Multiple Biomolecules Research Articles

Quantitative and Noninvasive Detection of SAH-Related MiRNA in Cerebrospinal Fluids In Vivo Using SERS Sensors Based on Acupuncture-Based Technology.

Quantitative analysis of microRNAs (miRNAs) in a noninvasive manner is of vital importance for disease diagnosis and prognosis evaluation. However, conventional strategies for realizing accurate, simple, and sensitive detection of target molecules are still a challenge, especially for miRNAs due to their low abundance and susceptibility in the complex biological environment. Here, a novel surface-enhanced Raman scattering (SERS) strategy was established for quantitative detection and monitoring of miRNA-21-5p (miR-21-5p) in living cells and in vivo cerebrospinal fluid (CSF) by applying hairpin DNA (hpDNA)-conjugated gold nanostars (GNSs) SERS probes combined with acupuncture-based technology. This strategy enabled ultrasensitive exploration toward miR-21-5p in a wide range from 1 fM to 100 pM in cell lysates. Moreover, SERS analysis facilitated the detection and long-term monitoring for in vivo miR-21-5p noninvasively. This developed strategy promises to offer a powerful method for the analysis of multiple biomolecules in single cells and living bodies.

Photochemical approach for multiplexed biofunctionalisation of gallium arsenide

The optoelectronic properties of gallium arsenide (GaAs) hold great promise in biosensing applications, currently being held back by the lack of methodologies reporting the spatially selective functionalisation of this material with multiple biomolecules. Here, we exploit the use of a photoreactive crosslinker - a diazirine derivative - for spatially selective covalent immobilisation of multiple bioreceptors on the GaAs surface. As a proof of principle we show the immobilisation of two proteins: neutravidin and endosulfine alpha protein. X-ray photoelectron spectroscopy results showed the presence of the biomolecules on the GaAs regions selectively exposed to ultraviolet light. The approach presented here is applicable to the covalent attachment of other biomolecules, paving the way for using GaAs as a platform for multiplexed biosensing.

Healing potential of neomycin-loaded electrospun nanofibers against burn wounds

BackgroundWound healing is a serious therapeutic and economic challenge in regenerative medicines. It is a complex interplay of multiple biomolecules and pathways. ObjectiveThe goal of this work was to fabricate neomycin (Ne)-loaded electrospun nanofibers of carrageenan, polyacrylonitrile, and pullulan (CG/PAN/PU) for skin wound healing, specifically burn wounds. CG/PAN/PU/Ne nanofibers were characterized structurally using scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and thermogravimetric analysis (TGA). As a wound dressing for a full-thickness skin lesion, the effect of CG/PAN/PU/Ne nanofibers on the wound-healing process was examined in the rabbits. ResultsNanofiber's surface was rough. TGA results showed that drug-loaded electrospun nanofibers were more stable as compared to drug-free nanofibers. The whole amount of drug release was achieved in 120–150 min. The visual inspection showed that the rabbit wounds treated with drug-loaded electrospun nanofibers were healed within 14 days and that drug-free nanofibers within 17 days. The histopathological results showed a greater healing potential of CG/PAN/PU/Ne nanofibers as compared to the positive control (Quench®) and negative control (Surgical gauze). The results of antibacterial activities of CG/PAN/PU fabricated nanofibers against Gram-negative Escherichia coli (E. coli) and Gram-positive Enterococcus faecalis revealed that the CG/PAN/PU/Ne nanofibers had more potential against Gram-negative bacteria as compared to Gram-positive bacteria. ConclusionThe CG/PAN/PU nanofibers have a promising potential for wound healing with and without antibiotic.

Simultaneous Multiplexed Imaging of Biomolecules in Transgenic Mouse Brain Tissues Using Mass Spectrometry Imaging: A Multi-omic Approach.

The importance of multi-omic-based approaches to better understand diverse pathological mechanisms including neurodegenerative diseases has emerged. Spatial information can be of great help in understanding how biomolecules interact pathologically and in elucidating target biomarkers for developing therapeutics. While various analytical methods have been attempted for imaging-based biomolecule analysis, a multi-omic approach to imaging remains challenging due to the different characteristics of biomolecules. Time-of-flight secondary ion mass spectrometry (ToF-SIMS) is a powerful tool due to its sensitivity, chemical specificity, and high spatial resolution in visualizing chemical information in cells and tissues. In this paper, we suggest a new strategy to simultaneously obtain the spatial information of various kinds of biomolecules that includes both labeled and label-free approaches using ToF-SIMS. The enzyme-assisted labeling strategy for the targets of interest enables the sensitive and specific imaging of large molecules such as peptides, proteins, and mRNA, a task that has been, to date, difficult for any MS analysis. Together with the strength of the analytical performance of ToF-SIMS in the label-free tissue imaging of small biomolecules, the proposed strategy allows one to simultaneously obtain integrated information of spatial distribution of metabolites, lipids, peptides, proteins, and mRNA at a high resolution in a single measurement. As part of the suggested strategy, we present a sample preparation method suitable for MS imaging. Because a comprehensive method to examine the spatial distribution of multiple biomolecules in tissues has remained elusive, our strategy can be a useful tool to support the understanding of the interactions of biomolecules in tissues as well as pathological mechanisms.

Molecular and Genetic Interactions between CCN2 and CCN3 behind Their Yin-Yang Collaboration.

Cellular communication network factor (CCN) 2 and 3 are the members of the CCN family that conduct the harmonized development of a variety of tissues and organs under interaction with multiple biomolecules in the microenvironment. Despite their striking structural similarities, these two members show contrastive molecular functions as well as temporospatial emergence in living tissues. Typically, CCN2 promotes cell growth, whereas CCN3 restrains it. Where CCN2 is produced, CCN3 disappears. Nevertheless, these two proteins collaborate together to execute their mission in a yin–yang fashion. The apparent functional counteractions of CCN2 and CCN3 can be ascribed to their direct molecular interaction and interference over the cofactors that are shared by the two. Recent studies have revealed the mutual negative regulation systems between CCN2 and CCN3. Moreover, the simultaneous and bidirectional regulatory system of CCN2 and CCN3 is also being clarified. It is of particular note that these regulations were found to be closely associated with glycolysis, a fundamental procedure of energy metabolism. Here, the molecular interplay and metabolic gene regulation that enable the yin–yang collaboration of CCN2 and CCN3 typically found in cartilage development/regeneration and fibrosis are described.

MXene-MoS2 carbon-fiber-based flexible electrochemical interface for multiple bioanalysis in biofluids

Flexible sensing technology has received widespread attention in the monitoring of human physiological indicators. Thus, it is of great significance to develop a universal flexible sensing interface for early diagnosis of multi-level biomarkers. Herein, we reported a multifunctional 3D carbon fiber paper (CFP)-MXene-MoS2 (CMM) sensing interface, which provided a flexible electrochemical biosensing platform for the ultrasensitive detection of multiple bioanalysis in biofluids. Notably, the CMM electrochemical interface was comprised of staggered carbon fibers on which the organ-like MXene-MoS2 heterostructure covered with MoS2 nanosheet arrays was uniformly allocated, permitting local enrichment of biomolecules for enhanced biomolecular capture efficiency, laying a solid foundation for the ultrasensitive detection of multiple biomolecules. In proof-of-concept experiments, we validated that the flexible CMM interface could be applied for ultrasensitive analyzing of quite a few biomolecules like ascorbic acid (AA), uric acid (UA), dopamine (DA) and microRNA (miRNA), with charming sensitivity and reproducibility. It was worth mentioned that an ultra-high sensitivity of 3.16 aM was acquired towards detecting microRNA. Under optimal parameters, the designed electrochemical interface worked well in complex biofluids such as human serum and urine samples. Overall, this flexible and disposable CMM interface with ultrahigh sensitivity provided insights for a more portable, accessible and valuable approach for monitoring biomolecules in clinical diagnosis.

Purification of the human cone‐rod homeobox protein

A “lock and key” is the most common depiction of protein binding, protein has a specific structure, and its binding molecule of interest can fit into it. What happens when a protein domain needs to bind to multiple biomolecules? The transcription factor CRX is one of those with an activation domain that can bind to various biomolecules. CRX regulates the gene expression of cone and rod photoreceptors. To effectively regulate gene expression, CRX’s activation domain interacts with multiple transcriptional co‐activators. Unlike the “lock and key” concept with defined structures, we propose that the activation domain of CRX is intrinsically disordered, allowing for interactions of multiple proteins in the same domain. In support of a disordered activation domain, we observe that proline and serine amino acids are abundant in the CRX activation domain. We also note an abundance of pathogenic frameshift mutations and a lack of point mutants in the activation domain. To characterize the structure of CRX, we attempted to purify full‐length CRX and CRX coexpressed with known transcriptional coactivator NRL. We hypothesize that this latter experiment will enhance the stability and expression of full‐length CRX. We have purified CRX using nickel chromatography and are currently optimizing the next steps to enhance the purity of CRX and NRL. The information from biochemical and structural experiments on purified CRX will provide insight into the transcriptional regulation of CRX, leading to the development of photoreceptor neurons.

Label-free DNAzyme for highly sensitive detection of multiple biomolecules in real samples through target-triggered catalytic cleavage reactions with auramine O's discriminated fluorescence emission.

A universal enzyme strand (E-DNA) recyclable L-histidine (L-His), melamine (MA), and cisplatin (CP) biosensor was fabricated on the basis of a target-specific RNA-cleaving DNAzyme with specific auramine O (AuO) dye instead of thioflavin T. In this strategy, the substrate strand (S-DNA) of the RNA-cleaving site was constructed as an intramolecular stem-loop structure, and a GT-rich sequence was imprisoned in the double-stranded stem which inhibits the formation of stable G-quadruplex (G4cpx) with AuO. The presence of L-His initiates a catalytic reaction for cleaving the RNA site of the S-DNA hydrolytically releasing the GT-rich portion, which subsequently combines with AuO and forms a G4cpx for enhanced fluorescent signal. The subsequent addition of MA uncoils the G4cpx to form T-MA-T dsDNA, or addition of CP unwound the G4cpx to form CP-DNA leading to an intensive decrease of AuO emission. Remarkably, the liberated L-His can ultimately cause several rounds of cleavage, and the liberated E-DNA can catalyze the subsequent reaction with the other S-DNA. The use of L-His and E-DNA repeatedly induces S-DNA cleavage and intensifies the emission signal. The results show that the proposed biosensor is extremely sensitive to L-His, MA, and CP with a detection limit of 0.98, 10, and 3.4nM respectively. To the best of our knowledge, the utilization of AuO as the G4cpx inducer and stabilizer for L-His, MA, and CP detection in real milk and urine samples has never been reported.

Survival analysis of pathway activity as a prognostic determinant in breast cancer.

High throughput biology enables the measurements of relative concentrations of thousands of biomolecules from e.g. tissue samples. The process leaves the investigator with the problem of how to best interpret the potentially large number of differences between samples. Many activities in a cell depend on ordered reactions involving multiple biomolecules, often referred to as pathways. It hence makes sense to study differences between samples in terms of altered pathway activity, using so-called pathway analysis. Traditional pathway analysis gives significance to differences in the pathway components' concentrations between sample groups, however, less frequently used methods for estimating individual samples' pathway activities have been suggested. Here we demonstrate that such a method can be used for pathway-based survival analysis. Specifically, we investigate the pathway activities' association with patients' survival time based on the transcription profiles of the METABRIC dataset. Our implementation shows that pathway activities are better prognostic markers for survival time in METABRIC than the individual transcripts. We also demonstrate that we can regress out the effect of individual pathways on other pathways, which allows us to estimate the other pathways' residual pathway activity on survival. Furthermore, we illustrate how one can visualize the often interdependent measures over hierarchical pathway databases using sunburst plots.

Dual-color imaging of DNA and RNA simultaneously with an aggregation/monomer-based deep-red fluorescent probe

Since many of the pivotal questions confronting chemical biology concern the interaction of more than one analyte, it is essential to investigate multiple analytes simultaneously. While commercial fluorescent dyes capable of detecting and quantitating DNA or RNA are widely used in biological research, few probes can be applied in DNA and RNA together. Distraction from each other is also a puzzling question for individual detection. Here, we have discovered a dual-responsive probe, namely, DR1, which can selectively recognize DNA by emitting out red fluorescence in the nucleus, while an entirely different stain pattern by labeling cytoplasm and nuclei with distinct cyanine fluorescence is observed owing to binding with RNA. Mechanism study found that DR1 binds to DNA in a monomer state with red fluorescence, and therefore the cyanine fluorescence may derive from its aggregated state while it binds to RNA. Then, DR1 was applied to quantify DNA and RNA contents and evaluate the effects of several inhibitors comprehensively on high-content screening platforms. Our study provided a new tool for simultaneously monitoring cellular DNA and RNA, and gaining new insights into developing dual-channel probes for multiple biomolecules.

“CORRELATION BETWEEN OXIDATIVE STRESS MARKERS AND DIABETIC CATARACT”

Reactive oxygen species causes oxidation that interferes with the physiological processes of the cell. Oxidative stress is dened as an “imbalance between reactive oxygen species production and breakdown by endogenous antioxidants. It causes hazardous events such as lipid peroxidation and oxidative DNAdamage. The reactive oxygen species (ROS), which consist principally of molecules like the superoxide anion (O2 −), hydrogen peroxide (H2O2), and hydroxyl radicals, are detoxied by enzymes such as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px). Different factors such as aging, drug actions and toxicity, inammation and chronic metabolic disorders like diabetes the production of ROS far exceed the capacity of antioxidants to neutralize. Due to oxidative stress, pancreatic β cell function may be affected, which, given the impaired expression of antioxidant enzymes, is outstandingly sensitive to reactive oxygen species. Oxidative stress in diabetes causes the over production of superoxide radicals in endothelial cells of large and small vessels, as well as in the myocardium and leads to many micro and macro vascular complications. The use of antioxidants, especially those with multiple antioxidant biomolecules like vegetables, fruits and seeds can be effective in preventing complications of diabetes.

Nanostructured plasmonic chips employing nanopillar and nanoring hole arrays for enhanced sensitivity of SPR-based biosensing.

We present a theoretical analysis of the different nanostructured plasmonic sensor chips—consisting of plasmonic nanostructures present on the surface of plasmonic thin films—interrogated using the Kretschmann configuration for highly sensitive localized sensing, with high tunability from the visible to the infrared regions. Rigorous coupled-wave analysis is performed to analyze all the proposed nanostructured sensor chips and compare their sensing performance. The sensitivity parameters are defined to focus on the detection of a thin layer of biomolecules on the surface of nanostructures. The dimensions of the nanostructures and the incident angle shift the plasmon resonance wavelengths and can be used to tune the operating wavelength. The nanostructured films create local regions of high electric fields, which results in enhanced sensitivity of the proposed structures. The proposed sensors can be used in surface plasmon resonance imaging to detect multiple biomolecules in a single measurement. An extremely high surface sensitivity and figure of merit (FOMS) of 91 nm nm−1 and 0.59 nm−1 has been found, respectively, for one of the proposed nanostructured sensing platforms. Moreover, we demonstrate a very high differential reflectance of 55% per nm thickness of the biolayer.

Multianalyte lateral flow immunoassay for simultaneous detection of protein-based inflammation biomarkers and pathogen DNA

The detection of multiple biomolecule classes in one go is highly desirable for a wide variety of areas, and in particular for point-of-care diagnostics. For example, wound infections are a major problem for patient’s health and cause huge efforts in our healthcare system. In this regard, monitoring infected wounds through simultaneous detection of pathogens via nucleic acid analysis and detection of local inflammation biomarkers is key in order to enable a personalized therapy, improve the clinical outcome and thus, leading to a reduction of overall healthcare costs. In this regard, wound exudate offers an attractive sample material which can be collected in a non-invasive manner. Here, we report the development of a Multianalyte-Assay detecting inflammation biomarkers and pathogen DNA simultaneously from one sample within 35 min. Protein-compatible amplification and labeling transforms nucleic acid information into the measurement principle for protein detection. The combination with rapid detection via lateral flow immunoassay enables a fast and straightforward analysis of multiple biomolecule classes using identical assay conditions. To demonstrate the feasibility of the Multianalyte-Assay, the proinflammatory cytokine interleukin-6 (IL-6) and gDNA of the opportunistic pathogen Pseudomonas aeruginosa (P. aeruginosa) are used. The detection limits of 4 ng/mL IL-6 and 70 copies/reaction P. aeruginosa gDNA meet the clinically relevant range and thus, having tremendous potential to improve the wound management at the point-of-care.

Simultaneous Targeted Analysis of GGT and Its H-Type mRNA in HepG2 Cells Based on Degradable Silicon Nanomaterials.

Most important physiological processes in live cells are usually maintained by the interaction of multiple related biomolecules; the multi-target simultaneous analysis of these related molecules can better reflect the dynamic changes of their biological regulatory processes, providing more comprehensive information for diseases diagnosis and research. Herein, we have constructed the degradable multifunctional silica nanomaterials from the prepared degradable organic silicon source and further established degradable composite nanoprobes (DCNPs). The low toxicity of DCNPs could reduce the impact on normal physiological processes in cells and achieve the needs of living cell analysis applications; by the loading of the gamma-glutamyltransferase (GGT) activity-identification probe (Cy-GGT) and surface nucleic acid-recognizing molecular beacon (hairpin) modification, the DCNP realized the simultaneous image analysis of GGT and its related H-type mutated GGT mRNA (H-mRNA) in HepG2 cells and their quantitative detection in vitro. Compared with the traditional multi-target analysis strategy, the lack of targets' physiological mechanism connection was improved, and the combined application of small molecular probes and nucleic acid analysis structures was realized under the control of the cross-influence. This strategy is expected to provide a new direction for the design of multi-target analysis in live cells and provide more accurate analytical tools for clinical research and cancer therapy.

Tetrahedral DNA mediated direct quantification of exosomes by contact-electrification effect

Exosomes are membrane-enclosed extracellular vesicles carrying multiple biomolecules for intercellular communications. Accurate detection of exosomes could provide critical clinical information and show great significance for early diagnosis and personalized therapy of cancer. In this work, we propose a triboelectric sensing strategy for direct quantification of exosomes based on the contact-electrification effect. The target exosomes can be selectively captured on the three-dimensional tetrahedral DNA (TDNA) monolayer. The electrons transfer between abundant amino groups from exosomes and the tribo-materials contribute to the measured signal. Due to the specific output characteristic, it is able to directly discriminate 3 exosomes /μL with a linear range from 20 to 1000 exosomes/μL, even without any signal amplification. The challenges for distinguishing different cell line-derived exosomes and anti-interference in complicated biological serum systems show good performances. The presence of target exosomes can also be easily determined by visual observation of LED lighted by the generated electric energy. The proposed method can be used as a powerful tool for ultrasensitive analysis of exosomes, which is expected to have broad biological and analytical applications.

Enzymatic Methods for Salivary Biomarkers Detection: Overview and Current Challenges.

Early detection is a key factor in patient fate. Currently, multiple biomolecules have been recognized as biomarkers. Nevertheless, their identification is only the starting line on the way to their implementation in disease diagnosis. Although blood is the biofluid par excellence for the quantification of biomarkers, its extraction is uncomfortable and painful for many patients. In this sense, there is a gap in which saliva emerges as a non-invasive and valuable source of information, as it contains many of the biomarkers found in blood. Recent technological advances have made it possible to detect and quantify biomarkers in saliva samples. However, there are opportunity areas in terms of cost and complexity, which could be solved using simpler methodologies such as those based on enzymes. Many reviews have focused on presenting the state-of-the-art in identifying biomarkers in saliva samples. However, just a few of them provide critical analysis of technical elements for biomarker quantification in enzymatic methods for large-scale clinical applications. Thus, this review proposes enzymatic assays as a cost-effective alternative to overcome the limitations of current methods for the quantification of biomarkers in saliva, highlighting the technical and operational considerations necessary for sampling, method development, optimization, and validation.

Label-free cell assays to determine compound uptake or drug action using MALDI-TOF mass spectrometry.

Cell-based assays for compound screening and profiling are fundamentally important in life sciences, chemical biology and pharmaceutical research. Most cell assays measure the amount of a single reporter molecule or cellular endpoint, and require the use of fluorescence or other labeled materials. Consequently, there is high demand for label-free technologies that enable multiple biomolecules or endpoints to be measured simultaneously. Here, we describe how to develop, optimize and validate MALDI-TOF mass spectrometry (MS) cell assays that can be used to measure cellular uptake of transporter substrates, to monitor cellular drug target engagement or to discover cellular drug-response markers. In uptake assays, intracellular accumulation of a transporter substrate and its inhibition by test compounds is measured. In drug response assays, changes to multiple cellular metabolites or to abundant posttranslational protein modifications are monitored as reporters of drug activity. We detail a ten-part optimization protocol with every part taking 1-2 d that leads to a final 2 d optimized procedure, which includes cell treatment, transfer, MALDI MS-specific sample preparation, quantification using stable-isotope-labeled standards, MALDI-TOF MS data acquisition, data processing and analysis. Key considerations for validation and automation of MALDI-TOF MS cell assays are outlined. Overall, label-free MS cell-based assays offer speed, sensitivity, accuracy and versatility in drug research.

Metal-Organic Framework Nanoparticles Power DNAzyme Logic Circuits for Aberrant MicroRNA Imaging.

At the molecular level, a large number of studies exist on the use of dynamic DNA molecular circuits for disease diagnosis and biomedicine. However, how to design programmable molecular circuit devices to autonomously and accurately diagnose multiple low-abundance biomolecules in complex cellular environments remains a challenge. Here, we constructed DNAzyme logic circuits for the analysis and imaging of multiple microRNAs in living cells using Cu/ZIF-8 NPs as a nanocarrier of the logic gate modules and the Cu2+ cofactor of the Cu2+-dependent DNAzyme. The logic gate modules of the logic operation system were adsorbed on the surface of Cu/ZIF-8 NPs via electrostatic interaction. After internalization, pH-responsive Cu/ZIF-8 NPs could efficiently release the logic gate modules and Cu2+, which allowed us to realize multiple logic computations initiated by endogenous miRNA, including one YES logic gate and two binary logic gates (OR and AND) in different living cells. Cu2+-DNAzyme logic circuits could quickly respond to multiple endogenous miRNAs in the complex cell environment, which also provided a new research method for the application of DNA biocomputing circuits in living cells.

An Electropolymerized Self Assembled Monolayer of Crystal Violet for Chemiresistive Hydrogen Peroxide Sensor

Chemiresistive sensors have been widely used for gas sensing but recently few studies have reported their use in liquids. Unlike electrochemical sensors, these sensors are simple to fabricate and operate with a single sensing film electrodes with two electrical contacts for monitoring change in current under a small potential bias, without the need of reference electrode. Hydrogen peroxide (H2O2) is an intermediate molecule generated in numerous peroxidase assays. Therefore, it is widely used for detecting multiple biomolecules like glucose, galactose etc. H2O2 detection is commonly performed using colorimetric and electrochemical sensors. However, colorimetric sensors require chemicals while electrochemical sensors need reference electrode for reliable measurements [1].Here, we demonstrated a reagent less H2O2 sensor fabricated using carbon nanotube (CNT) as the base substrate and an electropolymerized self assembled monolayer crystal violet as selective ligand. The functionalised 2D percolation network of CNT film transduces a selective response for H2O2. The chemiresistive sensor was fabricated using a low-cost xurography based fabrication process. The CNT dispersion was supplied by Nano-C, Inc. A diluted CNT dispersion (4% v/v) was prepared by sonicating the CNT ink in a mixture of methanol and water (1:1) for 15 minutes in a bath sonicator. The diluted CNT dispersion was drop casted on a frosted glass substrate and cured at 120 oC for 60 minutes. Patterned gold leaf was used as the contact electrodes and copper tape was used for the external electrode (Figure 1a). The sensor fabrication process is described in details elsewhere [2]. The electrodes were functionalized with a self assembled monolayer by immersing them in 0.6 mM crystal violet prepared in 0.1 M phosphate buffer for 90 minutes. Then, the self assembled monolayer of crystal violet was electropolymerized using cyclic voltammetry from 0 to 2V and scan rate of 0.1 V/s.The sensor response was tested in five different H2O2 concentrations (0.5 ppm, 5 ppm, 50 ppm, 500 ppm, and 1000 ppm). H2O2 solutions were prepared by diluting the stock solution in 39 mM acetate buffer similar to the buffer strength of blood. The sensor response for various H2O2 concentrations is shown in Figure 1b. The sensor response for each H2O2 solution was calculated with 39 mM acetate buffer as the baseline. Each bar graph represents the average of last-minute current measurements of a 30 minutes run. The sensor exhibited an increase in current value with increase in H2O2 concentration from 0.5 ppm to 1000 ppm. The crystal violet attached on the CNT film undergoes oxidation in presence of H2O2 concentration. The change in oxidation state of crystal violet resulted alterations in doping level of functionalized CNT network leading to a change in conductivity of the sensor. No significant interference was observed for common interferents like as glucose, galactose, urea, uric acid and gluconic acid. The sensor was also chemically reset using 0.1 M ascorbic acid by reducing the oxidized crystal violet.We demonstrated that an electropolymerized self assembled monolayer of crystal violet attached to CNT can measure H2O2 concentration within a range of 0.5 ppm to 1000 ppm. In addition, the low-cost fabrication process and the peroxide sensor enables this technology to a wide range of peroxidase based enzymatic assays that are biological relevant both in medical diagnostics biological research.

Research progress of DLX6-AS1 in human cancers.

Long non-coding RNAs (lncRNAs) are a kind of translational-repressor RNAs composed of more than 200 nucleotides and formerly considered as "transcriptional noise". Recently studies have shown that lncRNAs could bind to multiple biomolecules such as DNA, transcription factors, RNA, chromatin complexes and proteins, and regulate target gene expression at multi-levels, thus playing an essential role in human tumors. DLX6-AS1, a recently discovered oncogenic lncRNA, is highly expressed in various human tumors, including lung cancer, liver cancer and pancreatic cancer. This paper mainly reviewed the regulatory mechanism of DLX6-AS1 as a competitive endogenous RNA (ceRNA) in tumor cell proliferation, cell apoptosis, angiogenesis, epithelial-mesenchymal transformation, chemotherapy resistance and metabolic changes. Furthermore, the translational value of DLX6-AS1 in cancer was also elucidated, which suggested its potential as a diagnostic or prognostic biomarker in cancer. In summary, this present article not only makes an in-depth analysis of the expression changes and carcinogenic mechanism of DLX6-AS1 in various human cancers, but also provides a new breakthrough for the diagnosis and treatment of cancers.