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1.
  • Schulze, Yves, et al. (författare)
  • Chemical-genomic profiling identifies genes that protect yeast from aluminium, gallium, and indium toxicity
  • 2023
  • Ingår i: Metallomics. - : Oxford University Press. - 1756-5901 .- 1756-591X. ; 15:6
  • Tidskriftsartikel (refereegranskat)abstract
    • Aluminium, gallium, and indium are group 13 metals with similar chemical and physical properties. While aluminium is one of the most abundant elements in the Earth's crust, gallium and indium are present only in trace amounts. However, the increased use of the latter metals in novel technologies may result in increased human and environmental exposure. There is mounting evidence that these metals are toxic, but the underlying mechanisms remain poorly understood. Likewise, little is known about how cells protect themselves from these metals. Aluminium, gallium, and indium are relatively insoluble at neutral pH, and here we show that they precipitate in yeast culture medium at acidic pH as metal-phosphate species. Despite this, the dissolved metal concentrations are sufficient to induce toxicity in the yeast Saccharomyces cerevisiae. By chemical-genomic profiling of the S. cerevisiae gene deletion collection, we identified genes that maintain growth in the presence of the three metals. We found both shared and metal-specific genes that confer resistance. The shared gene products included functions related to calcium metabolism and Ire1/Hac1-mediated protection. Metal-specific gene products included functions in vesicle-mediated transport and autophagy for aluminium, protein folding and phospholipid metabolism for gallium, and chorismate metabolic processes for indium. Many of the identified yeast genes have human orthologues involved in disease processes. Thus, similar protective mechanisms may act in yeast and humans. The protective functions identified in this study provide a basis for further investigations into toxicity and resistance mechanisms in yeast, plants, and humans. © 2023 The Author(s). 
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3.
  • Urbancsok, János, et al. (författare)
  • Flexure wood formation via growth reprogramming in hybrid aspen involves jasmonates and polyamines and transcriptional changes resembling tension wood development
  • 2023
  • Ingår i: New Phytologist. - : John Wiley & Sons. - 0028-646X .- 1469-8137. ; 240:6, s. 2312-2334
  • Tidskriftsartikel (refereegranskat)abstract
    • Stem bending in trees induces flexure wood but its properties and development are poorly understood. Here, we investigated the effects of low-intensity multidirectional stem flexing on growth and wood properties of hybrid aspen, and on its transcriptomic and hormonal responses.Glasshouse-grown trees were either kept stationary or subjected to several daily shakes for 5 wk, after which the transcriptomes and hormones were analyzed in the cambial region and developing wood tissues, and the wood properties were analyzed by physical, chemical and microscopy techniques.Shaking increased primary and secondary growth and altered wood differentiation by stimulating gelatinous-fiber formation, reducing secondary wall thickness, changing matrix polysaccharides and increasing cellulose, G- and H-lignin contents, cell wall porosity and saccharification yields. Wood-forming tissues exhibited elevated jasmonate, polyamine, ethylene and brassinosteroids and reduced abscisic acid and gibberellin signaling. Transcriptional responses resembled those during tension wood formation but not opposite wood formation and revealed several thigmomorphogenesis-related genes as well as novel gene networks including FLA and XTH genes encoding plasma membrane-bound proteins.Low-intensity stem flexing stimulates growth and induces wood having improved biorefinery properties through molecular and hormonal pathways similar to thigmomorphogenesis in herbaceous plants and largely overlapping with the tension wood program of hardwoods.
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4.
  • Zhang, Heyang, et al. (författare)
  • Together is Better : mRNA Co-Encapsulation in Lipoplexes is Required to Obtain Ratiometric Co-Delivery and Protein Expression on the Single Cell Level
  • 2022
  • Ingår i: Advanced Science. - : John Wiley and Sons Inc. - 2198-3844. ; 9:4
  • Tidskriftsartikel (refereegranskat)abstract
    • Liposomes can efficiently deliver messenger RNA (mRNA) into cells. When mRNA cocktails encoding different proteins are needed, a considerable challenge is to efficiently deliver all mRNAs into the cytosol of each individual cell. In this work, two methods are explored to co-deliver varying ratiometric doses of mRNA encoding red (R) or green (G) fluorescent proteins and it is found that packaging mRNAs into the same lipoplexes (mingle-lipoplexes) is crucial to efficiently deliver multiple mRNA types into the cytosol of individual cells according to the pre-defined ratio. A mixture of lipoplexes containing only one mRNA type (single-lipoplexes), however, seem to follow the “first come – first serve” principle, resulting in a large variation of R/G uptake and expression levels for individual cells leading to ratiometric dosing only on the population level, but rarely on the single-cell level. These experimental observations are quantitatively explained by a theoretical framework based on the stochasticity of mRNA uptake in cells and endosomal escape of mingle- and single-lipoplexes, respectively. Furthermore, the findings are confirmed in 3D retinal organoids and zebrafish embryos, where mingle-lipoplexes outperformed single-lipoplexes to reliably bring both mRNA types into single cells. This benefits applications that require a strict control of protein expression in individual cells. © 2021 The Authors. 
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5.
  • Albers, Eva, 1966, et al. (författare)
  • Selective suppression of bacterial contaminants by process conditions during lignocellulose based yeast fermentations
  • 2011
  • Ingår i: Biotechnology for Biofuels. - : Springer Science and Business Media LLC. - 1754-6834 .- 1754-6834. ; 4
  • Tidskriftsartikel (refereegranskat)abstract
    • BackgroundContamination of bacteria in large-scale yeast fermentations is a serious problem and a threat to the development of successful biofuel production plants. Huge research efforts have been spent in order to solve this problem, but additional ways must still be found to keep bacterial contaminants from thriving in these environments. The aim of this project was to develop process conditions that would inhibit bacterial growth while giving yeast a competitive advantage.ResultsLactic acid bacteria are usually considered to be the most common contaminants in industrial yeast fermentations. Our observations support this view but also suggest that acetic acid bacteria, although not so numerous, could be a much more problematic obstacle to overcome. Acetic acid bacteria showed a capacity to drastically reduce the viability of yeast. In addition, they consumed the previously formed ethanol. Lactic acid bacteria did not show this detrimental effect on yeast viability. It was possible to combat both types of bacteria by a combined addition of NaCl and ethanol to the wood hydrolysate medium used. As a result of NaCl + ethanol additions the amount of viable bacteria decreased and yeast viability was enhanced concomitantly with an increase in ethanol concentration. The successful result obtained via addition of NaCl and ethanol was also confirmed in a real industrial ethanol production plant with its natural inherent yeast/bacterial community.ConclusionsIt is possible to reduce the number of bacteria and offer a selective advantage to yeast by a combined addition of NaCl and ethanol when cultivated in lignocellulosic medium such as wood hydrolysate. However, for optimal results, the concentrations of NaCl + ethanol must be adjusted to suit the challenges offered by each hydrolysate.
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6.
  • Ghiaci, Payam, et al. (författare)
  • Highly parallelized laboratory evolution of wine yeasts for enhanced metabolic phenotypes
  • 2024
  • Ingår i: MOLECULAR SYSTEMS BIOLOGY. - : Springer Nature. - 1744-4292.
  • Tidskriftsartikel (refereegranskat)abstract
    • Adaptive Laboratory Evolution (ALE) of microorganisms can improve the efficiency of sustainable industrial processes important to the global economy. However, stochasticity and genetic background effects often lead to suboptimal outcomes during laboratory evolution. Here we report an ALE platform to circumvent these shortcomings through parallelized clonal evolution at an unprecedented scale. Using this platform, we evolved 10(4) yeast populations in parallel from many strains for eight desired wine fermentation-related traits. Expansions of both ALE replicates and lineage numbers broadened the evolutionary search spectrum leading to improved wine yeasts unencumbered by unwanted side effects. At the genomic level, evolutionary gains in metabolic characteristics often coincided with distinct chromosome amplifications and the emergence of side-effect syndromes that were characteristic of each selection niche. Several high-performing ALE strains exhibited desired wine fermentation kinetics when tested in larger liquid cultures, supporting their suitability for application. More broadly, our high-throughput ALE platform opens opportunities for rapid optimization of microbes which otherwise could take many years to accomplish.
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7.
  • Sepehri, Sobhan, 1986, et al. (författare)
  • Characterization of Binding of Magnetic Nanoparticles to Rolling Circle Amplification Products by Turn-On Magnetic Assay
  • 2019
  • Ingår i: Biosensors-Basel. - : MDPI AG. ; 9:3
  • Tidskriftsartikel (refereegranskat)abstract
    • The specific binding of oligonucleotide-tagged 100 nm magnetic nanoparticles (MNPs) to rolling circle products (RCPs) is investigated using our newly developed differential homogenous magnetic assay (DHMA). The DHMA measures ac magnetic susceptibility from a test and a control samples simultaneously and eliminates magnetic background signal. Therefore, the DHMA can reveal details of binding kinetics of magnetic nanoparticles at very low concentrations of RCPs. From the analysis of the imaginary part of the DHMA signal, we find that smaller MNPs in the particle ensemble bind first to the RCPs. When the RCP concentration increases, we observe the formation of agglomerates, which leads to lower number of MNPs per RCP at higher concentrations of RCPs. The results thus indicate that a full frequency range of ac susceptibility observation is necessary to detect low concentrations of target RCPs and a long amplification time is not required as it does not significantly increase the number of MNPs per RCP. The findings are critical for understanding the underlying microscopic binding process for improving the assay performance. They furthermore suggest DHMA is a powerful technique for dynamically characterizing the binding interactions between MNPs and biomolecules in fluid volumes.
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8.
  • Warwas, Niklas, et al. (författare)
  • Marine yeast (Candida sake) cultured on herring brine side streams is a promising feed ingredient and omega-3 source for rainbow trout (Oncorhynchus mykiss)
  • 2023
  • Ingår i: Aquaculture. - : Elsevier BV. - 0044-8486 .- 1873-5622. ; 571
  • Tidskriftsartikel (refereegranskat)abstract
    • A major challenge for the aquaculture industry is the supply of sustainable feeds. A promising model to achieve this is to utilize circular flows where feed ingredients, such as single cell protein, are cultivated using side streams of the food industry. The aim of this study was to evaluate the marine yeast Candida sake, produced on herring brine side streams, as a source of protein and immune stimulant in feed for salmonid fish. The dry C. sake product contained 54% protein (3.3% lysine and 0.8% methionine) and 13% lipids (1.1% eicosapentaenoic, EPA, and 1% docosahexaenoic acid, DHA). Four experimental diets were designed and tested in a 9-week feeding trial using juvenile rainbow trout (Oncorhynchus mykiss). A control diet containing both fish and plant-based ingredients constituted the base feed to which 20% (to evaluate effects on digestibility, growth and intestinal physiology), 20% heat-treated (to evaluate effects of downstream processing) and 3% (to evaluate immune stimulatory properties, replacing 3% soy protein concentrate) C. sake was added. The apparent digestibility coefficient of C. sake for protein, fat and gross energy was above 80%, and for amino acids above 90% regardless of treatment, suggesting a high bioavailability of C. sake. All three yeast containing diets performed equally to the control regarding specific growth rate, feed conversion ratio and functional intestinal health. These results suggest that C. sake is a promising alternative protein source for circular feeds in the salmonid industry. The presence of EPA and DHA represents an added value. The heat treatment increased the apparent digestibility coefficient of dry matter by 8% but decreased amino acid digestibility by on average 3%, indicating that heat treatment may not be the optimal downstream processing technique. Furthermore, the inclusion of 3% C. sake increased the intestinal lamina propria width and TGF-beta transcription, indicating an immune stimulating effect. Future research is needed to understand these immune modulatory effects of C. sake supplementation.
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9.
  • Berta, Marco, et al. (författare)
  • Effect of zein protein and hydroxypropyl methylcellulose on the texture of model gluten-free bread
  • 2019
  • Ingår i: Journal of texture studies. - : Blackwell Publishing Ltd. - 0022-4901 .- 1745-4603. ; 50:4, s. 341-349
  • Tidskriftsartikel (refereegranskat)abstract
    • The influence of zein protein and hydroxypropyl methylcellulose (HPMC) on the texture and volume of gluten-free bread was investigated. The addition of HPMC to starch affected the dough viscoelasticity and it improved the bread volume during baking since it acts as an emulsifier. The addition of zein protein to gluten-free bread increased the crumb firmness and reduced the crust hardness within the range of concentrations investigated. No zein protein network could be observed in the bread crumb. The zein protein, cold mixed at low concentration, did not enhance the dough elasticity. Due to the lack of a protein network noncovalent interactions may stabilize the bubble structure stabilization within the crumb, rather than covalent links of the protein chain. With an optimized amount of zein protein and HPMC hydrocolloid, the gluten-free bread showed similar texture and staling behavior to that of model wheat bread. The optimized recipe, compiled into a spreadsheet, is available in the supporting information. The microstructural observations suggest that zein could be replaced with another protein for this recipe resulting in a similar bread texture.
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10.
  • Carlred, Louise, et al. (författare)
  • Imaging of amyloid-β in alzheimer’s disease transgenic mouse brains with ToF-SIMS using immunoliposomes
  • 2016
  • Ingår i: Biointerphases. - : American Institute of Physics (AIP). - 1934-8630 .- 1559-4106. ; 11:2, s. 1-11
  • Tidskriftsartikel (refereegranskat)abstract
    • Time-of-flight secondary ion mass spectrometry (ToF-SIMS) has been proven to successfully image different kinds of molecules, especially a variety of lipids, in biological samples. Proteins, however, are difficult to detect as specific entities with this method due to extensive fragmentation. To circumvent this issue, the authors present in this work a method developed for detection of proteins using antibody-conjugated liposomes, so called immunoliposomes, which are able to bind to the specific protein of interest. In combination with the capability of ToF-SIMS to detect native lipids in tissue samples, this method opens up the opportunity to analyze many different biomolecules, both lipids and proteins, at the same time, with high spatial resolution. The method has been applied to detect and image the distribution of amyloid-β (Aβ), a biologically relevant peptide in Alzheimer’s disease (AD), in transgenic mouse brain tissue. To ensure specific binding, the immunoliposome binding was verified on a model surface using quartz crystal microbalance with dissipation monitoring. The immunoliposome binding was also investigated on tissue sections with fluorescence microscopy, and compared with conventional immunohistochemistry using primary and secondary antibodies, demonstrating specific binding to Aβ. Using ToF-SIMS imaging, several endogenous lipids, such as cholesterol and sulfatides, were also detected in parallel with the immunoliposome-labeled Aβ deposits, which is an advantage compared to fluorescence microscopy. This method can thus potentially provide further information about lipid–protein interactions, which is important to understand the mechanisms of neurodegeneration in AD.
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