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The cellulosome of Ruminiclostridium thermocellum was one of the most efficient cellulase systems in nature. However, the product of cellulose degradation by R. thermocellum is cellobiose, which leads to the feedback inhibition of cellulosome, and it limits the R. thermocellum application in the field of cellulosic biomass consolidated bioprocessing (CBP) industry. In a previous study, R. thermocellum M3, which can hydrolyze cellulosic feedstocks into monosaccharides, was isolated from horse manure. In this study, the complete genome of R. thermocellum M3 was sequenced and assembled. The genome of R. thermocellum M3 was compared with the other R. thermocellum to reveal the mechanism of cellulosic saccharification by R. thermocellum M3. In addition, we predicted the key genes for the elimination of feedback inhibition of cellobiose in R. thermocellum. The results indicated that the whole genome sequence of R. thermocellum M3 consisted of 3.6 Mb of chromosomes with a 38.9% of GC%. To be specific, eight gene islands and 271 carbohydrate-active enzyme-encoded proteins were detected. Moreover, the results of gene function annotation showed that 2,071, 2,120, and 1,246 genes were annotated into the Clusters of Orthologous Groups (COG), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases, respectively, and most of the genes were involved in carbohydrate metabolism and enzymatic catalysis. Different from other R. thermocellum, strain M3 has three proteins related to β-glucosidase, and the cellobiose hydrolysis was enhanced by the synergy of gene BglA and BglX. Meanwhile, the GH42 family, CBM36 family, and AA8 family might participate in cellobiose degradation.

A diatomite-modified Fe2O3 (Fe2O3/Dia) catalyst was prepared to catalyze the ozonation degradation of sulfachloropyridazine sodium (SPDZ). The chemical oxygen demand (COD) was used as the index of pollutant degradation. The catalytic ozonation experiment showed that the COD removal rate of SPDZ was 87% under Fe2O3/Dia catalysis, which was much higher than that obtained when using Fe2O3 as the catalyst. The characteristics of the Fe2O3/Dia catalyst were investigated, and the successful synthesis of the Fe2O3/Dia composite catalyst was proved by XRD, XPS, SEM, FTIR, BET and other characterization methods. The catalytic mechanism of degradation by ozone with Fe2O3/Dia was analyzed. According to free-radical trapping experiments and an in situ electron paramagnetic spectrometer characterization analysis, the main oxidizing species in the catalytic Fe2O3/Dia ozone system is ·OH. The intermediates in the degradation process of SPDZ were detected and analyzed in detail by liquid chromatography-coupled mass spectrometry. The degradation mechanism and three degradation paths of SPDZ were proposed. [ABSTRACT FROM AUTHOR]

The cultivation of microalgae from municipal wastewater, while simultaneously removing nutrients from the water column, has the potential to aid biodiesel production and carbon dioxide fixation, thereby alleviating the pressure of energy shortages. In this research, different ratios of sodium bicarbonate and glucose were used to prepare simulated municipal wastewater. The results demonstrated that microalgae were most effectively treated under one-stage direct treatment conditions. During direct culture, the most effective treatment was observed for IAA-3, which exhibited a dry weight of 1.4363 g/L and a lipid content of 25.05% after stimulation with 0.0005 M NaHCO3. In contrast, NaHCO3-2 demonstrated optimal performance during the secondary culture, with a dry weight of 1.6844 g/L and a lipid content of 18.05%. Finally, the economic, social and environmental benefits of direct treatment (IAA-3) and secondary treatment NaHCO3-2 were analyzed. The benefits of direct treatment were found to be USD 0.50989/L, while those of secondary treatment were USD 0.43172/L. For each tonne of municipal wastewater treated, the carbon sequestration benefits of IAA-3 during direct treatment and NaHCO3-2 during secondary treatment were USD 0.45645 and USD 0.85725, respectively. [ABSTRACT FROM AUTHOR]

In this article, we describe an in-house polymerase chain reaction-sequence specific priming (PCR-SSP) assay designed for undergraduate medical students as part of the experimental pathogen biology and immunology (EPBI) course. It screens human leukocyte antigen (HLA)-DR2 allotype from genomic DNA samples using a rapid and single-tube PCR technique, yielding definitive typing result without conventional post-amplification step like probing or Sanger sequencing. This laboratory exercise offers the undergraduate medical students an opportunity to learn about current molecular biology techniques in HLA genotyping with limited effort and cost, in addition to a better understanding of concepts presented in the classroom lectures. Upon completing this experiment module, the students show statistically significant improvement in several key indexes, such as the knowledge about the mainstream HLA DNA typing techniques, awareness of the relevance of this knowledge for their future scientific research, immunogenetics-related basic laboratory skills they acquire, and interest and desire for mastering this assay (all p < 0.05). This easy to implement set of experiments is composed of a two-session lab module occupying eight teaching hours, and has been run successfully in our laboratory.

BackgroundMetagenomic next-generation sequencing (mNGS) technology has been central in detecting infectious diseases and helping to simultaneously reveal the complex interplay between invaders and their hosts immune response characteristics. However, it needs to be rigorously assessed for clinical utility. The present study is the first to evaluate the clinical characteristics of the host DNA-removed mNGS technology for detecting SARS-CoV-2, revealing host local immune signaling and assisting genomic epidemiology.Methods46 swab specimens collected from COVID-19 patients were assayed by two approved commercial RT-qPCR kits and mNGS. The evolutionary tree of SARS-CoV-2 was plotted using FigTree directly from one sample. The workflow of removing the host and retaining the host was compared to investigate the influence of host DNA removal on the performances of mNGS. Functional enrichment analysis of DEGs and xCell score were used to explore the characteristics of host local immune signaling.ResultsThe detection rate of mNGS achieved 92.9% (26/28) for 28 samples with a Ct value ≤ 35 and 81.1% (30/37) for all 46 samples. The genome coverage of SARS-CoV-2 could reach up to 98.9% when the Ct value is about 20 in swab samples. Removing the host could enhance the sensitivity of mNGS for detecting SARS-CoV-2 from the swab sample but does not affect the species abundance of microbes RNA. Improving the sequencing depth did not show a positive effect on improving the detection sensitivity of SARS-CoV-2. Cell type enrichment scores found multiple immune cell types were differentially expressed between patients with high and low viral load.ConclusionsThe host DNA-removed mNGS has great potential utility and superior performance on comprehensive identification of SARS-CoV-2 and rapid traceability, revealing the microbiome’s transcriptional profiles and host immune responses.