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1.
ACS Sens ; 8(11): 4071-4078, 2023 11 24.
Article in English | MEDLINE | ID: mdl-37889801

ABSTRACT

Hyaluronidase (HAase) is a biomarker for cancer, and its detection is of great significance for early diagnosis. However, the requirement of sophisticated instruments, tedious operation procedures, and labeled molecules of conventional HAase biosensing methods hampers their widespread applications. Herein, we report a portable slippery viscosity-sensing platform with time readout for the first time and demonstrate HAase and tannic acid (TA, HAase inhibitor) detection as a model system. HAase specifically cleaves hyaluronic acid (HA) and decreases HA solution viscosity, thereby shortening the aqueous droplet's sliding time on a slippery surface. Thus, the HA solution viscosity alteration due to enzymatic hydrolysis is used to quantify the HAase concentration through the difference in the sliding time of the aqueous droplets on a slippery surface. The developed HAase sensing platform exhibits high sensitivity with a minimum detection limit of 0.23 U/mL and excellent specificity without the use of specialized instruments and labeled molecules. HAase detection in actual urine samples by a standard addition method is performed as well. Moreover, the quantitative detection of TA with an IC50 value of 37.68 ± 1.38 µg/mL is achieved. As an equipment-free, label-free, and high-portability sensing platform, this method holds promise in developing a user-friendly and inexpensive point-of-care testing (POCT) device for HAase detection, and its use can be extended to analyze other analytes with different stimuli-responsive polymers for great universality and expansibility in biosensing applications.


Subject(s)
Hyaluronoglucosaminidase , Neoplasms , Humans , Hyaluronoglucosaminidase/urine , Viscosity , Biomarkers, Tumor/urine , Hyaluronic Acid/urine
2.
Anal Chem ; 95(35): 13385-13390, 2023 09 05.
Article in English | MEDLINE | ID: mdl-37622311

ABSTRACT

The development of liquid crystal (LC)-based sensors with superior performances such as high portability, excellent stability, great convenience, and remarkable sensitivity is highly demanded. This work proposes a new strategy for constructing the LC-based sensor using enzyme-linked dual-functional nucleic acid (d-FNA) on magnetic beads (MBs). The detection of kanamycin (KA) is demonstrated as a model. Acetylcholinesterase (AChE) is assembled onto the KA aptamer-modified MBs with a d-FNA strand that consists of an AChE aptamer and the complementary sequence of a KA aptamer. As the specific recognition of KA by its aptamer triggers the release of AChE from the MBs, the myristoylcholine (Myr) solution after incubation with the MBs causes the black image of the LCs due to the formation of the Myr monolayer at the aqueous/LC interface. Otherwise, in the absence of KA, AChE is still decorated on the MBs and causes the hydrolysis of Myr. Therefore, a bright image of LCs is obtained. The detection of KA is successfully achieved with a lower detection limit of 48.1 pg/mL. In addition, a thin polydimethylsiloxane (PDMS) layer-coated glass and a portable optical device are used to improve the stability and portability of the LC-based sensor to advance potential commercial applications. Furthermore, the detection of KA in milk with a portable device is demonstrated, showing the potential of the proposed enzyme-linked LC-based sensor.


Subject(s)
Liquid Crystals , Nucleic Acids , Optical Devices , Acetylcholinesterase , Kanamycin , Oligonucleotides , Magnetic Phenomena
3.
Biosensors (Basel) ; 12(6)2022 Jun 13.
Article in English | MEDLINE | ID: mdl-35735558

ABSTRACT

The SARS-CoV-2 coronavirus, also known as the disease-causing agent for COVID-19, is a virulent pathogen that may infect people and certain animals. The global spread of COVID-19 and its emerging variation necessitates the development of rapid, reliable, simple, and low-cost diagnostic tools. Many methodologies and devices have been developed for the highly sensitive, selective, cost-effective, and rapid diagnosis of COVID-19. This review organizes the diagnosis platforms into four groups: imaging, molecular-based detection, serological testing, and biosensors. Each platform's principle, advancement, utilization, and challenges for monitoring SARS-CoV-2 are discussed in detail. In addition, an overview of the impact of variants on detection, commercially available kits, and readout signal analysis has been presented. This review will expand our understanding of developing advanced diagnostic approaches to evolve into susceptible, precise, and reproducible technologies to combat any future outbreak.


Subject(s)
Biosensing Techniques , COVID-19 , Animals , Biosensing Techniques/methods , COVID-19/diagnosis , COVID-19 Testing , Humans , SARS-CoV-2
4.
Food Chem ; 390: 133202, 2022 Oct 01.
Article in English | MEDLINE | ID: mdl-35580521

ABSTRACT

Aflatoxin B1 (AFB1) is a common food mycotoxin that can cause various diseases. Therefore, reliable detection methods are required to ensure food safety against mycotoxins. In this study, we design a liquid-crystal (LC)-based assay for rapid detection of AFB1 in food samples. The surface-anchored LC droplets on glass (5CBSADrop) are obtained via a solvent evaporation method. The 5CBSADrop displays a four-leaf clover appearance that corresponds to an escape-radial configuration in a mixture of CTAB and AFB1 aptamer. Interestingly, they adopt a radial configuration in the mixture of CTAB, AFB1, and its aptamer. Using this approach, AFB1 can be detected using only 1 µL of the aqueous solution with a minimum detection concentration of 10 pg/mL. This LC-based sensing platform provides simple operation, remarkable sensitivity, high selectivity, low cost, and excellent portability without the use of any bulky instrument, which is very promising in rapid on-field detection of mycotoxins.


Subject(s)
Aptamers, Nucleotide , Biosensing Techniques , Liquid Crystals , Aflatoxin B1/analysis , Aptamers, Nucleotide/chemistry , Biosensing Techniques/methods , Cetrimonium , Food Contamination/analysis , Limit of Detection
5.
Anal Chim Acta ; 1205: 339756, 2022 May 01.
Article in English | MEDLINE | ID: mdl-35414397

ABSTRACT

Thrombin is a biomarker of blood-related diseases. Its detection and inhibitor screening are of great significance in the fields of medical and biological research. Herein, a novel paper-based lateral flow sensor for thrombin detection and inhibitors screening is developed. The formation of cross-links of fibrin from fibrinogen during the blood clotting process can efficiently capture water molecules. Based on the principle, in the presence of thrombin, the water of the plasma solution cannot flow along with the test paper as the water molecules are trapped in the fibrin. Upon the inactivation of thrombin by the inhibitors, water can flow along the test paper. The detection limit of thrombin reaches about 16.1 mU/mL and the sensing platform also exhibits excellent selectivity, reproducibility, and stability. Furthermore, the inhibition efficiency of argatroban, a clinical drug used as a thrombin inhibitor, is also successfully investigated using the assay. Overall, this strategy exhibits the advantages of the specific enzymatic reaction, low-cost, and label-free detection of thrombin with high sensitivity and remarkable specificity. This method is also competent to facilitate the screening of thrombin inhibitors, which is promising in the discovery of therapeutic drugs for thrombus-related diseases.


Subject(s)
Fibrin , Thrombin , Fibrinogen , Reproducibility of Results , Water
6.
Talanta ; 244: 123455, 2022 Jul 01.
Article in English | MEDLINE | ID: mdl-35397324

ABSTRACT

The abnormal levels of uric acid (UA) in body fluids are associated with gout, type (II) diabetes, leukemia, Lesch-Nyhan syndrome, uremia, kidney damage, and cardiovascular diseases. Also, the presence of uricase (UOx) symbolizes genetic disorders and corresponding complications. Therefore, the detection of UA and UOx in the body fluids is significant for clinical diagnosis. 4-Cyano-4'-pentylbiphenyl (5CB, a nematic liquid crystal (LC)) was doped with octadecyl trimethylammonium bromide (OTAB, a cationic surfactant), which formed a self-assembled monolayer at the aqueous/5CB interface. The UOx-catalyzed oxidation of UA yielded H2O2, releasing the single-strand deoxyribonucleic acid (ssDNA) from the nanoceria/ssDNA complex. The interaction of the released ssDNA with OTAB disrupted the monolayer at the aqueous/5CB interface, which resulted in a dark to bright change when observed through a polarized optical microscope. The LC-based sensor allowed the detection of UA with a linear range of 0.01-10 µM and a limit of detection (LOD) of 0.001 µM. The UA detection was also performed in human urine samples and the results were comparable to that of a standard commercial colorimetric method. Similarly, the detection of UOx was performed, with a noted linear range of 20-140 µg/mL. The LOD was as low as 0.34 µg/mL. The detection of UOx was also demonstrated in human serum samples with excellent performance. This method provides a robust sensing platform for the detection of UA and UOx and has potential for applications in clinical analysis.


Subject(s)
Biosensing Techniques , Body Fluids , Liquid Crystals , Body Fluids/chemistry , DNA, Single-Stranded , Humans , Hydrogen Peroxide/chemistry , Urate Oxidase/chemistry , Urate Oxidase/metabolism , Uric Acid , Water
7.
ACS Sens ; 7(2): 593-600, 2022 02 25.
Article in English | MEDLINE | ID: mdl-35050602

ABSTRACT

α-Amylase (AMS) in human serum is a critical biomarker for the early diagnosis of pancreatic damage. In addition, the inhibition of α-amylase has long been thought to decrease the occurrence of diabetes. Thus, it is critical to construct a facile and convenient method for the determination of AMS and its inhibitor. In this study, we demonstrate a novel amylase sensor based on translating the viscosity change of the aqueous solution into the difference of the water diffusion length on a pH paper strip. AMS can be quantitatively detected by measuring the viscosity change of the amylopectin solution in the presence of AMS with different concentrations. The paper-based AMS sensor has a very high sensitivity with a detection limit of 0.017 U/mL and also shows excellent specificity. In addition, the inhibitory effect of acarbose on AMS is demonstrated with the IC50 value determined to be 21.66 ± 1.13 µg/mL. Furthermore, it is also evaluated for the detection of AMS in human serum samples of healthy people and acute pancreatitis patients. The difference in amylase levels between the two groups is unambiguously distinguished. Overall, this study provides a very simple, cost-effective, equipment-free, high-throughput, and label-free method for rapid and quantitative detection of α-amylase and may have significant applications in the diagnosis of acute pancreatitis and the screening of AMS inhibitors.


Subject(s)
Pancreatitis , alpha-Amylases , Acute Disease , Amylases , Humans , Pancreatitis/diagnosis , Viscosity
8.
Anal Chim Acta ; 1150: 338247, 2021 Mar 15.
Article in English | MEDLINE | ID: mdl-33583545

ABSTRACT

Bleomycin (BLM) is a broadly used antibiotic to treat different types of cancer. It can be hydrolyzed by bleomycin hydrolase (BLMH), which eventually influences the anti-tumor efficacy of BLM. Therefore, it is particularly important to detect BLM and BLMH. Herein, we demonstrated highly sensitive detection of BLM and BLMH by a simple and convenient liquid crystal (LC)-based sensing platform for the first time. 5CB (a nematic LC) doped with the cationic surfactant OTAB was working as the sensing platform. When the OTAB-laden 5CB interface was in contact with an aqueous solution of ssDNA, LCs displayed a bright image due to disruption of the arrangement of OTAB monolayers by ssDNA, indicating the planar orientation of LCs at the aqueous/LC interface. When BLM·Fe(II) and ssDNA were both present in the aqueous solution, ssDNA underwent irreversible cleavage, which prevented disruption of the arrangement of OTAB monolayers. Accordingly, LCs showed a dark image, suggesting the homeotropic orientation of LCs at the aqueous/LC interface. However, when BLM·Fe(II) was enzymatically hydrolyzed by BLMH, LCs remained the bright image. This approach showed high sensitivity for the detection of BLM and BLMH with the limits of detection of 0.2 nM and 0.3 ng/mL, respectively. Besides, the detection of BLM and BLMH was successfully achieved in human serum. This method has the advantages of high sensitivity, robust stability, simple operation, low cost, and easy detection through naked eyes, which makes it a potential candidate for applications in clinical analysis.


Subject(s)
Liquid Crystals , Bleomycin , DNA, Single-Stranded , Humans , Hydrolases , Surface-Active Agents
9.
Anal Chem ; 92(11): 7900-7906, 2020 06 02.
Article in English | MEDLINE | ID: mdl-32366092

ABSTRACT

The pathological studies of cancer tissues and cell molecules could provide an early diagnosis for the treatment of cancer. In this work, we have designed a microfluidic surface extractor (MSE). The MSE has been coupled with electrospray mass spectrometry (extraction reagent, methanol; optimum flow rate, 0.5 mL/h) to analyze the phospholipid content of different tumor cells. Three types of cancer cell lines, including A549 cells, HepG2 cells, and U87 cells, were investigated, and the principle component analysis (PCA: linear discriminant analysis (LDA), PC1 97.2%; PC2, 2.8%) was carried out to analyze the difference in the lipid contents. The LDA revealed heterogeneity among the cancer cells. The designed MSE could have potential applications in the clinical analysis of cancer tissues.


Subject(s)
Lab-On-A-Chip Devices , Neoplasms/pathology , Phospholipids/analysis , Cell Adhesion , Humans , Spectrometry, Mass, Electrospray Ionization , Tumor Cells, Cultured
10.
Angew Chem Int Ed Engl ; 59(24): 9282-9287, 2020 06 08.
Article in English | MEDLINE | ID: mdl-32281727

ABSTRACT

Live-imaging of signaling molecules released from living cells is a fundamental challenge in life sciences. Herein, we synthesized liquid crystal elastomer microspheres functionalized with horse-radish peroxidase (LCEM-HRP), which can be immobilized directly on the cell membrane to monitor real-time release of H2 O2 at the single-cell level. LCEM-HRP could report H2 O2 through a concentric-to-radial (C-R) transfiguration, which is due to the deprotonation of LCEM-HRP and the break of inter or intra-chain hydrogen bonding in LCEM-HRP caused by HRP-catalyzed reduction of H2 O2 . The level of transfiguration of LCEM-HRP revealed the different amounts of H2 O2 released from cells. The estimated detection sensitivity was ≈2.2×10-7  µm for 10 min of detection time. The cell lines and cell-cell heterogeneity was explored from different configurations. LCEM-HRP presents a new approach for in situ real-time imaging of H2 O2 release from living cells and can be the basis for seeking more advanced chemical probes for imaging of various signaling molecules in the cellular microenvironment.


Subject(s)
Elastomers/chemistry , Hydrogen Peroxide/metabolism , Liquid Crystals/chemistry , Microspheres , Single-Cell Analysis , Cell Line, Tumor , Enzymes, Immobilized/chemistry , Enzymes, Immobilized/metabolism , Horseradish Peroxidase/chemistry , Horseradish Peroxidase/metabolism , Humans , Surface Properties
11.
Adv Sci (Weinh) ; 6(20): 1900778, 2019 Oct 16.
Article in English | MEDLINE | ID: mdl-31637159

ABSTRACT

Tumor cells exhibit prominent metabolic alterations through which they acclimatize to their stressful microenvironment. These cells have a high rate of glutaminolysis and release ammonia (NH3) as a byproduct, which may function as a diffusible signal among cancer cells and can reveal cellular heterogeneity. E7, a nematic liquid crystal (LC), is doped with 4-pentyl-4'-biphenyl carboxylic acid (PBA) and encapsulated in polymeric microcapsules (P-E7PBA), which are then immobilized on cells in a microfluidic channel. Normal human umbilical vein endothelial cells (HUVECs) and myeloma, human primary glioblastoma (U87), human colon carcinoma (Caco-2), and human breast adenocarcinoma (MCF-7) cells are investigated for the release of NH3. The P-E7PBA is able to visualize NH3 release from the cell via a radial-to-bipolar (R-B) orientation change, observed through a polarized optical microscope. The various cell lines significantly differ in their response time required for an R-B change. The mean response times for Caco-2, U87, and MCF-7 cells are 277, 155, and 121 s, respectively. NH3 release from a single cell captured in a microwell flow chip shows a similar R-B change. The P-E7PBA droplets technology could be applied to other multiple targets by functionalizing LCs with different probes.

12.
ACS Omega ; 4(12): 15004-15011, 2019 Sep 17.
Article in English | MEDLINE | ID: mdl-31552342

ABSTRACT

The hitherto unknown influence of 1,10-phenonthroline (1,10-phen) and its derivatives on the weak chemiluminescence (CL) of periodate-peroxide has been investigated, and a novel method for CL catalysis is described. Herein, we have deconvoluted the variation in CL intensity arising from the addition of various derivatives of 1,10-phen. Interestingly, similar derivatives of 1,10-phen show interesting differences in their reactivity toward CL. Electron-withdrawing substituents on 1,10-phen boosted the CL signals, indicating a negative charge buildup on 1,10-phen in the rate-determining step. The 1,10-phen derivatives having substitution at the C5=C6 position resulted in no CL signals due to the blockage of the reactive site. Mechanistic investigations are interpreted in terms of free radical (H2O2 reaction), followed by the oxygen atom transfer via an electrophilic attack of IO4 - (IO4 - reaction) on 1,10-phen resulting in dioxetane with enhanced CL emission. Additionally, the relationship between electronic structures and photophysical properties was investigated using density functional theory. Our results are expected to open up promising application of 1,10-phen as a molecular catalyst, providing a new strategy for metal-free catalytic CL enhancement reaction. We believe that this would foster in gleaning more detailed information on the nature of these reactions, thereby leading to a deeper understanding of the CL mechanism.

13.
Anal Chem ; 91(19): 12283-12289, 2019 10 01.
Article in English | MEDLINE | ID: mdl-31456388

ABSTRACT

Three-dimensional (3D) hydrogel microspheres have aroused increasing attention as an in vitro cell culture model. Yet the preservation of cells' original biological properties has been overlooked during model construction. Here we present an integrated microfluidic device to accomplish the overall process including cell-laden microsphere generation, online extraction, and dynamic-culture. The method extends the noninvasive and nonsuppression capabilities of the droplet preparation system and provides a constant microenvironment, which reduces intracellular oxidative stress damage and the accumulation of mitochondria. Compared to the conventional preparation method, the coculture model of tumor-endothelial construction on an integrated platform displays high-level angiogenic protein expression. We believe that this versatile and biocompatible platform will provide a more reliable analysis tool for tissue engineering and cancer therapy.


Subject(s)
Cell Culture Techniques/instrumentation , Lab-On-A-Chip Devices , Calcium/chemistry , Cell Culture Techniques/methods , Cell Line, Tumor , Cell Survival , Coculture Techniques/instrumentation , Coculture Techniques/methods , Edetic Acid/chemistry , Equipment Design , Glutamine/metabolism , Human Umbilical Vein Endothelial Cells , Humans , Hydrogels , Lactic Acid/metabolism , Microspheres , Vascular Endothelial Growth Factor A/metabolism
14.
ACS Sens ; 4(6): 1710-1715, 2019 06 28.
Article in English | MEDLINE | ID: mdl-31094503

ABSTRACT

Biochemical and physical factors affect the rolling of tumor cells across the blood vessel. The biochemical factors have been well studied, while the influence of physical factors such as fluid shear stress (FSS) remains poorly understood. Here, human glioma cells (U87 cells) in a straight microfluidic channel were exposed to FSS (0.12, 1.2, and 1.8 dyn/cm2); and their locomotion behaviors from crawling-to-rolling and changes in cellular morphology (concave, elongated, less elongated, and round) were observed. The adhesion strength and stiffness of the cells of different morphologies were analyzed using a live single-cell extractor and atomic force microscopy, respectively. In general, the FSS stimulated cells showed stronger adhesion strength than the cells not exposed to FSS. The cell not exposed to FSS always exhibited greater nuclear stiffness than cortex stiffness, while after FSS treatment the cortex hardened and nucleus softened, where the round-shaped cell had a cortex that was more rigid than its nucleus. These results indicated that FSS influenced the biomechanics of circulating tumor cells, and elucidation of the mechanical responses to FSS might provide a deeper insight for cancer metastasis.


Subject(s)
Cell Adhesion , Neoplastic Cells, Circulating/metabolism , Cell Line, Tumor , Equipment Design , Humans , Lab-On-A-Chip Devices , Microfluidics/instrumentation , Microfluidics/methods , Neoplastic Cells, Circulating/pathology , Shear Strength
15.
Chem Sci ; 10(7): 2081-2087, 2019 Feb 21.
Article in English | MEDLINE | ID: mdl-30881632

ABSTRACT

Single cells are increasingly recognized to be capable of wound repair that is important for our mechanistic understanding of cell biology. The lack of flexible, facile, and friendly subcellular treatment methods has hindered single-cell wound repair studies and organelle transport analyses. Here we report a laminar flow based approach, we call it fluid cell knife (Fluid CK), that is capable of precisely cutting off or treating a portion of a single cell from its remaining portion in its original adherent state. Local operations on portions of a living single cell in its adherent culture state were applied to various types of cells. Temporal wound repair was successfully observed. Moreover, we successfully stained portions of a living single cell to measure the organelle transport speed (mitochondria as a model) inside a cell. This technique opens up new avenues for cellular wound repair and subcellular behavior analyses.

16.
Chem Sci ; 10(2): 497-500, 2019 Jan 14.
Article in English | MEDLINE | ID: mdl-30713646

ABSTRACT

Reactive oxygen species (ROS) generation is of intense interest because of its crucial role in many fields. Here we demonstrate that MoS2-QDs exhibit a promising capability for the generation of reactive oxygen species, which leads to enhanced chemiluminescence. We discovered that the unique performance is due to hydroxyl radical activation increasing the active catalytic sites on molybdenum sulphide quantum dots (MoS2-QDs). The reactive oxygen species, such as hydroxyl radicals (˙OH), superoxide radicals (˙O2 -) and singlet oxygen (1O2) have been efficiently generated from H2O2 solution in alkaline conditions. In particular, the maximum ˙OH yield was enhanced significantly (9.18 times) compared to the Fe(ii)/H2O2 Fenton system under neutral conditions. These findings not only enrich our understanding of the fascinating performance of MoS2 QDs, but also provide a new pathway for ROS generation in all kinds of pH environment.

17.
Chem Commun (Camb) ; 55(15): 2166-2169, 2019 Feb 14.
Article in English | MEDLINE | ID: mdl-30698595

ABSTRACT

The non-uniform deposition of matrix-analyte cocrystals and poor ionization are major obstacles in quantitative analysis through MALDI-MS. An Au-nanobowl array was prepared and applied to overcome these limitations, which enabled the quantitative detection of oligonucleotides and polypeptides.

18.
Talanta ; 196: 370-375, 2019 May 01.
Article in English | MEDLINE | ID: mdl-30683378

ABSTRACT

The nitrogen doped carbon dots (N-CDs) produces strong chemiluminescence (CL)-emission due to hydroxyl radical (•OH) induced electron-hole transition in N-CDs. The Fe2+ has the ability to generate •OH from available hydrogen peroxide (H2O2). Therefore, a pre-mixed N-CDs/H2O2 solution was utilized for selective quantification of Fe2+ in solution via CL-emission. A linear increase in the CL-emission intensity was observed within increase in Fe2+ concentration. The N-CDs/H2O2 system enabled the detection of Fe2+ up to lower concentration of 0.2 × 10-9 M with a linear dynamic range of 1.0 × 10-9-1.0 × 10-6 M. Significantly, no CL-emission was observed when other divalent cations, Al3+, Fe3+, or Cr3+ were injected to this system. Moreover, no interference was observed when a mixed solution of Fe2+ and other cations were introduced to N-CDs/H2O2. The practical evaluation of N-CDs/H2O2 system was demonstrated for detection of Fe2+ in tap, lotus pond, and canal water samples. The easy detection, high sensitivity, and selectivity make this method a significant tool for analysis of Fe2+ in solution.

19.
Chem Sci ; 11(1): 253-256, 2019 Nov 11.
Article in English | MEDLINE | ID: mdl-34040719

ABSTRACT

This work describes a microfluidic system for in situ extraction of a single-cell and its phosphatidylcholine analysis through mass spectrometry. This approach uncovered cellular heterogeneity among seemingly identical cells and provided a new platform for identification and classification of cells.

20.
Analyst ; 144(3): 808-823, 2019 Jan 28.
Article in English | MEDLINE | ID: mdl-30177979

ABSTRACT

Advances in microfluidic techniques have prompted researchers to study the inherent heterogeneity of single cells in cell populations. This would be helpful in the identification of major diseases and the design of personalized medicine. Different microfluidic approaches provide a variety of functions in the process of single-cell analysis. In this review, we take a broad overview of various microfluidic-based approaches for single-cell isolation, single-cell lysis, and single-cell analysis. Up-to-date flagship techniques and the pros and cons of these methods are discussed in detail.


Subject(s)
Microfluidics/methods , Single-Cell Analysis/methods , Humans , Precision Medicine
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