RESUMO
DNA methylation has been considered an essential epigenetic biomarker for diagnosing various diseases, such as cancer. A simple and sensitive way for DNA methylation level detection is necessary. Inspired by the label-free and ultra-high sensitivity of solid-state nanopores to double-stranded DNA (dsDNA), we proposed a nanopore counter for evaluating DNA methylation by integrating a dual-restriction endonuclease digestion strategy coupled with polymerase chain reaction (PCR) amplification. Simultaneous application of BstUI/HhaI endonucleases can ensure the full digestion of the unmethylated target DNA but shows no effect on the methylated ones. Therefore, only the methylated DNA remains intact and can trigger the subsequent PCR reaction, producing a large quantity of fixed-length PCR amplicons, which can be directly detected through glassy nanopores. By simply counting the event rate of the translocation signals, the concentration of methylated DNA can be determined to range from 1 aM to 0.1 nM, with the detection limit as low as 0.61 aM. Moreover, a 0.01% DNA methylation level was successfully distinguished. The strategy of using the nanopore counter for highly sensitive DNA methylation evaluation would be a low-cost but reliable alternative in the analysis of DNA methylation.
Assuntos
Metilação de DNA , Nanoporos , DNA/análise , Reação em Cadeia da Polimerase , Enzimas de Restrição do DNARESUMO
Glass nanopore is an ideal candidate for biosensors due to its unique advantages such as label-free analysis, single-molecule sensitivity, and easy operation. Previous studies have shown that glass nanopores can distinguish different lengths of double-stranded DNA (dsDNA) at the same time with the length-resolution ability. Based on this, we proposed a novel design of a dsDNA block containing a programmable sensing site inside, which can be programmed to respond to different target molecules and cleaved into two smaller DNA blocks. When programming the sensing site with different sequences, for example, programming it as the substrate of GR-5 DNAzyme and CRISPR-Cas12a system, the DNA block could realize Pb2+ and cfDNA detection with the length-resolution ability of the glass nanopore. This strategy achieved a Pb2+ detection range from 0.5 nM to 100 nM, with a detection limit of 0.4 nM, and a BRCA-1 detection range from 1 pM to 10 pM, with a detection limit of 1 pM. The programable sensing site is easy to design and has strong expandability, which gives full play to the advantages of glass nanopore in length-resolution ability for dsDNA, and is expected to become an optional design for biosensing strategy for the glass nanopore as a biosensing platform.
Assuntos
Técnicas Biossensoriais , Nanoporos , Chumbo , Leitura , DNA/química , NanotecnologiaRESUMO
Solid-state nanopores have been proven as a powerful platform for label-free single-molecule analysis. However, due to its relatively low resolution and selectivity, developing biosensors with good translocation signals faces two significant challenges: (1) small-sized chemical or biological targets show difficulty in producing recognizable translocation signals because of their weak interaction with the nanopore and (2) protein interferents that widely exist in biological samples or buffers would considerably deteriorate the noise level of the nanopore, submerging the translocation signal. Herein, we demonstrate an effective way to overcome both the challenges. DNA cubes were used as signal transducers that can achieve an ultra-high (>50 : 1) signal-to-noise ratio (SNR) translocation signal, which is maintained even in protein interferent-rich buffers. A sensing strategy was constructed via hepatitis B virus (HBV) target-triggered cleavage of the component elements of the DNA cube with the assistance of the CRISPR-Cas12a technology, which caused a great drop in the translocation rate. The elements to cleave were optimized, and the sensor performance was tested in different protein stabilizer-rich buffers and human serum. Coupling with the polymerase chain reaction (PCR) pre-amplification technology, HBV-positive or -negative classification was achieved with the detection limit reaching 5 aM. It is worth noting that in our method, all reaction buffers were directly used without further optimization, which is of great help for the practical application of solid-state nanopores.
Assuntos
Nanoporos , Humanos , Vírus da Hepatite B/genética , Sistemas CRISPR-Cas , DNA/química , DigestãoRESUMO
The fabrication of nanopores with a matched pore size, and the existence of multiple interferents make the reproducible detection of small-sized molecules by means of solid-state nanopores still challenging. A useful method to solve these problems is based on the detection of large DNA nanostructures related to the existence of small-sized targets. In particular, a DNA tetrahedron with a well-defined 3D nanostructure is the ideal candidate for use as a signal transducer. Here, we demonstrate the detection of an L1-encoding gene of HPV18 as a test DNA target sequence in a reaction buffer solution, where long single-stranded DNA linking DNA tetrahedra onto the surface of the magnetic beads is cleaved by a target DNA-activated CRISPR-cas12 system. The DNA tetrahedra are subsequently released and can be detected by the current pulse in a glassy nanopore. This approach has several advantages: (1) one signal transducer can be used to detect different targets; (2) a glassy nanopore with a pore size much larger than the target DNA fragment can boost the tolerance of the contaminants and interferents which often degrade the performance of a nanopore sensor.
Assuntos
Nanoporos , Sistemas CRISPR-Cas/genética , DNA/química , DNA/genética , DNA de Cadeia Simples/genéticaRESUMO
Despite numerous clinical trials and pre-clinical developments, the diagnosis of cracked tooth, especially in the early stages, remains a challenge. Cracked tooth syndrome is often accompanied by dramatic painful responses from occlusion and temperature stimulation, which has become one of the leading causes for tooth loss in adults. Current clinical diagnostical approaches for cracked tooth have been widely investigated based on X-rays, optical light, ultrasound wave, etc. Advances in artificial intelligence (AI) development have unlocked the possibility of detecting the crack in a more intellectual and automotive way. This may lead to the possibility of further enhancement of the diagnostic accuracy for cracked tooth disease. In this review, various medical imaging technologies for diagnosing cracked tooth are overviewed. In particular, the imaging modality, effect and the advantages of each diagnostic technique are discussed. What's more, AI-based crack detection and classification methods, especially the convolutional neural network (CNN)-based algorithms, including image classification (AlexNet), object detection (YOLO, Faster-RCNN), semantic segmentation (U-Net, Segnet) are comprehensively reviewed. Finally, the future perspectives and challenges in the diagnosis of the cracked tooth are lighted.
Assuntos
Síndrome de Dente Quebrado , Dente , Adulto , Algoritmos , Inteligência Artificial , Síndrome de Dente Quebrado/diagnóstico , Humanos , Redes Neurais de Computação , Dente/diagnóstico por imagemRESUMO
Although immigrant mothers from some Latinx subgroups initially achieve healthy birth outcomes despite lower socioeconomic status, this advantage deteriorates across generations in the United States. Interpersonal discrimination and acculturative stress may interact with economic hardship to predict an intergenerational cascade of emotional and biological vulnerabilities, particularly perinatal depression. Network analyses may elucidate not only how and which psychosocial experiences relate to depressive symptoms, but which symptom-to-symptom relationships emerge. This study aims to understand (1) how economic, acculturative, and discrimination stressors relate to prenatal depression and low birth weight and (2) how Latinas may respond to and cope with stressors by exploring symptom-symptom and symptom-experience relationships. A sample of 151 pregnant Latinas (predominantly foreign-born and Mexican and Central American descent) completed the EPDS and psychosocial questionnaires (discrimination, acculturation, acculturative stress, economic hardship) during pregnancy (24-32 weeks). Birth weights were recorded from postpartum medical records. We created network models using the Extended Bayesian Information Criterion Graphical Least Absolute Shrinkage and Selection Operator to estimate the relationship between variables. Discrimination exposure connected psychosocial stressors to depressive symptoms, particularly worry, crying, sadness, and self-blame. Discrimination also revealed a connection between acculturation and low birth weight. Furthermore, younger age of migration and greater acculturation levels were correlated to greater discrimination stress and low birth weights. Perinatal research in Latinas must account not only for measures of cultural adaptation but recognize how developmental exposures across the life span, including discrimination, may be associated with adverse health trajectories for a mother and her child.
Assuntos
Aculturação , Depressão , Teorema de Bayes , Criança , Depressão/psicologia , Feminino , Hispânico ou Latino , Humanos , Recém-Nascido de Baixo Peso , Recém-Nascido , Mães/psicologia , Gravidez , Estresse Psicológico , Estados UnidosRESUMO
Osteosarcoma is predominant in the adolescent and the elderly population, but few studies have described the characteristics and prognostic factors of patients older than 60 years. In this study, the Surveillance, Epidemiology, and End Results registry database was used to identify all patients diagnosed with primary osteosarcoma from 1973 to 2014. We utilized Cox proportional hazards regression analysis to evaluate the association between patient overall survival and relevant characteristics, including gender, race, disease stage, treatment methods, primary tumor site, differentiation grade, and histologic subtype. In the data set, a total of 1139 patients with osteosarcoma older than 60 years old were identified. The overall rate of distant metastatic cases was 28.6%. Osteosarcoma occurred equally in men and women (49.5% vs 50.5%). Of all, 41.3% of tumors were located in axial location (pelvis, spine, and ribs), 34.1% of tumors were located in extremity (long or short bones of the upper or lower extremity), and 24.6% in other location (mandible, skull, and other atypical locations). Male (hazard ratio [HR] = 1.201; 95% confidence interval [CI]: 1.056-1.366), axial location (HR = 1.342; 95% CI: 1.157-1.556), distant metastasis (HR = 2.369; 95% CI: 2.015-2.785), non-surgery perform (HR = 2.108; 95% CI: 1.814-2.451) were independent risk factors for 5-year overall survival. This study revealed distinct clinicopathological features of patients with osteosarcoma older than 60 years. Male gender, tumor in axial site, nonsurgery perform, and distant metastasis indicated worse prognosis survival. Performing surgery is still an effective and reliable treatment method for patients older than 60 years.
Assuntos
Osteossarcoma/diagnóstico , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Osteossarcoma/mortalidade , Prognóstico , Fatores de Risco , Programa de SEER , Análise de SobrevidaRESUMO
Dibutyl phthalate (DBP) is a widely used plasticizer which has been found to be a reproductive and developmental toxicant and ubiquitously existing in the air. A highly sensitive method for DBP monitoring in the environment is urgently needed. A DBP sensor based on a homemade wireless-electrodeless quartz crystal microbalance with dissipation (QCM-D) coated with nano-structured nickel hydroxide is presented. With the noncontact configuration, the sensing system could work at a higher resonance frequency (the 3rd overtone) and the response of the system was even more stable compared with a conventional quartz crystal microbalance (QCM). The sensor achieved a sensitivity of 7.3 Hz/ppb to DBP in a concentration range of 0.4-40 ppb and an ultra-low detection limit of 0.4 ppb of DBP has also been achieved.
RESUMO
In this study, it is confirmed that without addition of organic solvent and embedding polymer hydrogel into glass nanopore, bare glass nanopore can faithfully measure various lengths of DNA duplexes from 200 to 3000 base pairs with 200 base pairs resolution, showing well-separated peak amplitudes of blockage currents. Furthermore, motivated by this readout capability of duplex DNA, amplicons from Polymerase Chain Reaction (PCR) amplification are straightforwardly discriminated by bare glassy nanopore without fluorescent labeling. Except simultaneous discrimination of up to 7 different segments of the same lambda genome, various pathogenic bacteria and viruses including SARS-CoV-2 and its mutants in clinical samples can be discriminated at high resolution. Moreover, quantitative measurement of PCR amplicons is obtained with detection range spanning from 0.75 aM to 7.5 pM and detection limit of 7.5 aM, which reveals that bare glass nanopore can faithfully disclose PCR results without any extra labeling.
Assuntos
COVID-19 , Nanoporos , Humanos , SARS-CoV-2/genética , Leitura , Reação em Cadeia da Polimerase , DNA/genética , Bactérias , Teste para COVID-19RESUMO
Collecting water from fog flow is emerging as a promising solution to the water shortage problem. This work demonstrated a novel environmentally responsive water collector made from a self-prepared Janus polyvinyl alcohol sponge in combination with a two-way shape memory alloy spring, which transforms the traditional manner of static water collection into a dynamic one. The unidirectional water transport of the Janus structure together with the dynamic collection approach correspond to a 30.8% increase in the water-collection rate (WCR). The resultant WCR is up to 5.1 g/h, which ranks relatively high compared to similar studies. The light- and thermal-response capability, easy fabrication, and good cycling performance indicate that our devices could be utilized in a variety of applications. In this work, an efficient, intelligent adaptive, simple-preparation, precision-guided, and economical fog-collecting devices are recommended. Our work provides new insights on the design of high-efficient water collectors with practicability.
RESUMO
The abuse of adamantanamine (ADA) and its derivatives as veterinary drugs in the poultry industry could cause severe health problems for humans. It is of great need to develop a rapid, cheap and ultrasensitive method for ADA detection. In this study, a sensitive conical nanochannel sensor was established for the rapid quantitative detection of ADA with the distinctive design of the host-guest competition. The sensor was constructed by functionalizing the nanochannel surface with p-toluidine and was then assembled with Cucurbit [7]uril (CB [7]). When ADA is added, it could occupy the cavity of CB [7] due to the host-guest competition and makes CB [7] to release from the CB [7]-p-toluidine complex, resulting in a distinct change of hydrophobicity of the nanochannel, which could be determined by the ionic current. Under the optimal conditions, the strategy permitted sensitive detection of ADA in a linear range of 10-1000 nM. The nanochannel based ADA sensing platform showed both high sensitivity and excellent reproducibility and the limit of detection was 4.54 nM. For the first time, the rapid and sensitive recognition of an illegal medicine was realized based on the host-guest competition method with the nanochannel system and the principle and feasibility of this method were described at length. This strategy provides a simple, reliable, and effective way to apply host-guest system in the development of nanochannel sensor for small-molecule drug detection.