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1.
Materials (Basel) ; 16(23)2023 Nov 29.
Artículo en Inglés | MEDLINE | ID: mdl-38068178

RESUMEN

Graphene oxide (GO) is an oxidized form of graphene accommodating various oxygen-containing functional groups such as hydroxyl, epoxy, and carboxyl groups on its surface. GO has been extensively utilized in various biomedical applications including the delivery of biomolecules and the development of biosensors owing to its beneficial properties such as high surface area, nucleic acid adsorption, and fluorescence quenching through fluorescence resonance energy transfer (FRET). However, despite these favorable properties, the direct utilization of GO in these applications is often limited by low dispersibility in a physiological medium, cytotoxicity, low biocompatibility, and a strong binding affinity of nucleic acids to GO surface. The large surface area of GO and the presence of various functional groups on its surface make it highly amenable to facile surface modifications, offering scope for GO surface functionalization to overcome these limitations. When polyethylene glycol (PEG), which is a biocompatible polymer, is conjugated to GO, the PEGylated GO enhances the biocompatibility and dispersibility, reduces cytotoxicity, and allows controlled drug delivery with controllable binding affinity towards nucleic acid. PEG-engrafted GO retains the beneficial properties of GO while effectively addressing its limitations, rendering it suitable for various biomedical applications. In this review, we present the recent advancements of PEGylated GO in gene/drug delivery and the facilitation of nucleic acid amplification techniques, which aid in the development of therapeutic and diagnostic tools, respectively.

2.
Nanomaterials (Basel) ; 13(8)2023 Apr 12.
Artículo en Inglés | MEDLINE | ID: mdl-37110934

RESUMEN

Quantitative real-time polymerase chain reaction (qPCR) is an important and extensively utilized technique in medical and biotechnological applications. qPCR enables the real-time detection of nucleic acid during amplification, thus surpassing the necessity of post-amplification gel electrophoresis for amplicon detection. Despite being widely employed in molecular diagnostics, qPCR exhibits limitations attributed to nonspecific DNA amplification that compromises the efficiency and fidelity of qPCR. Herein, we demonstrate that poly(ethylene glycol)-engrafted nanosized graphene oxide (PEG-nGO) can significantly improve the efficiency and specificity of qPCR by adsorbing single-stranded DNA (ssDNA) without affecting the fluorescence of double-stranded DNA binding dye during DNA amplification. PEG-nGO adsorbs surplus ssDNA primers in the initial phase of PCR, having lower concentrations of DNA amplicons and thus minimizing the nonspecific annealing of ssDNA and false amplification due to primer dimerization and erroneous priming. As compared to conventional qPCR, the addition of PEG-nGO and the DNA binding dye, EvaGreen, in the qPCR setup (dubbed as PENGO-qPCR) significantly enhances the specificity and sensitivity of DNA amplification by preferential adsorption of ssDNA without inhibiting DNA polymerase activity. The PENGO-qPCR system for detection of influenza viral RNA exhibited a 67-fold higher sensitivity than the conventional qPCR setup. Thus, the performance of a qPCR can be greatly enhanced by adding PEG-nGO as a PCR enhancer as well as EvaGreen as a DNA binding dye to the qPCR mixture, which exhibits a significantly improved sensitivity of the qPCR.

3.
Biosensors (Basel) ; 12(8)2022 Aug 20.
Artículo en Inglés | MEDLINE | ID: mdl-36005057

RESUMEN

Loop-mediated isothermal amplification (LAMP) is a nucleic acid amplification method that allows the simple, quick, and low-cost detection of various viral genes. LAMP assays are susceptible to generating non-specific amplicons, as high concentrations of DNA primers can give rise to primer dimerization and mismatched hybridizations, resulting in false-positive signals. Herein, we reported that poly(ethylene glycol)-engrafted nanosized graphene oxide (PEG-nGO) can significantly enhance the specificity of LAMP, owing to its ability to adsorb single-stranded DNA (ssDNA). By adsorbing surplus ssDNA primers, PEG-nGO minimizes the non-specific annealing of ssDNAs, including erroneous priming and primer dimerization, leading to the enhanced specificity of LAMP. The detection of complementary DNAs transcribed from the hepatitis C virus (HCV) RNA was performed by the PEG-nGO-based LAMP. We observed that the inclusion of PEG-nGO significantly enhances the specificity and sensitivity of the LAMP assay through the augmented difference in fluorescence signals between the target and non-target samples. The PEG-nGO-based LAMP assay greatly facilitates the detection of HCV-positive clinical samples, with superior precision to the conventional quantitative real-time PCR (RT-qPCR). Among the 20 clinical samples tested, all 10 HCV-positive samples are detected as positive in the PEG-nGO-based LAMP, while only 7 samples are detected as HCV-positive in the RT-qPCR. In addition, the PEG-nGO-based LAMP method significantly improves the detection precision for the false-positive decision by 1.75-fold as compared to the LAMP without PEG-nGO. Thus, PEG-nGO can significantly improve the performance of LAMP assays by facilitating the specific amplification of target DNA with a decrease in background signal.


Asunto(s)
Hepatitis C , Polietilenglicoles , Genes Virales , Grafito , Humanos , Técnicas de Diagnóstico Molecular , Técnicas de Amplificación de Ácido Nucleico/métodos , Sensibilidad y Especificidad
4.
Toxicon ; 194: 53-62, 2021 Apr 30.
Artículo en Inglés | MEDLINE | ID: mdl-33610634

RESUMEN

The Gram-negative bacteria Photorhabdus lives in a symbiotic relationship with the insect-pathogenic Heterorhabditis nematodes and produces numerous hydrolytic enzymes, secondary metabolites and protein toxins. Seven Photorhabdus strains were previously isolated from the Heterorhabditis nematodes collected from different geographical regions of India. The strains IARI-SGMG3, IARI-SGHR2, IARI-SGHR4, IARI-SGMS1 and IARI-SGGJ2 were identified as P. akhurstii, whereas IARI-SGLDK1 and IARI-SGHP1 were identified as P. laumondii subsp. laumondii and P. laumondii subsp. clarkeii, respectively. A new and previously unreported 35 kDa molecular weight protein toxin 'Galtox' was identified from these Photorhabdus strains. The nucleotide sequences of the toxin gene from seven Photorhabdus strains were PCR amplified, sequenced, cloned into pET protein expression vector, and the protein toxin was expressed and purified. The Galtox sequence from various strains showed variations in sequence and toxicity against Galleria mellonella. The injection of purified Galtox protein into the 4th instar larvae showed median lethal dose (LD50) values of 2.39-26.08 ng toxin/g G. mellonella bodyweight after 48 h. The protein injection killed the insects quickly and exhibited a median lethal time (LT50) of 12-60 h when injected at the rate of 3.1-31.2 ng toxin/g G. mellonella bodyweight. Galtox protein sequence analysis indicated similarity to several bacterial toxin-related protein domains, such as 6rgnA domain of Bordetella membrane targeting toxin BteA, 6gy6 domain of Xenorhabdus α-Xenorhabdolysins, 4mu6A and 4xa9a domains similar to effector protein LegC3 from Legionella pneumophila and 1cv8.1 domain of staphylococcal cysteine proteinase staphopain B. The mode of action of Galtox needs to be understood to enable its use for the management of agricultural insect-pests.


Asunto(s)
Toxinas Bacterianas/toxicidad , Nematodos , Photorhabdus , Animales , Toxinas Bacterianas/aislamiento & purificación , India , Mariposas Nocturnas , Xenorhabdus
5.
Data Brief ; 28: 104856, 2020 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-31871996

RESUMEN

In this data in brief (DIB) article, major photodetector (PD) characteristics of anisotype (Ag/n-TiO2/p-Si/Al), isotype (Ag/n-TiO2/n-Si/Ag) and M-S-M type (Ag/p-Si/Al) structures under reverse bias conditions (-1 to -5 V) over a broad spectral region (300-800 nm) have been presented. Critical figures of merit like current-voltage (IV), responsivity (R), detectivity (D), gain, sensitivity (S), linear dynamic range (LDR), normalized photo to dark current ratio (NPDR) and noise equivalent power (NEP) of TiO2 embedded Si PDs are presented in graphical forms. I-V characteristics of PDs under dark and monochromatic illuminations (365, 425, 515 and 600 nm) were acquired by using source measure unit (Kithley). Internal gain was deduced from photoresponse spectra which were recorded with the help of Potentiostat/Galvanostat (PGSTAT302N, Autolab) under monochromatic illumination at 100 Hz chopping frequency. Quantum efficiency instrument supplied by Optosolar was utilized to accurately measure the spectral responsivity and detectivity of PDs in wide spectral region (300-1100 nm). Please refer our main article [1] to understand the role of functional nanocrystalline TiO2 films on the performance of the photodetectors.

6.
J Nematol ; 51: 1-11, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31115203

RESUMEN

A new amphimictic species Oscheius indicus n. sp. is described and illustrated with morphological and molecular data. The species is characterized by a medium-sized and slender body (female: L = 1.1 to 1.5 mm; a = 16.8 to 20.6; b = 5.7 to 7.1; c = 7.5 to 10.4; c' = 5.0 to 7.6; V = 45 to 51%), presence of four incisures each in the lateral fields with three minute warts, long rectum (2 to 3 anal body diameters), nine pairs of papillae arranged as 1+1+1/3+3 pattern, a prominent double-flapped epipytigma on vulval opening, presence of open leptoderan bursa and crochet needle-shaped spicules place it in the insectivora group. Morphologically, O. indicus n. sp. closely resembles O. carolinensis, O. chongmingensis, O. colombiana, and O. nadarajani. Molecular phylogenetic analysis carried out using ITS and D2/D3 expansion region of 28S rDNA sequences suggests that O. indicus n. sp. is closer to O. chongmingensis and O. rugaonensis. In summary, the morphometrical data, morphological observations and molecular phylogenetic analysis suggested that O. indicus n. sp. is sufficiently different from any known species and is therefore proposed as a new species within the insectivora group.A new amphimictic species Oscheius indicus n. sp. is described and illustrated with morphological and molecular data. The species is characterized by a medium-sized and slender body (female: L = 1.1 to 1.5 mm; a = 16.8 to 20.6; b = 5.7 to 7.1; c = 7.5 to 10.4; c' = 5.0 to 7.6; V = 45 to 51%), presence of four incisures each in the lateral fields with three minute warts, long rectum (2 to 3 anal body diameters), nine pairs of papillae arranged as 1+1+1/3+3 pattern, a prominent double-flapped epipytigma on vulval opening, presence of open leptoderan bursa and crochet needle-shaped spicules place it in the insectivora group. Morphologically, O. indicus n. sp. closely resembles O. carolinensis, O. chongmingensis, O. colombiana, and O. nadarajani. Molecular phylogenetic analysis carried out using ITS and D2/D3 expansion region of 28S rDNA sequences suggests that O. indicus n. sp. is closer to O. chongmingensis and O. rugaonensis. In summary, the morphometrical data, morphological observations and molecular phylogenetic analysis suggested that O. indicus n. sp. is sufficiently different from any known species and is therefore proposed as a new species within the insectivora group.

7.
Chemphyschem ; 16(8): 1750-6, 2015 Jun 08.
Artículo en Inglés | MEDLINE | ID: mdl-25820185

RESUMEN

An intensive electrochemical impedance study was carried out to understand the charge-transfer processes in photoelectrochemical (PEC) cells based on ionic liquid (IL) electrolytes. Three different electrolytes were utilized to understand the role of redox species as well as the medium on the charge-transfer mechanism. The negligible diffusion resistance, despite the presence of two different redox species in the case of Fe(CN)(6) (-4/-3) in IL, was explained on the basis of charge transfer between species of two different redox couples. Accordingly, the redox species are not required to travel through the bulk of the electrolyte for the removal of accumulated charges, as short-range charge transfer between the IL and the Fe(CN)(6) (-4/-3) species facilitates the removal of accumulated charges. It is also shown that PEC cells utilizing dual redox couples are highly stable with larger photoelectrochmeical windows, >3 V.

8.
Phys Chem Chem Phys ; 16(41): 22735-44, 2014 Nov 07.
Artículo en Inglés | MEDLINE | ID: mdl-25234329

RESUMEN

Ionic liquid based electrolytes are gaining great interest in the field of photoenergy conversion. We have found that the ionic liquids namely BMIm Cl, BMIm PF6 and BMIm Tf2N inherently offer redox activity. The device performance of the photoelectrochemical (PEC) cells of the configuration PbOx (0.25 cm(2))|blank ionic liquids|platinum (2 cm(2)) was analyzed in detail to get insights into the working principle of such systems. It was found that partially reversible redox ion pairs diminish the performance of such cells as power generating devices. The partial redox activity of the ionic liquids was confirmed by a number of observations derived from the PEC spectra. The important parameter, Vredox, which determines the performance of any PEC cell was also calculated for all the ionic liquids. The difficulties that arise in high frequency C-V measurements for ionic liquid systems were overcome by choosing the appropriate probing frequency. The evaluated Vredox of BMIm Cl, BMIm PF6 and BMIm Tf2N ionic liquids was found to be -0.30, -0.20 and -0.78 V (vs. NHE), respectively. This study will be beneficial to understand the role of ionic liquids as redox active electrolyte media in several applications.

9.
Phys Chem Chem Phys ; 16(38): 20900-8, 2014 Oct 14.
Artículo en Inglés | MEDLINE | ID: mdl-25170591

RESUMEN

In this paper, we have carefully investigated the operation of a photoelectrochemical (PEC) cell configured of PbOx|Fe(CN)6(-4/-3)|Pt in the accumulation, flat band, depletion, inversion and deep-depletion regions using impedance measurements. The increases in the photocurrent for the different regions differ in their nature: a logarithmic increase in the depletion region, a linear increase in the inversion region along with a linear increase in the dark current and an exponential increase in the photo- and dark current in the deep-depletion region. All these variations are studied in detail to correlate these observations to the charge transfer mechanisms. The characteristics of the impedance spectrum itself can be assigned to the mentioned regions. We have found that the maximum photocurrent of the PEC cell, in the present investigation, can be extracted when the cell is working in the inversion region, while the maximum rate of the increase in photocurrent is found when the junction behaves as an ideal Schottky diode with a single RC element. Systematic experiments are suggested to establish a correlation between the observations obtained from the photocurrent, impedance, conductance, low frequency and high frequency capacitance measurements. It was found that light induced trap states in the semiconductor limit the photocurrent which has a linear dependency on the irradiance. A detailed investigation with A.C. conductivity measurements showed that the trap states actively participate in the current mechanism via a hopping phenomenon with small activation energies of 0.2 and 0.8 meV. The hopping rate increased exponentially with the applied bias under dark and illumination conditions. We also show a new way of finding the potential at which the maximum photocurrent will be extracted from the PEC cell, wherein the hopping via trap states is a dominating charge transfer mechanism. This study will help in pin pointing the key affecting parameters which limit the charge transfer process in the cell.

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