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1.
Virus Genes ; 54(6): 840-845, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30311179

RESUMO

Turncurtoviruses (family: Geminiviridae; genus: Turncurtovirus) appear to have a high degree of genetic variation in Iran. Leafhoppers of the species Circulifer haematoceps (Mulsant and Rey, 1855) (family: Cicadellidae) were collected in 2014 from three geographical regions in south-eastern Iran (Orzoeyeh, Jiroft and Sirjan; Kerman province) and screened for the presence of turncurtoviruses using a combination of PCR and rolling circle amplification (RCA) methods. Eleven genomes of turncurtovirus were recovered and sequenced. Leafhoppers were sampled off sesame (S. indicum L.) and turnip (Brassica rapa sub sp. rapa). Thus, we identified three symptomatic sesame plants (yellowing, boat-shaped leaf curling, vein swelling on the lower leaf surfaces) from sesame farms in Jiroft. In these samples, we identified the same turncurtovirus as in the leafhoppers and have named it sesame curly top virus (SeCTV). Collectively, these SeCTV share > 98% genome-wide pairwise identity and ~ 87.3% to a recently identified turncurtovirus (sesame yellow mosaic virus; SeYMV) from sesame in Pakistan (GenBank accession MF344550). The SeCTV and SeYMV sequences share < 70% genome-wide pairwise identity with isolates of Turnip curly top virus and Turnip leaf roll virus, the two species in the genus Turncurtovirus. Based on the pairwise identities and phylogenetic analysis, SeCTV (n = 12) and SeYMV (n = 1) represent two strains of a new species in the genus Turncurtovirus.


Assuntos
Geminiviridae/genética , Hemípteros/virologia , Sesamum/virologia , Animais , Geminiviridae/isolamento & purificação , Genes Virais , Genoma Viral , Genômica/métodos , Filogenia
2.
Virus Genes ; 53(2): 323-327, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28004232

RESUMO

Beet curly top Iran virus (BCTIV) is a distinct geminivirus which has been reported from sugar-beet-growing farms in Iran. In this study, the role of the splicing in expression of complementary-sense genes of BCTIV was studied. Total RNA was extracted from BCTIV-infected tissue, and the predicted intron position of complementary-sense mRNA transcripts was amplified by RT-PCR followed by cloning of the amplicons. Sequence confirmed that both spliced and unspliced mRNAs are synthesized by the same transcription unit. Sequence comparison showed that a 155-nt segment (intron) corresponding to nucleotides 1890-2044 of the viral genome has been removed from the latter transcript and therefore fusion of the C1:C2 genes resulted creation of a continuous reading frame for potential production of intact replication initiator protein (Rep). BCTIV intron comprises of most consensus splicing signals required for splicing in eukaryotes and several plant viruses including mastre- and capulaviruses.


Assuntos
Geminiviridae/genética , Filogenia , Splicing de RNA/genética , Proteínas Virais/genética , Beta vulgaris/virologia , Geminiviridae/patogenicidade , Genoma Viral , Irã (Geográfico) , Dados de Sequência Molecular , Doenças das Plantas/genética , Doenças das Plantas/virologia , Vírion/genética
3.
Differentiation ; 92(4): 148-158, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27575952

RESUMO

Tissue engineering has attracted a great deal of interest by combining fibrous scaffolds and stem cells regarding bone regeneration applications. In the present study, polymeric fibrous polyethersulphone-polyethylene glycol (PES-PEG) was fabricated by electrospinning. It was then treated with NH3 plasma to enhance surface hydrophilicity, cell attachment, growth and differentiation potential. X-ray photoelectron spectroscopy (XPS) measurements were used to evaluate the modification of the scaffold's surface chemistry. Electrospun scaffolds were coated with willemite (Zn2SiO4) bioceramic nanoparticles. Scaffold characterization was done by scanning electron microscope (SEM), differential scanning calorimetry (DSC), contact angle measurements and tensile analysis. MTT assay was used to assess the biocompatibility of fibrous scaffolds loaded with Zn2SiO4 regarding proliferation support. Osteogenic differentiation of cultured human mesenchymal stem cells (hMSCs) on fibers was evaluated using common osteogenic markers such as alkaline phosphatase (ALP) activity, calcium mineral deposition, quantitative real-time PCR (qPCR) and immunocytochemical analysis (ICC). According to the results, proliferation and osteogenic differentiation of hMSCs were significantly enhanced after coating Zn2SiO4 on fibrous scaffolds. These results were detected by higher ALP activity, biomineralization and expression of osteogenic related genes and proteins in differentiated hMSCs. In conclusion, our results indicated that the combination of Zn2SiO4 nanoparticles and electrospun fibers is able to provide a new, suitable and more efficient matrix to support stem cells differentiation for bone tissue engineering applications.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Células-Tronco Mesenquimais/efeitos dos fármacos , Nanopartículas/administração & dosagem , Osteogênese/efeitos dos fármacos , Fosfatase Alcalina/biossíntese , Regeneração Óssea/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Humanos , Nanopartículas/química , Osteoblastos/efeitos dos fármacos , Polietilenoglicóis/administração & dosagem , Polietilenoglicóis/química , Polímeros/administração & dosagem , Polímeros/química , Dióxido de Silício/administração & dosagem , Dióxido de Silício/química , Sulfonas/administração & dosagem , Sulfonas/química , Engenharia Tecidual , Alicerces Teciduais , Zinco/administração & dosagem , Zinco/química
4.
Arch Virol ; 161(3): 551-61, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26611911

RESUMO

Turnip curly top virus (TCTV) is the only member of the newly established genus Turncurtovirus (family Geminiviridae). As part of an ongoing study to identify additional plant hosts and the diversity of turncurtoviruses, between 2012 and 2014, we sampled symptomatic turnip plants and other crops in the provinces Fars and Khorasan Razavi (southern and northeastern Iran, respectively). Infection by turncurtoviruses was tested by PCR and/or rolling-circle amplification (RCA) coupled with restriction enzyme digests. Turncurtoviruses were identified in turnip as well as seven other field crops, including eggplant, basil, radish, lettuce, sugar beet, red beet and spinach. Full turncurtovirus genomes were recovered from 25 of these samples, leading to the identification of TCTV and a new putative turncurtovirus, turnip leaf roll virus (TLRV; 13 isolates), which shares <80% genome-wide pairwise identity with TCTV. Agroinoculation of plants with an infectious clone of TLRV demonstrated that this virus could infect several plant hosts under greenhouse conditions and could be transmitted by the leafhopper Circulifer haematoceps (Mulsant and Rey, 1855) from agroinoculated to healthy plants.


Assuntos
Produtos Agrícolas/virologia , Geminiviridae/classificação , Geminiviridae/genética , Variação Genética , Doenças das Plantas/virologia , Análise por Conglomerados , Geminiviridae/isolamento & purificação , Genoma Viral , Irã (Geográfico) , Dados de Sequência Molecular , Técnicas de Amplificação de Ácido Nucleico , Filogenia , RNA Viral/genética , Mapeamento por Restrição , Análise de Sequência de DNA , Homologia de Sequência
5.
Cell Mol Biol Lett ; 21: 6, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-28536609

RESUMO

Naturally occurring antisense RNAs are small, diffusible, untranslated transcripts that pair to target RNAs at specific regions of complementarity to control their biological function by regulating gene expression at the post-transcriptional level. This review focuses on known cases of antisense RNA control in prokaryotes and provides an overview of some natural RNA-based mechanisms that bacteria use to modulate gene expression, such as mRNA sensors, riboswitches and antisense RNAs. We also highlight recent advances in RNA-based technology. The review shows that studies on both natural and synthetic systems are reciprocally beneficial.


Assuntos
Bactérias/genética , Regulação Bacteriana da Expressão Gênica , RNA Antissenso , RNA Mensageiro
6.
RSC Adv ; 14(24): 16801-16808, 2024 May 22.
Artigo em Inglês | MEDLINE | ID: mdl-38784415

RESUMO

Cobalt ferrite (CoFe) nanoparticles (NPs) with appropriate physicochemical and biological properties have attracted great attention for biomedical applications. In the present study, chitosan-coated mesoporous CoFe (CoFeCH) NPs were synthesized using a facile one-step hydrothermal method and fully characterized using FE-SEM, EDS, BET, FTIR spectroscopy, DLS, TGA, XRD, and VSM. The spherical, highly colloidal, and monodispersed CoFeCH NPs with an average hydrodynamic size of 177.9 nm, PDI of 0.238 and zeta potential value of -33 represented a high saturation magnetization value of 59.37 emu g-1. N2 adsorption-desorption analysis confirmed the mesoporous structure of CoFeCH NPs with a type IV isotherm, calculated specific surface area of 89.583 m2 g-1 and total pore volume of 0.3668 cm3 g-1. CoFeCH NPs exhibited high antibacterial effects on S. aureus and E. coli, comparable with standard antibiotics, while CH-coating led to higher biocompatibility of CoFe NPs on human cells in vitro. CoFeCH NPs also showed significant peroxidase activity with a Km value of 14.37 and specific activity of 0.632 mmol min-1. CoFeCH NPs were successfully used as a MRI contrast agent with an R2 value of 91.3 mM-1 s-1. The overall results indicated the high potential of synthesized CoFeCH NPs by the present method for biomedical applications, especially as an antibacterial and MRI contrast agent.

7.
Virus Genes ; 46(2): 345-53, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23225113

RESUMO

Turnip curly top virus (TCTV) is a unique geminivirus that has recently been characterised as infecting turnips in Iran. The genome of TCTV shares <68 % pairwise identity with other geminiviruses and has a genome organisation similar to that of curtoviruses and topocuvirus. The replication-associated protein (Rep) bears the highest similarity to curtovirus Reps (48.5-69.0 %); however, in the case of the capsid protein (CP), the extent of similarity is only 39.5-44.5 %. We constructed an agroinfectious clone of TCTV and undertook host range studies on ten plant species; in three species (turnip, sugar beet and cowpea), we detected infection which presents curly top symptoms in turnip and sugar beet. The efficiency of TCTV infection in agroinoculated turnip plants was 71.7 %, and the infection was successfully transmitted to 80 % of the healthy turnip plants used in the insect transmission studies by Circulifer haematoceps under greenhouse conditions. We also determined the genome sequence of 14 new TCTV isolates from southern Iran isolated from turnips. We observed ~13 % diversity amongst all the TCTV isolates and found evidence of recombination in the CP- and Rep-coding regions of the genomes.


Assuntos
Geminiviridae/fisiologia , Variação Genética , Especificidade de Hospedeiro , Doenças das Plantas/virologia , Beta vulgaris/virologia , Brassica napus/virologia , Brassica rapa/virologia , Fabaceae/virologia , Geminiviridae/classificação , Geminiviridae/genética , Geminiviridae/isolamento & purificação , Dados de Sequência Molecular , Filogenia
8.
Spectrochim Acta A Mol Biomol Spectrosc ; 229: 117891, 2020 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-31818642

RESUMO

An Immuno-Nano-Biosensor with high sensitivity was designed based on iron and silica nanoparticles to detect B. abortus. Briefly explain, primary polyclonal antibody (IgG1) was conjugated on surface magnetic nanoparticles (MNPs) to form MNP-IgG1. Secondary polyclonal antibody (IgG2) and Horseradish Peroxidase enzyme were conjugated on silica nanoparticles (SNPs) to form HRP-SNP-IgG2. HRP-SNP-IgG2. MNP-IgG1 and HRP-SNP-IgG2 were added to B. abortus. The MNP-IgG1-B.abortus-IgG2-SNP-HRP complex was isolated from the reaction mixture using a magnet. After that, tetramethylbenzidine was added to the complex. The reaction was stopped with HCl and investigated using UV-Vis spectrophotometry. The nanoparticles' structure and size were investigated using SEM and DLS. Immuno-Nano-Biosensor sensitivity and specificity were determined. The SEM and DLS results indicated that the SNPs, MNPs, HRP-SNP-IgG2 and MNP-IgG1 size and structure were 35, 44, 60 and 56 nm, respectively. In addition, a good linear correlation was observed at 102-107 CFU mL-1 concentrations, which their linear equation and regression were Y = 0.3× + 0.18 and R2 0.982, respectively. The limitation of detecting B. abortus was 160 CFU mL-1. Finally, the results demonstrated that those designed Immuno-Nano-Biosensor could be specifically detected B. abortus and B. melitensis in real samples.


Assuntos
Anticorpos Antibacterianos/química , Técnicas Biossensoriais , Brucella abortus , Brucella melitensis , Peroxidase do Rábano Silvestre/química , Imunoglobulina G/química , Nanopartículas de Magnetita/química
9.
J Microbiol Methods ; 162: 42-49, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31100315

RESUMO

Early detection of infectious bacteria is a necessity for combating infectious diseases. Due to low infectious dose of Shigella, rapid and sensitive detection is needed. Compared to the presented genes, Spa gene can be introduced as a novel sequence for all species of Shigella detection. Herein, the possibility of Spa genes for detection of four species of Shigella was investigated for the first time by AuNPs-based optical genosensing system. In this method, AuNP-DNA probes were hybridized with Spa gene sequence. When the complementary target is present, it prevents the aggregation of the complex under acid environment and the solution remains red whereas in the absence of the specific sequence, it turns to purple. Therefore, visual detection is possible with bare eye. The comparison of this Optical DNA biosensor and PCR-based method showed that the proposed method is simple, cost-effective, rapid operation, with high or comparable detection limit of (LOD and LOQ: 8.14 and 26.6 ng mLl-1, respectively), without need of any expensive techniques, and equipments compared to the conventional methods. In conclusion, the described method may develop into a platform that could be utilized for detection of various bacterial species with high accuracy and prompt screening of samples.


Assuntos
Antígenos de Bactérias/genética , Proteínas da Membrana Bacteriana Externa/genética , Técnicas Biossensoriais/métodos , Colorimetria/métodos , DNA Bacteriano/isolamento & purificação , Genes Bacterianos , Shigella/isolamento & purificação , Sondas de DNA/química , Ouro/química , Nanopartículas Metálicas/química , Reação em Cadeia da Polimerase , Shigella/genética
10.
Mater Sci Eng C Mater Biol Appl ; 105: 110113, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31546438

RESUMO

The highly sensitive and specificity detection are very important in diagnosis of foodborne pathogens and prevention of spread diseases. Therefore, in the present study, a highly sensitive fluorescence Nano-biosensors was designed for detection of Shigella species. For achieved this purpose, DNA probes and gold nanoparticles (AuNPs) were designed and synthesized, respectively. Then, two DNA probes as signal reporter were immobilized on surface of AuNPs. On the other hand, Iron nanoparticles (MNPs) were synthesized and modified with SMCC (Sulfosuccinimidyl 4-Nmaleimidomethyl cyclohexane-1- carboxylate). The 3th DNA probe was immobilized on surface of MNPs for separation of target DNA. The MNP-DNA probe and DNA probe-AuNP-fluorescence DNA probe were added to target DNA. The MNP- DNA probe-target DNA-DNA probe-AuNP-fluorescence DNA probe complex was isolated by a magnet. The fluorescence DNA probe was released on surface of AuNPs and the fluorescence intensity was read by fluorescence spectrophotometry. Sensitivity and specificity of designed Nano-biosensor was determined. The results showed that the fluorescence intensity was increased with increasing of target DNA concentration. Linear related between target DNA and fluorescence intensity was observed in 2.3 × 102 up to 2.3 × 107 CFU mL-1. The linear equation and regression were Y = 1.8 X + 23.4 and R2 0.9953. Limit of detection (LOD) were determined 90 CFUmL-1. The specificity of Nano-biosensor in present of other bacteria was confirmed.


Assuntos
Técnicas Biossensoriais/métodos , Ouro/química , Ferro/química , Nanopartículas Metálicas/química , Shigella/isolamento & purificação , Sondas de DNA/química , Difusão Dinâmica da Luz , Fluorescência , Concentração de Íons de Hidrogênio , Limite de Detecção , Nanopartículas Metálicas/ultraestrutura , Espectroscopia de Infravermelho com Transformada de Fourier , Temperatura , Fatores de Tempo
11.
Int J Biol Macromol ; 124: 1256-1263, 2019 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-30445082

RESUMO

S. aureus is one of important causes of disease, food poisoning in humans and animals. The generally methods for detection of S. aureus is time consuming. Therefore, a new method is necessary for rapid, sensitive and specific diagnosis of S. aureus. In the present study, two probes and a Bio-barcode DNA were designed for detection of S. aureus (Protein A). Firstly, magnetic nanoparticle (MNPs) and gold nanoparticle (AuNPs) were synthesized at 80 °C and 100 °C, respectively. The AuNPs and the MNPs were functionalized with probe1, Bio-barcode DNA and probe2, respectively. Target DNA was added into the nanomaterial's system containing bio-barcode DNA-AuNPs-probe1 and probe2-MNPs to formed bio-barcode DNA-AuNPs-probe1-target DNA-probe2-MNPs complex. The bio-barcode DNA-AuNPs-probe1-target DNA-probe2-MNPs complex was separated with magnetic field. Finally, the bio-barcode DNA was released from surface of complex using DTT (0.8 M) and there was isolated of nanoparticles by magnetic field and centrifuge. The fluorescence intensity of bio-barcode DNA was measured in different concentrations of S. aureus (101 to 108 CFU mL-1) by fluorescence spectrophotometry. The results showed that standard curve was linearly from 102 to 107 CFU mL-1. Limit of detection of bio-barcode assay for both PBS and real samples was 86 CFU mL-1.


Assuntos
Técnicas de Tipagem Bacteriana , Técnicas Biossensoriais , Código de Barras de DNA Taxonômico/métodos , DNA Bacteriano/química , Nanopartículas Metálicas/química , Proteína Estafilocócica A/análise , Staphylococcus aureus/genética , Sondas de DNA/síntese química , Sondas de DNA/química , DNA Bacteriano/metabolismo , Ouro/química , Humanos , Ferro/química , Limite de Detecção , Imãs , Espectrometria de Fluorescência , Staphylococcus aureus/classificação , Staphylococcus aureus/isolamento & purificação
12.
Mar Pollut Bull ; 56(4): 751-7, 2008 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-18241891

RESUMO

A reconnaissance study has been made of the distribution of 238U, 232Th, 40K and 137Cs and geochemical features in soils and sediments samples at various locations in the northwestern coast of Persian Gulf. Activity concentration levels due to radionuclides were measured in 30 soil and sediment samples collected from this region. From the measured spectra, activity concentrations were determined for 40K (range from 146 to 500 Bq kg(-1)), 137Cs (from 5 to 20 Bq kg(-1)), 238U (from 21 to 65 Bq kg(-1)) and 232Th (from 15 to 45 Bq kg(-1)) with lowest limit detection (LLD) of 68, 3.2, 4.3 and 4.3 Bq kg(-1), respectively. The dose rate from ambient air at the soil ranges was between 19 and 58 nGy h(-1) with an average of 37.41+/-9.66 nGy h(-1).


Assuntos
Monitoramento Ambiental , Metais/análise , Radioisótopos/análise , Poluentes Radioativos da Água/análise , Ar/análise , Sedimentos Geológicos/química , Oceano Índico , Irã (Geográfico) , Padrões de Referência , Solo/análise
13.
Talanta ; 184: 537-556, 2018 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-29674080

RESUMO

Recent advances in nanotechnology are as a result of the development of engineered nanoparticles. Efficiently, metallic nanoparticles have been widely exploited for biomedical application and among them, gold nanoparticles (AuNPs) are highly remarkable. Consequent upon their significant nature, spherical and gold nanorods (Au NRs) nanoparticles attract extreme attention. Their intrinsic features such as optical, electronic, physicochemical and, surface plasmon resonance (SPR); which can be altered by changing the characterizations of particles such as shape, size, aspect ratio, or environment; ease of synthesis and functionalization properties have resulted to various applications in different fields of biomedicine such as sensing, targeted drug delivery, imaging, photothermal and photodynamic therapy as well as the modulation of two or three applications. This article reviewed the popular AuNPs synthesis methods and mentioned their established applications in various demands, especially in biological sensing.

14.
Spectrochim Acta A Mol Biomol Spectrosc ; 199: 421-429, 2018 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-29649678

RESUMO

Colorimetric DNA detection is preferred over other methods for clinical molecular diagnosis because it does not require expensive equipment. In the present study, the colorimetric method based on gold nanoparticles (GNPs) and endonuclease enzyme was used for the detection of P. aeruginosa ETA gene. Firstly, the primers and probe for P. aeruginosa exotoxin A (ETA) gene were designed and checked for specificity by the PCR method. Then, GNPs were synthesized using the citrate reduction method and conjugated with the prepared probe to develop the new nano-biosensor. Next, the extracted target DNA of the bacteria was added to GNP-probe complex to check its efficacy for P. aeruginosa ETA gene diagnosis. A decrease in absorbance was seen when GNP-probe-target DNA cleaved into the small fragments of BamHI endonuclease due to the weakened electrostatic interaction between GNPs and the shortened DNA. The right shift of the absorbance peak from 530 to 562nm occurred after adding the endonuclease. It was measured using a UV-VIS absorption spectroscopy that indicates the existence of the P. aeruginosa ETA gene. Sensitivity was determined in the presence of different concentrations of target DNA of P. aeruginosa. The results obtained from the optimized conditions showed that the absorbance value has linear correlation with concentration of target DNA (R: 0.9850) in the range of 10-50ngmL-1 with the limit detection of 9.899ngmL-1. Thus, the specificity of the new method for detection of P. aeruginosa was established in comparison with other bacteria. Additionally, the designed assay was quantitatively applied to detect the P. aeruginosa ETA gene from 103 to 108CFUmL-1 in real samples with a detection limit of 320CFUmL-1.


Assuntos
ADP Ribose Transferases/genética , Toxinas Bacterianas/genética , Queimaduras/microbiologia , Sondas de DNA/química , DNA Bacteriano/genética , Endonucleases/metabolismo , Exotoxinas/genética , Ouro/química , Nanopartículas Metálicas/química , Pseudomonas aeruginosa/genética , Fatores de Virulência/genética , ADP Ribose Transferases/análise , Toxinas Bacterianas/análise , Técnicas Biossensoriais/métodos , Calorimetria/métodos , DNA Bacteriano/análise , Exotoxinas/análise , Humanos , Infecções por Pseudomonas/diagnóstico , Infecções por Pseudomonas/microbiologia , Espectrofotometria/métodos , Fatores de Virulência/análise , Exotoxina A de Pseudomonas aeruginosa
15.
Biosens Bioelectron ; 102: 226-233, 2018 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-29149688

RESUMO

The increasing progress in using nano-biomaterials for medical purposes has opened new horizons toward researchers around the globe. To investigate the presence of these nanomaterials and the impacts they might have, a comparative enhanced-electrochemiluminescence immunosensing study has been designed. The effects of utilizing graphene oxide, silica, and gold nanoparticles in cancer diagnosis were evaluated during the quantification of two major cancer biomarkers (CEA and AFP) in different approaches. In other words, first and second approaches were designed to employ nanomaterials while third and fourth approaches were developed in absence of those. Accordingly, resulted LODs experienced dramatic amplification when nano-biomaterials were included in the immunosensor modification (for AFP: 1st and 3rd approaches: 1.36fg/ml in comparison with 0.39ng/ml, and for CEA: 2nd and 4th approaches: 1.90fg/ml versus 0.46ng/ml, respectively). Correspondingly, capability of nano-biomaterials for developing highly sensitive and more efficient immunosensors was validated through selectivity, stability, reproducibility, and feasibility examinations.


Assuntos
Biomarcadores Tumorais/isolamento & purificação , Técnicas Biossensoriais/métodos , Antígeno Carcinoembrionário/isolamento & purificação , Neoplasias/diagnóstico , Biomarcadores Tumorais/genética , Antígeno Carcinoembrionário/química , Ouro/química , Grafite/química , Humanos , Medições Luminescentes , Nanopartículas Metálicas/química , Dióxido de Silício/química
16.
Artif Cells Nanomed Biotechnol ; 46(sup2): 727-736, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29873516

RESUMO

In the present research, we assumed that reducing the amounts of E6 and E7 oncoproteins by a specific siRNA sequence and recovering p53 and RB proteins, along with the recovery of the FOXO1 protein by applying anti-miR-182, would increase apoptosis and reduce proliferation rate in cancer cells. The HPV16-positive CaSki cervical cancer cell line was used. 48 hours after transfection of siRNA for targeting E6 and E7 oncoproteins and anti-miR-182, expression of its cellular targets p53, p21 and FOXO1 was assessed by real-time PCR, western blot analysis and immunocytofluorescence staining. In all treatments, apoptosis rate and viability were evaluated using Annexin-V-FITC apoptosis detection kits and MTT assays, respectively. Among the designed siRNAs, E6-1 and E7-2 proved the most effective in reducing E6 and E7 expressions by increasing the apoptotic rates to 12.4% and 16%, respectively, after 48 hours. Also, using anti-miR-182 increased apoptotic rate to 12.7% 48 hours after transfection of cervical cancer cells. The combinational use of either E6-1 or E7-2 siRNAs with anti-miR-182 resulted in a rise in apoptosis to 19.3% and 26%, respectively, higher than those obtained from the individual application of either without anti-miR-182. The simultaneous use of siRNA E6-1 and siRNA E7-2 with cisplatin increased sensitivity to cisplatin and reduced the viability of the cancer cells as compared to the use of cisplatin alone. The simultaneous use of cisplatin and anti-miR-182 had no considerable effect on viability or apoptosis rate compared to cisplatin alone.


Assuntos
Apoptose/genética , Papillomavirus Humano 16/fisiologia , MicroRNAs/genética , Proteínas Oncogênicas Virais/genética , Proteínas E7 de Papillomavirus/genética , RNA Interferente Pequeno/genética , Proteínas Repressoras/genética , Neoplasias do Colo do Útero/patologia , Linhagem Celular Tumoral , Proliferação de Células/genética , Sobrevivência Celular/genética , Cisplatino/farmacologia , Feminino , Proteína Forkhead Box O1/genética , Regulação Neoplásica da Expressão Gênica/genética , Técnicas de Silenciamento de Genes , Humanos , Proteínas Oncogênicas Virais/deficiência , Proteínas E7 de Papillomavirus/deficiência , Interferência de RNA , Proteínas Repressoras/deficiência
17.
Ultrason Sonochem ; 39: 676-685, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-28732994

RESUMO

Cadmium selenide/graphene quantum dots (CdSe/GQDs) nanocatalyst with small band gap energy and a large specific surface area was produced via a facile three-step sonochemical-hydrothermal process. The features of the as-prepared CdSe, GQDs and CdSe/GQDs samples were characterized by photoluminescence spectroscopy (PL), scanning electron microscopy (SEM), energy dispersive X-ray (EDX), X-ray diffraction (XRD), Fourier transformed infrared (FT-IR), diffuse-reflectance spectrophotometer (DRS), and Brunauer-Emmett-Teller (BET) analysis. The sonocatalytic activity of the synthesized CdSe/GQDs was effectively accelerated compared with that of pure CdSe nanoparticles in degradation of methylene blue (MB). The influence of the CdSe/GQDs dosage (0.25-1.25g/L), initial MB concentration (20-30mg/L), initial solution pH (3-12), and ultrasonic output power (200-600W/L) were examined on the sonocatalytic treatment of MB aqueous solutions. The degradation efficiency (DE%) of 99% attained at 1g/L of CdSe/GQDs, 20mg/L of MB, pH of 9, and an output power of 200W/L at 90min of ultrasonic irradiation. Furthermore, DE% increased with addition of K2S2O8 and H2O2 as the enhancers via producing more free radicals. However, addition of sulfate, carbonate, and chloride as radical sweeper decreased DE%. Furthermore, well-reusability of the CdSe/GQDs sonocatalyst was demonstrated for 5 successive runs and some of the sonocatalytic generated intermediates were indicated by GC-MS analysis.

18.
Biosens Bioelectron ; 92: 679-686, 2017 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-27838203

RESUMO

Bio-barcode DNA based on gold nanoparticle (bDNA-GNPs) as a new generation of biosensor based detection tools, holds promise for biological science studies. They are of enormous importance in the emergence of rapid and sensitive procedures for detecting toxins of microorganisms. Exotoxin A (ETA) is the most toxic virulence factor of Pseudomonas aeruginosa. ETA has ADP-ribosylation activity and decisively affects the protein synthesis of the host cells. In the present study, we developed a fluorescence bio-barcode technology to trace P. aeruginosa ETA. The GNPs were coated with the first target-specific DNA probe 1 (1pDNA) and bio-barcode DNA, which acted as a signal reporter. The magnetic nanoparticles (MNPs) were coated with the second target-specific DNA probe 2 (2pDNA) that was able to recognize the other end of the target DNA. After binding the nanoparticles with the target DNA, the following sandwich structure was formed: MNP 2pDNA/tDNA/1pDNA-GNP-bDNA. After isolating the sandwiches by a magnetic field, the DNAs of the probes which have been hybridized to their complementary DNA, GNPs and MNPs, via the hydrogen, electrostatic and covalently bonds, were released from the sandwiches after dissolving in dithiothreitol solution (DTT 0.8M). This bio-barcode DNA with known DNA sequence was then detected by fluorescence spectrophotometry. The findings showed that the new method has the advantages of fast, high sensitivity (the detection limit was 1.2ng/ml), good selectivity, and wide linear range of 5-200ng/ml. The regression analysis also showed that there was a good linear relationship (∆F=0.57 [target DNA]+21.31, R2=0.9984) between the fluorescent intensity and the target DNA concentration in the samples.


Assuntos
ADP Ribose Transferases/genética , Toxinas Bacterianas/genética , DNA Bacteriano/genética , Exotoxinas/genética , Ouro/química , Nanopartículas de Magnetita/química , Nanopartículas Metálicas/química , Pseudomonas aeruginosa/genética , Fatores de Virulência/genética , Técnicas Biossensoriais/métodos , Sondas de DNA/química , Sondas de DNA/genética , DNA Bacteriano/análise , Humanos , Limite de Detecção , Nanopartículas de Magnetita/ultraestrutura , Nanopartículas Metálicas/ultraestrutura , Infecções por Pseudomonas/diagnóstico , Infecções por Pseudomonas/microbiologia , Espectrometria de Fluorescência/métodos , Exotoxina A de Pseudomonas aeruginosa
19.
Genome Announc ; 5(8)2017 Feb 23.
Artigo em Inglês | MEDLINE | ID: mdl-28232449

RESUMO

Implementation of a vector-enabled metagenomics approach resulted in the identification of various geminiviruses. We identified the genome sequences of Beet curly top Iran virus, Turnip curly top viruses, Oat dwarf viruses, the first from Iran, and Wheat dwarf virus from leafhoppers feeding on beet, parsley, pumpkin, and turnip plants.

20.
Mater Sci Eng C Mater Biol Appl ; 78: 1195-1202, 2017 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-28575957

RESUMO

In the present study, for the first time, polycaprolactone (PCL) and gelatin (GEL) were used for neural differentiation of human induced pluripotent stem cells (hiPSCs) in the form of bi-electrospun nanofibers. The electrospun fibers were evaluated by FTIR and tensile analysis. MTT assay was used to evaluate the toxicity on the scaffolds. The hiPSCs were seeded on the fibers and after 14days in neural differentiation medium. To confirm the differentiation, real-time PCR and immunocytochemistry (ICC) analyses were performed. For morphological studies of fibers and cultured cells on them, scanning electron microscopy (SEM) and optical microscopy (OM) were used. Our results indicated that hiPSCs had differentiated to neural cells completely after incubation time. Our study demonstrates that PCL/GEL bi-electrospun nanofibers not only have the capability to support hiPSCs differentiation to neural cells, but they also are able to enhance and improve such process. Overall, PCL/GEL scaffolds seem to be a feasible, reliable and easily accessed composite for further tissue engineering experiments.


Assuntos
Células-Tronco Pluripotentes Induzidas , Diferenciação Celular , Gelatina , Humanos , Nanofibras , Neurônios , Poliésteres , Engenharia Tecidual , Alicerces Teciduais
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