Biosensors for Rapid Detection of Avian Influenza

The scope of this chapter was to review the advancements made in the area of biosensors for rapid detection of avian influenza viruses (AIVs). It is intended to provide general background about biosensor technology and to discuss important aspects for developing biosensors, such as selection of the suitable biological recognition elements (anti-AIV bioreceptors) as well as their immobilization strategies. A major concern of this chapter is also to critically review the biosensors’ working principles and their applications in AIV detection. A table containing the types of biosensor, bioreceptors, target AIVs, methods, etc. is given in this chapter. A number of papers for the different types of biosensors give hints on the current trends in the field of biosensor research for its application on AIV detection. By discussing recent research and future trends based on many excellent publications and reviews, it is hoped to give the readers a comprehensive view on this fast-growing field

Metastable state of gas hydrate during decomposition: a novel phenomenon

Natural gas hydrates are solid compounds with cage-like structures formed by gas and water. An intriguing phenomenon that gas hydrates can dissociate at a low rate below the ice freezing point has been viewed as the metastability of hydrate. The mechanisms of hydrate metastability have been widely studied, and many mechanisms were proposed involving the self-preservation effect, supercooled water-gas-hydrate metastable equilibrium, and supersaturated liquid–gas-hydrate system etc. The metastable state of hydrate could be of crucial significance in the kinetics of hydrate formation and decomposition, heat and mass transfer during gas production processes, and the application of hydrate-based technique involving desalination, energy storage and transportation, and gas separation and sequestration. Few researches have systematically considered this phenomenon, and its mechanism remains unclear. In this work, various mechanisms and hypothesis explaining the metastable state of gas hydrates were introduced and discussed. Further studies are still required to reveal the intrinsic nature of this metastable state of gas hydrate, and this work could give some implications on the existing theory and current status of relevant efforts

Selection, characterization, and application of DNA aptamers for detection of Mycobacterium tuberculosis secreted protein MPT64

Abstract Rapid detection of Mycobacterium tuberculosis (Mtb), an etiological agent of tuberculosis (TB), is important for global control of this disease. Aptamers have emerged as a potential rival for antibodies in therapeutics, diagnostics and biosensing due to their inherent characteristics. The aim of the current study was to select and characterize single-stranded DNA aptamers against MPT64 protein, one of the predominant secreted proteins of Mtb pathogen. Aptamers specific to MPT64 protein were selected in vitro using systematic evolution of ligands through exponential enrichment (SELEX) method. The selection was started with a pool of ssDNA library with randomized 40-nucleotide region. A total of 10 cycles were performed and seventeen aptamers with unique sequences were identified by sequencing. Dot Blot analysis was performed to monitor the SELEX process and to conduct the preliminary tests on the affinity and specificity of aptamers. Enzyme linked oligonucleotide assay (ELONA) showed that most of the aptamers were specific to the MPT64 protein with a linear correlation of R2 = 0.94 for the most selective. Using Surface Plasmon Resonance (SPR), dissociation equilibrium constant KD of 8.92 nM was obtained. Bioinformatics analysis of the most specific aptamers revealed the existence of a conserved as well as distinct sequences and possible binding site on MPT64. The specificity was determined by testing non-target ESAT-6 and CFP-10. Negligible cross-reactivity confirmed the high specificity of the selected aptamer. The selected aptamer was further tested on clinical sputum samples using ELONA and had sensitivity and specificity of 91.3% and 90%, respectively. Microscopy, culture positivity and nucleotide amplification methods were used as reference standards. The aptamers studied could be further used for the development of medical diagnostic tools and detection assays for Mtb

Surveying pediatric caregivers’ readiness for dyad isolation in the hospital during COVID-19

The onset of any emerging outbreak is stressful for everyone. Singapore was one of many countries affected early by COVID-19. In response, many precautionary measures were quickly initiated, including the isolation of suspected COVID-19 pediatric cases, and their caregivers were isolated together with their hospitalized children as a result. Caregivers play an important role in facilitating their child’s health in the hospital. Rooming in with their children during hospitalization promotes the benefits of parental presence and reduces separation effects. However, sudden admission with strict movement restrictions poses stress to these caregivers too. This study ran a 3-part paper-based survey to understand the stresses and concerns which caregivers faced when suddenly entering dyad isolation. The survey polled caregivers’ general perception of the situation, and also used questions adapted from the SARS Fear Scale and the Hospital Anxiety & Depression Scale (HADS). Caregivers in the COVID-19 isolation units did not expect their child to be isolated and were not prepared for dyad isolation with their children. They were found to be more dejected and were concerned that they themselves might have possibly infected their family and friends. Caregivers of children suspected of COVID-19 should be pre-empted to prepare for the possibility of isolation. This may include bringing in toys and personal entertainment to reduce boredom, as well as other essential needs. Patient mental wellness programs may consider extending their services to these caregivers too. Experience Framework This article is associated with the Patient, Family & Community Engagement lens of The Beryl Institute Experience Framework (https://www.theberylinstitute.org/ExperienceFramework). Access other PXJ articles related to this lens. Access other resources related to this lens

Misregulation of Alternative Splicing in a Mouse Model of Rett Syndrome

Mutations in the human MECP2 gene cause Rett syndrome (RTT), a severe neurodevelopmental disorder that predominantly affects girls. Despite decades of work, the molecular function of MeCP2 is not fully understood. Here we report a systematic identification of MeCP2-interacting proteins in the mouse brain. In addition to transcription regulators, we found that MeCP2 physically interacts with several modulators of RNA splicing, including LEDGF and DHX9. These interactions are disrupted by RTT causing mutations, suggesting that they may play a role in RTT pathogenesis. Consistent with the idea, deep RNA sequencing revealed misregulation of hundreds of splicing events in the cortex of Mecp2 knockout mice. To reveal the functional consequence of altered RNA splicing due to the loss of MeCP2, we focused on the regulation of the splicing of the flip/flop exon of Gria2 and other AMPAR genes. We found a significant splicing shift in the flip/flop exon toward the flop inclusion, leading to a faster decay in the AMPAR gated current and altered synaptic transmission. In summary, our study identified direct physical interaction between MeCP2 and splicing factors, a novel MeCP2 target gene, and established functional connection between a specific RNA splicing change and synaptic phenotypes in RTT mice. These results not only help our understanding of the molecular function of MeCP2, but also reveal potential drug targets for future therapies

ChIP-seq identifies McSLC35E2 as a novel target gene of McNrf2 in Mytilus coruscus, highlighting its role in the regulation of oxidative stress response in marine mollusks

NF-E2-related factor 2 (Nrf2) plays a crucial role in the oxidative regulatory process, which could trigger hundreds of antioxidant elements to confront xenobiotics. In the previous study, we identified Nrf2 from the marine mussel Mytilus coruscus, and the findings demonstrated that McNrf2 effectively protected the mussels against oxidative stress induced by benzopyrene (Bap). In order to delve deeper into the underlying mechanism, we utilized Chromatin Immunoprecipitation followed by sequencing (ChIP-seq) technology to systematically identify potential novel target genes of McNrf2. A total of 3,465 potential target genes were screened, of which 219 owned binding sites located within the promoter region. During subsequent experimental verification, it was found that McSLC35E2, a candidate target gene of McNrf2, exhibited negative regulation by McNrf2, as confirmed through dual luciferase and qRT-PCR detection. Further, the enzyme activity tests demonstrated that McNrf2 could counteract Bap induced oxidative stress by inhibiting McSLC35E2. The current study provides valuable insights into the application of ChIP-seq technology in the research of marine mollusks, advancing our understanding of the key role of Nrf2 in antioxidant defense mechanisms, and highlighting the significance of SLC35E2 in the highly sophisticated regulation of oxidative stress response in marine invertebrates

Downregulated expression of HSP27 in human low-grade glioma tissues discovered by a quantitative proteomic analysis

<p>Abstract</p> <p>Background</p> <p>Heat shock proteins (HSPs), including mainly HSP110, HSP90, HSP70, HSP60 and small HSP families, are evolutionary conserved proteins involved in various cellular processes. Abnormal expression of HSPs has been detected in several tumor types, which indicates that specific HSPs have different prognostic significance for different tumors. In the current studies, the expression profiling of HSPs in human low-grade glioma tissues (HGTs) were investigated using a sensitive, accurate SILAC (stable isotope labeling with amino acids in cell culture)-based quantitative proteomic strategy.</p> <p>Results</p> <p>The five HSP family members were detected and quantified in both HGTs and autologous para-cancerous brain tissues (PBTs) by the SILAC-based mass spectrometry (MS) simultaneously. HSP90 AB1, HSP A5(70 KDa), and especially HSP27 were significantly downregulated in HGTs, whereas the expression level of HSPA9 (70 KDa) was little higher in HGTs than that in PBTs. It was noted that the downregulation ratio of HSP27 was 0.48-fold in HGTs <it>versus </it>PBTs, which was further validated by results from RT-PCR, western blotting and immunohistochemistry. Furthermore, we detected HSP27 expression changes along with cell growth under heat shock treatment in glioma H4 cells.</p> <p>Conclusion</p> <p>The SILAC-MS technique is an applicable and efficient novel method, with a high-throughput manner, to quantitatively compare the relative expression level of HSPs in brain tumors. Different HSP family members have specific protein expression levels in human low-grade glioma discovered by SILAC-MS analysis. HSP27 expression was obviously downregulated in HGTs <it>versus </it>PBTs, and it exhibited temporal and spatial variation under heat shock treatment (43°C/0-3 h) <it>in vitro</it>. HSP27's rapid upregulation was probably correlated with the temporary resistance to heat shock in order to maintain the survival of human glioma cells.</p