| 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. |
- Klionsky, Daniel J., et al.
(författare)
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Guidelines for the use and interpretation of assays for monitoring autophagy
- 2012
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Ingår i: Autophagy. - : Informa UK Limited. - 1554-8635 .- 1554-8627. ; 8:4, s. 445-544
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Forskningsöversikt (refereegranskat)abstract
- In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. A key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process vs. those that measure flux through the autophagy pathway (i.e., the complete process); thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from stimuli that result in increased autophagic activity, defined as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (in most higher eukaryotes and some protists such as Dictyostelium) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the field understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field.
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| 3. |
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| 4. |
- Bäcklund, Tomas, et al.
(författare)
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Trunk sway in idiopathic normal pressure hydrocephalus : quantitative assessment in clinical practice
- 2017
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Ingår i: Gait & Posture. - : Elsevier. - 0966-6362 .- 1879-2219. ; , s. 62-70
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Tidskriftsartikel (refereegranskat)abstract
- Background: In diagnosis and treatment of patients with idiopathic normal pressure hydrocephalus (iNPH), there is need for clinically applicable, quantitative assessment of balance and gait. Using a body worn gyroscopic system, the aim of this study was to assess postural stability of iNPH patients in standing, walking and during sensory deprivation before and after cerebrospinal fluid (CSF) drainage and surgery. A comparison was performed between healthy elderly (HE) and patients with various types of hydrocephalus (ventriculomegaly (VM)).Methods: Trunk sway was measured in 31 iNPH patients, 22 VM patients and 58 HE. Measurements were performed at baseline in all subjects, after CSF drainage in both patient groups and after shunt surgery in the iNPH group.Results: Preoperatively, the iNPH patients had significantly higher trunk sway compared to HE, specifically for the standing tasks (p < 0.001). Compared to VM, iNPH patients had significantly lower sway velocity during gait in three of four cases on firm support (p < 0.05). Sway velocity improved after CSF drainage and in forward-backward direction after surgery (p < 0.01). Compared to HE both patient groups demonstrated less reliance on visual input to maintain stable posture.Conclusions: INPH patients had reduced postural stability compared to HE, particularly during standing, and for differentiation between iNPH and VM patients sway velocity during gait is a promising parameter. A reversible reduction of visual incorporation during standing was also seen. Thus, the gyroscopic system quantitatively assessed postural deficits in iNPH, making it a potentially useful tool for aiding in future diagnoses, choices of treatment and clinical follow-up.
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| 5. |
- Frankel, Jennifer, et al.
(författare)
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Assessing exposures to Magnetic resonance imaging's complex Mixture of Magnetic Fields for In Vivo, In Vitro, and epidemiologic studies of Health effects for staff and Patients
- 2018
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Ingår i: Frontiers In Public Health. - : Frontiers Media S.A.. - 2296-2565. ; 6
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Tidskriftsartikel (refereegranskat)abstract
- A complex mixture of electromagnetic fields is used in magnetic resonance imaging (MRI): static, low-frequency, and radio frequency magnetic fields. Commonly, the static magnetic field ranges from one to three Tesla. The low-frequency field can reach several millitesla and with a time derivative of the order of some Tesla per second. The radiofrequency (RF) field has a magnitude in the microtesla range giving rise to specific absorption rate values of a few Watts per kilogram. Very little attention has been paid to the case where there is a combined exposure to several different fields at the same time. Some studies have shown genotoxic effects in cells after exposure to an MRI scan while others have not demonstrated any effects. A typical MRI exam includes muliple imaging sequences of varying length and intensity, to produce different types of images. Each sequence is designed with a particular purpose in mind, so one sequence can, for example, be optimized for clearly showing fat water contrast, while another is optimized for high-resolution detail. It is of the utmost importance that future experimental studies give a thorough description of the exposure they are using, and not just a statement such as "An ordinary MRI sequence was used." Even if the sequence is specified, it can differ substantially between manufacturers on, e.g., RF pulse height, width, and duty cycle. In the latest SCENIHR opinion, it is stated that there is very little information regarding the health effects of occupational exposure to MRI fields, and long-term prospective or retrospective cohort studies on workers are recommended as a high priority. They also state that MRI is increasingly used in pediatric diagnostic imaging, and a cohort study into the effects of MRI exposure on children is recommended as a high priority. For the exposure assessment in epidemiological studies, there is a clear difference between patients and staff and further work is needed on this. Studies that explore the possible differences between MRI scan sequences and compare them in terms of exposure level are warranted.
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| 6. |
- Frankel, Jennifer, 1981-
(författare)
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Characterization of the MRI patient exposure environment and exposure assessment methods for magnetic fields in MRI scanners
- 2021
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Magnetic resonance imaging (MRI) has become one of the most common imaging modalities available in modern medicine, and it is an indispensable diagnostic tool thanks to the unparalleled soft-tissue contrast and high image resolution. It is also a unique exposure environment consisting of a complex mix of magnetic fields. During an MRI scan, the patient is simultaneously exposed to a strong static magnetic field, a fast-switching gradient magnetic field, and a pulsed radiofrequency (RF) magnetic field. Transient acute effects, such as nerve excitation and tissue heating, are well known and limited by universal safety guidelines. Long-term health effects related to MRI exposure have, however, not been scientifically established, and no interaction mechanisms have been verified, despite a growing body of research on electromagnetic field exposure. Further epidemiological and experimental research on MRI exposure has been recommended but the lack of a common definition of dose or exposure metric makes evaluation of past research and the design of future experiments difficult.The objectives of this thesis were to characterize the MRI patient exposure environment in terms of the magnetic fields involved, suggest relevant exposure metrics, and introduce exposure assessment methods suitable for epidemiological and experimental research on MRI and long-term health effects.In Paper I, we discussed the MRI exposure environment and its complexity and gave an overview of the current scientific situation. In Paper II, we investigated the exposure variability between different MRI sequences and suggested patient-independent exposure metrics that describe different characteristics of the magnetic field exposure, including mean, peak, and threshold values. In Paper III, we presented three exposure assessment methods, specifically suited to the complex MRI exposure environment: a measurement-based method, a calculation-based method, and a proxy method.Papers I and II showed that MRI exams are not homogenous in terms of exposure, and exposure variability exists between the individual sequences that comprise an exam. Differences in mean exposure between sequences were several-fold, peak exposure differences up to 30-fold, and differences in threshold exposure were in some cases more than 100-fold. Furthermore, within-sequence exposure variability, related to the parameter adjustments that can be made at the scanner console before the start of a scan, gave rise to 5-to-8-fold exposure increases. Paper III showed that magnetic field models could be used to approximate the exposure at arbitrary locations inside the scanner, with slight underestimation of gradient field metrics and large variability in some RF field metrics. With improvements in accuracy and efficiency, the method could become a useful exposure assessment tool for in vitro and in vivo research as well as clinical work on medical implant safety. Our search for suitable exposure metric proxies resulted in a limited selection with low correlation between proxies and their counterpart metrics, but, with further development, the proxy method has the potential to allow for much needed exposure classification relevant to large-scale epidemiological research.The work in this thesis has contributed to increased awareness of the unique MRI exposure environment, the characteristics of the magnetic fields involved, and the inherent exposure variability in MRI exams. The metrics and methods presented are specifically suited to exposure assessment of the unique MRI environment, and may contribute to improved research quality by allowing for meaningful comparisons between study results and for experimental conditions to be easily replicated in future studies.
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| 7. |
- Frankel, Jennifer, et al.
(författare)
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EMF exposure variation among MRI sequences from pediatric examination protocols
- 2019
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Ingår i: Bioelectromagnetics. - : Wiley-Blackwell. - 0197-8462 .- 1521-186X. ; 40:1, s. 3-15
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Tidskriftsartikel (refereegranskat)abstract
- The magnetic resonance imaging (MRI) exposure environment is unique due to the mixture and intensity of magnetic fields involved. Current safety regulations are based on well-known acute effects of heating and neuroexcitation while the scientific grounds for possible long-term effects from MRI exposure are lacking. Epidemiological research requires careful exposure characterization, and as a first step toward improved exposure assessment we set out to characterize the MRI-patient exposure environment. Seven MRI sequences were run on a 3-Tesla scanner while the radiofrequency and gradient magnetic fields were measured inside the scanner bore. The sequences were compared in terms of 14 different exposure parameters. To study within-sequence variability, we varied sequence settings such as flip angle and slice thickness one at a time, to determine if they had any impact on exposure endpoints. There were significant differences between two or more sequences for all fourteen exposure parameters. Within-sequence differences were up to 60% of the corresponding between-sequence differences, and a 5-8 fold exposure increase was caused by variations in flip angle, slice spacing, and field of view. MRI exposure is therefore not only sequence-specific but also patient- and examination occurrence-specific, a complexity that requires careful consideration for an MRI exposure assessment in epidemiological studies to be meaningful.
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| 8. |
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| 9. |
- Frankel, Jennifer, et al.
(författare)
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Measurements of the switched gradient magnetic field in MRI : A closer look at some unintuitive spatial characteristics
- 2023
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Ingår i: iRADIOLOGY. - : John Wiley & Sons. - 2834-2860 .- 2834-2879. ; 1:4, s. 390-396
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Tidskriftsartikel (refereegranskat)abstract
- Concomitant fields are the unwanted transverse components that arise when spatial encoding gradients are applied in MRI. We measured the changing gradient magnetic field at multiple locations inside the scanner and examined the internal distribution and linearity of the three vector components of the field. Our results illustrate some not-so-obvious spatial characteristics of the gradient field, which can seem unintuitive at first glance, but are quite reasonable when considering electromagnetic theory and MRI-scanner physics constraints.
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| 10. |
- Mild, Kjell Hansson, et al.
(författare)
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Exposure to the magnetic field from an induction loop pad for a hearing aid system
- 2017
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Ingår i: International Journal of Occupational Safety and Ergonomics. - : Informa UK Limited. - 1080-3548 .- 2376-9130. ; 23:1, s. 143-145
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Tidskriftsartikel (refereegranskat)abstract
- As a case study we have measured the magnetic field from an induction loop pad designed for hearing aid assistance. The magnitude of the field was high, although well below international guidelines. We recorded values up to 70% of the recommended standard in some instances. However, in view of the many reports indicating health effects of low-level exposure, we recommend that the precautionary principle is applied when such pads are given to people who might be especially vulnerable, such as children, pregnant women and women on breast cancer medication.
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