Rapid approach for cloning bacterial single-genes directly from soils

Obtaining functional genes of bacteria from environmental samples usually depends on library-based approach which is not favored as its large amount of work with small possibility of positive clones. A kind of bacterial single-gene encoding glutamine synthetase (GS) was selected as example to detect the efficiency of cloning strategy in this study. Five GS genes were directly cloned from soils using degenerate primers with two steps of nested polymerase chains reactions. The genes showed 94 to 99% amino acid identities to the homologs in the known database, and encoded proteins affiliated to GS I and GS II families, respectively. All the five genes could rescue the growth of Escherichia coli glutamine auxotroph mutant ET6017 in minimum medium (ammonium chloride was sole nitrogen source in this medium). This study develops one rapid approach for cloning bacterial single-genes directly from soils. Comparing with the conventional strategies for gene cloning from complex environmental samples, this method did not need making genomic library and isolating target genes from large amount of library clones. This approach distinctively demonstrates its advantages of rapidity and effectiveness particularly when it aims at cloning short single-genes that had known homologs in all kinds of nucleic acid databases.Keywords: Gene cloning, soil, glutamine synthetase, nested PCR, single-geneAfrican Journal of Biotechnology Vol. 12(32), pp. 5029-503

Pathologically Activated Neuroprotection via Uncompetitive Blockade of \u3cem\u3eN\u3c/em\u3e-Methyl-d-aspartate Receptors with Fast Off-rate by Novel Multifunctional Dimer Bis(propyl)-cognitin

Uncompetitive N-methyl-d-aspartate (NMDA) receptor antagonists with fast off-rate (UFO) may represent promising drug candidates for various neurodegenerative disorders. In this study, we report that bis(propyl)-cognitin, a novel dimeric acetylcholinesterase inhibitor and γ-aminobutyric acid subtype A receptor antagonist, is such an antagonist of NMDA receptors. In cultured rat hippocampal neurons, we demonstrated that bis(propyl)-cognitin voltage-dependently, selectively, and moderately inhibited NMDA-activated currents. The inhibitory effects of bis(propyl)-cognitin increased with the rise in NMDA and glycine concentrations. Kinetics analysis showed that the inhibition was of fast onset and offset with an off-rate time constant of 1.9 s. Molecular docking simulations showed moderate hydrophobic interaction between bis(propyl)-cognitin and the MK-801 binding region in the ion channel pore of the NMDA receptor. Bis(propyl)-cognitin was further found to compete with [3H]MK-801 with a Ki value of 0.27 μm, and the mutation of NR1(N616R) significantly reduced its inhibitory potency. Under glutamate-mediated pathological conditions, bis(propyl)-cognitin, in contrast to bis(heptyl)-cognitin, prevented excitotoxicity with increasing effectiveness against escalating levels of glutamate and much more effectively protected against middle cerebral artery occlusion-induced brain damage than did memantine. More interestingly, under NMDA receptor-mediated physiological conditions, bis(propyl)-cognitin enhanced long-term potentiation in hippocampal slices, whereas MK-801 reduced and memantine did not alter this process. These results suggest that bis(propyl)-cognitin is a UFO antagonist of NMDA receptors with moderate affinity, which may provide a pathologically activated therapy for various neurodegenerative disorders associated with NMDA receptor dysregulation

Peregrine and saker falcon genome sequences provide insights into evolution of a predatory lifestyle

As top predators, falcons possess unique morphological, physiological and behavioral adaptations that allow them to be successful hunters: for example, the peregrine is renowned as the world's fastest animal. To examine the evolutionary basis of predatory adaptations, we sequenced the genomes of both the peregrine (Falco peregrinus) and saker falcon (Falco cherrug), and we present parallel, genome-wide evidence for evolutionary innovation and selection for a predatory lifestyle. The genomes, assembled using Illumina deep sequencing with greater than 100-fold coverage, are both approximately 1.2 Gb in length, with transcriptome-assisted prediction of approximately 16,200 genes for both species. Analysis of 8,424 orthologs in both falcons, chicken, zebra finch and turkey identified consistent evidence for genome-wide rapid evolution in these raptors. SNP-based inference showed contrasting recent demographic trajectories for the two falcons, and gene-based analysis highlighted falcon-specific evolutionary novelties for beak development and olfaction and specifically for homeostasis-related genes in the arid environment–adapted saker

An Optimized Error Compensation Method for Phase Measurement Profilometry

Phase measurement profilometry (PMP) is primarily employed to analyze the morphology of a functional surface with precision. Historically, one of the most complex and persistent challenges in PMP has been reducing errors stemming from inconsistent indicators at the edges of a surface. In response to this challenge, we propose an optimized error compensation methodology specifically designed to handle edge artefacts. This methodology introduces the Hilbert transform and object surface albedo as tools to detect the edges of the artefact region that need to be compensated. Moreover, we analyze the characteristics of the sinusoidal fringe waveform propagation direction and investigate the reconstruction results of the fringe vertical to the current directions to compensate for edge artefacts. The experimental results for various objects show that the optimized approach can compensate for edge artefacts by projecting in two directions and reducing the projection by half. The compensated root mean square error (RMSE) for planar objects can be reduced by over 45%

Effects of omalizumab on lung function in patients with moderate-to-severe allergic asthma: a systematic review and meta-analysis

Background: With the rise of targeted treatments for asthma, treatment with omalizumab is a new option. Objectives: To assess the improvement of pulmonary function with additional omalizumab treatment in patients (⩾6 years old) with moderate-to-severe allergic asthma. Data sources and methods: Observational studies of randomized controlled trials of add-on omalizumab for the treatment of patients with moderate-to-severe allergic asthma, published from the establishment till August 2022, were retrieved from WAN FANG DATA, PubMed, CNKI, Embase, Cochrane, and Web of Science databases. Data extraction and quality evaluation were performed on the literature that met the inclusion criteria, using RevMan 5.3 to analyze the data. Results: A total of 11 randomized controlled clinical trials were included, involving a total of 3578 patients with asthma, 1856 patients in the omalizumab group, and 1722 patients in the control group. The improvement in Forced expiratory volume in 1 s as a percentage of predicted normal and Forced expiratory volume in 1 s was more pronounced in the omalizumab-treated group [MD = 3.91, 95% confidence interval (CI): 1.89–5.94, p  = 0.0002; MD = 0.09, 95% CI: 0.05–0.13, p  < 0.0001], while the improvement in Morning Peak expiratory flow rate was not statistically different between the two groups (MD = 3.64, 95% CI: −22.17–29.45, p  = 0.78). Conclusion: Additional omalizumab treatment showed some improvement in lung function in patients with moderate-to-severe asthma. Trial registration: PROSPERO ID:CRD42022378498

The better growth phenotype of DvGS1-transgenic arabidopsis thaliana is attributed to the improved efficiency of nitrogen assimilation

The overexpression of the algal glutamine synthetase (GS) gene DvGS1 in Arabidopsis thaliana resulted in higher plant biomass and better growth phenotype. The purpose of this study was to recognize the biological mechanism for the growth improvement of DvGS1-transgenic Arabidopsis. A series of molecular and biochemical investigations related to nitrogen and carbon metabolism in the DvGS1-transgenic line was conducted. Analysis of nitrogen use efficiency (NUE)-related gene transcription and enzymatic activity revealed that the transcriptional level and enzymatic activity of the genes encoding GS, glutamate synthase, glutamate dehydrogenase, alanine aminotransferase and aspartate aminotransferase, were significantly upregulated, especially from leaf tissues of the DvGS1-transgenic line under two nitrate conditions. The DvGS1-transgenic line showed increased total nitrogen content and decreased carbon: nitrogen ratio compared to wild-type plants. Significant reduced concentrations of free nitrate, ammonium, sucrose, glucose and starch, together with higher concentrations of total amino acids, individual amino acids (glutamate, aspartate, asparagine, methionine), soluble proteins and fructose in leaf tissues confirmed that the DvGS1-transgenic line demonstrated a higher efficiency of nitrogen assimilation, which subsequently affected carbon metabolism. These improved metabolisms of nitrogen and carbon conferred the DvGS1-transgenic Arabidopsis higher NUE, more biomass and better growth phenotype compared with the wild-type plants

A Y-shaped network based single-shot absolute phase recovery method for fringe projection profilometry

Fringe projection profilometry (FPP) is a widely used non-contact 3D measurement method. Though maturing in the last decade, it remains a significant challenge when facing the phase unwrapping of measured object surfaces in a single-shot measurement setting. With the rapid development of deep learning techniques, the adoption of a data-driven approach is gaining popularity in the field of optical metrology. This study proposes a new absolute phase recovery method based on the devised single-stage deep learning network. The aim is to ensure high quality absolute phase recovery from a single-shot fringe projection measurement. Unlike most existing approaches, where the numerators and denominators of the wrapped phases and the fringe orders are predicted in various stages, the proposed method acquires the wrapped phases and the corresponding fringe orders within a single network, i.e. it can predict both wrapped phases and the corresponding fringe orders directly and simultaneously from the single fringe pattern projected in the single-shot mode based on a unified Y-shaped network. Experiments on benchmark datasets and models have demonstrated the effectiveness and efficiency of the technique, especially in terms of high-quality recovery of absolute phase information by using the lightweight single-stage network, and enabling the FPP-based phase 3D measurements in an online manner