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Graphene-Based Steganographicly Aptasensing System for Information Computing, Encryption and Hiding, Fluorescent Sensing and In Vivo Imaging of Fish Pathogens
摘要： Inspired by information processing and communication of life based on complex molecular interactions, some artificial (bio)chemical systems have been developed for applications in molecular information processing or chemo/biosensing and imaging. However, little attention has been paid to simultaneously and comprehensively utilize the information computing, encoding and molecular recognition capabilities of molecular-level systems (such as DNA-based systems) for multifunctional applications. Herein, a graphene-based steganographicly aptasensing system was constructed for multifunctional application, which relies on specific molecular recognition and information encoding abilities of DNA aptamers (Aeromonas hydrophila and Edwardsiella tarda-binding aptamers as models) and the selective adsorption and fluorescence quenching capacities of graphene oxide (GO). Although graphene-DNA systems have been widely used in biosensors and diagnostics, our proposed graphene-based aptasensing system can not only be utilized for fluorescent sensing and in vivo imaging of fish pathogens (Aeromonas hydrophila and Edwardsiella tarda), but can also function as a molecular-level logic computing system where the combination of matters (specific molecules or materials) as inputs produces the resulting product (matter level) or fluorescence (energy level) changes as two outputs. More importantly and interestingly, our graphene-based steganographicly aptasensing system can also be served as a generally doubly cryptographic and steganographic system for sending different secret messages by using pathogen-binding DNA aptamers as information carriers, GO as a cover, a pair of keys: target pathogen as a public key, the encryption key used to encode or decode a message in DNA as a private key. Our study not only provides a novel nano-biosensing assay for rapid and effective sensing and in vivo imaging fish pathogens, but also demonstrates a prototype of (bio)molecular steganography as an important and interesting extension direction of molecular information technology, which is helpful in probably promoting the development of multifunctional molecular-level devices or machines.
关键词： aptasensing,steganography,graphene oxide,DNA aptamer,encryption,fish pathogens,in vivo imaging,information hiding
更新于2025-11-21 11:24:58
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Graphene oxide-quenching-based fluorescence in situ hybridization (G-FISH) to detect RNA in tissue: Simple and fast tissue RNA diagnostics
摘要： FISH-based RNA detection in paraffin-embedded tissue can be challenging, with complicated procedures producing uncertain results and poor image quality. Here, we developed a robust RNA detection method based on graphene oxide (GO) quenching and recovery of fluorescence in situ hybridization (G-FISH) in formalin-fixed paraffin-embedded (FFPE) tissues. Using a fluorophore-labeled peptide nucleic acid (PNA) attached to GO, the endogenous long noncoding RNA BC1, the constitutive protein β-actin mRNA, and miR-124a and miR-21 could be detected in the cytoplasm of a normal mouse brain, primary cultured hippocampal neurons, an Alzheimer’s disease model mouse brain, and glioblastoma multiforme tumor tissues, respectively. Coding and non-coding RNAs, either long or short, could be detected in deparaffinized FFPE or frozen tissues, as well as in clear lipid-exchanged anatomically rigid imaging/immunostaining-compatible tissue hydrogel (CLARITY)-transparent brain tissues. The fluorescence recovered by G-FISH correlated highly with the amount of miR-21, as measured by quantitative real time RT-PCR. We propose G-FISH as a simple, fast, inexpensive, and sensitive method for RNA detection, with a very low background, which could be applied to a variety of research or diagnostic purposes.
关键词： glioblastoma multiforme tumor,tissue RNA diagnostics,Graphene oxide-quenching-based fluorescence in situ hybridization (G-FISH),Alzheimer’s disease,formalin-fixed paraffin-embedded (FFPE) tissue
更新于2025-09-23 15:23:52
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Preparation of fluorescent in situ hybridisation probes without the need for optimisation of fragmentation
摘要： DNA-fluorescence in situ hybridisation (DNA-FISH) allows visualisation of chromosome organisation and fluorescently labelled DNA fragments that are often produced from rearrangement. FISH probes are pools of short template plasmids that contain large genomic inserts. For effective sample penetration and target hybridisation it is critical that probe fragments are between 200 and 500bp. Production of these short probes requires significant optimisation and can be confounded access to expensive sonication equipment or inherent sequence features that influence enzymatic fragmentation or amplification. Here we demonstrate that effective FISH probes can be prepared without the need for optimisation of fragmentation using a cocktail of two the 4bp recognition sequence restriction enzymes CviQI and AluI.
关键词： Cancer,Fluorescence in situ hybridization,Translocation,FISH,Probe
更新于2025-09-23 15:23:52
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Peptide Nucleic Acid–Fluorescence In Situ Hybridization for Detection of Staphylococci From Endophthalmitis Isolates: A Proof-of-Concept Study
摘要： PURPOSE. Rapid identi?cation of pathogens causing endophthalmitis may improve treatment outcomes through early administration of species-speci?c medication. The current study reports a new molecular application of peptide nucleic acid–?uorescence in situ hybridization (PNA-FISH) with Staphylococcus-speci?c molecular PNA probes for the potential rapid detection of common pathogens causing endophthalmitis. METHODS. An experimental study was designed to evaluate the proof of concept at the microbiology laboratory of the Bascom Palmer Eye Institute. Stored culture-positive staphylococci endophthalmitis isolates obtained from prior vitreous samples (n ? 15), along with broth as negative controls (n ? 5) were used. Inoculum was prepared to a ?nal concentration of 1 3 105 colony-forming units/mL to ensure that the isolates were viable. Smears of samples were ?xed and hybridized using QuickFISH protocol with probes for Staphylococcus. RESULTS. With PNA-FISH technique, Staphylococcus aureus was identi?ed in 9 of 10 samples and coagulase-negative staphylococci were identi?ed in 10 of 10 samples. Detection time was 20 minutes. CONCLUSIONS. This study serves a proof of concept using a new microbial detection system with FISH probes, and may have the potential for clinical use in the rapid and accurate identi?cation of isolates from patients with endophthalmitis.
关键词： peptide nucleic acid (PNA),endophthalmitis,rapid identification,fluorescence in situ hybridization (FISH)
更新于2025-09-23 15:22:29
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Fish oil preparation inhibits leukocyte recruitment and bands that characterize inflamed tissue in a model of phenol-induced skin inflammation: percutaneous penetration of a topically applied preparation demonstrated by photoacoustic spectroscopy
摘要： Fish oil (FO) is a natural source of omega-3 fatty acids, with well-established beneficial effects in inflammatory diseases when FO is orally administered. This study investigated the effects of a topically applied FO preparation (FOP) on phenol-induced ear edema and evaluated the percutaneous penetration of FOP in ear tissue. After applying phenol, groups of mice received FOP on the ear. After 1 h, ear tissue was collected to determine the percent inhibition of edema, myeloperoxidase activity, and to perform photoacoustic spectroscopy (PAS). Treatment with FOP did not reduce edema, but reduced myeloperoxidase activity. The FOP decreased the area of bands that characterize inflamed tissue and penetrated into the tissue. These results indicated an inhibitory effect of FOP on leukocyte recruitment in phenol-induced ear edema. These data support the applicability of PAS as a non-destructive method for evaluating the inflammatory response, percutaneous penetration and antiinflammatory activity of compounds.
关键词： ear edema,Fish oil,inflammation,phenol,skin,photoacoustic spectroscopy
更新于2025-09-23 15:22:29
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Detection of Alkali-Labile Sites on Satellite DNA by DNA Breakage Coupled with Fluorescence in Situ Hybridization (DNA-FISH) Monitor DNA Damage in Cervical Epithelial Cells
摘要： Satellite DNA is the main component of functional centromeres and forms the main structural constituent of heterochromatin. The presence of alkali-labile sites (ALSs) is a feature inherent to chromatin structure. Here we aimed to characterize ALSs in different satellite DNA loci in cervical epithelial cells using DNA breakage detection coupled with fluorescence in situ hybridization (DBD–FISH). Cervical epithelial cells embedded in an agarose matrix were deproteinized and exposed to alkaline denaturation, which generated single-stranded DNA (ssDNA) starting from the ends of spontaneous basal DNA breaks and ALSs. The amount of ssDNA produced within a specific sequence area could be detected by DBD–FISH using specific probes. The DBD–FISH signals, which were corrected for respective FISH signals during metaphase, were remarkably stronger in the 5 bp classical satellite DNA domains analyzed (D1Z1, D9Z3, and D16Z3) compared with alphoid satellite regions (D3Z1, D8Z2, and DXZ1). D1Z1 locus of chromome-1 being the most affected by alkali denaturation, and contrary, D3Z1 locus of chromosome–3 was the least sensitive to alkali treatment. These findings suggest a high density of constitutive ALSs—probably abasic sites—within the 5 bp satellite DNA sequences in cervical epithelial cells. The presence and relative abundance of ALSs might help explain the high frequency of spontaneous breakage and rearrangements in the pericentromeric heterochromatin of chromosomes 1, 9, and 16 when this chromatin region is undercondensed spontaneously or via induction, such as following viral infections. ALSs in these sequences could be useful tools to monitor DNA damage in cases of cervical carcinogenesis. ALSs on these sequences could be useful tools to monitor DNA damage in cases of cervical carcinogenesis.
关键词： DBD–FISH,cervical epithelium,Alkali-labile sites,DNA breakage detection,ALS
更新于2025-09-23 15:21:01
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Determination of citalopram in fish brain tissue: benefits of coupling laser diode thermal desorption with low- and high-resolution mass spectrometry
摘要： Recent state-of-the-art methods developed for the analysis of polar xenobiotics from different types of biological matrices usually employ liquid chromatography with mass spectrometry. However, there are limitations when a small amount of sample mass is available. For example, individual benthic invertebrates or fish tissue samples often weigh less than 100 mg (e.g., brain, liver) but are necessary to understand environmental fate and bioaccumulation dynamics. We developed ultra-fast methods based on a direct sample introduction technique. This included coupling laser diode thermal desorption with atmospheric pressure chemical ionization mass spectrometry (LDTD-APCI-MS). We then quantitated a common selective serotonin reuptake inhibitor (citalopram) in brain tissues of individual juvenile fish after in vivo exposure to environmentally relevant concentration. Two mass spectrometric methods based on low (LDTD-APCI-triple quadrupole (QqQ)-MS/MS) and high (LDTD-APCI-high-resolution product scan (HRPS)) resolutions were developed and evaluated. Individual instrument conditions were optimized to achieve an accurate and robust analytical method with minimum sample preparation requirements. We achieved very good recovery (97–108%) across the range of 1–100 ng g?1 for LDTD-APCI-HRPS. LDTD-APCI-QqQ-MS/MS showed poorer performance due to interferences from the matrix at the lowest concentration level. LDTD-APCI ionization was successfully validated for analysis of non-filtered sample extracts. Evaluation of final methods was performed for a set of real fish brain samples, including comparison of LDTD-APCI-HRPS with a previously validated LC-heated electrospray ionization-HRPS method. This new LDTD-APCI-HRPS method avoids the chromatographic step and provides important benefits such as analysis of limited sample masses, lower total sample volume (typically μL), and reduction in analysis time per sample run to a few seconds.
关键词： Ambient ionization,Green chemistry,Laser diode thermal desorption,Psychoactive pharmaceutical,Juvenile fish
更新于2025-09-23 15:21:01
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The prognostic impact of loss of chromosome 7 material detected by fluorescence in situ hybridization (FISH) in myeloid malignancies
摘要： Background: Monosomy 7 ((cid:1)7) or deletion in its long arm [del(7q)] is among the most common chromosomal abnormalities in myeloid malignancies. There are prognostic variations between (cid:1)7 and del(7q) in acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS). Aim: To describe the clinical characteristics, response to treatment, and survival of patients with primary AML and MDS having (cid:1)7 or del(7q) detected by fluorescence in situ hybridization (FISH). Patients and methods: The study was conducted on 53 patients with primary AML and MDS. They were tested for chromosome 7 abnormality using FISH technique. Results: Thirty-one patients had chromosome 7 abnormality and 22 did not. Lower complete remission and higher death rates were observed in patients with (cid:1)7 (47.6% and 62%, respectively) when compared to patients with del(7q) (70% and 40%, respectively) with no significant difference (p = 0.218 and 0.101, respectively). The median overall survival (OS) of patients with (cid:1)7, del(7q) and normal chromosome 7 were 32.0, 43.0 and 50.0 months, respectively, with significant statistical difference (p = 0.001). This difference was evident between patients with (cid:1)7 and those with normal chromosome 7 (p = 0.001), and less evident between patients with (cid:1)7 and those with del(7q) (p = 0.021). Conclusion: Chromosome 7 analysis has clear impact on the outcome of myeloid malignancies. The prognostic variations between (cid:1)7 and del(7q) is attributed to multiple factors. Cases with del(7q) have better outcome than cases with (cid:1)7. FISH provides a powerful tool for detecting and monitoring patients with chromosome 7 abnormalities.
关键词： del(7q),Myeloid malignancies,Chromosome 7,Monosomy 7,FISH
更新于2025-09-23 15:21:01
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Infrared Imagery and Inert Media Used in Treating Upwelling Groundwater with Rotenone
摘要： Untreated upwelling groundwater from seeps and springs in and adjacent to surface water bodies has been long suspected of causing failed rotenone treatments by providing a refugia of nontoxic water. A possible solution involves the use of an inert media to carry the liquid rotenone to the source of upwelling groundwater and release rotenone over an extended period of time suf?cient to affect the mortality of the target ?sh. In our initial study to address this problem, we used thermal infrared imagery (FLIR One) on a smartphone to locate groundwater that was subsequently treated with mixtures of the liquid rotenone formulation CFT Legumine (3.3% rotenone) utilizing two commercially available inert carriers: (1) CatSan Hygiene Litter (mixture of quartz sand and calcite) and (2) Vectocarb (?ne powder of modi?ed CaCO3). Trials on the mixtures were conducted in 2015 in upwelling groundwater areas of the Skibotn River drainage, Troms County, Norway, the site of previously failed eradication efforts. Following application, mean concentrations of 75.6 to 131 μg/L rotenone were present at 0.5 h in the pools and the brooks downstream of the upwelling groundwater that decreased and stabilized to 11.5 to 16.8 μg/L rotenone at 3 h. Both carriers have large surface areas (porosity) that transport (through sorption) the rotenone liquid to the source of upwelling groundwater and release (through desorption) concentrations of rotenone over at least 3 h. Both mixtures show promise in treating upwelling groundwater to eradicate ?sh from those areas and were used successfully in the 2016 retreatment of Skibotn River for the eradication of Atlantic Salmon Salmo salar infested with the ectoparasite Gyrodactylus salaris.
关键词： fish eradication,infrared imagery,groundwater,inert media,rotenone
更新于2025-09-23 15:21:01
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Construction of novel xanthine biosensor by using Zinc Oxide (ZnO) by biotemplate method for fish freshness detection
摘要： In the food industry, fish is a product with a short shelf life, and xanthine attracts much attention as an indicator of fish freshness. In this study, we demonstrated a simple assay system containing Zinc Oxide (ZnO) and xanthine oxidase (XOD) for specific and quantitative detection of xanthine. ZnO nanomaterial have been successfully synthesized using biotemplated method. A mechanism is based on the fluorescence quenching of ZnO via electron transfer mechanism, which is caused by the hydrogen peroxide (H2O2) that is produced from the XOD-catalyzed oxidation of xanthine. By virtue of the specific response, the present assay allowed for the selective determination of xanthine in the range of 3.30×10-10-6.67×10-7 mol/L and 2.67×10-6-2.67×10-4 mol/L with a detection limit of 1.30×10-10 mol/L. The proposed method was successfully applied to the determination of xanthine in real fish samples with satisfactory results.
关键词： Biotemplated method,Xanthine oxidase,Fish Biosensor,ZnO,Xanthine
更新于2025-09-19 17:15:36