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1.
Langmuir ; 40(28): 14724-14737, 2024 Jul 16.
Artigo em Inglês | MEDLINE | ID: mdl-38956832

RESUMO

Superhydrophobic coatings are increasingly recognized as a promising approach to enhancing power generation efficiency and prolonging the operational lifespan of wind turbines. In this research, a durable superhydrophobic perfluoroalkoxy alkane (PFA) coating was developed and specifically designed for spray application onto the surface of wind turbine blades. The PFA coating features a micronano hierarchical structure, exhibiting a high water contact angle of 167.0° and a low sliding angle of 1.7°. The optimal PFA coating exhibits stability and maintains a superhydrophobic performance during mechanical and chemical tests. The findings of this study establish a positive association between the surface energy of the coating and its effectiveness in anti-icing. The delayed icing time for the PFA-coated surface is 46.83 times longer than that of an uncoated surface, and the ice adhesion strength is only 1.875 kPa. Additionally, the PFA coating demonstrates remarkably high ice suppression efficiencies of 94.7 and 99.5% in anti-icing experiments at ambient temperatures of -6 and -10 °C, respectively. It is anticipated that this stable superhydrophobic PFA coating will be a candidate for anti-icing applications in wind turbine blades.

2.
Appl Microbiol Biotechnol ; 108(1): 190, 2024 Feb 02.
Artigo em Inglês | MEDLINE | ID: mdl-38305911

RESUMO

Metabolic engineering frequently makes use of point mutation and saturation mutation library creation. At present, sequencing is the only reliable and direct technique to detect point mutation and screen saturation mutation library. In this study, mismatch amplification mutation assay (MAMA) PCR was used to detect point mutation and screen saturation mutation library. In order to fine-tune the expression of odhA encoding 2-oxoglutarate dehydrogenase E1 component, a saturating mutant library of the RBS of odhA was created in Corynebacterium glutamicum P12 based on the CRISPR-Cas2a genome editing system, which increased the L-proline production by 81.3%. MAMA PCR was used to filter out 42% of the non-mutant transformants in the mutant library, which effectively reduced the workload of the subsequent fermentation test and the number of sequenced samples. The rapid and sensitive MAMA-PCR method established in this study provides a general strategy for detecting point mutations and improving the efficiency of mutation library screening. KEY POINTS: • MAMA PCR was optimized and developed to detect point mutation. • MAMA PCR greatly improves the screening efficiency of point mutation. • Attenuation of odhA expression in P12 effectively improves proline production.


Assuntos
Corynebacterium glutamicum , Mutação Puntual , Mutação , Sequência de Bases , Corynebacterium glutamicum/genética , Reação em Cadeia da Polimerase/métodos
3.
Int J Mol Sci ; 24(14)2023 Jul 24.
Artigo em Inglês | MEDLINE | ID: mdl-37511626

RESUMO

L-tryptophan (L-trp), produced through bio-manufacturing, is widely used in the pharmaceutical and food industries. Based on the previously developed L-trp-producing strain, this study significantly improved the titer and yield of L-trp, through metabolic engineering of the shikimate pathway and the L-tryptophan branch. First, the rate-limiting steps in the shikimate pathway were investigated and deciphered, revealing that the combined overexpression of the genes aroE and aroD increased L-trp production. Then, L-trp synthesis was further enhanced at the shaking flask level by improving the intracellular availability of L-glutamine (L-gln) and L-serine (L-ser). In addition, the transport system and the competing pathway of L-trp were also modified, indicating that elimination of the gene TnaB contributed to the extracellular accumulation of L-trp. Through optimizing formulas, the robustness and production efficiency of engineered strains were enhanced at the level of the 30 L fermenter. After 42 h of fed-batch fermentation, the resultant strain produced 53.65 g/L of L-trp, with a yield of 0.238 g/g glucose. In this study, the high-efficiency L-trp-producing strains were created in order to establish a basis for further development of more strains for the production of other highly valuable aromatic compounds or their derivatives.


Assuntos
Proteínas de Escherichia coli , Escherichia coli , Escherichia coli/genética , Escherichia coli/metabolismo , Triptofano/metabolismo , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Engenharia Metabólica , Fermentação
4.
Langmuir ; 34(8): 2734-2740, 2018 02 27.
Artigo em Inglês | MEDLINE | ID: mdl-29384379

RESUMO

Coalescence-induced self-propelled jumping of droplets on superhydrophobic surfaces has potential applications for condensation heat transfer enhancement, anti-icing, self-cleaning, antidew, and so forth. However, most of the previous studies focused on two identical droplets which are not commonly encountered in the nature. In this work, coalescence-induced jumping phenomena of two unequal-sized droplets on superhydrophobic surfaces were investigated theoretically and numerically. First, by introducing modified inertial-capillary velocity (uic*) and Ohnesorge number (Oh*) with consideration of radius ratio (r*) of two coalescing droplets, we proposed a generalized inertial-capillary scaling law for the jumping velocity of coalesced droplets, which is expected to be applicable for both two identical droplets and two unequal-sized droplets coalescing on superhydrophobic surfaces. Subsequently, we employed molecular dynamics simulations to investigate the coalescence-induced jumping process of two unequal-sized nanodroplets. Our simulations showed that the dimensionless jumping velocity (vj/uic*) well follows the generalized inertial-capillary scaling law with vj/uic* ≈ 0.127 in a specific Oh* range; however, it rapidly reduces and finally vanishes when the radius ratio of large droplet to small droplet is larger than a certain threshold value. Our simulations also revealed that nonjumping of two unequal-sized droplets with a very large radius ratio is due to that the larger droplet swallows the small one, so that the liquid bridge has no chance to impact the solid surface, and hence the "liquid bridge impacting substrate" mechanism fails in this circumstance.

5.
Plant Cell ; 26(9): 3472-87, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-25248552

RESUMO

Alternative splicing enhances transcriptome diversity in all eukaryotes and plays a role in plant tissue identity and stress adaptation. To catalog new maize (Zea mays) transcripts and identify genomic loci that regulate alternative splicing, we analyzed over 90 RNA-seq libraries from maize inbred lines B73 and Mo17, as well as Syn10 doubled haploid lines (progenies from B73 × Mo17). Transcript discovery was augmented with publicly available data from 14 maize tissues, expanding the maize transcriptome by more than 30,000 and increasing the percentage of intron-containing genes that undergo alternative splicing to 40%. These newly identified transcripts greatly increase the diversity of the maize proteome, sometimes coding for entirely different proteins compared with their most similar annotated isoform. In addition to increasing proteome diversity, many genes encoding novel transcripts gained an additional layer of regulation by microRNAs, often in a tissue-specific manner. We also demonstrate that the majority of genotype-specific alternative splicing can be genetically mapped, with cis-acting quantitative trait loci (QTLs) predominating. A large number of trans-acting QTLs were also apparent, with nearly half located in regions not shown to contain genes associated with splicing. Taken together, these results highlight the currently underappreciated role that alternative splicing plays in tissue identity and genotypic variation in maize.


Assuntos
Processamento Alternativo/genética , Regulação da Expressão Gênica de Plantas , Genoma de Planta , Zea mays/genética , Mapeamento Cromossômico , Perfilação da Expressão Gênica , Genes de Plantas , Variação Genética , Genótipo , MicroRNAs/genética , MicroRNAs/metabolismo , Especificidade de Órgãos/genética , Proteínas de Plantas/metabolismo , Reação em Cadeia da Polimerase , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Proteoma/metabolismo , Locos de Características Quantitativas/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reprodutibilidade dos Testes , Análise de Sequência de RNA
6.
Med Sci Monit ; 23: 5630-5636, 2017 Nov 26.
Artigo em Inglês | MEDLINE | ID: mdl-29176539

RESUMO

BACKGROUND The aim of this study was to investigate the efficacy and safety of chemotherapy (CT) combined with stereotactic radiotherapy (SRT) in the treatment of nasopharyngeal carcinoma (NPC). MATERIAL AND METHODS A total of 329 NPC patients without any previous treatment were included in this study between January 2009 and November 2013. These patients were divided into three groups: CT group (n=114), SRT group (n=109), and CT + SRT group (n=106). Contrast-enhanced nasopharyngeal computed tomography (CT)/magnetic resonance (MR) scan was performed on the third month after treatment. Short-term efficacy was evaluated according to the Response Evaluation Criteria in Solid Tumors (RECIST). Toxicity was graded according to the Acute Radiation Morbidity Scoring Criteria (RTOG) and the World Health Organization (WHO) toxicity grading scale. Overall survival (OS), progression free survival (PFS), and incidence rate of acute toxicity (grade ≥3) were calculated after a 24 month follow-up. RESULTS Total response rate of all patients was 85.41%. Compared with the CT group and the SRT group, the CT + SRT group showed a substantially improved efficacy in NPC treatment. The incidence rate of the acute toxicity in the CT + SRT group was slightly higher than in the CT group and the SRT group, but the difference was not statistically significant. No treatment-related deaths were observed. The CT + SRT group had the highest two-year OS and PFS, followed by the CT group and the SRT group. CONCLUSIONS It was shown that NPC patients treated with CT + SRT had better short- and long-term efficacy than those treated with CT or SRT alone.


Assuntos
Carcinoma/terapia , Quimiorradioterapia/métodos , Neoplasias Nasofaríngeas/terapia , Radiocirurgia/métodos , Adulto , Idoso , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , China , Cisplatino/administração & dosagem , Intervalo Livre de Doença , Tratamento Farmacológico/métodos , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Carcinoma Nasofaríngeo , Estadiamento de Neoplasias , Prognóstico , Dosagem Radioterapêutica , Resultado do Tratamento
7.
Molecules ; 21(4): 429, 2016 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-27043512

RESUMO

Twelve new steroidal saponins, including eleven furostanol saponins, terrestrinin J-T (1-11), and one spirostanol saponin, terrestrinin U (12), together with seven known steroidal saponins 13-19 were isolated from T. terrestris. The structures of the new compounds were established on the basis of spectroscopic data, including 1D and 2D NMR and HRESIMS, and comparisons with published data.


Assuntos
Extratos Vegetais/química , Saponinas/química , Espirostanos/química , Esteróis/química , Tribulus/química , Medicamentos de Ervas Chinesas/química , Estrutura Molecular , Saponinas/isolamento & purificação , Espirostanos/isolamento & purificação , Esteróis/isolamento & purificação
8.
Langmuir ; 31(27): 7457-62, 2015 Jul 14.
Artigo em Inglês | MEDLINE | ID: mdl-26087117

RESUMO

When an electric field with various strengths is applied to two adjacent conducting droplets, the droplets may completely coalesce, partially coalesce, or bounce off one another. To reveal an atom-scale mechanism of coalescence or non-coalescence, dynamic behaviors of two conducting nanodroplets at a homogeneous electric field are studied via molecular dynamics simulations in this work. The results show that there is a critical field strength and a critical cone angle above which the two droplets partially coalesce or bounce off. Charge transfer between the two droplets is observed when the droplets are brought into contact. The partial coalescence and the bounce-off of the two droplets at strong field strengths are found to be due to the high charge transfer rate, which leads to the breakup of the coalescing droplet at different locations.

9.
Int Immunopharmacol ; 132: 112024, 2024 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-38608475

RESUMO

Ulcerative colitis (UC) is a recurrent intestinal disease with an increasing incidence worldwide that seriously affects the life of patients. Turtle peptide (TP) is a bioactive peptide extracted from turtles that has anti-inflammatory, antioxidant and anti-aging properties. However, studies investigating the effect of TP on the progression of UC are lacking. The aim of this study was to investigate effects and underlying mechanisms of TP and its derivative peptide GPAGPIGPV (GP-9) in alleviating UC in mice. The results showed that 500 mg/kg TP treatment significantly ameliorated colitis symptoms and oxidative stress in UC mice. TP alleviated intestinal barrier damage in UC mice by promoting mucosal repair and increasing the expression of tight junction proteins (ZO1, occludin and claudin-1). TP also modulated the composition of the gut microbiota by increasing the abundance of the beneficial bacteria Anaerotignum, Prevotellaceae_UCG-001, Alistipes, and Lachno-spiraceae_NK4A136_group and decreasing the abundance of the harmful bacteria Prevotella_9 and Parasutterella. Furthermore, we characterized the peptide composition of TP and found that GP-9 ameliorated the symptoms of dextran sodium sulfate (DSS)-induced colitis in mice by inhibiting the TLR4/NF-κB signaling pathway. In conclusion, TP and its derivative peptides ameliorated DSS-induced ulcerative colitis by inhibiting the expression of inflammatory factors and modulating the composition of the intestinal microbiota; this study provides a theoretical basis for the application of TP and its derivative peptides for their anti-inflammatory activity.


Assuntos
Anti-Inflamatórios , Colite Ulcerativa , Sulfato de Dextrana , Microbioma Gastrointestinal , Camundongos Endogâmicos C57BL , Peptídeos , Tartarugas , Animais , Colite Ulcerativa/tratamento farmacológico , Colite Ulcerativa/induzido quimicamente , Colite Ulcerativa/patologia , Colite Ulcerativa/imunologia , Microbioma Gastrointestinal/efeitos dos fármacos , Camundongos , Peptídeos/uso terapêutico , Peptídeos/farmacologia , Anti-Inflamatórios/uso terapêutico , Anti-Inflamatórios/farmacologia , Tartarugas/microbiologia , Tartarugas/imunologia , Masculino , Receptor 4 Toll-Like/metabolismo , NF-kappa B/metabolismo , Modelos Animais de Doenças , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/patologia , Mucosa Intestinal/metabolismo , Mucosa Intestinal/microbiologia , Colo/patologia , Colo/efeitos dos fármacos , Humanos , Estresse Oxidativo/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos
10.
Bioorg Med Chem Lett ; 22(24): 7518-22, 2012 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-23122867

RESUMO

High throughput screening of the Roche compound collection led to the identification of diaminopyrroloquinazoline series as a novel class of PTP1B inhibitors. Structural modification of diaminopyrroloquinazoline series resulted in pyrido[2,3-d]pyrimidine-2,4-diamine series which was further optimized to give compounds 5 and 24 as potent, selective (except T-cell phosphatase) PTP1B inhibitors with good mouse PK properties.


Assuntos
Diaminas/farmacologia , Inibidores Enzimáticos/farmacologia , Proteína Tirosina Fosfatase não Receptora Tipo 1/antagonistas & inibidores , Pirimidinas/farmacologia , Animais , Diaminas/síntese química , Diaminas/química , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/administração & dosagem , Inibidores Enzimáticos/química , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Estrutura Molecular , Proteína Tirosina Fosfatase não Receptora Tipo 1/metabolismo , Pirimidinas/síntese química , Pirimidinas/química , Relação Estrutura-Atividade
11.
DNA Repair (Amst) ; 118: 103389, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-36030574

RESUMO

DNA recombination repair systems are essential for organisms to maintain genomic stability. In recent years, we have improved our understanding of the mechanisms of RecBCD/AddAB family-mediated DNA double-strand break repair. In E. coli, it is RecBCD that plays a central role, and in Firmicute Bacillus subtilis it is the AddAB complex that functions. However, there are open questions about the mechanism of DNA repair in bacteria. For example, how bacteria containing crossover hotspot instigator (Chi) sites regulate the activity of proteins. In addition, we still do not know the exact process by which the RecB nuclease or AddA nuclease structural domains load RecA onto DNA. We also know little about the mechanism of DNA repair in the industrially important production bacterium Corynebacterium glutamicum (C. glutamicum). Therefore, exploring DNA repair mechanisms in bacteria may not only deepen our understanding of the DNA repair process in this species but also guide us in the targeted treatment of diseases associated with recombination defects, such as cancer. In this paper, we firstly review the classical proteins RecBCD and AddAB involved in DNA recombination repair, secondly focus on the novel helical nuclease AdnAB found in the genus Mycobacterium.


Assuntos
Escherichia coli , Exodesoxirribonucleases , Bacillus subtilis , DNA/metabolismo , Reparo do DNA , Enzimas Reparadoras do DNA/metabolismo , DNA Bacteriano/metabolismo , Escherichia coli/genética , Exodesoxirribonuclease V/metabolismo , Exodesoxirribonucleases/metabolismo
12.
Opt Express ; 19(18): 17408-12, 2011 Aug 29.
Artigo em Inglês | MEDLINE | ID: mdl-21935106

RESUMO

High order harmonics generation from argon gas was comprehensively investigated by using multi-cycle and few-cycle laser pulses. Non-odd order harmonics were observed for sub-5-fs pulses, compare to the normal odd-order harmonics in the multi-cycle case. Theoretic analysis shows that the new spectral structure origins from the asymmetry of laser field in few-cycle pulses. This asymmetry induced both amplitude and phase difference between attosecond pulses from consecutive half-cycle of the laser field, which change the interference property of attosecond pulses and result in complex spectrum.

13.
Onco Targets Ther ; 12: 2911-2920, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31114235

RESUMO

Objective: Neuropilin-1 has been reported to be a valuable diagnostic biomarker in patients with cervical intraepithelial neoplasia (CIN) and early cervical cancer. The aim of this study was to investigate the association between Neuropilin-1 and the prognosis of cervical cancer in Henan Chinese population. Methods: Tissues were collected in The Third Affiliated Hospital of Zhengzhou University between 2010 and 2012, determining the level and expression of Neuropilin-1 in different cervical lesions by immunohistochemistry. The cell proliferation assay, wound-healing assays and Transwell assay were performed to explore the ability of proliferation, migration and invasion for Hela and Caski cells after NRP-1 was knocked down by shRNA transfection. Western blotting was performed to investigate the role of NRP-1 in endothelial-to-mesenchymal transition (EndMT). Tumor xenografts model was used to evaluate the effect of NRP-1 on the tumor growth. Results: The expression of NRP-1 was upregulated in the tumor tissues compared with the CIN and normal tissues (P<0.0001). The overall survival time of the high NRP-1 expression group was significantly shorter than that of the low NRP-1 expression group (P<0.0001); NRP-1-depleted cells had dramatically lower rate of proliferation, migration and invasion compared to control cells (all P<0.05). Depletion of NRP-1 significantly suppressed the growth of CaSki xenograft tumor in nude mice. Conclusions: The current study demonstrated that NRP-1 expression is significantly correlated with the progression of CC. Notably, high NRP-1 expression is correlated with a poorer survival in patients with CC, and has been shown to be an independent prognostic factor.

14.
BMC Plant Biol ; 8: 17, 2008 Feb 19.
Artigo em Inglês | MEDLINE | ID: mdl-18282305

RESUMO

BACKGROUND: Although originally thought to be less frequent in plants than in animals, alternative splicing (AS) is now known to be widespread in plants. Here we report the characteristics of AS in legumes, one of the largest and most important plant families, based on EST alignments to the genome sequences of Medicago truncatula (Mt) and Lotus japonicus (Lj). RESULTS: Based on cognate EST alignments alone, the observed frequency of alternatively spliced genes is lower in Mt (approximately 10%, 1,107 genes) and Lj (approximately 3%, 92 genes) than in Arabidopsis and rice (both around 20%). However, AS frequencies are comparable in all four species if EST levels are normalized. Intron retention is the most common form of AS in all four plant species (~50%), with slightly lower frequency in legumes compared to Arabidopsis and rice. This differs notably from vertebrates, where exon skipping is most common. To uncover additional AS events, we aligned ESTs from other legume species against the Mt genome sequence. In this way, 248 additional Mt genes were predicted to be alternatively spliced. We also identified 22 AS events completely conserved in two or more plant species. CONCLUSION: This study extends the range of plant taxa shown to have high levels of AS, confirms the importance of intron retention in plants, and demonstrates the utility of using ESTs from related species in order to identify novel and conserved AS events. The results also indicate that the frequency of AS in plants is comparable to that observed in mammals. Finally, our results highlight the importance of normalizing EST levels when estimating the frequency of alternative splicing.


Assuntos
Processamento Alternativo/genética , Sequência Conservada/genética , Etiquetas de Sequências Expressas , Lotus/genética , Medicago truncatula/genética , Genes de Plantas , Lotus/metabolismo , Medicago truncatula/metabolismo , Alinhamento de Sequência , Especificidade da Espécie
15.
Chem Commun (Camb) ; (41): 4233-5, 2007 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-18217590

RESUMO

A photoreversible organogel made from dendrons was reported.

16.
J Colloid Interface Sci ; 314(1): 289-96, 2007 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-17588598

RESUMO

Three novel series of amphiphiles based on poly(amidoamine) dendrons (from G1 to G3) and having different aromatic chromophores (Cz I, Cz II, and Py) at the focal point were synthesized and studied for their self-assembly behavior in aqueous solution by using electronic microscopies (i.e., SEM and TEM), UV-vis, fluorescence, IR, and (1)H NMR spectroscopy. It was found that the generation of dendrons affected significantly the self-assembly of these amphiphiles in aqueous solution and the morphological structures of the resulting assemblies depended greatly on the architecture of the focal chromophores. As a result, the first generation of dendrons assembled readily into vesicles at low concentrations. These vesicular structures subsequently fused to form a stable tubular structure. Similar tubular structures could also be directly obtained through self-assembly of these amphiphilic dendrons at high concentrations. X-ray investigations showed that the resulting tubules possessed a lamellar structure. A head-to-head packing model of amphiphilic dendrons in the assemblies was proposed.


Assuntos
Dendrímeros/química , Modelos Moleculares , Nylons/química , Colorimetria , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Difração de Raios X
17.
Guang Pu Xue Yu Guang Pu Fen Xi ; 27(12): 2427-31, 2007 Dec.
Artigo em Zh | MEDLINE | ID: mdl-18330277

RESUMO

In the present work, 20 patients with salivary pleomorphic adenoma were recruited for FTIR spectroscopic measurement. These obtained FTIR spectra were analyzed and compared. It was found that there were significant differences in the spectral features of the skin covering normal salivary gland, pleomorphic adenoma, and carcinoma change of pleomorphic adenoma, such as the changes in peak position, band shape and relative intensity of the bands in the ranges of 1000-1800 cm(-1) and 2800-3000 cm(-1). Pathological diagnosis demonstrated that 2 of the 20 patients suffered actually carcinoma change of pleomorphic adenoma, which is in good agreement with the result of FTIR spectroscopicmeasurement. FTIR spectroscopic m ethodsuggested that pleomorphic adenoma is the intermediate between normal salivary gland and carcinoma change of pleomorphic adenoma.


Assuntos
Adenoma Pleomorfo/química , Neoplasias das Glândulas Salivares/química , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Adenoma Pleomorfo/diagnóstico , Adenoma Pleomorfo/patologia , Humanos , Neoplasias das Glândulas Salivares/diagnóstico , Neoplasias das Glândulas Salivares/patologia
18.
Cancer Biother Radiopharm ; 32(6): 193-203, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28820634

RESUMO

AIM: The study investigated the ability of ataxia-telangiectasia mutated (ATM)/Rad3-related (ATR) signaling pathway to influence the proliferation, apoptosis, and radiosensitivity of nasopharyngeal carcinoma (NPC) cells. MATERIALS AND METHODS: NPC tissues and corresponding adjacent normal tissues were collected from 143 NPC patients. The NPC CNE2 cells were assigned into a control group, X-ray group, CGK-733 group, and X-ray+CGK-733 group. The mRNA levels of ATM and ATR were evaluated using quantitative real-time polymerase chain reaction (qRT-PCR) and the protein levels of ATM and ATR using western blotting. The positive expression of ATM and ATR in tissues and nude mouse tumor tissues was determined by immunohistochemistry. Cell proliferation, migration, invasion, and apoptosis rates were analyzed by the 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di- phenytetrazoliumromide (MTT) assay, scratch test, transwell assay, and flow cytometry, respectively. A nude mouse model of NPC was established to observe tumor volume and growth. RESULTS: The mRNA levels of ATR and ATM and the expression of ATR and ATM protein in NPC tissues were significantly higher than those in adjacent normal tissues. The colony formation assay showed that the colony-forming rate decreased, showing radiation dose-dependent and CGK-733 concentration-dependent manners. Expression of ATM, ATR, Chk1, and Chk2 was evidently increased in the X-ray, CGK-733, and X-ray+CGK-733groups compared with the control group, and the aforementioned expression was highest in the X-ray+CGK-733 group among the four groups. The cell proliferation, invasion, and migration were decreased, tumor volume decreased and cell apoptosis increased in the X-ray, CGK-733, and X-ray+CGK-733 groups compared with the control group; the X-ray+CGK-733 group exhibited lowest cell proliferation, invasion and migration, smallest tumor volume, and highest cell apoptosis among the four groups. CONCLUSIONS: Inhibition of ATM/ATR signaling pathway reduces proliferation and enhances apoptosis and radiosensitivity of NPC cells.


Assuntos
Proteínas Mutadas de Ataxia Telangiectasia/metabolismo , Carcinoma/metabolismo , Carcinoma/radioterapia , Neoplasias Nasofaríngeas/metabolismo , Neoplasias Nasofaríngeas/radioterapia , Idoso , Animais , Apoptose/fisiologia , Proteínas Mutadas de Ataxia Telangiectasia/antagonistas & inibidores , Proteínas Mutadas de Ataxia Telangiectasia/biossíntese , Proteínas Mutadas de Ataxia Telangiectasia/genética , Benzenoacetamidas/farmacologia , Carcinoma/genética , Carcinoma/patologia , Linhagem Celular Tumoral , Feminino , Humanos , Imuno-Histoquímica , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Pessoa de Meia-Idade , Carcinoma Nasofaríngeo , Neoplasias Nasofaríngeas/genética , Neoplasias Nasofaríngeas/patologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Tolerância a Radiação , Distribuição Aleatória , Transdução de Sinais , Tioureia/análogos & derivados , Tioureia/farmacologia , Ensaios Antitumorais Modelo de Xenoenxerto
19.
Biochim Biophys Acta ; 1728(3): 105-14, 2005 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-15780972

RESUMO

The serine-arginine (SR)-rich splicing proteins are highly conserved RNA binding nuclear phosphor-proteins that play important roles in both regular and alternative splicing. Here we describe two novel putative SR genes from maize, designated zmRSp31A and zmRSp31B. Both genes contain characteristic RNA binding motifs RNP-1 and RNP-2, a serine/arginine-rich (RS) domain and share significant sequence similarity to the Arabidopsis atRSp31 family of SR proteins. Both zmRSp31A and zmRSp31B produce multiple transcripts by alternative splicing, of which majority of the alternatively spliced transcripts utilize non-canonical splice sites. zmRSp31A and zmRSp31B produce at least six and four transcripts, respectively, of which only one corresponds to the wild type proteins for each gene. All the alternatively spliced transcripts of both the genes, with one exception, are predicted to encode small truncated proteins containing only the RNP-2 domain of their first RNA recognition motif and completely lack the carboxyl terminal RS domain. We provide evidence that some of the alternatively spliced transcripts of both genes are associated with polysomes and interact with the translational machinery.


Assuntos
Processamento Alternativo/genética , Regulação da Expressão Gênica de Plantas , Proteínas Nucleares/genética , Fosfoproteínas/genética , Splicing de RNA/genética , Proteínas de Ligação a RNA/genética , Zea mays/genética , Sequência de Aminoácidos , Proteínas de Arabidopsis/genética , Sequência de Bases , Northern Blotting , Primers do DNA , Dados de Sequência Molecular , Polirribossomos/genética , Estrutura Terciária de Proteína , Sítios de Splice de RNA/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Alinhamento de Sequência , Análise de Sequência de DNA , Fatores de Processamento de Serina-Arginina
20.
Methods Enzymol ; 395: 400-18, 2005.
Artigo em Inglês | MEDLINE | ID: mdl-15984049

RESUMO

Expressed sequence tag (EST) data are a major contributor to the known plant sequence space. Organization of the data into non-redundant clusters representing tentative unique genes provides snapshots of the gene repertoires of a species. This chapter reviews availability of sequences and sequence analysis results and describes several resources and tools that should facilitate broad-based utilization of EST data for gene structure annotation, gene discovery, and comparative genomics.


Assuntos
Etiquetas de Sequências Expressas , Plantas/genética , Processamento Alternativo , Sequência de Bases , Sequência Conservada , DNA Complementar/genética , DNA de Plantas/genética , Bases de Dados Genéticas , Biblioteca Gênica , Genes de Plantas , Genômica/métodos , Dados de Sequência Molecular , Família Multigênica , Alinhamento de Sequência/métodos
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