| 1. |
- Beal, Jacob, et al.
(författare)
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Robust estimation of bacterial cell count from optical density
- 2020
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Ingår i: Communications Biology. - : Springer Science and Business Media LLC. - 2399-3642. ; 3:1
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Tidskriftsartikel (refereegranskat)abstract
- Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data.
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| 2. |
- Beier, Sebastian, et al.
(författare)
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Construction of a map-based reference genome sequence for barley, Hordeum vulgare L.
- 2017
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Ingår i: Scientific Data. - : Springer Science and Business Media LLC. - 2052-4463. ; 4
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Tidskriftsartikel (refereegranskat)abstract
- Barley (Hordeum vulgare L.) is a cereal grass mainly used as animal fodder and raw material for the malting industry. The map-based reference genome sequence of barley cv. â € Morex' was constructed by the International Barley Genome Sequencing Consortium (IBSC) using hierarchical shotgun sequencing. Here, we report the experimental and computational procedures to (i) sequence and assemble more than 80,000 bacterial artificial chromosome (BAC) clones along the minimum tiling path of a genome-wide physical map, (ii) find and validate overlaps between adjacent BACs, (iii) construct 4,265 non-redundant sequence scaffolds representing clusters of overlapping BACs, and (iv) order and orient these BAC clusters along the seven barley chromosomes using positional information provided by dense genetic maps, an optical map and chromosome conformation capture sequencing (Hi-C). Integrative access to these sequence and mapping resources is provided by the barley genome explorer (BARLEX).
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| 3. |
- Borthakur, Dulal, et al.
(författare)
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Current status and trends in forest genomics
- 2022
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Ingår i: Forestry Research. - : Maximum Academic Press. - 2767-3812. ; 2
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Forskningsöversikt (refereegranskat)abstract
- Forests are not only the most predominant of the Earth's terrestrial ecosystems, but are also the core supply for essential products for human use. However, global climate change and ongoing population explosion severely threatens the health of the forest ecosystem and aggravtes the deforestation and forest degradation. Forest genomics has great potential of increasing forest productivity and adaptation to the changing climate. In the last two decades, the field of forest genomics has advanced quickly owing to the advent of multiple high-throughput sequencing technologies, single cell RNA-seq, clustered regularly interspaced short palindromic repeats (CRISPR)-mediated genome editing, and spatial transcriptomes, as well as bioinformatics analysis technologies, which have led to the generation of multidimensional, multilayered, and spatiotemporal gene expression data. These technologies, together with basic technologies routinely used in plant biotechnology, enable us to tackle many important or unique issues in forest biology, and provide a panoramic view and an integrative elucidation of molecular regulatory mechanisms underlying phenotypic changes and variations. In this review, we recapitulated the advancement and current status of 12 research branches of forest genomics, and then provided future research directions and focuses for each area. Evidently, a shift from simple biotechnology-based research to advanced and integrative genomics research, and a setup for investigation and interpretation of many spatiotemporal development and differentiation issues in forest genomics have just begun to emerge.
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| 4. |
- Cao, Hongru, et al.
(författare)
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Unsupervised domain-share CNN for machine fault transfer diagnosis from steady speeds to time-varying speeds
- 2022
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Ingår i: Journal of manufacturing systems. - : Elsevier. - 0278-6125 .- 1878-6642. ; 62, s. 186-198
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Tidskriftsartikel (refereegranskat)abstract
- The existing deep transfer learning-based intelligent fault diagnosis studies for machinery mainly consider steady speed scenarios, and there exists a problem of low diagnosis efficiency. In order to overcome these limitations, an unsupervised domain-share convolutional neural network (CNN) is proposed for efficient fault transfer diagnosis of machines from steady speeds to time-varying speeds. First, a Cauchy kernel-induced maximum mean discrepancy based on unbiased estimation is developed for improving the efficiency and robustness of feature adaptation. Secondly, an unsupervised domain-share CNN is constructed to simultaneously extract the domain-invariant features from the source domain and the target domain. Finally, adjustable and segmented balance factors are designed to flexibly weigh the distribution-adaptation loss and cross-entropy loss to improve diagnosis accuracy and transferability. The proposed method analyzes raw vibration signals collected from bearings and gears under different rotating speeds. Results of case studies show that the proposed method can achieve higher diagnosis accuracy, faster convergence, and better robustness than the reported methods, which demonstrates its potential applications in machine fault transfer diagnosis from a steady speed condition to a time-varying speed condition.
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| 5. |
- Cao, Xuan, et al.
(författare)
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Engineering yeast for high-level production of diterpenoid sclareol
- 2023
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Ingår i: Metabolic Engineering. - : Elsevier BV. - 1096-7176 .- 1096-7184. ; 75, s. 19-28
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Tidskriftsartikel (refereegranskat)abstract
- The diterpenoid sclareol is an industrially important precursor for alternative sustainable supply of ambergris. However, its current production from plant extraction is neither economical nor environmental-friendly, since it requires laborious and cost-intensive purification procedures and plants cultivation is susceptible to environmental factors. Engineering cell factories for bio-manufacturing can enable sustainable production of natural products. However, stringent metabolic regulation poses challenges to rewire cellular metabolism for overproduction of compounds of interest. Here we used a modular approach to globally rewire the cellular metabolism for improving sclareol production to 11.4 g/L in budding yeast Saccharomyces cerevisiae, the highest reported diterpenoid titer in microbes. Metabolic flux analysis showed that modular balanced metabolism drove the metabolic flux toward the biosynthesis of targeted molecules, and transcriptomic analysis revealed that the expression of central metabolism genes was shaped for a new balanced metabolism, which laid a foundation in extensive metabolic engineering of other microbial species for sustainable bio-production.
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| 6. |
- Chen, Jie, et al.
(författare)
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Bidirectional Mendelian Randomisation Analysis Provides Evidence for the Causal Involvement of Dysregulation of CXCL9, CCL11 and CASP8 in the Pathogenesis of Ulcerative Colitis
- 2023
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Ingår i: Journal of Crohn's & Colitis. - : Oxford University Press. - 1873-9946 .- 1876-4479. ; 17:5, s. 777-785
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Tidskriftsartikel (refereegranskat)abstract
- Background and Aims Systemic inflammation is well recognised to be associated with ulcerative colitis [UC], but whether these effects are causal or consequential remains unclear. We aimed to define potential causal relationship of cytokine dysregulation with different tiers of evidence. Methods We first synthesised serum proteomic profiling data from two multicentred observational studies, in which a panel of systemic inflammatory proteins was analysed to examine their associations with UC risk. To further dissect observed associations, we then performed a bidirectional two-sample Mendelian randomisation [TSMR] analysis from both forward and reverse directions using five genome-wide association study [GWAS] summary level data for serum proteomic profiles and the largest GWAS of 28 738 European-ancestry individuals for UC risk. Results Pooled analysis of serum proteomic data identified 14 proteins to be associated with the risk of UC. Forward MR analysis using only cis-acting protein quantitative trait loci [cis-pQTLs] or trans-pQTLs further validated causal associations of two chemokines and the increased risk of UC: C-X-C motif chemokine ligand 9 [CXCL9] [OR 1.45, 95% CI 1.08, 1.95, p = 0.012] and C-C motif chemokine ligand 11 [CCL11] [OR 1.14, 95% CI 1.09, 1.18, p = 3.89 x 10(-10)]. Using both cis- and trans-acting pQTLs, an association of caspase-8 [CASP8] [OR 1.04, 95% CI 1.03, 1.05, p = 7.63 x 10(-19)] was additionally identified. Reverse MR did not find any influence of genetic predisposition to UC on any of these three inflammation proteins. Conclusion Pre-existing elevated levels of CXCL9, CCL11 and CASP8 may play a role in the pathogenesis of UC.
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| 7. |
- Huang, Qi, et al.
(författare)
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Effective photocatalytic sterilization based on composites of Ag/InVO4/BiOBr : Factors, mechanism and application
- 2023
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Ingår i: Separation and Purification Technology. - 1383-5866 .- 1873-3794. ; 327
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Tidskriftsartikel (refereegranskat)abstract
- We hypothesized that photocatalysts with a low band gap could be useful in the sterilization of ceramic tiles in the natural environments of toilets using natural light in those settings. Certain photocatalysts can produce reactive oxygen species (ROS) under light illumination, which in turn are bactericidal. The properties of the BiOBr-containing photocatalysts were tuned by creating junctions and heterostructures with Ag and InVO4 and studied with respect to their bactericidal effect in dispersion. The bactericidal mechanism was studied through experiments in which active species were captured and via electron paramagnetic resonance (EPR) spectroscopy. At an optimal dosage of 0.5 g/L, the Ag/InVO4/BiOBr composite had a sterilization efficacy of 99.9999 % in 30 min under visible light illumination of 1000 W. It retained a sterilization efficacy of 99.999 % after four cycles. Anions such as Cl−, SO42−, and NO3− were shown to have no negative impact on sterilization efficacy. It was shown that the holes in the composite photocatalyst and hydroxyl radicals (·OH) were mechanistically critical for the sterilization. The photocatalysts were also studied in the field in the natural environment of a restroom, where they were loaded on ceramic tiles. Samples were collected from the surface of the ceramic tiles and analyzed for bacterial cultures and microbial diversity. The results were compared in the scope of the sterilization ability of various agents at the microbial level. The ceramic tiles loaded with Ag/InVO4/BiOBr showed the least amount of bacteria on their surfaces, and the microbial community richness was also the lowest.
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| 8. |
- Hüttner, Silvia, 1984, et al.
(författare)
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Specific xylan activity revealed for AA9 Lytic Polysaccharide Monooxygenases of the thermophilic fungus Malbranchea cinnamomea by functional characterization
- 2019
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Ingår i: Applied and Environmental Microbiology. - 1098-5336 .- 0099-2240. ; 85:23
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Tidskriftsartikel (refereegranskat)abstract
- The thermophilic biomass-degrader Malbranchea cinnamomea exhibits poor growth on cellulose but excellent growth on hemicelluloses as the sole carbon source. This is surprising considering that its genome encodes eight lytic polysaccharide monooxygenases (LPMOs) from auxiliary activity family 9 (AA9), enzymes known for their high potential in accelerating cellulose depolymerization. We characterized four of the eight ( M. cinnamomea AA9s) Mc AA9s, namely, Mc AA9A, Mc AA9B, Mc AA9F, and Mc AA9H, to gain a deeper understanding about their roles in the fungus. The characterized Mc AA9s were active on hemicelluloses, including xylan, glucomannan, and xyloglucan, and furthermore, in accordance with transcriptomics data, differed in substrate specificity. Of the Mc AA9s, Mc AA9H is unique, as it preferentially cleaves residual xylan in phosphoric acid-swollen cellulose (PASC). Moreover, when exposed to cellulose-xylan blends, Mc AA9H shows a preference for xylan and for releasing (oxidized) xylooligosaccharides. The cellulose dependence of the xylan activity suggests that a flat conformation, with rigidity similar to that of cellulose microfibrils, is a prerequisite for productive interaction between xylan and the catalytic surface of the LPMO. Mc AA9H showed a similar trend on xyloglucan, underpinning the suggestion that LPMO activity on hemicelluloses strongly depends on the polymers’ physicochemical context and conformation. Our results support the notion that LPMO multiplicity in fungal genomes relates to the large variety of copolymeric polysaccharide arrangements occurring in the plant cell wall.
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| 9. |
- Mascher, Martin, et al.
(författare)
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A chromosome conformation capture ordered sequence of the barley genome
- 2017
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Ingår i: Nature. - : Springer Science and Business Media LLC. - 0028-0836 .- 1476-4687. ; 544:7651, s. 427-433
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Tidskriftsartikel (refereegranskat)abstract
- Cereal grasses of the Triticeae tribe have been the major food source in temperate regions since the dawn of agriculture. Their large genomes are characterized by a high content of repetitive elements and large pericentromeric regions that are virtually devoid of meiotic recombination. Here we present a high-quality reference genome assembly for barley (Hordeum vulgare L.). We use chromosome conformation capture mapping to derive the linear order of sequences across the pericentromeric space and to investigate the spatial organization of chromatin in the nucleus at megabase resolution. The composition of genes and repetitive elements differs between distal and proximal regions. Gene family analyses reveal lineage-specific duplications of genes involved in the transport of nutrients to developing seeds and the mobilization of carbohydrates in grains. We demonstrate the importance of the barley reference sequence for breeding by inspecting the genomic partitioning of sequence variation in modern elite germplasm, highlighting regions vulnerable to genetic erosion.
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| 10. |
- Wishart, David S., et al.
(författare)
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HMDB 5.0 : the Human Metabolome Database for 2022
- 2022
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Ingår i: Nucleic Acids Research. - : Oxford University Press (OUP). - 0305-1048 .- 1362-4962. ; 50:D1, s. D622-D631
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Tidskriftsartikel (refereegranskat)abstract
- The Human Metabolome Database or HMDB (https://hmdb.ca) has been providing comprehensive reference information about human metabolites and their associated biological, physiological and chemical properties since 2007. Over the past 15 years, the HMDB has grown and evolved significantly to meet the needs of the metabolomics community and respond to continuing changes in internet and computing technology. This year's update, HMDB 5.0, brings a number of important improvements and upgrades to the database. These should make the HMDB more useful and more appealing to a larger cross-section of users. In particular, these improvements include: (i) a significant increase in the number of metabolite entries (from 114 100 to 217 920 compounds); (ii) enhancements to the quality and depth of metabolite descriptions; (iii) the addition of new structure, spectral and pathway visualization tools; (iv) the inclusion of many new and much more accurately predicted spectral data sets, including predicted NMR spectra, more accurately predicted MS spectra, predicted retention indices and predicted collision cross section data and (v) enhancements to the HMDB's search functions to facilitate better compound identification. Many other minor improvements and updates to the content, the interface, and general performance of the HMDB website have also been made. Overall, we believe these upgrades and updates should greatly enhance the HMDB's ease of use and its potential applications not only in human metabolomics but also in exposomics, lipidomics, nutritional science, biochemistry and clinical chemistry.
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