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1.
ACS Nano ; 2024 May 08.
Artículo en Inglés | MEDLINE | ID: mdl-38717114

RESUMEN

Facing the escalating threat of viruses worldwide, the development of efficient sensor elements for rapid virus detection has never been more critical. Traditional point-of-care (POC) sensors struggle due to their reliance on fragile biological receptors and limited adaptability to viral strains. In this study, we introduce a nanosensor design for receptor-free virus recognitions using near-infrared (NIR) fluorescent single-walled carbon nanotubes (SWCNTs) functionalized with a poly(ethylene glycol) (PEG)-phospholipid (PEG-lipid) array. Three-dimensional (3D) corona interfaces of the nanosensor array enable selective and sensitive detection of diverse viruses, including Ebola, Lassa, H3N2, H1N1, Middle East respiratory syndrome (MERS), severe acute respiratory syndrome coronavirus 1 (SARS-CoV-1), and SARS-CoV-2, even without any biological receptors. The PEG-lipid components, designed considering chain length, fatty acid saturation, molecular weight, and end-group moieties, allow for precise quantification of viral recognition abilities. High-throughput automated screening of the array demonstrates how the physicochemical properties of the PEG-lipid/SWCNT 3D corona interfaces correlate with viral detection efficiency. Utilizing molecular dynamics and AutoDock simulations, we investigated the impact of PEG-lipid components on 3D corona interface formation, such as surface coverage and hydrodynamic radius and specific molecular interactions based on chemical potentials. Our findings not only enhance detection specificity across various antigens but also accelerate the development of sensor materials for promptly identifying and responding to emerging antigen threats.

2.
J Microbiol Biotechnol ; 34(4): 804-811, 2024 Apr 28.
Artículo en Inglés | MEDLINE | ID: mdl-38379304

RESUMEN

Foamy viruses (FVs) are generally recognized as non-pathogenic, often causing asymptomatic or mild symptoms in infections. Leveraging these unique characteristics, FV vectors hold significant promise for applications in gene therapy. This study introduces a novel platform technology using a pseudo-virus with single-round infectivity. In contrast to previous vector approaches, we developed a technique employing only two vectors, pcHFV lacking Env and pCMV-Env, to introduce the desired genes into target cells. Our investigation demonstrated the efficacy of the prototype foamy virus (PFV) dual-vector system in producing viruses and delivering transgenes into host cells. To optimize viral production, we incorporated the codon-optimized Env (optEnv) gene in pCMV-Env and the Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element (WPRE) at the 3' end of the transgene in the transfer vector. Consequently, the use of optEnv led to a significant enhancement in transgene expression in host cells. Additionally, the WPRE exhibited an enhancing effect. Furthermore, the introduced EGFP transgene was present in host cells for a month. In an effort to expand transgene capacity, we further streamlined the viral vector, anticipating the delivery of approximately 4.3 kbp of genes through our PFV dual-vector system. This study underscores the potential of PFVs as an alternative to lentiviruses or other retroviruses in the realm of gene therapy.


Asunto(s)
Técnicas de Transferencia de Gen , Vectores Genéticos , Spumavirus , Transgenes , Spumavirus/genética , Vectores Genéticos/genética , Humanos , Terapia Genética/métodos , Animales , Células HEK293 , Proteínas Fluorescentes Verdes/genética , Línea Celular
3.
Cornea ; 2024 Jan 19.
Artículo en Inglés | MEDLINE | ID: mdl-38251908

RESUMEN

PURPOSE: The purpose of this study was to describe a new surgical technique for deep anterior lamellar keratoplasty. METHODS: All pupils in the recipient eyes were dilated preoperatively. Vertical grooving was performed using a crescent blade with a width of 5 mm and a depth of one-third to half corneal thickness on the temporal side of the limbus. Stromal dissection was performed as close as possible to Descemet membrane by observing the gap between the gold line by retinal reflex and the front edge of the crescent blade. Lamellar dissection was performed along the lamellar plane using corneal dissectors. The ophthalmic viscoelastic device was injected into the intrastromal pocket to separate the anterior and posterior stroma and an anterior corneal lamella was excised. A donor cornea was sutured into the recipient bed. RESULTS: In 18 eyes, none of the patients had Descemet membrane rupture during surgery. The mean postoperative residual stromal thickness was 80 ± 31 µm. The mean central corneal thickness after surgery was 660 ± 69 µm. At the last follow-up, the cornea was cleared in all 18 eyes on slit-lamp examination. CONCLUSIONS: We estimated the residual stromal thickness based on the gap between the gold line by the retinal reflex and crescent blade, and intrastromal lamellar dissection was performed using a smooth corneal dissector. Consequently, the surface of stromal dissection was smooth, and the residual stromal thickness was even.

4.
ACS Meas Sci Au ; 3(6): 393-403, 2023 Dec 20.
Artículo en Inglés | MEDLINE | ID: mdl-38145025

RESUMEN

With the definition of therapeutics now encompassing transplanted or engineered cells and their molecular products, there is a growing scientific necessity for analytics to understand this new category of drugs. This Perspective highlights the recent development of new measurement science on label-free single cell analysis, nanosensor chemical cytometry (NCC), and their potential for cellular therapeutics and precision medicine. NCC is based on microfluidics integrated with fluorescent nanosensor arrays utilizing the optical lensing effect of a single cell to real-time extract molecular properties and correlate them with physical attributes of single cells. This new class of cytometry can quantify the heterogeneity of the multivariate physicochemical attributes of the cell populations in a completely label-free and nondestructive way and, thus, suggest the vein-to-vein conditions for the safe therapeutic applications. After the introduction of the NCC technology, we suggest the technological development roadmap for the maturation of the new field: from the sensor/chip design perspective to the system/software development level based on hardware automation and deep learning data analytics. The advancement of this new single cell sensing technology is anticipated to aid rich and multivariate single cell data setting and support safe and reliable cellular therapeutics. This new measurement science can lead to data-driven personalized precision medicine.

5.
ACS Sens ; 8(11): 4207-4215, 2023 11 24.
Artículo en Inglés | MEDLINE | ID: mdl-37874627

RESUMEN

There has been considerable interest in detecting atmospheric and process-associated methane (CH4) at low concentrations due to its potency as a greenhouse gas. Nanosensor technology, particularly fluorescent single-walled carbon nanotube (SWCNT) arrays, is promising for such applications because of their chemical sensitivities at single-molecule detection limits. However, the methodologies for connecting the stochastic molecular fluctuations from gas impingement on such sensors require further development. In this work, we synthesize Pd-conjugated ss(GT)15-DNA-wrapped SWCNTas near-infrared (nIR) fluorescent, single-molecule sensors of CH4. The complexes are characterized using X-ray photoelectron spectroscopy (XPS) and spectrophotometry, demonstrating spectral changes between the Pd2+ and Pd0 oxidation states. The nIR fluctuations generated upon exposure from 8 to 26 ppb of CH4 were separated into high- and low-frequency components. Aggregating the low-frequency components for an array of sensors showed the most consistent levels of detection with a limit of 0.7 ppb. These results advance the hardware and computational methods necessary to apply this approach to the challenge of environmental methane sensing.


Asunto(s)
Nanotubos de Carbono , Nanotubos de Carbono/química , Paladio , Metano , Nanotecnología , Colorantes
6.
Transl Vis Sci Technol ; 12(7): 5, 2023 07 03.
Artículo en Inglés | MEDLINE | ID: mdl-37405796

RESUMEN

Purpose: To use the revised model eye to observe and compare how the world is perceived by patients with monofocal intraocular lens (IOL), Eyhance, bifocal IOL, and Symfony, and check its performance. Methods: The new mobile model eye consists of an artificial cornea, an IOL, a wet cell, an adjustable lens tube, a lens tube, an objective lens, a tube lens, and a digital single-lens reflex camera. We collected photographs of distant buildings and streets at night, videos of the focusing process, and videos of United States Air Force resolution target from 6 m to 15 cm and analyzed them quantitatively. Results: In this revised model eye using an objective lens, an artificial cornea similar to the human cornea could be used. Using a digital single-lens reflex camera, high-resolution imaging was possible without an additional computer. Fine focusing was possible using an adjustable lens tube. For monofocal IOL, the contrast modulation was 0.39 at 6 m and decreased consistently. It was nearly 0 as the model eye got closer than 1.6 m. For Eyhance, the contrast modulation was 0.40 at 6 m. It then decreased and increased again. At 1.3 m, it was 0.07 and then decreased again. For Symfony, the contrast modulation was 0.18 at 6 m. Symfony showed the characteristics of a bifocal IOL with low add diopter. Halos (234 pixels) were observed around lights, although smaller than those seen with bifocal IOL (432 pixels). Conclusions: We could objectively observe and compare how patients with monofocal IOL, Eyhance, bifocal IOL, and Symfony perceived the world using this revised model eye. Translational Relevance: Data obtained by this new mobile model eye can be used to help patients select their IOLs before cataract surgeries.


Asunto(s)
Lentes Intraoculares , Humanos , Agudeza Visual , Visión Ocular
7.
Anesth Pain Med (Seoul) ; 18(2): 139-147, 2023 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-37183282

RESUMEN

BACKGROUND: Perioperative hyperglycemia can occur in surgical patients and may increase postoperative morbidity and mortality, especially in patients with diabetes. Therefore, we conducted the present study to evaluate whether the administration of 6% hydroxyethyl starch (HES)-130/0.4 increases blood glucose levels in patients with diabetes. METHODS: Forty patients undergoing lower limb surgery under spinal anesthesia were randomly allocated into two groups according to the fluids administered 20 min before spinal anesthesia (Group L, lactated Ringer's solution; Group H, 6% HES-130/0.4). Patient characteristics, intraoperative variables, blood glucose levels, mean blood pressure (MBP), and heart rate (HR) were recorded at five time-points (0, 20, 60, 120, and 240 min). RESULTS: A total of 39 patients were analyzed (Group L, n = 20; Group H, n = 19). The amount of intraoperative fluid was significantly higher in Group L than in Group H (718.2 ml vs. 530.0 ml, P = 0.010). There were no significant differences in the changes in blood glucose levels, HR, or MBP between the two groups (P = 0.737, P = 0.896, and P = 0.141, respectively). Serial changes in mean blood glucose levels from baseline also showed no significant differences between the groups (P = 0.764). CONCLUSIONS: There were no significant changes in blood glucose levels when lactated Ringer's solution or 6% HES-130 was used. When compared to the lactated Ringer's solution, no evidence that 6% HES-130/0.4 produces hyperglycemia in diabetic patients could be found. Further evaluation of larger populations is needed.

8.
ACS Nano ; 17(11): 10898-10905, 2023 Jun 13.
Artículo en Inglés | MEDLINE | ID: mdl-37222273

RESUMEN

Developing a methodology to enhance long-term stability is one of the most important issues in MXene research, since they are prone to oxidation in the ambient environment. Although various approaches have been suggested to improve the stability of MXene, they have suffered from complicated processes and limited applicability to various types of MXene nanostructures. Herein, we report a simple and versatile technique to enhance the environmental stability of MXenes. Ti3C2Tx MXene films were decorated with a highly hydrophobic polymer, 1H,1H,2H,2H-perfluorodecyl methacrylate (PFDMA), using initiated chemical vapor deposition (iCVD) where iCVD allows the facile postdeposition of polymer films of desired thickness on MXene films. The oxidation resistance was evaluated by fabricating MXene gas sensors and measuring the change in signal-to-noise ratio (SNR) of volatile organic compound (VOC) gases under harsh conditions (RH 100% at 50 °C) for several weeks where the performance in the absence and presence of PFDMA was compared. The results show that while the SNR of PFDMA-Ti3C2Tx sensors was retained, a dramatic increase of the noise level and a decrease in the SNR were observed in pristine Ti3C2Tx. We believe that this simple and nondestructive method will offer great potential to enhance the stability of a wide range of MXenes.

9.
ACS Sens ; 8(4): 1371-1372, 2023 04 28.
Artículo en Inglés | MEDLINE | ID: mdl-37114432
10.
PLoS One ; 18(4): e0283261, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-37058518

RESUMEN

The increased interest in outdoor activities has prompted the demand for water-repellent fabrics that can withstand various environmental factors. In this study, the water repellency and physical properties, namely thickness, weight, tensile strength, elongation, and stiffness, of cotton woven fabrics were analyzed according to various treatments with different types of household water-repellent agents and number of coating layers. Fluorine-, silicone-, and wax-based water-repellent agents were coated on cotton woven fabrics once, thrice, and five times. Thickness, weight, and stiffness increased with the number of coating layers, which may reduce comfort. These properties increased minimally for the fluorine- and silicone-based water-repellent agents, whereas they considerably increased for the wax-based water-repellent agent. The fluorine-based water-repellent agent had a low water repellency rating of 2.2 even after five coating layers, and the silicone-based water-repellent agent had a higher rating of 3.4 with the same five coating layers. Meanwhile, the wax-based water-repellent agent had the highest water repellency rating of 5 even with only one coating layer, which was maintained with repeated coatings. Therefore, fluorine- and silicone-based water-repellent agents minimally altered the fabric properties even with repeated coatings; multiple coating layers, especially five or more layers for the fluorine-based water-repellent agent, are recommended to attain excellent water repellency. Conversely, one coating layer of the wax-based water-repellent agent is recommended to retain the comfort of the wearer.


Asunto(s)
Flúor , Textiles , Fenómenos Físicos , Siliconas , Agua
11.
ACS Sens ; 8(4): 1676-1683, 2023 04 28.
Artículo en Inglés | MEDLINE | ID: mdl-37018205

RESUMEN

Label-free single-cell analytics have been developed for understanding the collective immune response mechanism of immune cells. However, it remains difficult to analyze the physicochemical properties of a single cell in high spatiotemporal resolution for an immune cell having dynamic morphological changes and significant molecular heterogeneities. It is deemed due to the absence of a sensitive molecular sensing construct and single-cell imaging analytic program. In this study, we developed a deep learning integrated nanosensor chemical cytometry (DI-NCC) platform, which combines a fluorescent nanosensor array in microfluidics and a deep learning model for cell feature analysis. The DI-NCC platform possesses the capability to collect rich, multivariate data sets for each individual immune cell (e.g., macrophage) within the population. We obtained LPS+ (n = 25) and LPS- (n = 61) near-infrared images and analyzed 250 cells/mm2 in 1 µm spatial resolution and 0 to 1.0 confidence level even with overlapped or adhered cell configurations. This enables automatic quantification of the activation and nonactivation levels of a single macrophage upon instantaneous immune stimulations. Furthermore, we support the activation level quantified by deep learning with heterogeneities analysis of both biophysical (cell size) and biochemical (nitric oxide efflux) properties. The DI-NCC platform can be promising for activation profiling of dynamic heterogeneity variations of cell populations.


Asunto(s)
Aprendizaje Profundo , Lipopolisacáridos , Macrófagos
12.
ACS Sens ; 8(3): 1151-1160, 2023 03 24.
Artículo en Inglés | MEDLINE | ID: mdl-36799655

RESUMEN

The design of new nanomaterials for rapid and reversible detection of molecules in existence is critical for real-world sensing applications. Current nanomaterial libraries such as carbon nanotubes, graphene, MoS2, and MXene are fundamentally limited by their slow detection speed and small signals; thus, the atomic-level material design of molecular transport pathways and active binding sites must be accompanied. Herein, we fully explore the chemical and physical properties of a hydrogen-substituted graphdiyne (HsGDY) for its molecular sensing properties. This new carbon framework comprises reactive sp carbons in acetylenic linkages throughout the 16.3 Å nanopores and allows for detecting target molecules (e.g., H2) with an exceptionally high sensitivity (ΔR/Rb = 542%) and fast response/recovery time (τ90 = 8 s and τ10 = 38 s) even without any postmodification process. It possesses 2 orders of magnitude higher sensing ability than that of existing nanomaterial libraries. We demonstrate that rapid and reversible molecular binding is attributed to the cooperative interaction with adjacent double sp carbon in the layered nanoporous structure of HsGDY. This new class of carbon framework provides fundamental solutions for nanomaterials in reliable sensor applications that accelerate real-world interfacing.


Asunto(s)
Grafito , Nanoporos , Nanotubos de Carbono , Hidrógeno
13.
Anal Chem ; 95(4): 2356-2365, 2023 01 31.
Artículo en Inglés | MEDLINE | ID: mdl-36645297

RESUMEN

Alkaline phosphatase (ALP) and interleukin-1beta (IL-1ß) are crucial salivary biomarkers for the diagnosis of periodontal disease that harms the periodontal tissue along with tooth loss. However, there has been no way of sensitive and portable detection of both biomarkers in saliva with multivariate signal readout. In this work, we design the multicolorimetric ALP and IL-1ß sensing platform based on geometrical transformation of silver nanoplate transducer. By utilizing enzymatic activity of ALP that dephosphorylates p-aminophenol phosphate (p-APP) to p-aminophenol (p-AP), localized surface plasmon resonance properties of silver nanoplate vary with ALP and show a distinct color change from blue to yellow based on a controlled seed transformation from triangular to hexagonal, rounded pentagonal, and spherical shape. The multicolor sensor shows an ALP detection range of 0-25 U/L with a limit of detection (LOD) of 0.0011 U/L, which is the lowest range of LOD demonstrated to date for state-of-the-art ALP sensor. Furthermore, we integrate the sensor with the conventional ELISA to detect IL-1ß for multicolor signaling and it exhibits a linear detection range of 0-250 pg/mL and an LOD of 0.066 pg/mL, which is 2 orders of magnitude lower than the monochromic conventional ELISA (LOD of 3.8 pg/mL). The ALP multicolor sensor shows high selectivity with a recovery of 100.9% in real human saliva proving its reliability and suitability for the readily accessible periodontal diagnosis with multivariate signal readout.


Asunto(s)
Enfermedades Periodontales , Plata , Humanos , Reproducibilidad de los Resultados , Fosfatasa Alcalina/análisis , Enfermedades Periodontales/diagnóstico , Colorantes , Biomarcadores , Límite de Detección
14.
Anal Chem ; 94(47): 16393-16401, 2022 11 29.
Artículo en Inglés | MEDLINE | ID: mdl-36378652

RESUMEN

Colloidal single-walled carbon nanotubes (SWCNTs) offer a promising platform for the nanoscale engineering of molecular recognition. Optical sensors have been recently designed through the modification of noncovalent corona phases (CPs) of SWCNTs through a phenomenon known as corona phase molecular recognition (CoPhMoRe). In CoPhMoRe constructs, DNA CPs are of great interest due to the breadth of the design space and our ability to control these molecules with sequence specificity at scale. Utilizing these constructs for metal ion sensing is a natural extension of this technology due to DNA's well-known coordination chemistry. Additionally, understanding metal ion interactions of these constructs allows for improved sensor design for use in complex aqueous environments. In this work, we study the interactions between a panel of 9 dilute divalent metal cations and 35 DNA CPs under the most controlled experimental conditions for SWCNT optical sensing to date. We found that best practices for the study of colloidal SWCNT analyte responses involve mitigating the effects of ionic strength, dilution kinetics, laser power, and analyte response kinetics. We also discover that SWCNT with DNA CPs generally offers two unique sensing states at pH 6 and 8. The combined set of sensors in this work allowed for the differentiation of Hg2+, Pb2+, Cr2+, and Mn2+. Finally, we implemented Hg2+ sensing in the context of portable detection within fish tissue extract, demonstrating nanomolar level detection.


Asunto(s)
Mercurio , Nanotubos de Carbono , Nanotubos de Carbono/química , ADN/química , Cationes Bivalentes , Cationes
15.
ACS Nano ; 16(10): 17210-17219, 2022 Oct 25.
Artículo en Inglés | MEDLINE | ID: mdl-36223595

RESUMEN

Metal oxide semiconductors (MOS) have proven to be most powerful sensing materials to detect hydrogen sulfide (H2S), achieving part per billion (ppb) level sensitivity and selectivity. However, there has not been a way of extending this approach to the top-down H2S sensor fabrication process, completely limiting their commercial-level productions. In this study, we developed a top-down lithographic process of a 10 nm-scale SnO2 nanochannel for H2S sensor production. Due to high-resolution (15 nm thickness) and high aspect ratio (>20) structures, the nanochannel exhibited highly sensitive H2S detection performances (Ra/Rg = 116.62, τres = 31 s at 0.5 ppm) with selectivity (RH2S/Racetone = 23 against 5 ppm acetone). In addition, we demonstrated that the nanochannel could be efficiently sensitized with the p-n heterojunction without any postmodification or an additional process during one-step lithography. As an example, we demonstrated that the H2S sensor performance can be drastically enhanced with the NiO nanoheterojunction (Ra/Rg = 166.2, τres = 21 s at 0.5 ppm), showing the highest range of sensitivity demonstrated to date for state-of-the-art H2S sensors. These results in total constitute a high-throughput fabrication platform to commercialize the H2S sensor that can accelerate the development time and interface in real-life situations.

16.
ACS Sens ; 7(8): 2225-2234, 2022 Aug 26.
Artículo en Inglés | MEDLINE | ID: mdl-35838305

RESUMEN

It is highly important to implement various semiconducting, such as n- or p-type, or conducting types of sensing behaviors to maximize the selectivity of gas sensors. To achieve this, researchers so far have utilized the n-p (or p-n) two-phase transition using doping techniques, where the addition of an extra transition phase provides the potential to greatly increase the sensing performance. Here, we report for the first time on an n-p-conductor three-phase transition of gas sensing behavior using Mo2CTx MXene, where the presence of organic intercalants and film thickness play a critical role. We found that 5-nm-thick Mo2CTx films with a tetramethylammonium hydroxide (TMAOH) intercalant displayed a p-type gas sensing response, while the films without the intercalant displayed a clear n-type response. Additionally, Mo2CTx films with thicknesses over 700 nm exhibited a conductor-type response, unlike the thinner films. It is expected that the three-phase transition was possible due to the unique and simultaneous presence of the intrinsic metallic conductivity and the high-density of surface functional groups of the MXene. We demonstrate that the gas response of Mo2CTx films containing tetramethylammonium (TMA) ions toward volatile organic compounds (VOCs), NH3, and NO2 is ∼30 times higher than that of deintercalated films, further showing the influence of intercalants on sensing performance. Also, DFT calculations show that the adsorption energy of NH3 and NO2 on Mo2CTx shifts from -0.973, -1.838 eV to -1.305, -2.750 eV, respectively, after TMA adsorption, demonstrating the influence of TMA in enhancing sensing performance.

18.
BMC Ophthalmol ; 21(1): 444, 2021 Dec 27.
Artículo en Inglés | MEDLINE | ID: mdl-34961511

RESUMEN

BACKGROUND: A polarization-directed flat (PDF) lens acts as a converging lens with a focal length (f) > 0 and a diverging lens with f < 0, depending on the polarization state of the incidental light. To produce a multifocal lens with two focal lengths, a PDF and a converging lens having shorter focal length were combined. In this study, we tested a bifocal PDF to determine its potential as a new multifocal intraocular lens (IOL). METHODS: Constructed a multifocal lens with a PDF lens (f = +/- 100 mm) and a converging lens (f = + 25 mm). In an optical bench test, we measured the defocus curve to test the multifocal function. The multifocal function and optical quality of the lens in various situations were tested. An Early Treatment Diabetic Retinopathy Study (ETDRS) chart as a near target and a building as a distant target were photographed using a digital single-lens reflex (DSLR) camera. Both lenses (multifocal and monofocal) were tested under the same conditions. RESULTS: For the 0 D and - 20 D focal points, the multifocal lens showed sharp images in the optical bench test. In the DSLR test using the multifocal lens, the building appeared slightly blurry compared with the results using the monofocal lens. With the multifocal lens, the ETDRS chart's images became blurry as the ETDRS chart's distance decreased, but became very clear again at a certain position. CONCLUSIONS: We confirmed the multifocal function of the multifocal lens using a PDF lens. This lens can be used as a multifocal IOL in the future.


Asunto(s)
Lentes Intraoculares , Lentes Intraoculares Multifocales , Facoemulsificación , Sensibilidad de Contraste , Humanos , Refracción Ocular , Visión Ocular
19.
J Microbiol ; 59(12): 1075-1082, 2021 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-34705258

RESUMEN

Aconitase, a highly conserved protein across all domains of life, functions in converting citrate to isocitrate in the tricarboxylic acid cycle. Cytosolic aconitase is also known to act as an iron regulatory protein in mammals, binding to the RNA hairpin structures known as iron-responsive elements within the untranslated regions of specific RNAs. Aconitase-2 (Aco2) in fission yeast is a fusion protein consisting of an aconitase and a mitochondrial ribosomal protein, bL21, residing not only in mitochondria but also in cytosol and the nucleus. To investigate the role of Aco2 in the nucleus and cytoplasm of fission yeast, we analyzed the transcriptome of aco2ΔN mutant that is deleted of nuclear localization signal (NLS). RNA sequencing revealed that the aco2ΔN mutation caused increase in mRNAs encoding iron uptake transporters, such as Str1, Str3, and Shu1. The half-lives of mRNAs for these genes were found to be significantly longer in the aco2ΔN mutant than the wild-type strain, suggesting the role of Aco2 in mRNA turnover. The three conserved cysteines required for the catalytic activity of aconitase were not necessary for this role. The UV cross-linking RNA immunoprecipitation analysis revealed that Aco2 directly bound to the mRNAs of iron uptake transporters. Aco2-mediated degradation of iron-uptake mRNAs appears to utilize exoribonuclease pathway that involves Rrp6 as evidenced by genetic interactions. These results reveal a novel role of non-mitochondrial aconitase protein in the mRNA turnover in fission yeast to fine-tune iron homeostasis, independent of regulation by transcriptional repressor Fep1.


Asunto(s)
Aconitato Hidratasa/metabolismo , Proteínas de Transporte de Catión/genética , Regulación Fúngica de la Expresión Génica , Hierro/metabolismo , ARN de Hongos/metabolismo , Proteínas de Schizosaccharomyces pombe/metabolismo , Schizosaccharomyces/genética , Proteínas de Transporte de Catión/metabolismo , Núcleo Celular/enzimología , Citoplasma/enzimología , Exorribonucleasas/genética , Exorribonucleasas/metabolismo , Factores de Transcripción GATA/genética , Factores de Transcripción GATA/metabolismo , Genes Fúngicos , Proteínas Reguladoras del Hierro/genética , Proteínas Reguladoras del Hierro/metabolismo , Proteínas de la Membrana/genética , Proteínas de la Membrana/metabolismo , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Estabilidad del ARN , ARN Mensajero/metabolismo , Regulón , Ribonucleasas/genética , Ribonucleasas/metabolismo , Schizosaccharomyces/enzimología , Proteínas de Schizosaccharomyces pombe/genética
20.
Anal Chem ; 93(44): 14685-14693, 2021 11 09.
Artículo en Inglés | MEDLINE | ID: mdl-34698489

RESUMEN

To develop better analytical approaches for future global pandemics, it is widely recognized that sensing materials are necessary that enable molecular recognition and sensor assay development on a much faster scale than currently possible. Previously developed severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) point-of-care devices are based on the specific molecular recognition using subunit protein antibodies and protein receptors that selectively capture the viral proteins. However, these necessarily involve complex and lengthy development and processing times and are notoriously prone to a loss of biological activity upon sensor immobilization and device interfacing, potentially limiting their use in applications at scale. Here, we report a synthetic strategy for nanoparticle corona interfaces that enables the molecular recognition of SARS-CoV-2 proteins without any antibody and receptor design. Our nanosensor constructs consist of poly(ethylene glycol) (PEG)─phospholipid heteropolymers adsorbed onto near-infrared (nIR) fluorescent single-walled carbon nanotubes (SWCNTs) that recognize the nucleocapsid (N) and spike (S) protein of SARS-CoV-2 using unique three-dimensional (3D) nanosensor interfaces. This results in rapid and label-free nIR fluorescence detection. This antibody-free nanosensor shows up to 50% sensor responses within 5 min of viral protein injections with limit of detection (LOD) values of 48 fM and 350 pM for N and S proteins, respectively. Finally, we demonstrate instrumentation based on a fiber-optic platform that interfaces the advantages of antibody-free molecular recognition and biofluid compatibility in human saliva conditions.


Asunto(s)
COVID-19 , Nanotubos de Carbono , Anticuerpos Antivirales , Humanos , Pandemias , SARS-CoV-2 , Glicoproteína de la Espiga del Coronavirus
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