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1.
Anal Methods ; 14(8): 781-788, 2022 02 24.
Artigo em Inglês | MEDLINE | ID: mdl-35083987

RESUMO

Glove-based wearable sensors can offer the potential ability to a fast and on-site environmental threat assessment, which is crucial for timely and informed incident management. In this study, an on-demand surface-enhanced Raman scattering (SERS) array sensor has been patterned on fire-retardant fibre gloves via the screen-printing technique in large batches. The screen-printed ink contains one-pot synthesized silver nanoparticle and molybdenum disulfide nanocomposite (Ag/MoS2), and polyanionic cellulose (PAC) as a new adhesive agent. Rhodamine 6G (R6G) was employed as an initial probe molecule to systematically evaluate the performance of the resulting sensor. The results suggest that the fabricated fire-retardant screen-printed SERS array sensor displays high reproducibility and stability at 250 °C, with the lower detection limit of 10-13 M for R6G. The spot-to-spot SERS signals show that the intensity variation was less than 10%. Besides, the SERS signals can be maintained over 7 weeks. Further investigation was then successfully carried out to detect polycyclic aromatic hydrocarbons (PAHs), which are commonly used as flammable chemicals. In our perception, this wearable fire-retardant screen-printed SERS array sensor would be an ideal candidate for practical on-site environmental emergency monitoring due to its fire-retardant capability and timely measurement on a portable carrier.


Assuntos
Retardadores de Chama , Nanopartículas Metálicas , Dispositivos Eletrônicos Vestíveis , Nanopartículas Metálicas/química , Reprodutibilidade dos Testes , Prata/química
2.
J Integr Plant Biol ; 64(1): 39-55, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34796654

RESUMO

Cotton which produces natural fiber materials for the textile industry is one of the most important crops in the world. Class II KNOX proteins are often considered as transcription factors in regulating plant secondary cell wall (SCW) formation. However, the molecular mechanism of the KNOX transcription factor-regulated SCW synthesis in plants (especially in cotton) remains unclear in details so far. In this study, we show a cotton class II KNOX protein (GhKNL1) as a transcription repressor functioning in fiber development. The GhKNL1-silenced transgenic cotton produced longer fibers with thicker SCWs, whereas GhKNL1 dominant repression transgenic lines displayed the opposite fiber phenotype, compared with controls. Further experiments revealed that GhKNL1 could directly bind to promoters of GhCesA4-2/4-4/8-2 and GhMYB46 for modulating cellulose synthesis during fiber SCW development in cotton. On the other hand, GhKNL1 could also suppress expressions of GhEXPA2D/4A-1/4D-1/13A through binding to their promoters for regulating fiber elongation of cotton. Taken together, these data revealed GhKNL1 functions in fiber elongation and SCW formation by directly repressing expressions of its target genes related to cell elongation and cellulose synthesis. Thus, our data provide an effective clue for potentially improving fiber quality by genetic manipulation of GhKNL1 in cotton breeding.


Assuntos
Fibra de Algodão , Gossypium , Parede Celular/metabolismo , Regulação da Expressão Gênica de Plantas/genética , Gossypium/genética , Gossypium/metabolismo , Melhoramento Vegetal , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo
3.
Neurol India ; 69(1): 115-118, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33642281

RESUMO

CONTEXT: A host of microRNAs have been reported to suppress tumor growth, invasion, and metastasis and play roles in neurodegeneration disorders. Moreover, microRNA changes are found in the peripheral blood, cerebrospinal fluid (CSF), and brain tissues of central nervous system diseases, including glioma, Alzheimer's disease (AD), Parkinson's disease (PD), multiple sclerosis, and depression. Compared with other body fluids, CSF can reflect the brain pathological processes more accurately. AIMS: To understand whether microRNA expression may be misregulated in patients with PD, and further discover potential diagnostic biomarkers and promising therapeutic targets for PD. MATERIALS AND METHODS: Here, through real-time reverse-transcription polymerase chain reaction (RT-PCR), we compared CSF microRNA from 15 PD patients, 11 AD patients, and 16 controls with other neurologic disorders, such as encephalitis and Guillain-Barre syndrome. RESULTS: Finally, we identified hsa-miR-626 changes in the CSF of PD patients. The mean expression level of hsa-miR-626 was significantly reduced in the CSF of PD patients compared with AD patients and controls. CONCLUSIONS: Our approach provides a preliminary research for identifying biomarkers in the CSF that could be used for the detection, diagnosis, and monitoring of PD.


Assuntos
Doença de Alzheimer , MicroRNAs , Doença de Parkinson , Biomarcadores , Humanos , MicroRNAs/genética , Doença de Parkinson/genética
4.
Acta Pharmacol Sin ; 42(1): 45-54, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32451415

RESUMO

Lifestyle factors may affect mental health and play a critical role in the development of neurodegenerative diseases including Alzheimer's disease (AD). However, whether the temperatures of daily beverages have any impact on cognitive function and AD development has never been studied. In this study, we investigated the effects of daily drinking water temperatures on cognitive function and AD development and progression in mice and the underlying mechanisms. Cognitive function of mice was assessed using passive avoidance test, open field test, and Morris water maze. Wild-type Kunming mice receiving intragastric water (IW, 10 mL/kg, 2 times/day) at 0 °C for consecutive 15 days displayed significant cognitive defects accompanied by significant decrease in gain of body weight, gastric emptying rate, pepsin activity, and an increase in the energy charge in the cortex when compared with mice receiving the same amount of IW at 25 °C (a temperature mimicking most common drinking habits in human), suggesting the altered neuroenergetics may cause cognitive decline. Similarly, in the transgenic APPwse/PS1De9 familial AD mice and their age- and gender-matched wild-type C57BL/6 mice, receiving IW at 0 °C, but not at 25 °C, for 35 days caused a significant time-dependent decrease in body weight and cognitive function, accompanied by a decreased expression of PI3K, Akt, the glutamate/GABA ratio, as well as neuropathy with significant amyloid lesion in the cortex and hippocampus. All of these changes were significantly aggravated in the APPwse/PS1De9 mice than in the control C57BL/6 mice. These data demonstrate that daily beverage at 0 °C may alter brain insulin-mediated neuroenergetics, glutamate/GABA ratio, cause cognitive decline and neuropathy, and promote AD progression.


Assuntos
Doença de Alzheimer/fisiopatologia , Cognição/fisiologia , Temperatura Baixa , Água Potável/administração & dosagem , Doença de Alzheimer/metabolismo , Doença de Alzheimer/patologia , Animais , Encéfalo/metabolismo , Encéfalo/patologia , Progressão da Doença , Água Potável/química , Ácido Glutâmico/metabolismo , Insulina/metabolismo , Masculino , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Teste do Labirinto Aquático de Morris/fisiologia , Neurotransmissores/metabolismo , Teste de Campo Aberto/fisiologia , Transdução de Sinais/fisiologia , Ácido gama-Aminobutírico/metabolismo
5.
Brain ; 143(1): 222-233, 2020 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-31819945

RESUMO

Essential tremor is one of the most common movement disorders. Despite its high prevalence and heritability, the genetic aetiology of essential tremor remains elusive. Up to now, only a few genes/loci have been identified, but these genes have not been replicated in other essential tremor families or cohorts. Here we report a genetic study in a cohort of 197 Chinese pedigrees clinically diagnosed with essential tremor. Using a comprehensive strategy combining linkage analysis, whole-exome sequencing, long-read whole-genome sequencing, repeat-primed polymerase chain reaction and GC-rich polymerase chain reaction, we identified an abnormal GGC repeat expansion in the 5' region of the NOTCH2NLC gene that co-segregated with disease in 11 essential tremor families (5.58%) from our cohort. Clinically, probands that had an abnormal GGC repeat expansion were found to have more severe tremor phenotypes, lower activities of daily living ability. Obvious genetic anticipation was also detected in these 11 essential tremor-positive families. These results indicate that abnormal GGC repeat expansion in the 5' region of NOTCH2NLC gene is associated with essential tremor, and provide strong evidence that essential tremor is a family of diseases with high clinical and genetic heterogeneities.


Assuntos
Povo Asiático/genética , Tremor Essencial/genética , Expansão das Repetições de Trinucleotídeos/genética , Adulto , Idoso , Feminino , Sequência Rica em GC , Ligação Genética , Humanos , Corpos de Inclusão Intranuclear/genética , Corpos de Inclusão Intranuclear/ultraestrutura , Masculino , Microscopia Eletrônica , Pessoa de Meia-Idade , Doenças Neurodegenerativas/genética , Linhagem , Reação em Cadeia da Polimerase , Pele/ultraestrutura , Sequenciamento do Exoma , Sequenciamento Completo do Genoma
6.
J Clin Neurosci ; 70: 198-201, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31492481

RESUMO

microRNAs have been reported to suppress tumor growth, invasion, and metastasis and play roles in neurodegeneration disorders. Moreover, changes in microRNAs are found in the peripheral blood, cerebrospinal fluid (CSF), and brain tissues in patients of central nervous system diseases, including glioma, Alzheimer's disease (AD), Parkinson's disease (PD), multiple sclerosis and depression. Compared with other bodily fluids, CSF is the most accurate at representing the pathological processes of the brain. To understand whether microRNA expression may be dysregulated in the patients of PD, and to further discover potential diagnostic biomarkers and promising therapeutic targets for PD, we used real-time polymerase chain reaction (RT-PCR) to compare CSF microRNAs from 20 PD patients, 13 AD patients and 27 controls with other neurologic disorders such as encephalitis and Guillain-Barre syndrome. Finally, we found that the mean expression level of hsa-miR-626 was significantly reduced in the CSF of patients with PD compared with AD and controls. Our approach potentially identified a biomarker in CSF that upon further investigation, could be used for the detection, diagnosis, and monitoring of PD in combination with other PD biomarkers.


Assuntos
Biomarcadores/líquido cefalorraquidiano , MicroRNAs/líquido cefalorraquidiano , Doença de Parkinson/líquido cefalorraquidiano , Doença de Parkinson/diagnóstico , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
7.
Oxid Med Cell Longev ; 2017: 5094934, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28348719

RESUMO

Loss-of-function mutations in gene encoding DJ-1 contribute to the pathogenesis of autosomal recessive early-onset familial forms of Parkinson's disease (PD). DJ-1 is a multifunctional protein and plays a protective role against oxidative stress-induced mitochondrial damage and cell death, but the exact mechanism underlying this is not yet clearly understood. Here, using coimmunoprecipitation (Co-IP) and immunofluorescence methods, we prove that Bcl-2-associated athanogene 5 (BAG5), a BAG family member, interacts with DJ-1 in mammalian cells. Moreover, we show that BAG5 could decrease stability of DJ-1 and weaken its role in mitochondrial protection probably by influencing dimerization in stress condition. Our study reveals the relationship of BAG5 and DJ-1 suggesting a potential role for BAG5 in the pathogenesis of PD through its functional interactions with DJ-1.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Mitocôndrias/metabolismo , Fármacos Neuroprotetores/metabolismo , Estresse Oxidativo , Proteína Desglicase DJ-1/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/antagonistas & inibidores , Proteínas Adaptadoras de Transdução de Sinal/genética , Apoptose/efeitos dos fármacos , Células HEK293 , Proteínas de Choque Térmico HSP70/antagonistas & inibidores , Proteínas de Choque Térmico HSP70/genética , Proteínas de Choque Térmico HSP70/metabolismo , Humanos , Imunoprecipitação , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Microscopia de Fluorescência , Proteína Desglicase DJ-1/genética , Interferência de RNA , RNA Interferente Pequeno/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Rotenona/farmacologia
8.
Plant J ; 89(5): 957-971, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-27888523

RESUMO

Arabinogalactan proteins (AGPs) are highly glycosylated proteins that play pivotal roles in diverse developmental processes in plants. Type-II AG glycans, mostly O-linked to the hydroxyproline residues of the protein backbone, account for up to 95% w/w of the AGP, but their functions are still largely unclear. Cotton fibers are extremely elongated single-cell trichomes on the seed epidermis; however, little is known of the molecular basis governing the regulation of fiber cell development. Here, we characterized the role of a CAZy glycosyltransferase 31 (GT31) family member, GhGalT1, in cotton fiber development. The fiber length of the transgenic cotton overexpressing GhGalT1 was shorter than that of the wild type, whereas in the GhGalT1-silenced lines there was a notable increase in fiber length compared with wild type. The carbohydrate moieties of AGPs were altered in fibers of GhGalT1 transgenic cotton. The galactose: arabinose ratio of AG glycans was higher in GhGalT1 overexpression fibers, but was lower in GhGalT1-silenced lines, compared with that in the wild type. Overexpression of GhGalT1 upregulates transcript levels of a broad range of cell wall-related genes, especially the fasciclin-like AGP (FLA) backbone genes. An enzyme activity assay demonstrated that GhGalT1 is a ß-1,3-galactosyltransferase (ß-1,3-GalT) involved in biosynthesis of the ß-1,3-galactan backbone of the type-II AG glycans of AGPs. We also show that GhGalT1 can form homo- and heterodimers with other cotton GT31 family members to facilitate AG glycan assembly of AGPs. Thus, our data demonstrate that GhGalT1 influences cotton fiber development via controlling the glycosylation of AGPs, especially FLAs.


Assuntos
Galactosiltransferases/metabolismo , Gossypium/enzimologia , Proteínas de Plantas/metabolismo , Parede Celular/metabolismo , Fibra de Algodão , Galactosiltransferases/genética , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Gossypium/genética , Gossypium/metabolismo , Mucoproteínas/genética , Mucoproteínas/metabolismo , Proteínas de Plantas/genética
9.
Neural Regen Res ; 11(3): 427-34, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-27127481

RESUMO

GIT1, a G-protein-coupled receptor kinase interacting protein, has been reported to be involved in neurite outgrowth. However, the neurobiological functions of the protein remain unclear. In this study, we found that GIT1 was highly expressed in the nervous system, and its expression was maintained throughout all stages of neuritogenesis in the brain. In primary cultured mouse hippocampal neurons from GIT1 knockout mice, there was a significant reduction in total neurite length per neuron, as well as in the average length of axon-like structures, which could not be prevented by nerve growth factor treatment. Overexpression of GIT1 significantly promoted axon growth and fully rescued the axon outgrowth defect in the primary hippocampal neuron cultures from GIT1 knockout mice. The GIT1 N terminal region, including the ADP ribosylation factor-GTPase activating protein domain, the ankyrin domains and the Spa2 homology domain, were sufficient to enhance axonal extension. Importantly, GIT1 bound to many tubulin proteins and microtubule-associated proteins, and it accelerated microtubule assembly in vitro. Collectively, our findings suggest that GIT1 promotes neurite outgrowth, at least partially by stimulating microtubule assembly. This study provides new insight into the cellular and molecular pathogenesis of GIT1-associated neurological diseases.

10.
Sci Rep ; 6: 20371, 2016 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-26829353

RESUMO

Di19 (drought-induced protein 19) family is a novel type of Cys2/His2 zinc-finger proteins. In this study, we demonstrated that cotton Di19-1 and Di19-2 (GhDi19-1/-2) proteins could be phosphorylated in vitro by the calcium-dependent protein kinase (CDPK). Mutation of Ser to Ala in N-terminus of GhDi19-1/-2 led to the altered subcellular localization of the two proteins, but the constitutively activated form (Ser was mutated to Asp) of GhDi19-1/-2 still showed the nuclear localization. GhDi19-1/-2 overexpression transgenic Arabidopsis seedlings displayed the hypersensitivity to high salinity and abscisic acid (ABA). However, Ser site-mutated GhDi19-1(S116A) and GhDi19-2(S114A), and Ser and Thr double sites-mutated GhDi19-1(S/T-A/A) and GhDi19-2(S/T-A/A) transgenic Arabidopsis did not show the salt- and ABA-hypersensitive phenotypes. In contrast, overexpression of Thr site-mutated GhDi19-1(T114A) and GhDi19-2(T112A) in Arabidopsis still resulted in salt- and ABA-hypersensitivity phenotypes, like GhDi19-1/-2 transgenic lines. Overexpression of GhDi19-1/-2 and their constitutively activated forms in Atcpk11 background could recover the salt- and ABA-insensitive phenotype of the mutant. Thus, our results demonstrated that Ser phosphorylation (not Thr phosphorylation) is crucial for functionally activating GhDi19-1/-2 in response to salt stress and ABA signaling during early plant development, and GhDi19-1/-2 proteins may be downstream targets of CDPKs in ABA signal pathway.


Assuntos
Ácido Abscísico/metabolismo , Gossypium/fisiologia , Proteínas de Plantas/metabolismo , Salinidade , Serina/metabolismo , Transdução de Sinais , Estresse Fisiológico , Sequência de Aminoácidos , Arabidopsis/genética , Cálcio/metabolismo , Proteínas de Ligação ao Cálcio/química , Proteínas de Ligação ao Cálcio/genética , Proteínas de Ligação ao Cálcio/metabolismo , Secas , Regulação da Expressão Gênica de Plantas , Germinação/genética , Mutação , Fenótipo , Fosforilação , Proteínas de Plantas/química , Proteínas de Plantas/genética , Proteínas Quinases/química , Proteínas Quinases/genética , Proteínas Quinases/metabolismo , Transporte Proteico , Plântula/genética , Plântula/metabolismo
11.
Chin Med J (Engl) ; 128(15): 2097-101, 2015 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-26228225

RESUMO

OBJECTIVE: To summarize the clinical presentation, pathogenesis, neuroimaging, treatment, and outcome of stroke-like migraine attacks after radiation therapy (SMART) syndrome, and to propose diagnostic criteria for this disorder. DATA SOURCES: We searched the PubMed database for articles in English published from 1995 to 2015 using the terms of "stroke-like AND migraine AND radiation." Reference lists of the identified articles and reviews were used to retrieve additional articles. STUDY SELECTION: Data and articles related to late-onset effects of cerebral radiation were selected and reviewed. RESULTS: SMART is a rare condition that involves complex migraines with focal neurologic deficits following cranial irradiation for central nervous system malignancies. The recovery, which ranges from hours to days to weeks, can be partial or complete. We propose the following diagnostic criteria for SMART: (1) Remote history of therapeutic external beam cranial irradiation for malignancy; (2) prolonged, reversible clinical manifestations mostly years after irradiation, which may include migraine, seizures, hemiparesis, hemisensory deficits, visuospatial defect, aphasia, confusion and so on; (3) reversible, transient, unilateral cortical gadolinium enhancement correlative abnormal T2 and fluid-attenuated inversion recovery signal of the affected cerebral region; (4) eventual complete or partial recovery, the length of duration of recovery ranging from hours to days to weeks; (5) no evidence of residual or recurrent tumor; (6) not attributable to another disease. To date, no specific treatment has been identified for this syndrome. CONCLUSIONS: SMART is an extremely rare delayed complication of brain irradiation. However, improvements in cancer survival rates have resulted in a rise in its frequency. Hence, awareness and recognition of the syndrome is important to make a rapid diagnosis and avoid aggressive interventions such as brain biopsy and cerebral angiography.


Assuntos
Transtornos de Enxaqueca/diagnóstico , Transtornos de Enxaqueca/etiologia , Lesões por Radiação/complicações , Lesões por Radiação/diagnóstico , Acidente Vascular Cerebral/diagnóstico , Neoplasias do Sistema Nervoso Central/terapia , Feminino , Humanos , Masculino
12.
Eur Spine J ; 24 Suppl 4: S636-43, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25906379

RESUMO

PURPOSE: To report an unusual case of idiopathic hypertrophic spinal pachymeningitis (IHSP) with a review of relevant literature and to discuss the etiology, clinical features, imaging, treatment and prognosis of IHSP. METHODS: The case of a 44-year-old woman is reported. MEDLINE was used to search relevant literatures written in English since 2004. RESULTS: The patient suffered from progressive mild thoracic backache followed by truncal and lower extremity weakness, numbness and urinary retention. The diagnosis was confirmed by magnetic resonance (MR) imaging and histopathologic examination. Although she received corticosteroid therapy and decompressive surgery, the patient suffered a rapid relapse probably because of the withdrawal of postoperative steroid therapy. CONCLUSIONS: IHSP is a rare disease characterized by inflammatory hypertrophy of the dura mater without identifiable cause and featured clinical progress of radiculalgia to myelopathy. It is a diagnosis of exclusion. In our view, surgical decompression with postoperative steroid therapy may be optimal. Furthermore,we speculated that increased levels of protein and cell count in cerebrospinal fluid (CSF) might be positively related to the disease progression. High inflammatory signs or CSF protein and cell levels before surgery or postoperative residual lesions are possible reasons of poor prognosis in patients with IHSP.


Assuntos
Meningite/diagnóstico , Adulto , Dor nas Costas/etiologia , Terapia Combinada , Descompressão Cirúrgica/métodos , Dura-Máter/patologia , Feminino , Glucocorticoides/uso terapêutico , Humanos , Hipertrofia/complicações , Hipertrofia/diagnóstico , Hipertrofia/terapia , Hipestesia/etiologia , Imageamento por Ressonância Magnética , Meningite/complicações , Meningite/terapia , Prognóstico , Doenças Raras/patologia , Recidiva
13.
Plant Mol Biol ; 86(6): 609-25, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25218132

RESUMO

Di19 (drought-induced protein19) family is a novel type of Cys2/His2 zinc-finger proteins. In this study, Arabidopsis Di19-3 was functionally characterized. The experimental results revealed that AtDi19-3 is a transcriptional activator, and could bind to the TACA(A/G)T sequence. AtDi19-3 expression in plants was remarkably induced by NaCl, mannitol and abscisic acid (ABA). T-DNA insertion mutation of AtDi19-3 results in an increase in plant tolerance to drought and high salinity stresses and ABA, whereas overexpression of AtDi19-3 leads to a drought-, salt- and ABA-sensitive phenotype of the transgenic plants. In the presence of NaCl, mannitol or ABA, rates of seed germination and cotyledon greening in Atdi19-3 mutant were higher, but in AtDi19-3 overexpression transgenic plants were lower than those in wild type. Roots of Atdi19-3 mutant seedlings were longer, but those of AtDi19-3 overexpression transgenic seedlings were shorter than those of wild type. Chlorophyll and proline contents in Atdi19-3 mutant were higher, but in AtDi19-3 overexpression seedlings were lower than those in wild type. Atdi19-3 mutant showed greater drought-tolerance, whereas AtDi19-3 overexpression transgenic plants exhibited more drought-sensitivity than wild type. Furthermore, expression of the genes related to ABA signaling pathway was altered in Atdi19-3 mutant and AtDi19-3 transgenic plants. These data suggest that AtDi19-3 may participate in plant response to drought and salt stresses in an ABA-dependent manner.


Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/fisiologia , Proteínas de Transporte/genética , Regulação da Expressão Gênica de Plantas , Transdução de Sinais , Estresse Fisiológico , Ácido Abscísico/farmacologia , Arabidopsis/efeitos dos fármacos , Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Proteínas de Transporte/metabolismo , Secas , Genes Reporter , Germinação/efeitos dos fármacos , Manitol/farmacologia , Mutagênese Insercional , Fenótipo , Reguladores de Crescimento de Plantas/farmacologia , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/genética , Raízes de Plantas/fisiologia , Plantas Geneticamente Modificadas , Ligação Proteica , Salinidade , Plântula/efeitos dos fármacos , Plântula/genética , Plântula/fisiologia , Cloreto de Sódio/farmacologia , Ativação Transcricional/efeitos dos fármacos , Dedos de Zinco
14.
J Exp Bot ; 65(15): 4133-47, 2014 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24831118

RESUMO

In this study, the GhKNL1 (KNOTTED1-LIKE) gene, encoding a classical class II KNOX protein was identified in cotton (Gossypium hirsutum). GhKNL1 was preferentially expressed in developing fibres at the stage of secondary cell wall (SCW) biosynthesis. GhKNL1 was localized in the cell nucleus, and could interact with GhOFP4, as well as AtOFP1, AtOFP4, and AtMYB75. However, GhKNL1 lacked transcriptional activation activity. Dominant repression of GhKNL1 affected fibre development of cotton. The expression levels of genes related to fibre elongation and SCW biosynthesis were altered in transgenic fibres of cotton. As a result, transgenic cotton plants produced aberrant, shrunken, and collapsed fibre cells. Length and cell-wall thickness of fibres of transgenic cotton plants were significantly reduced compared with the wild type. Furthermore, overexpression and dominant repression of GhKNL1 in Arabidopsis resulted in a reduction in interfascicular fibre cell-wall thickening of basal stems of transgenic plants. Complementation revealed that GhKNL1 rescued the defective phenotype of Arabidopsis knat7 mutant in some extent. These data suggest that GhKNL1, as a transcription factor, participates in regulating fibre development of cotton.


Assuntos
Fibra de Algodão , Gossypium/metabolismo , Proteínas de Plantas/metabolismo , Fatores de Transcrição/metabolismo , Gossypium/crescimento & desenvolvimento , Fenótipo , Desenvolvimento Vegetal , Proteínas de Plantas/isolamento & purificação , Ativação Transcricional
15.
Anal Chem ; 85(20): 9549-55, 2013 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-24047198

RESUMO

A novel surface-enhanced Raman scattering (SERS) nanosensor was developed by modifying oxidized cytochrome c (Cyt c) on gold nanoparticles (Au NPs) for the sensitive and selective determination of intracellular superoxide anion radical (O2(•-)). On the basis of the differences in the SERS spectra between the oxidized and reduced form of Cyt c, this nanosensor could be employed to investigate O2(•-) concentration by measuring the SERS spectra of the reduced Cyt c. Using this SERS nanosensor, a detection limit of 1.0 × 10(-8) M for O2(•-) could be attained. Additionally, the selectivity of the SERS nanosensor for O2(•-) was examined, showing that other reactive oxygen species (ROS) and biologically relevant species did not influence the detection of O2(•-). More importantly, the nanosensor could be delivered to the living HeLa and normal human liver cells and permitted the concentration of O2(•-) to be monitored in real time and in a noninvasive manner, which indicates that this nanosensor will be suitable for the qualitative and quantitative analysis of O2(•-) in biosystems, thus leading to a greater understanding of oxidative-stress-related diseases at a cellular level.


Assuntos
Técnicas Biossensoriais/métodos , Citocromos c/metabolismo , Ouro/química , Espaço Intracelular/química , Nanotecnologia/métodos , Análise Espectral Raman , Superóxidos/análise , Citocromos c/química , Células HeLa , Humanos , Fígado/citologia , Nanopartículas Metálicas/química , Modelos Moleculares , Conformação Proteica
16.
Plant Mol Biol ; 82(4-5): 353-65, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23625445

RESUMO

Proline-rich proteins contribute to cell wall structure of specific cell types and are involved in plant growth and development. In this study, a fiber-specific gene, GhPRP5, encoding a proline-rich protein was functionally characterized in cotton. GhPRP5 promoter directed GUS expression only in trichomes of both transgenic Arabidopsis and tobacco plants. The transgenic Arabidopsis plants with overexpressing GhPRP5 displayed reduced cell growth, resulting in smaller cell size and consequently plant dwarfs, in comparison with wild type plants. In contrast, knock-down of GhPRP5 expression by RNA interference in cotton enhanced fiber development. The fiber length of transgenic cotton plants was longer than that of wild type. In addition, some genes involved in fiber elongation and wall biosynthesis of cotton were up-regulated or down-regulated in the transgenic cotton plants owing to suppression of GhPRP5. Collectively, these data suggested that GhPRP5 protein as a negative regulator participates in modulating fiber development of cotton.


Assuntos
Fibra de Algodão , Regulação da Expressão Gênica de Plantas/fisiologia , Gossypium/metabolismo , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Prolina/genética , Arabidopsis/genética , Arabidopsis/metabolismo , Regulação da Expressão Gênica de Plantas/genética , Gossypium/genética , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/genética
17.
Sci Rep ; 3: 1537, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23524384

RESUMO

Quantum dots (QDs) have attracted increasing interest in bioimaging and sensing. Here, we report a biosensor of complex I using ubiquinone-terminated disulphides with different alkyl spacers (QnNS, n = 2, 5 and 10) as surface-capping ligands to functionalise CdSe/ZnS QDs. The enhancement or quenching of the QD bioconjugates fluorescence changes as a function of the redox state of QnNS, since QDs are highly sensitive to the electron-transfer processes. The bioconjugated QnNS-QDs emission could be modulated by complex I in the presence of NADH, which simulates an electron-transfer system part of the mitochondrial respiratory chain, providing an in vitro and intracellular complex I sensor. Epidemiological studies suggest that Parkinson's patients have the impaired activity of complex I in the electron-transfer chain of mitochondria. We have demonstrated that the QnNS-QDs system could aid in early stage Parkinson's disease diagnosis and progression monitoring by following different complex I levels in SH-SY5Y cells.


Assuntos
Técnicas Biossensoriais , Complexo I de Transporte de Elétrons/metabolismo , Mitocôndrias/metabolismo , Pontos Quânticos , Ubiquinona/química , Cádmio/química , Técnicas Eletroquímicas , Transporte de Elétrons , Humanos , NAD/metabolismo , Oxirredução , Doença de Parkinson/diagnóstico , Doença de Parkinson/metabolismo , Respiração , Selênio/química , Zinco/química
18.
PLoS One ; 8(3): e59115, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23527103

RESUMO

Arabinogalactan proteins (AGPs), are a group of highly glycosylated proteins that are found throughout the plant kingdom. To date, glycosyltransferases that glycosylate AGP backbone have remained largely unknown. In this study, a gene (GhGalT1) encoding a putative ß-1,3-galactosyltransferase (GalT) was identified in cotton. GhGalT1, belonging to CAZy GT31 family, is the type II membrane protein that contains an N-terminal transmembrane domain and a C-terminal galactosyltransferase functional domain. A subcellular localization assay demonstrated that GhGalT1 was localized in the Golgi apparatus. RT-PCR analysis revealed that GhGalT1 was expressed at relatively high levels in hypocotyls, roots, fibers and ovules. Overexpression of GhGalT1 in Arabidopsis promoted plant growth and metabolism. The transgenic seedlings had much longer primary roots, higher chlorophyll content, higher photosynthetic efficiency, the increased biomass, and the enhanced tolerance to exogenous D-arabinose and D-galactose. In addition, gas chromatography (GC) analysis of monosaccharide composition of cell wall fractions showed that pectin was changed in the transgenic plants, compared with that of wild type. Three genes (GAUT8, GAUT9 and xgd1) involved in pectin biosynthesis were dramatically up-regulated in the transgenic lines. These data suggested that GhGalT1 may be involved in regulation of pectin biosynthesis required for plant development.


Assuntos
Parede Celular/metabolismo , Galactosiltransferases/genética , Galactosiltransferases/metabolismo , Regulação da Expressão Gênica de Plantas , Gossypium/genética , Gossypium/metabolismo , Pectinas/biossíntese , Adaptação Biológica/genética , Sequência de Aminoácidos , Arabidopsis/genética , Arabidopsis/metabolismo , Arabinose/metabolismo , Galactose/metabolismo , Galactosiltransferases/química , Espaço Intracelular , Dados de Sequência Molecular , Especificidade de Órgãos/genética , Filogenia , Plantas Geneticamente Modificadas , Transporte Proteico , Plântula/genética , Plântula/crescimento & desenvolvimento , Alinhamento de Sequência
19.
Nat Protoc ; 8(3): 439-50, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23391888

RESUMO

Here we report a protocol to investigate the electron-transfer processes of redox-active biomolecules in biological membranes by electrochemistry using biomimetic hybrid bilayer membranes (HBMs) assembled on gold electrodes. Redox-active head groups, such as the ubiquinone moiety, are embedded in HBMs that contain target molecules, e.g., nicotinamide adenine dinucleotide (NADH). By using this approach, the electron-transfer processes between redox molecules and target biomolecules are mediated by mimicking the redox cycling processes in a natural membrane. Also included is a procedure for in situ surface-enhanced Raman scattering (SERS) to confirm the electrochemically induced conformational changes of the target biomolecules in the HBMs. In addition, each step in constructing the HBMs is characterized by electrochemical impedance spectroscopy (EIS), high-resolution X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM). The time required for the entire protocol is ∼12 h, whereas the electrochemical measurement of electron-transfer processes takes less than 1 h to complete.


Assuntos
Biomimética/métodos , Transporte de Elétrons , Bicamadas Lipídicas/química , Espectroscopia Dielétrica/métodos , Eletroquímica/métodos , Eletrodos , Ouro/química , Membranas Artificiais , Microscopia de Força Atômica/métodos , NAD/química , Oxirredução , Espectroscopia Fotoeletrônica/métodos , Análise Espectral Raman/métodos
20.
Chem Commun (Camb) ; 49(17): 1741-3, 2013 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-23340583

RESUMO

SP1 protein as a new type of biological nanopore is described and is utilized to distinguish single-stranded DNA at the single-molecule level. Using the SP1 nanopore to investigate single molecule detection broadens the existing research areas of pore-forming biomaterials from unsymmetrical biological nanopores to symmetrical biological nanopores. This novel nanopore could provide a good candidate for single-molecule detection and characterization of biomaterial applications.


Assuntos
DNA de Cadeia Simples/análise , Técnicas Eletroquímicas , Nanoporos , Proteínas/química , Proteínas Hemolisinas/química , Concentração de Íons de Hidrogênio , Bicamadas Lipídicas/química
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