Efficient Screening of nNOS-PSD95 Uncoupling Agents Based on Radiometric Fluorescent Molecularly Imprinted Sensors

Novel and efficient ratiometric fluorescent molecularly imprinted sensors (RFMIS) based on epitopes were developed, which can be used for the sensitive detection of neuronal nitric oxide synthase in the screening of neuronal nitric oxide synthase-postsynaptic density95 (nNOS-PSD95) coupling inhibitors. Under appropriate conditions, the fluorescence of the carbon dots quenched with the increasing concentration of nNOS1-133, while the fluorescence of the quantum dots remained unchanged. The fluorescence ratio had a good linearity in the concentration range of 0-500 ng mL-1 for nNOS1-133 and the determination limit was 0.14 ng mL-1. Using the classical nNOS-PSD95 coupling inhibitor (ZL006) as a control, the RFMIS were used as the detector to detect the free nNOS released by Gnetol and 2,3,5,4ʹ-tetrahydroxystilbene-2-O-β-Dglucoside from natural medicine after inhibition of nNOS-PSD95. The results have shown that the uncoupling efficiencies was consistent with co-immunoprecipitation experiments. The study provides a new idea and a new way for efficient screening of nature nNOS-PSD95 coupling inhibitors from natural medicine with the advantages of high efficiency, sensitivity and traceability

Preparation and Characterization of Epitope-Based Ratiometric Fluorescent Molecularly Imprinted Polymers

In order to solve the problem of difficult detection of neuronal nitric oxide synthase in the screening of neuronal nitric oxide synthase-postsynaptic density95 (nNOS-PSD95) uncoupling agent, this study used 133 amino acids (nNOS1-133) at the nitrogen terminal of nNOS as template molecules, carbon dots and quantum dots as ratio fluorescence recognition elements, SiO2 as matrix for the first time, combined with surface molecular imprinting technology and antigen-determining principle, to prepare ratiometric flurescent molecularly imprinted polymers (RFMIPs). The resulting RFMIPs were characterized by Fourier transform infrared spectroscopy, scanning electron microscopy, transmission electron microscopy and thermogravimetric analysis，exhibiting uniform spherical morphology, which unambiguously confirmed the successful formation of the nanosensor. The result indicates that the synthesized sensors have promising potential for the assay of trace peptides in complex matrices

Mapping the potential distribution of major tick species in China

Ticks are known as the vectors of various zoonotic diseases such as Lyme borreliosis and tick-borne encephalitis. Though their occurrences are increasingly reported in some parts of China, our understanding of the pattern and determinants of ticks’ potential distribution over the country remain limited. In this study, we took advantage of the recently compiled spatial dataset of distribution and diversity of ticks in China, analyzed the environmental determinants of ten frequently reported tick species and mapped the spatial distribution of these species over the country using the MaxEnt model. We found that presence of urban fabric, cropland, and forest in a place are key determents of tick occurrence, suggesting ticks were likely inhabited close to where people live. Besides, precipitation in the driest month was found to have a relatively high contribution in mapping tick distribution. The model projected that theses ticks could be widely distributed in the Northwest, Central North, Northeast, and South China. Our results added new evidence on the potential distribution of a variety of major tick species in China and pinpointed areas with a high potential risk of tick bites and tick-borne diseases for raising public health awareness and prevention response

The stress model of neuroticism and anxiety symptoms in fishermen

Background: Marine fishermen experience high levels of environmental and relationship stress and anxiety.The current study explored the role of stress in the relationship between neuroticism and anxiety symptomsamong marine fishermen. Materials and methods: Participants (fishermen from Tanmen in Qionghai city, Hainan Province) completedthree questionnaires: the NEO-Five-Factor Inventory-Neuroticism Subscale (NEO-FFI-N); the Mental StressorInvestigation Questionnaire (MSIQ); and the Mood and Anxiety Symptoms Questionnaire-30-item-AnxiousArousal Subscale (MASQ-D30-AA) within 1 week before embarking on a fishing trip and then again within1 week after their return to port. The data were subjected to correlational analyses and structural equationmodelling. Results: Positive correlations were found between NEO-FF-N (neuroticism) score, MSIQ score (total stress),MSIQ work-relationship score, ship environmental stress score, and MASQ score (anxiety symptoms). Regressionanalyses showed environmental stress had a significant moderating effect on the relationshipbetween neuroticism and anxiety symptoms, and further analysis showed a mediating effect of work-relationshipstress on the relationship between neuroticism and anxiety symptoms. Conclusions: Marine fishermen with high environmental stress had greater anxiety symptoms than thosewith low environmental stress. Neuroticism in marine fishermen further affects anxiety symptoms by affectingthe level of work-relationship stress

RNA sequencing analysis to capture the transcriptome landscape during skin ulceration syndrome progression in sea cucumber Apostichopus japonicus

Complement and coagulation cascades pathways (tif). Red boxes represent up-regulated genes, and green boxes represent down-regulated genes. (TIF 627 kb

The Molecular Mechanism Of Alpha-Synuclein Dependent Regulation Of Protein Phosphatase 2A Activity

Background/Aims: Alpha-synuclein (α-Syn) is a neuronal protein that is highly implicated in Parkinson\u27s disease (PD), and protein phosphatase 2A (PP2A) is an important serine/threonine phosphatase that is associated with neurodegenerative diseases, such as PD. α-Syn can directly upregulate PP2A activity, but the underling mechanism remains unclear. Therefore, we investigated the molecular mechanism of α-Syn regulating PP2A activity. Methods: α-Syn and its truncations were expressed in E.coli, and purified by affinity chromatography. PP2A Cα and its mutants were expressed in recombinant baculovirus, and purified by affinity chromatography combined with gel filtration chromatography. The interaction between α-Syn and PP2A Cα was detected by GST pull-down assay. PP2A activity was investigated by the colorimetric assay. Results: The hydrophobic non-amyloid component (NAC) domain of α-Syn interacted with PP2A Cα and upregulated its activity. α-Syn aggregates reduced its ability to upregulate PP2A activity, since the hydrophobic domain of α-Syn was blocked during aggregation. Furthermore, in the hydrophobic center of PP2A Cα, the residue of I123 was responsible for PP2A to interact with α-Syn, and its hydrophilic mutation blocked its interaction with α-Syn as well as its activity upregulation by α-Syn. Conclusions: α-Syn bound to PP2A Cα by the hydrophobic interaction and upregulated its activity. Blocking the hydrophobic domain of α-Syn or hydrophilic mutation on the residue I123 in PP2A Cα all reduced PP2A activity upregulation by α-Syn. Overall, we explored the mechanism of α-Syn regulating PP2A activity, which might offer much insight into the basis underlying PD pathogenesis

Effects of water stress on nutrients and enzyme activity in rhizosphere soils of greenhouse grape

In grape cultivation, incorrect water regulation will lead to significant water wastage, which in turn will change soil structure and disrupt soil nutrient cycling processes. This study aimed to investigate the effects of different water regulation treatments [by setting moderate water stress (W1), mild water stress (W2), and adequate water availability (CK)] on soil physical–chemical properties and enzyme activity in greenhouse grape during the growing season. The result showed that the W2 treatment had a negative impact on the build-up of dissolved organic carbon (DOC), nitrate nitrogen (NO3-N), and available phosphorus (AP). Throughout the reproductive period, the W1 and W2 treatments decreased the soil’s microbial biomass carbon (MBC) and microbial biomass nitrogen (MBN) contents, and MBC was more vulnerable to water stress. During the growth period, the trends of urease, catalase, and sucrase activities in different soil depth were ranked as 10–20 cm > 0–10 cm > 20–40 cm. The urease activity in 0–10 cm soil was suppressed by both W1 and W2 treatments, while the invertase activity in various soil layers under W1 treatment differed substantially. The W1 treatment also reduced the catalase activity in the 20–40 cm soil layer in the grape growth season. These findings suggested that W2 treatment can conserve water and enhance microbial ecology of greenhouse grape soils. Therefore, W2 treatment was the most effective water regulation measure for local greenhouse grape cultivation

Study on the bearing characteristics of overrun hydraulic support under impact loading

In order to study the effects of stress change and adaptability of the overrun hydraulic stent under impact load, the rigid–flexible coupling numerical model of the stent is established using multi-body dynamics simulation software ADAMS. In the model, the stent is flexibilized using the HyperMesh module, and the column and jack are equivalently replaced with a spring damping system. By applying impact loads to different positions above the front roof beam of the stent, the dynamic response characteristics of the column and the articulation point are obtained at different positions with different strengths. The results show that when the impact load is applied to different positions of the front roof beam, the steady-state response force of the column shows an overall “M" distribution, and along the length direction of the roof beam, the steady-state response force of the column shows an upward trend, and the maximum steady-state response force of the column is 547 kN at point (1, 6); when the impact load is applied to both sides of the front roof beam, the response at the articulation point is greater, resulting in a higher maximum simple harmonic response at that articulation point. When the impact load acts on both sides of the front roof beam, the response of the articulation point is greater, with a maximum simple harmonic response coefficient of 0.75 and a maximum excitation response coefficient of 0.28; when the impact loads of different strengths act on the whole front roof beam, the force acting on the column and pin shaft at the articulation point will produce a large impact, reducing its adaptability. To address this, we can consider changing the top plate stress conditions using methods like high-pressure injection of water and control the gradual and slow release of the top plate stresses. The results of the study provide a reference for the structural strength design and reliability analysis of the hydraulic support, which is of practical significance and value for improving the safe mining of the coal mining face