| 1. |
- Lieske, Julia, et al.
(författare)
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On-chip crystallization for serial crystallography experiments and on-chip ligand-binding studies
- 2019
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Ingår i: IUCrJ. - 2052-2525. ; 6:4, s. 714-728
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Tidskriftsartikel (refereegranskat)abstract
- Efficient and reliable sample delivery has remained one of the bottlenecks for serial crystallography experiments. Compared with other methods, fixed-target sample delivery offers the advantage of significantly reduced sample consumption and shorter data collection times owing to higher hit rates. Here, a new method of on-chip crystallization is reported which allows the efficient and reproducible growth of large numbers of protein crystals directly on micro-patterned silicon chips for in-situ serial crystallography experiments. Crystals are grown by sitting-drop vapor diffusion and previously established crystallization conditions can be directly applied. By reducing the number of crystal-handling steps, the method is particularly well suited for sensitive crystal systems. Excessive mother liquor can be efficiently removed from the crystals by blotting, and no sealing of the fixed-target sample holders is required to prevent the crystals from dehydrating. As a consequence, 'naked' crystals are obtained on the chip, resulting in very low background scattering levels and making the crystals highly accessible for external manipulation such as the application of ligand solutions. Serial diffraction experiments carried out at cryogenic temperatures at a synchrotron and at room temperature at an X-ray free-electron laser yielded high-quality X-ray structures of the human membrane protein aquaporin 2 and two new ligand-bound structures of thermolysin and the human kinase DRAK2. The results highlight the applicability of the method for future high-throughput on-chip screening of pharmaceutical compounds.
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| 2. |
- Andersson, Pontus, 1971, et al.
(författare)
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Radiative lifetimes of metastable states of negative ions
- 2006
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Ingår i: Physical Review A. Atomic, Molecular, and Optical Physics. - 1050-2947 .- 1094-1622 .- 2469-9926 .- 2469-9934. ; 73, s. 032705-
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Tidskriftsartikel (refereegranskat)abstract
- We present a technique for measuring the radiative lifetimes of metastable states of negative ions that involves the use of a heavy-ion storage ring. The method has been applied to investigate the radiative decay of the np(3) P-2(1/2) levels of Te-(n=5) and Se-(n=4) and the 3p(3) D-2 state of Si- for which the J=3/2 and 5/2 levels were unresolved. All of these states are metastable and decay primarily by emission of E2 and M1 radiation. Multi Configuration Dirac-Hartree-Fock calculations of rates for the transitions in Te- and Se- yielded lifetimes of 0.45 s and 4.7 s, respectively. The measured values agree well with these predicted values. In the case of the D-2 state of Si-, however, our measurement was only able to set a lower limit on the lifetime. The upper limit of the lifetime that can be measured with our apparatus is set by how long the ions can be stored in the ring, a limit determined by the rate of collisional detachment. Our lower limit of 1 min for the lifetime of the D-2 state is consistent with both the calculated lifetimes of 162 s for the D-2(3/2) level and 27.3 h for the D-2(5/2) level reported by O'Malley and Beck and 14.5 h and 12.5 h, respectively, from our Breit-Pauli calculations.
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| 3. |
- Beyerlein, Kenneth R., et al.
(författare)
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Mix-and-diffuse serial synchrotron crystallography
- 2017
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Ingår i: IUCrJ. - : INT UNION CRYSTALLOGRAPHY. - 2052-2525. ; 4:6, s. 769-777
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Tidskriftsartikel (refereegranskat)abstract
- Unravelling the interaction of biological macromolecules with ligands and substrates at high spatial and temporal resolution remains a major challenge in structural biology. The development of serial crystallography methods at X-ray free-electron lasers and subsequently at synchrotron light sources allows new approaches to tackle this challenge. Here, a new polyimide tape drive designed for mix-and-diffuse serial crystallography experiments is reported. The structure of lysozyme bound by the competitive inhibitor chitotriose was determined using this device in combination with microfluidic mixers. The electron densities obtained from mixing times of 2 and 50 s show clear binding of chitotriose to the enzyme at a high level of detail. The success of this approach shows the potential for high-throughput drug screening and even structural enzymology on short timescales at bright synchrotron light sources.
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| 4. |
- Ellmann, A, et al.
(författare)
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Radiative lifetime of a bound excited state of Te-
- 2004
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Ingår i: Physical Review Letters. - 0031-9007 .- 1079-7114. ; 92:25, s. 253002-
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Tidskriftsartikel (refereegranskat)abstract
- We report on the first experimental study of the lifetime of a bound excited state of a negative ion. A new experimental technique was developed and used to measure the radiative lifetime of the 5p(5) P-2(1/2) level of Te-. The experiment was performed in a magnetic storage ring, where a laser beam was applied along one of the straight sections. In the experiment the population of the excited J=1/2 level was probed each time the Te- ions passed through the laser field. A decay curve was built up by sampling the population of the excited level of the Te- ions as a function of time after injection into the ring. A multiconfiguration Dirac-Hartree-Fock calculation was performed in conjunction with the experiment. The calculation yielded a radiative lifetime of 0.45 s, in excellent agreement with the measured value of 0.42(5) s.
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| 5. |
- Hammarsten, Ola, et al.
(författare)
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The ratio of cardiac troponin T to troponin I may indicate non-necrotic troponin release among COVID-19 patients
- 2022
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Ingår i: Clinica Chimica Acta. - : Elsevier. - 0009-8981 .- 1873-3492. ; 527, s. 33-37
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Tidskriftsartikel (refereegranskat)abstract
- Background: Although cardiac troponin T (cTnT) and troponin I(cTnI) are expressed to similar amount in cardiac tissue, cTnI often reach ten-times higher peak levels compared to cTnT in patients with myocardial necrosis such as in acute myocardial infarction (MI). In contrast, similar levels of cTnT and cTnI are observed in other situations such as stable atrial fibrillation and after strenuous exercise.Objective: Examine cTnT and cTnI levels in relation to COVID-19 disease and MI. Methods: Clinical and laboratory data from the local hospital from an observational cohort study of 27 patients admitted with COVID-19 and 15 patients with myocardial infarction (MI) that were analyzed with paired cTnT and cTnI measurement during hospital care.Results: Levels of cTnI were lower than cTnT in COVID-19 patients (TnI/TnT ratio 0.3, IQR: 0.1-0.6). In contrast, levels of cTnI were 11 times higher compared to cTnT in 15 patients with MI (TnI/TnT ratio 11, IQR: 7-14). The peak cTnI/cTnT ratio among the patients with MI following successful percutaneous intervention were 14 (TnI/ TnT ratio 14, IQR: 12-23). The 5 COVID-19 patient samples collected under possible necrotic events had a cTnI/ cTnT ratio of 5,5 (IQR: 1,9-8,3).Conclusions: In patients with COVID-19, cTnT is often elevated to higher levels than cTnI in sharp contrast to patients with MI, indicating that the release of cardiac troponin has a different cause in COVID-19 patients.
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