 12(2)2020 02 15.
Artigo em Inglês | MEDLINE | ID: mdl-32075318

RESUMO

Black spot caused by Alternaria alternata is one of the important diseases of pear fruit during storage. Isothiocyanates are known as being strong antifungal compounds in vitro against different fungi. The aim of this study was to assess the antifungal effects of the volatile compound 2-phenylethyl isothiocyanate (2-PEITC) against A. alternata in vitro and in pear fruit, and to explore the underlying inhibitory mechanisms. The in vitro results showed that 2-PEITC significantly inhibited spore germination and mycelial growth of A. alternata-the inhibitory effects showed a dose-dependent pattern and the minimum inhibitory concentration (MIC) was 1.22 mM. The development of black spot rot on the pear fruit inoculated with A. alternata was also significantly decreased by 2-PEITC fumigation. At 1.22 mM concentration, the lesion diameter was only 39% of that in the control fruit at 7 days after inoculation. Further results of the leakage of electrolyte, increase of intracellular OD260, and propidium iodide (PI) staining proved that 2-PEITC broke cell membrane permeability of A. alternata. Moreover, 2-PEITC treatment significantly decreased alternariol (AOH), alternariolmonomethyl ether (AME), altenuene (ALT), and tentoxin (TEN) contents of A. alternata. Taken together, these data suggest that the mechanisms underlying the antifungal effect of 2-PEITC against A. alternata might be via reduction in toxin content and breakdown of cell membrane integrity.

5.
Food Chem ; 302: 125288, 2020 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-31419774

RESUMO

The effects of benzothiadiazole (BTH) on Penicillium expansum development, mitochondria energy metabolism, and changes in the number and structure of mitochondria in apple fruit were investigated after the fruit were immersed in 100 mg L-1 BTH for 10 min and then stored at 22 °C. The results indicated that BTH treatment significantly decreased the lesion diameter of fruit challenged with P. expansum; further, treatment enhanced the activities of mitochondrial respiratory metabolism-related enzymes, such as succinate dehydrogenase, cytochrome oxidase, H+-ATPase and Ca2+-ATPase, along with high ATP level and energy status in apple fruit during storage. Moreover, transmission electron microscopy results indicated that BTH treatment was beneficial for maintaining the number and structure of mitochondria during storage. The results suggested that BTH treatment enhanced ATP levels via mitochondrial energy metabolism, which might contribute to the induced resistance in apple fruit during storage.


Assuntos
Metabolismo Energético/efeitos dos fármacos , Armazenamento de Alimentos , Frutas/metabolismo , Malus/efeitos dos fármacos , Malus/metabolismo , Mitocôndrias/efeitos dos fármacos , Tiadiazóis/farmacologia , Frutas/microbiologia , Malus/microbiologia , Mitocôndrias/metabolismo , Penicillium/fisiologia
6.
Food Chem ; 309: 125608, 2020 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-31678673

RESUMO

Benzo-(1,2,3)-thiadiazole-7-carbothioic acid S-methyl ester (BTH) can improve wound healing of potato tubers; however, how the chemical regulates reactive oxygen species (ROS) generation and scavenging during wound healing is not completely understood. BTH at 100 mg·L-1 regulated changes in ROS generation and scavenging in healing tissues of potato tubers. A higher H2O2 content was presented in healing tissues of potato tubers, while cell membrane permeability and malondialdehyde content declined due to BTH treatment. Additionally, the activities and transcript level of enzymes related with ROS generation, including NADPH oxidase, peroxidase and polyamine oxidase, as well as enzymes involved in ROS scavenging, such as superoxide dismutase, catalase, ascorbate peroxidase, and glutathione reductase, were significantly enhanced by BTH treatment. It is suggested that ROS metabolism might play a crucial role in wound healing of potato tubers mediated by BTH during postharvest.


Assuntos
Tubérculos/efeitos dos fármacos , Tubérculos/metabolismo , Solanum tuberosum/efeitos dos fármacos , Solanum tuberosum/metabolismo , Tiadiazóis/farmacologia , Membrana Celular/efeitos dos fármacos , Permeabilidade da Membrana Celular/efeitos dos fármacos , Enzimas/genética , Enzimas/metabolismo , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Peróxido de Hidrogênio/metabolismo , Malondialdeído/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Espécies Reativas de Oxigênio/metabolismo
7.
In Vitro Cell Dev Biol Anim ; 55(9): 713-722, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31515718

RESUMO

Hepatic progenitor cells (HPCs) are potential seed cells for hepatocyte transplantation treatment of liver diseases. ATRA can induce the differentiation and mature function of hepatic progenitor cells, but the mechanism is still poorly understood. Here, by using microRNA array to analyze the expression profiles of microRNA (miR), we found that miR-200 family molecules in HPCs were upregulated after ATRA treatment, especially miR-200a-3p, 200c-3p, and 141-3p. ATRA induction could downregulate the expression of hepatic stem markers Oct4 and AFP, and improve the expression of hepatic markers ALB, CK18, and TAT, and the activity of ALB-GLuc, as well as indocyanine green uptake and glycogen storage function of HPCs. These above effects of ATRA on HPC differentiation were almost inhibited by blocking of miR-200a-3p, but not miR-200c-3p and 141-3p using antagomir. Cell autophagy is associated with ATRA regulation in HPCs, compared with control group, the expression of LC3 and Beclin1 increased in ATRA-treated HPCs, and orange and red fluorescent spot, which represents autophagy flow, also enhanced after ATRA treatment. However, ATRA-induced cell autophagy level was inhibited in antagomir-200a-3p+ATRA-treated cells. Therefore, the present study indicates that antagomir-200a-3p is related to ATRA-induced hepatic differentiation of HPCs through regulating cell autophagy, supporting the possible use of ATRA as a key inducer in HPC-based therapy of liver diseases.


Assuntos
Hepatócitos/efeitos dos fármacos , Hepatopatias/genética , MicroRNAs/genética , Tretinoína/farmacologia , Animais , Antagomirs/genética , Autofagia/efeitos dos fármacos , Biomarcadores/metabolismo , Diferenciação Celular/efeitos dos fármacos , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Hepatócitos/metabolismo , Fígado/efeitos dos fármacos , Fígado/crescimento & desenvolvimento , Hepatopatias/metabolismo , Camundongos , Células-Tronco/efeitos dos fármacos , Ativação Transcricional/efeitos dos fármacos , Tretinoína/metabolismo
8.
Food Chem ; 301: 125303, 2019 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-31387045

RESUMO

The effects of exogenous polyamines treatment on reactive oxygen species (ROS) metabolism in apricot fruits were systematically analyzed through the investigation of their curative and preventive effects on black spot disease. Results showed that 1.5 mM spermine (Spm), 1.5 mM spermidine (Spd) and 10 mM putrescine (Put) treatment significantly inhibited black spot development, additionally, the efficacy of this control was dependent upon the type of polyamines used and concentration level applied. Further studies have shown that exogenous polyamines treatments significantly improved production of O2- and H2O2, and increased the activities and gene expression levels of NADPH oxidase (NOX), super oxide dismutase (SOD), catalase (CAT) ascorbate peroxidase (AXP) and glutathione reductase (GR) in apricot fruit. Increased ascorbic acid (AsA) and reduced glutathione (GSH) content were also observed after exogenous polyamines treatment. These results have revealed that postharvest polyamines treatment effectively enhanced disease resistance through the maintenance of homeostasis in apricot fruits.


Assuntos
Alternaria/patogenicidade , Frutas/efeitos dos fármacos , Doenças das Plantas/microbiologia , Poliaminas/farmacologia , Prunus armeniaca/efeitos dos fármacos , Ascorbato Peroxidases/metabolismo , Catalase/metabolismo , Resistência à Doença/efeitos dos fármacos , Frutas/metabolismo , Frutas/microbiologia , Glutationa/metabolismo , Glutationa Redutase/metabolismo , Homeostase/efeitos dos fármacos , Peróxido de Hidrogênio/metabolismo , Oxirredução , Prunus armeniaca/metabolismo , Prunus armeniaca/microbiologia , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/metabolismo
9.
Biochem Biophys Res Commun ; 515(1): 24-30, 2019 Jul 12.
Artigo em Inglês | MEDLINE | ID: mdl-31122700

RESUMO

Human umbilical cord-derived mesenchymal stromal cells (hUC-MSCs) in vitro expansion for long term may undergo epigenetic and genetic alterations that subsequently induce cellular senescence and associated growth inhibition. Increasing evidence implicated that aberrant histone acetylation modulates gene expression responsible for MSCs aging. Whether the dysregulation of p300 and its KAT activity is involved in the aging process of MSCs was still unexplored. In this study, we found a significant decrease of p300 but elevated p53/p21 levels in senescent hUC-MSCs at late-passage. Then we used two different approaches: (i) downregulation of p300 by siRNA and (ii) inhibition of the acetyltransferase(KAT) activity by C646 to determine the role of p300 in regulating MSCs senescence. We showed that inhibition of p300 induce premature senescence and decrease proliferation potential in hUC-MSCs. Moreover, upregulations of p53 and p21 expressions were confirmed in p300 knockdown and C646-treated hUC-MSCs. Taken together, these results suggest that p300 plays an important role in aging process of MSCs associated with activation of p53/p21 signaling pathway.

10.
World J Gastroenterol ; 25(16): 1936-1949, 2019 Apr 28.
Artigo em Inglês | MEDLINE | ID: mdl-31086462

RESUMO

BACKGROUND: Study shows that signal transducer and activator of transcription 3 (STAT3) can increase the Warburg effect by stimulating hexokinase 2 in breast cancer and upregulate lactate dehydrogenase A and pyruvate dehydrogenase kinase 1 in myeloma. STAT3 and pyruvate kinase M2 (PKM2) can also be activated and enhance the Warburg effect in hepatocellular carcinoma. Precancerous lesions are critical to human and rodent hepatocarcinogenesis. However, the underlying molecular mechanism for the development of liver precancerous lesions remains unknown. We hypothesized that STAT3 promotes the Warburg effect possibly by upregulating p-PKM2 in liver precancerous lesions in rats. AIM: To investigate the mechanism of the Warburg effect in liver precancerous lesions in rats. METHODS: A model of liver precancerous lesions was established by a modified Solt-Farber method. The liver pathological changes were observed by HE staining and immunohistochemistry. The transformation of WB-F344 cells induced with N-methyl-N'-nitro-N-nitrosoguanidine and hydrogen peroxide was evaluated by the soft agar assay and aneuploidy. The levels of glucose and lactate in the tissue and culture medium were detected with a spectrophotometer. The protein levels of glutathione S-transferase-π, proliferating cell nuclear antigen (PCNA), STAT3, and PKM2 were examined by Western blot and immunofluorescence. RESULTS: We found that the Warburg effect was increased in liver precancerous lesions in rats. PKM2 and p-STAT3 were upregulated in activated oval cells in liver precancerous lesions in rats. The Warburg effect, p-PKM2, and p-STAT3 expression were also increased in transformed WB-F344 cells. STAT3 activation promoted the clonal formation rate, aneuploidy, alpha-fetoprotein expression, PCNA expression, G1/S phase transition, the Warburg effect, PKM2 phosphorylation, and nuclear translocation in transformed WB-F344 cells. Moreover, the Warburg effect was inhibited by stattic, a specific inhibitor of STAT3, and further reduced in transformed WB-F344 cells after the intervention for PKM2. CONCLUSION: The Warburg effect is initiated in liver precancerous lesions in rats. STAT3 activation promotes the Warburg effect by enhancing the phosphorylation of PKM2 in transformed WB-F344 cells.


Assuntos
Transformação Celular Neoplásica/patologia , Neoplasias Hepáticas/patologia , Lesões Pré-Cancerosas/patologia , Piruvato Quinase/metabolismo , Fator de Transcrição STAT3/metabolismo , Animais , Linhagem Celular , Transformação Celular Neoplásica/efeitos dos fármacos , Óxidos S-Cíclicos/farmacologia , Modelos Animais de Doenças , Glicólise/efeitos dos fármacos , Hepatócitos , Humanos , Peróxido de Hidrogênio/toxicidade , Fígado/citologia , Fígado/patologia , Masculino , Metilnitronitrosoguanidina/toxicidade , Fosforilação/efeitos dos fármacos , Lesões Pré-Cancerosas/induzido quimicamente , Ratos , Ratos Wistar , Fator de Transcrição STAT3/antagonistas & inibidores , Células-Tronco , Regulação para Cima
11.
Front Plant Sci ; 10: 338, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30949192

RESUMO

Blue mold and core rot caused by Penicillium expansum and Trichothecium roseum are major diseases of apple fruit in China; however, their differential aggressiveness in apples and effect on fruit postharvest physiology are unclear. The effects of colonization of apples cv. Red Delicious by both pathogens were compared to physiological parameters of ripening and release of volatile compounds (VOCs). P. expansum colonization showed increased aggressiveness compared to T. roesum colonization of apple fruits. P. expansum enhanced colonization occurred with differential higher ethylene production and respiratory rate evolution, lower membrane integrity and fruit firmness in correspondence with the colonization pattern of inoculated apples. Moreover, P. expansum caused lower contents of total soluble solid and titratable acid, and higher malondialdehyde compared with T. roesum colonization. While both pathogen infections enhanced VOCs release, compared with T. roseum inoculated apples, P. expansum inoculated apple showed a higher total VOCs production including alcohols, aldehydes and esters, being the C6 alcohols, aldehydes and esters amount. PLS-DA analysis indicated that hexanoic acid was the most important factor to distinguish the inoculated fruits from the controls. Interestingly, propyl acetate and hexyl benzoate, and undecylenic acid and hexadecane were only identified in the P. expansum and T. roseum inoculated fruits, respectively. Taken together, our findings indicate that both fungi inoculations promote apple fruit ripening and release specific VOCs; moreover, apple fruits are more susceptible to P. expansum colonization than T. roesum.

12.
Food Chem ; 289: 278-284, 2019 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-30955613

RESUMO

Fusarium rot of muskmelon is a common and frequently-occurring postharvest disease, which leads to quality deterioration and neosolaniol (NEO) contamination. New strategies to control postharvest decay and reduce NEO contamination are of paramount importance. The effects of acetylsalicylic acid (ASA) treatment on the growth of Fusarium sulphureum in vitro, and Fusarium rot development and NEO accumulation in fruits inoculated with F. sulphureum in vivo were investigated. The results showed that ASA inhibited the growth of F. sulphureum, evident morphological and major cellular changes were observed under the microscope. In vivo testing showed that 3.2 mg/mL ASA significantly suppressed Fusarium rot development and NEO accumulation after 6 and 8 d of pathogen inoculation. Meanwhile, Tri gene expressions involved in NEO biosynthesis were down-regulated after treatment. Taken together, ASA treatment not only reduced Fusarium rot development by inhibiting the growth of F. sulphureum, but decreased NEO accumulation by suppressing NEO biosynthesis pathway.


Assuntos
Aspirina/farmacologia , Cucurbitaceae/química , Tricotecenos/metabolismo , Cucurbitaceae/metabolismo , Cucurbitaceae/microbiologia , Frutas/química , Frutas/metabolismo , Frutas/microbiologia , Fusarium/efeitos dos fármacos , Fusarium/crescimento & desenvolvimento , Fusarium/metabolismo , Fusarium/ultraestrutura , Doenças das Plantas/prevenção & controle , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , RNA de Plantas/genética , RNA de Plantas/metabolismo , Tricotecenos/química
13.
Food Chem ; 289: 369-376, 2019 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-30955625

RESUMO

The effects of postharvest treatment with sodium silicate (Si) (100 mM) on mitochondrial ROS production and energy metabolism of the muskmelon fruits (cv. Yujinxiang) on development of defense responses to Trichothecium roseum were studied. Si treatment decreased decay severity of inoculated muskmelons, enhanced the activities of energy metabolism of key enzymes and kept the intracellular ATP at a higher level; meanwhile, Si also induced the mtROS accumulation such as H2O2 and superoxide anion. TMT-based quantitative proteomics analysis revealed that a total of 24 proteins with significant differences in abundance involved in energy metabolism, defense and stress responses, glycolytic and TCA cycle, and oxidation-reduction process. It is suggested by our study that melon fruit mitochondria, when induced by Si treatments, play a key role in priming of host resistance against T. roseum infection through the regulation of energy metabolism and ROS production in the pathogen infected muskmelon fruits.


Assuntos
Cucumis , Metabolismo Energético/efeitos dos fármacos , Frutas/metabolismo , Mitocôndrias/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Silicatos/farmacologia , Trifosfato de Adenosina/análise , Frutas/efeitos dos fármacos , Peróxido de Hidrogênio/metabolismo , Oxirredução , Doenças das Plantas/prevenção & controle , Proteínas/metabolismo , Superóxidos/metabolismo
14.
Nanoscale Res Lett ; 14(1): 90, 2019 Mar 12.
Artigo em Inglês | MEDLINE | ID: mdl-30874913

RESUMO

This study aimed to improve the magnetofection of MG-63 osteoblasts by integrating the use of a novel uniform magnetic field with low molecular weight polyethylenimine modified superparamagnetic iron oxide nanoparticles (PEI-SPIO-NPs). The excellent characteristics of PEI-SPIO-NPs such as size, zeta potential, the pDNA binding and protective ability were determined to be suitable for gene delivery. The novel uniform magnetic field enabled polyethylenimine-modified superparamagnetic iron oxide nanoparticles/pDNA complexes (PEI-SPIO-NPs/pDNA complexes) to rapidly and uniformly distribute on the surface of MG-63 cells, averting local transfection and decreasing disruption of the membrane caused by the centralization of positively charged PEI-SPIO-NPs, thereby increasing the effective coverage of magnetic gene carriers during transfection, and improving magnetofection efficiency. This innovative uniform magnetic field can be used to determine the optimal amount between PEI-SPIO-NPs and pDNA, as well as screen for the optimal formulation design of magnetic gene carrier under the homogenous conditions. Most importantly, the novel uniform magnetic field facilitates the transfection of PEI-SPIO-NPs/pDNA into osteoblasts, thereby providing a novel approach for the targeted delivery of therapeutic genes to osteosarcoma tissues as well as a reference for the treatment of other tumors.

15.
J Agric Food Chem ; 67(15): 4337-4345, 2019 Apr 17.
Artigo em Inglês | MEDLINE | ID: mdl-30865450

RESUMO

Mitochondria play an essential part in fighting against pathogen infection in the defense responses of fruits. In this study, we investigated the reactive oxygen species (ROS) production, energy metabolism, and changes of mitochondrial proteins in harvested muskmelon fruits ( Cucumis melo cv. Yujinxiang) inoculated with Trichothecium roseum. The results indicated that the fungal infection obviously induced the H2O2 accumulation in mitochondria. Enzyme activities were inhibited in the first 6 h postinoculation (hpi), including succinic dehydrogenase, cytochrome c oxidase, H+-ATPase, and Ca2+-ATPase. However, the activities of Ca2+-ATPase and H+-ATPase and the contents of intracellular adenosine triphosphate (ATP) were improved to a higher level at 12 hpi. A total of 42 differentially expressed proteins were identified through tandem mass tags-based proteomic analyses, which are mainly involved in energy metabolism, stress responses and redox homeostasis, glycolysis and tricarboxylic acid cycle, and transporter and mitochondria dysfunction. Taken together, our results suggest that mitochondria play crucial roles in the early defense responses of muskmelons against T. roseum infection through regulation of ROS production and energy metabolism.


Assuntos
Cucumis melo/metabolismo , Cucumis melo/microbiologia , Metabolismo Energético , Hypocreales/fisiologia , Doenças das Plantas/microbiologia , Espécies Reativas de Oxigênio/metabolismo , Trifosfato de Adenosina/metabolismo , Cucumis melo/enzimologia , Cucumis melo/genética , Complexo IV da Cadeia de Transporte de Elétrons/genética , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Mitocôndrias/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , ATPases Translocadoras de Prótons/genética , ATPases Translocadoras de Prótons/metabolismo , Succinato Desidrogenase/genética , Succinato Desidrogenase/metabolismo
16.
Toxins (Basel) ; 11(2)2019 01 28.
Artigo em Inglês | MEDLINE | ID: mdl-30696025

RESUMO

A simple and sensitive label-free impedimetric aptasensor for rapid determination of ochratoxin A (OTA) has been developed, which was based on the combination between thiolated aptamer and gold nanoparticles by layer-by-layer self-assembly. Because of the interaction between aptamer and OTA, the relative normalized electron-transfer resistance (ΔRct) values obtained by electrochemical impedance spectroscopy (EIS) was proportional to the concentration of OTA and showed a good linear relationship from 0.1 to 10.0 ng/mL, with a lower detection limit (0.030 ng/mL) than one-step thiolated DNA aptasensor. The established method was successfully applied to detect and analyze OTA in table wine and grape juice, and the recovery was 90.56%⁻104.21% when PVP effective removed of phenolic substances. The label-free impedimetric aptasensor was used for rapid detection and quantitation of OTA in the inoculated grapes with the Aspergillus Nigri (H1), and the production of OTA (62.4 µg/kg, 20 µg/kg) far exceeded the maximum levels of 2 µg/kg after inoculation for three days. The developed method exhibited a good specificity, high sensitivity, time-efficient, and it could be applied to detect the OTA concentration in grape and its commodities.


Assuntos
Técnicas Biossensoriais , Contaminação de Alimentos/análise , Ocratoxinas/análise , Aptâmeros de Nucleotídeos/química , Aspergillus , Espectroscopia Dielétrica , Eletrodos , Sucos de Frutas e Vegetais/análise , Ouro/química , Nanopartículas Metálicas/química , Ocratoxinas/química , Vitis/química , Vitis/microbiologia , Vinho/análise
17.
Am J Transplant ; 19(8): 2306-2317, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-30664327

RESUMO

For many patients with end-stage kidney disease, transplantation improves survival and quality of life compared with dialysis. However, complications and side effects in kidney transplant recipients can limit their ability to participate in activities of daily living including work, study, and recreational activities. The aim of this study was to identify the characteristics, content, and psychometric properties of the outcome measures used to assess life participation in kidney transplant recipients. We searched MEDLINE, Embase, PsycINFO, and CINAHL from inception to July 2018 for all studies that reported life participation in kidney transplant recipients. Two authors identified instruments measuring life participation and reviewed for characteristics. In total, 230 studies were included: 19 (8%) randomized trials, 17 (7%) nonrandomized trials, and 194 (85%) observational studies. Across these studies, we identified 29 different measures that were used to assess life participation. Twelve (41%) measures specifically assessed aspects of life participation (eg, disability assessment, daily activities of living), while 17 (59%) assessed other constructs (eg, quality of life) that included questions on life participation. Validation data to support the use of these measures in kidney transplant recipients were available for only 7 measures. A wide range of measures have been used to assess life participation in kidney transplant recipients, but validation data supporting the use of these measures in this population are sparse. A content relevant and validated measure to improve the consistency and accuracy of measuring life participation in research may inform strategies for transplant recipients to be better able to engage in their life activities.

18.
Sci Rep ; 8(1): 13014, 2018 08 29.
Artigo em Inglês | MEDLINE | ID: mdl-30158652

RESUMO

The response of plant root development to nutrient deficiencies is critical for crop production. Auxin, nitric oxide (NO), and strigolactones (SLs) are important regulators of root growth under low-nitrogen and -phosphate (LN and LP) conditions. Polar auxin transport in plants, which is mainly dependent on auxin efflux protein PINs, creates local auxin maxima to form the basis for root initiation and elongation; however, the PIN genes that play an important role in LN- and LP-modulated root growth remain unclear. qRT-PCR analysis of OsPIN family genes showed that the expression of OsPIN1b is most abundant in root tip and is significantly downregulated by LN, LP, sodium nitroprusside (SNP, NO donor), and GR24 (analogue of SLs) treatments. Seminal roots in ospin1b mutants were shorter than those of the wild type; and the seminal root, [3H]IAA transport, and IAA concentration responses to LN, LP, SNP, and GR24 application were attenuated in ospin1b-1 mutants. pCYCB1;1::GUS expression was upregulated by LN, LP, SNP, and GR24 treatments in wild type, but not in the ospin1b-1 mutant, suggesting that OsPIN1b is involved in auxin transport and acts as a downstream mediator of NO and SLs to induce meristem activity in root tip in rice under LN and LP.


Assuntos
Proteínas de Membrana Transportadoras/metabolismo , Meristema/enzimologia , Meristema/crescimento & desenvolvimento , Nitrogênio/metabolismo , Oryza/enzimologia , Oryza/crescimento & desenvolvimento , Fosfatos/metabolismo , Perfilação da Expressão Gênica , Proteínas de Membrana Transportadoras/genética , Meristema/metabolismo , Oryza/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raízes de Plantas/enzimologia , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Reação em Cadeia da Polimerase em Tempo Real
19.
J Cell Biochem ; 119(11): 8872-8886, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-30076626

RESUMO

Human mesenchymal stem cells (MSCs) are a heterogeneous subset of nonhematopoietic multipotent stromal stem cells and can differentiate into mesodermal lineage, such as adipocytes, osteocytes, and chondrocytes, as well as ectodermal and endodermal lineages. Human umbilical cord (UC) is one of the most promising sources of MSCs. However, the molecular and cellular characteristics of UC-derived MSCs (UC-MSCs) require extensive investigations, which are hampered by the limited lifespan and the diminished potency over passages. Here, we used the piggyBac transposon-based simian virus 40 T antigen (SV40T) immortalization system and effectively immortalized UC-MSCs, yielding the iUC-MSCs. A vast majority of the immortalized lines are positive for MSC markers but not for hematopoietic markers. The immortalization phenotype of the iUC-MSCs can be effectively reversed by flippase recombinase-induced the removal of SV40T antigen. While possessing long-term proliferation capability, the iUC-MSCs are not tumorigenic in vivo. Upon bone morphogenetic protein 9 (BMP9) stimulation, the iUC-MSC cells effectively differentiate into osteogenic, chondrogenic, and adipogenic lineages both in vitro and in vivo, which is indistinguishable from that of primary UC-MSCs, indicating that the immortalized UC-MSCs possess the characteristics similar to that of their primary counterparts and retain trilineage differentiation potential upon BMP9 stimulation. Therefore, the engineered iUC-MSCs should be a valuable alternative cell source for studying UC-MSC biology and their potential utilities in immunotherapies and regenerative medicine.


Assuntos
Adipogenia/fisiologia , Diferenciação Celular/fisiologia , Fator 2 de Diferenciação de Crescimento/metabolismo , Células-Tronco Mesenquimais/metabolismo , Osteogênese/fisiologia , Cordão Umbilical/citologia , Análise de Variância , Animais , Antígenos Transformantes de Poliomavirus/metabolismo , Técnicas de Cultura de Células/métodos , Proliferação de Células , Condrogênese/fisiologia , Feminino , Vetores Genéticos , Células HEK293 , Humanos , Recém-Nascido , Camundongos Nus , Transposon Resolvases/metabolismo
20.
Int J Mol Med ; 42(4): 1977-1986, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30085335

RESUMO

A large number of functional hepatocytes is required for bioartificial liver therapy. Simian virus 40 T­antigen (SV40T) has been previously reported to improve the immortalized proliferation of primary hepatocytes to generate a sufficient number of cells; however, these long­term immortalized hepatocytes may induce further malignant transformation in vivo. In the present study, the SV40T immortalization gene and two suicide genes, herpes simplex virus thymidine kinase (HSV­tk) and cytosine deaminase (CD), were transducted into primary hepatocytes to construct a novel type of Cre/LoxP­mediated reversible immortalized hepatocyte line. Polymerase chain reaction analysis and western blotting confirmed that the SV40T, HSV­tk and CD genes were successfully inserted into hepatic progenitor cells and their expression was controlled by Cre/LoxP recombination. Total removal of SV40T could be achieved via the ganciclovir (GCV)/HSV­tk suicide system. Cells maintained their biosafety in vivo with CD gene expression and 5­fluorocytosine (5­FC) induced cell death. Following transplantation into the carbon tetrachloride (CCl4) model group, the majority of cells had survived after 14 days post­implantation and a number of the cells had transported into the liver parenchyma. When compared with the CCl4 model group, the transplanted cells repaired the liver biochemical index and pathological structure markedly. Thus, the present study reports a novel reversible immortalized hepatocyte with double suicide genes, which exhibited the cellular phenotype and recovery function of normal liver cells. This method maximally guaranteed the biological safety of immortalized hepatocytes for in vivo application, providing a reliable, safe and ideal cell material for the artificial liver technique.


Assuntos
Hepatócitos/citologia , Hepatócitos/metabolismo , Fígado/citologia , Animais , Western Blotting , Linhagem Celular , Sobrevivência Celular/genética , Sobrevivência Celular/fisiologia , Humanos , Falência Hepática/metabolismo , Camundongos , Camundongos Nus , Plasmídeos/genética
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