"DSN-mismatched CRISPR″sensor for highly selective and sensitive detection of under-expressed miR-let-7a.

Several microRNAs (miRNAs) are expressed at lower levels in specific tumors, e.g., miR-let-7a in non-small cell lung cancer (NSCLC). This makes it challenging to analyze their lower abundance versus specifically elevated miRNAs. Here, we describe a novel fluorescent biosensor for the highly selective and sensitive detection of miR-let-7a constructed by combining miRNA screening assisted by a duplex-specific nuclease (DSN) with CRISPR-Cas12a system signal amplification. We meticulously designed a mismatch in the first three to four bases at the 5'-end of the capture DNA to improve the signal-to-noise ratio of the CRISPR-Cas12a system. Within this "DSN-mismatched CRISPR" fluorescence strategy, miR-let-7a was accurately screened by DSN-assisted cleavage, and the mismatched capture DNA unbound to target miRNA could trigger the CRISPR-Cas12a system to produce a mass of trans-cleave fluorescence signals. This "turn-off" approach was suitable for detecting decreased levels of miRNAs. This approach can not only discriminate the single-base mismatched let-7 family but also reach a limit of detection at 64.17 fM as well as be quantified from 100 fM to 500 pM. The miR-let-7a levels were then measured in clinical serum samples from healthy volunteers and patients with NSCLC. This study holds promise for the development of a universal under-expressed miRNA assay for early diagnosis and treatment of cancers.

Flushing control strategies to improve the stability of a biogas slurry drip irrigation system: Behavioral characteristics and mechanisms.

Biogas slurry drip irrigation can mitigate environmental pollution and reduce the use of chemical fertilizers to enable sustainable development. However, the stability of the biogas slurry drip irrigation system (BSDIS) is disrupted by emitter clogging; hence, it is essential to explore the flushing control strategy of BSDIS. By means of combining actual measurement and simulation, this study investigates the BSDIS stability based on the three technical parameters of the flushing control strategy. Appropriate flushing control strategies can improve system stability and cause spatial differences on the drip irrigation tape. Under various flushing control strategies, the system stability primarily undergoes delays, sensitivity, and ineffectiveness of flushing with time. Compared with the without flushing and emitter outlet downward-oriented treatment, the optimal flushing combination (the high frequency flushing + emitter outlet upward-oriented treatment) reduces the emitter clogging content by approximately 70.97% and increases system stability by 189.1%. In the internal hydrodynamics, the laying direction of emitters does not change the movement characteristics of water flow, although the clogging particles do not completely follow the water flow, with some particles settling owing to gravity, thereby clotting the emitters. When clogging occurs, the increase in flushing speed is conducive to the increase in turbulent kinetic energy on the inlet surface of the emitter, which facilitate the flushing of clogged substances. This study proposes optimal flushing strategy parameters along with a new management mode for the waste liquid represented by biogas slurry.

Characterization of labyrinth emitter-clogging substances in biogas slurry drip irrigation systems.

Drip irrigation is important for efficiently returning biogas slurry to fields. Elucidating the characteristics and components of clogging substances produced by labyrinth emitters in biogas slurry drip irrigation systems will help to develop various clogging substance-remediation strategies. However, previous studies were unable to characterize the clogging substances in emitters. Thus, we aimed to characterize and quantify the substances clogging emitters in a biogas slurry drip irrigation system and determine the micromorphology and dominance of microbial communities. Here, emitter discharge changes and the micromorphologies, phase compositions, and biological communities of clogging substances were studied via hydraulic performance tests, scanning electron microscopy-energy depressive spectra (SEM-EDS), and high-throughput sequencing. The degree of emitter-clogging increased over time (first quickly, then slowly) and was deeper at the end of the drip irrigation tape than at the head. The clogging substances were viscous agglomerations primarily comprising 0.3-1.5-µm particles. Their formation was affected by settlement with gravity, water pressure adhesion, and mobile biological adhesion. The dominant microbial communities in the clogging substances included Firmicutes (29.7%) and Proteobacteria (19%); the emitter-clogging substances primarily comprised water (85%) and composite dry matter. The water, dry matter, and extracellular polymer substance (EPS) weights in the clogging substances increased over time, but their relative proportions remained stable. In the composite dry matter, typical physical (organic carbon, Al2O3, and SiO2), chemical (CaCO3 and MgCO3), and biological (EPS) clogging substances accounted for >50, 9, and 5.62% of the total dry matter mass, respectively. This study provides a good foundation and reference idea and will be very helpful to propose targeted solutions for solving the clogging of biogas slurry drip irrigation system.

Design, Synthesis, and Functional Evaluation of 1, 5-Disubstituted Tetrazoles as Monoamine Neurotransmitter Reuptake Inhibitors.

Tetrazoles were designed and synthesized as potential inhibitors of triple monoamine neurotransmitters (dopamine, norepinephrine, serotonin) reuptake based on the functional and docking simulation of compound 6 which were performed in a previous study. The compound structure consisted of a tetrazole-linker (n)-piperidine/piperazine-spacer (m)-phenyl ring, with tetrazole attached to two phenyl rings (R1 and R2). Altering the carbon number in the linker (n) from 3 to 4 and in the spacer (m) from 0 to 1 increased the potency of serotonin reuptake inhibition. Depending on the nature of piperidine/piperazine, the substituents at R1 and R2 exerted various effects in determining their inhibitory effects on monoamine reuptake. Docking study showed that the selectivity of tetrazole for different transporters was determined based on multiple interactions with various residues on transporters, including hydrophobic residues on transmembrane domains 1, 3, 6, and 8. Co-expression of dopamine transporter, which lowers dopamine concentration in the biophase by uptaking dopamine into the cells, inhibited the dopamine-induced endoctytosis of dopamine D2 receptor. When tested for compound 40 and 56, compound 40 which has more potent inhibitory activity on dopamine reuptake more strongly disinhibited the inhibitory activity of dopamine transporter on the endocytosis of dopamine D2 receptor. Overall, we identified candidate inhibitors of triple monoamine neurotransmitter reuptake and provided a theoretical background for identifying such neurotransmitter modifiers for developing novel therapeutic agents of various neuropsychiatric disorders.

Enhanced decomposition of laminated ammonium perchlorate composite.

In order to improve the thermal decomposition performances of ammonium perchlorate (AP), the laminated AP composite was prepared by ice-template induced self-assembly method. In this study, Iron-Konjac glucomannan (Fe3+-KGM) hydrosol rich in AP was selected as the freezing precursor. Through directional freezing of precursor and recrystallization of AP molecules, the laminated AP composite was obtained. The results showed that the thickness of the lamellar composite structure is about 10 to 30 µm, and the recrystallized AP particles are uniformly dispersed in the gel system. The oxygen bomb test results show that the micro-/nano-layered structure can significantly improve the sample's combustion heat value. Thermal analyses indicated that with the increasing Fe3+ content, the peak exothermic temperature of lamellar AP composite at different heating rates both showed a decreasing trend. With 10 wt% Fe(NO3)3·9H2O added, the decomposition peak temperature decreased from 433.0 to 336.2 °C at a heating rate of 5 °C/min, and the apparent activation energy (Ea) decreased dramatically from 334.1 kJ/mol to 255.4 kJ/mol. A possible catalytic thermal decomposition mechanism of lamellar AP composite catalyzed by Fe3+ was proposed. This work is beneficial to the structural design of other energetic materials.