Design of small volume HX and triple-resonance probes for improved limits of detection in protein NMR experiments

2003 ◽  
Vol 164 (1) ◽  
pp. 128-135 ◽  
Author(s):  
Yu Li ◽  
Timothy M Logan ◽  
Arthur S Edison ◽  
Andrew Webb
Keyword(s):  
Small Volume ◽  
Protein Nmr ◽  
Triple Resonance ◽  
Limits Of Detection

Quantification of Furanylfentanyl and its Metabolites in Human and Rat Plasma Using LC–MS-MS

2020 ◽  
Vol 44 (6) ◽  
pp. 589-595 ◽  
Author(s):  
Kaitlyn B Palmquist ◽  
Madeleine J Swortwood
Keyword(s):  
Human Plasma ◽  
Small Volume ◽  
Matrix Effects ◽  
Rat Plasma ◽  
Opioid Epidemic ◽  
Limits Of Detection ◽  
Low Concentration ◽  
The World ◽  
Fentanyl Analogs ◽  
Lower Limits

Abstract Fentanyl analogs (novel and traditional) continue to impact the ever-growing opioid epidemic. Furanylfentanyl (FuF) is one analog equipotent to fentanyl that has documented involvement in thousands of intoxication and fatality cases around the world. Due to its prevalence, toxicologists need to improve detection and understanding of this analog. A method for the quantification of FuF and its metabolites (4-ANPP, furanyl norfentanyl (FuNorF)) in a small volume (100 μL) of human plasma by LC–MS-MS was developed and validated according to ANSI/ASB Standard. The method was cross validated in rat plasma for a future pharmacokinetic (PK)/pharmacodynamic (PD) study. In human plasma, calibration ranges were 0.025–25 ng/mL (FuF and 4-ANPP) and 0.5–25 ng/mL (FuNorF). Limits of detection were 0.0125 ng/mL (FuF and 4-ANPP) and 0.25 ng/mL (FuNorF). Lower limits of quantification coincided with lowest calibrator concentrations of 0.025 ng/mL (FuF and 4-ANPP) and 0.5 ng/mL (FuNorF). Precision and bias values were determined to be acceptable for all analytes. Matrix effects were acceptable for all analytes (−8.6–25.0%), except FuNorF with suppression >25%. Extraction recoveries ranged from 84.5 to 98.1%. No carryover or endogenous interferences were observed. Qualitative interferences with 4-ANPP were observed from some n-acyl substituted fentanyl analogs predicted to be low-concentration standard impurities. Analytes were stable under all conditions and dilution integrity was sustained. The method was successfully cross validated in rat plasma with acceptable bias (−7.4–8.4%), precision (within-run < 19%CV and between-run < 12.6%CV), matrix effects (−9.3–17.2%, except FuNorF with >25% suppression), recoveries (79.2–94.5%) and dilution integrity (1/2 and 1/10).

scholarly journals Complete protein assignment from sets of spectra recorded overnight

2019 ◽  
Vol 73 (1-2) ◽  
pp. 59-70
Author(s):  
Jonas Fredriksson ◽  
Wolfgang Bermel ◽  
Martin Billeter
Keyword(s):  
Input Data ◽  
Protein Nmr ◽  
Pulse Sequences ◽  
Side Chain ◽  
Sequential Assignment ◽  
Triple Resonance ◽  
Recording Time ◽  
Protein Assignment ◽  
Complete Protein ◽  
Joint Decomposition

Abstract A flexible and scalable approach for protein NMR is introduced that builds on rapid data collection via projection spectroscopy and analysis of the spectral input data via joint decomposition. Input data may originate from various types of spectra, depending on the ultimate goal: these may result from experiments based on triple-resonance pulse sequences, or on TOCSY or NOESY sequences, or mixtures thereof. Flexible refers to the free choice of spectra for the joint decompositions depending on the purpose: assignments, structure, dynamics, interactions. Scalable means that the approach is open to the addition of similar or different experiments, e.g. larger proteins may require a wider selection of triple-resonance based experiments. Central to the proposed approach is the mutual support among the different spectra during the spectral analysis: for example, sparser triple-resonance spectra may help decomposing (separating) spin systems in a TOCSY or identifying unique NOEs. In the example presented, backbone plus side chain assignments of ubiquitin were obtained from the combination of either two or three of the following projection experiments: a 4D HCCCONH, a 4D HNCACO and a 3D HNCACB. In all cases, TOCSY data (4D HCCCONH) proved crucial not only for the side chain assignments, but also for the sequential assignment. Even when total recording time was reduced to about 10 h, nearly complete assignments were obtained, with very few missing assignments and even fewer differences to a reference.

Protamine-DNA interaction

1978 ◽  
Vol 36 (2) ◽  
pp. 200-201
Author(s):  
D.P. Bazett-Jones ◽  
F.P. Ottensmeyer
Keyword(s):  
Small Volume ◽  
Double Helix ◽  
Dna Interaction ◽  
Dark Field ◽  
Sperm Head ◽  
Nuclear Proteins ◽  
Field Electron Microscopy ◽  
Dark Field Electron ◽  
Dna Double Helix ◽  
Positively Charged

Dark field electron microscopy has been used for the study of the structure of individual macromolecules with a resolution to at least the 5Å level. The use of this technique has been extended to the investigation of structure of interacting molecules, particularly the interaction between DNA and fish protamine, a class of basic nuclear proteins of molecular weight 4,000 daltons.Protamine, which is synthesized during spermatogenesis, binds to chromatin, displaces the somatic histones and wraps up the DNA to fit into the small volume of the sperm head. It has been proposed that protamine, existing as an extended polypeptide, winds around the minor groove of the DNA double helix, with protamine's positively-charged arginines lining up with the negatively-charged phosphates of DNA. However, viewing protamine as an extended protein is inconsistent with the results obtained in our laboratory.

The Theoretical Contrast in Scanning and Conventional High Resolution Microscopes

1969 ◽  
Vol 27 ◽  
pp. 170-171
Author(s):  
E. Zeitler ◽  
M. G. R. Thomson
Keyword(s):  
Small Volume ◽  
Volume Element ◽  
Optical Processing ◽  
Image Construction ◽  
Object Interaction ◽  
Transmission Electron ◽  
Scanning Transmission ◽  
Optical Treatment ◽  
Electron Microscopes ◽  
Intensity Contrast

In the formation of an image each small volume element of the object is correlated to an areal element in the image. The structure or detail of the object is represented by changes in intensity from element to element, and this variation of intensity (contrast) is determined by the interaction of the electrons with the specimen, and by the optical processing of the information-carrying electrons. Both conventional and scanning transmission electron microscopes form images which may be considered in this way, but the mechanism of image construction is very different in the two cases. Although the electron-object interaction is the same, the optical treatment differs.

Planning Micronutrient-Dense Menus in Ontario Long-Term Care Homes: Strategies and Challenges

2020 ◽  
Vol 81 (4) ◽  
pp. 198-203
Author(s):  
Lisa M. Duizer ◽  
Heather H. Keller
Keyword(s):  
Small Volume ◽  
Long Term Care ◽  
Term Care ◽  
Dietary Reference Intakes ◽  
Portion Sizes ◽  
Calcium Magnesium ◽  
Micronutrient Malnutrition ◽  
Low Levels ◽  
Food Consumed

Prevalence of micronutrient malnutrition is high in individuals living in long-term care (LTC) homes with many individuals consuming low levels of vitamins B6, D, and E; folate; calcium; magnesium; and zinc. The focus of this research was to identify strategies and challenges encountered during development of micronutrient-dense menus for use in Ontario LTC homes and to examine costs associated with development of a menu with acceptable micronutrients. Semi-structured open-ended interviews were conducted with 13 menu planners (7 dietitians, 6 nutrition managers) in diverse LTC homes in Ontario. Data were thematically analyzed. A 7-day hypothetical menu meeting all nutrient requirements was developed and costed. Analysis of the interview data showed that menus are planned according to the Canada’s Food Guide (2007) and focus placed on Dietary Reference Intakes of protein, fibre, calcium, and sodium. Little focus is placed on micronutrients. Flexibility in foods offered was important to accommodate the small volume of food consumed. Resident preferences were balanced against nutritional requirements. Challenges included planning for diverse populations, managing portion sizes, and balancing the budget. A hypothetical menu planned to contain adequate levels of all micronutrients is 49% higher in food costs than the amount currently provided to Ontario LTC homes.

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