| 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. |
- Margos, Gabriele, et al.
(författare)
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Rejection of the name Borreliella and all proposed species comb. nov. placed therein
- 2020
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Ingår i: International Journal of Systematic and Evolutionary Microbiology. - : MICROBIOLOGY SOC. - 1466-5026 .- 1466-5034. ; 70:5, s. 3577-3581
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Tidskriftsartikel (refereegranskat)abstract
- Rejection (nomen rejiciendum) of the name Borreliella and all new combinations therein is being requested on grounds of risk to human health and patient safety (Principle 1, subprinciple 2 and Rule 56a) and violation to aim for stability of names, to avoid useless creation of names (Principle 1, subprinciple 1 and 3) and that names should not be changed without sufficient reason (Principle 9 of the International Code of Nomenclature of Prokaryotes).
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| 3. |
- Marciano, Sabina, et al.
(författare)
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Combining CRISPR-Cas9 and brain imaging to study the link from genes to molecules to networks
- 2022
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Ingår i: Proceedings of the National Academy of Sciences of the United States of America. - : Proceedings of the National Academy of Sciences. - 0027-8424. ; 119:40
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Tidskriftsartikel (refereegranskat)abstract
- Receptors, transporters, and ion channels are important targets for therapy development in neurological diseases, but their mechanistic role in pathogenesis is often poorly understood. Gene editing and in vivo imaging approaches will help to identify the molecular and functional role of these targets and the consequence of their regional dysfunction on the whole-brain level. We combine CRISPR-Cas9 gene editing with in vivo positron emission tomography (PET) and functional MRI (fMRI) to investigate the direct link between genes, molecules, and the brain connectome. The extensive knowledge of the Slc18a2 gene encoding the vesicular monoamine transporter (VMAT2), involved in the storage and release of dopamine, makes it an excellent target for studying the gene network relationships while structurally preserving neuronal integrity and function. We edited the Slc18a2 in the substantia nigra pars compacta of adult rats and used in vivo molecular imaging besides behavioral, histological, and biochemical assessments to characterize the CRISPR-Cas9–mediated VMAT2 knockdown. Simultaneous PET/fMRI was performed to investigate molecular and functional brain alterations. We found that stage-specific adaptations of brain functional connectivity follow the selective impairment of presynaptic dopamine storage and release. Our study reveals that recruiting different brain networks is an early response to the dopaminergic dysfunction preceding neuronal cell loss. Our combinatorial approach is a tool to investigate the impact of specific genes on brain molecular and functional dynamics, which will help to develop tailored therapies for normalizing brain function.
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| 4. |
- Margos, Gabriele, et al.
(författare)
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Borrelia Ecology, Evolution, and Human Disease : A Mosaic of Life
- 2023. - 2
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Ingår i: Zoonoses. - Cham : Springer International Publishing. - 9783030858773 - 9783030858773 ; , s. 1087-1151
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Bokkapitel (refereegranskat)abstract
- Lyme borreliosis (LB) and relapsing fever (RF) are zoonotic diseases that are caused by spirochetal bacteria belonging to the genus Borrelia. The agents are generally maintained in natural transmission cycles by vector ticks (exception: body louse) and reservoir hosts. Lyme borreliosis (synonym in North America: Lyme disease, LD) is the most frequently reported tick-borne disease in Europe and North America. It mainly affects skin, large joints, nervous system or heart and is considered a multi-system disorder. Relapsing fever manifests as recurrent febrile attacks accompanied by headaches, muscle and joint aches, interrupted by afebrile intervals. It mainly occurs in tropical and subtropical regions including North and South America, Africa, Asia, and South European countries. In this chapter we describe the genus Borrelia, the huge diversity that has become apparent in recent years, the geographical distribution of its species, and the complex genome that is reflected in the complex ecology and disease symptoms. We also give information on diagnostics and its challenges, therapy, and prophylactic measures.
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| 5. |
- Margos, Gabriele, et al.
(författare)
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Evidence of taxonomic bias in public databases: The example of the genus Borrelia
- 2022
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Ingår i: Ticks and Tick-borne Diseases. - : ELSEVIER GMBH. - 1877-959X .- 1877-9603. ; 13:5
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Tidskriftsartikel (refereegranskat)abstract
- The taxon names used in public databases are of critical importance in all areas of biology because they are needed for linking organisms to sequence data and other information. Since most users of taxonomic classifications may be unprepared for dealing with synonyms, the names that are preferred in such databases are of high impact. Using the genus Borrelia as an example, we here show how simplistic approaches for determining the preferred synonym may lead to biases regarding the preferences for taxonomic opinions. We highlight that in this and other cases where genera were split, for reverting to the previous "merged" genus it is neither possible nor necessary to generate validly published and legitimate names that are newer than those that were proposed as new combinations when the genus was split. The policy to always prefer the latest validly published name in a public database may thus render this database oblivious to reversals in taxonomic opinion. We emphasize that users of public databases should be aware of such potential shortcomings, and that curators of databases which provide nomenclatural information should be open-minded about taxonomic views expressed in the literature.
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