| 1. |
|
|
| 2. |
- Beal, Jacob, et al.
(författare)
-
Robust estimation of bacterial cell count from optical density
- 2020
-
Ingår i: Communications Biology. - : Springer Science and Business Media LLC. - 2399-3642. ; 3:1
-
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.
|
|
| 3. |
- Klionsky, Daniel J., et al.
(författare)
-
Guidelines for the use and interpretation of assays for monitoring autophagy
- 2012
-
Ingår i: Autophagy. - : Informa UK Limited. - 1554-8635 .- 1554-8627. ; 8:4, s. 445-544
-
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.
|
|
| 4. |
- Ulyanov, A.N., et al.
(författare)
-
La0.7Ca0.3-xSrxMnO3 manganites: Effect of structure on the magnetic and transport properties.
- 2002
-
Ingår i: Journal of the Physical Society of Japan. - : The Physical Society of Japan. - 0031-9015 .- 1347-4073. ; 71:3, s. 927-929
-
Tidskriftsartikel (refereegranskat)abstract
- The effect of lattice structure on the magnetic and transport properties of La0.7Ca0.3-xSrxMnO3 (x= 0; 0.03; 0.06; ...; 0.3) lanthanum manganites have been studied. Curie temperature pressure coefficient, dTc/dP, showed the dependence on x value. Temperature Tc increased almost linearly under applied pressure as dTc/dP≈14 K/GPa and as dTc/dP≈7.5 K/GPa in the orthorhombic (Pbnm) and rhombohedral (R3c) phases, respectively. The value of dTc/dP(≈3.5 K/GPa) showed a minimum and a temperature of the resistance maximum, Tp(x), showed a change of slope at x=0.15, corresponding to the concentration structural phase transition. Differences between the values of dTc/dP and slopes of Tp versus x in Pbnm and R3c phases are explained by the different effect of external pressure on the Mn–O bond length and Mn–O–Mn bond angle, and by the different internal pressure effect in those phases, respectively
|
|
| 5. |
- Ulyanov, A. N., et al.
(författare)
-
Structure and pressure effects on the properties of La0.7Ca0.3-xSrxMnO3 manganites.
- 2002
-
Ingår i: Journal of Applied Physics vol. 91. - : American Institute of Physics. ; , s. 7739-7741
-
Konferensbidrag (refereegranskat)abstract
- We report a study of the structure and pressure effects on magnetic and transport properties of a lanthanum manganite, La0.7Ca0.3–xSrxMnO3 (0<=x<=0.3, Deltax = 0.03). The pressure coefficient of the Curie temperature (Tc), dTc/dP, is shown to depend on x. The temperature Tc increases approximately linearly under applied pressure as dTc/dP[approximate]14 K/GPa in the orthorhombic (Pbnm) phase and as dTc/dP[approximate]7.5 K/GPa in the rhombohedral (R3-bar c). The value of dTc/dP shows a minimum ([approximate]3.5 K/GPa) and the temperature of the resistance maximum, Tp(x), shows a change of slope at x = 0.15, corresponding to a concentration structural phase transition. Differences between the values of dTc/dP and the slopes of Tp vs x in Pbnm and R3-bar c phases are explained by the different effect of external pressure on the Mn–O bond length and the Mn–O–Mn bond angle, and by the different internal pressure effect (depending on the value of x) in those phases, respectively.
|
|
| 6. |
- Cheon, Hwanju, et al.
(författare)
-
Role of JNK activation in pancreatic β-cell death by streptozotocin
- 2010
-
Ingår i: Molecular and Cellular Endocrinology. - 0303-7207. ; 321:2, s. 131-137
-
Tidskriftsartikel (refereegranskat)abstract
- c-Jun N-terminal kinase (JNK) is activated by cellular stress and plays critical roles in diverse types of cell death. However, role of JNK in β-cell injury is obscure. We investigated the role for JNK in streptozotocin (STZ)-induced β-cell death. STZ induced JNK activation in insulinoma or islet cells. JNK inhibitors attenuated insulinoma or islet cell death by STZ. STZ-induced JNK activation was decreased by PARP inhibitors, suggesting that JNK activation is downstream of PARP-1. Phosphatase inhibitors induced activation of JNK and abrogated the suppression of STZ-induced JNK activation by PARP inhibitors, suggesting that the inhibition of phosphatases is involved in the activation of JNK by STZ. STZ induced production of reactive oxygen species (ROS) as potential inhibitors of phosphatases, which was suppressed by PARP inhibitors. PARP-1 siRNA attenuated insulinoma cell death and JNK activation after STZ treatment, which was reversed by MKP (MAP kinase phosphatase)-1 siRNA. These results suggest that JNK is activated by STZ downstream of PARP-1 through inactivation of phosphatases such as MKP, which plays important roles in STZ-induced β-cell death.
|
|
| 7. |
- Ulyanov, A.N., et al.
(författare)
-
Crystal symmetry and pressure effects on the properties of mixed-valence manganites
- 2003
-
Ingår i: Journal of Magnetism and Magnetic Materials vol. 258-259, Special Issue SI. - Amsterdam : Elsevier B.V.. ; , s. 312-314
-
Konferensbidrag (refereegranskat)abstract
- A lattice structure effect on the magnetic and magnetotransport properties of La0.7Ca(0.3-x)SrxMnO3 lanthanum manganites have been studied. The Curie temperature pressure coefficient increased almost linearly under an applied pressure as dTc/dP=14K/GPa and as dTc/dP=7.5K/GPa in the orthorhombic and rhombohedral structures,respectively. This is caused by the different pressure effect on the Mn–O bond distances and on the Mn–O–Mn bond angles in those phases. The anomalous low value of dTc/dP(=3.5K/GPa) for La0.7Ca0.15Sr0.15MnO3 composition and a jump of the magnetoresistance maximum value at the concentrational Pbnm2R%3c phase transition were observed and discussed.
|
|
