More accurate prediction of the extent of drug brain exposure in early drug discovery and understanding potential species differences could help to guide medicinal chemistry and avoid unnecessary animal studies. Hence, the aim of the current study was to validate the use of a P-gp transfected LLC-PK1 model to predict the unbound brain-to-plasma concentration ratio (Kp
  ) in rats and humans.
 
  MOCK-, Mdr1a- and MDR1-transfected LLC-PK1 monolayers were applied in a transwell setup to quantify the bidirectional transport for 12 specific P-gp substrates, 48 UCB drug discovery compounds, 11 compounds with reported rat in situ brain perfusion data and 6 compounds with reported human Kp
  values. The in vitro transport data were introduced in a minimal PBPK model (SIVA®) to determine the transport parameters. These parameters were combined with the differences between in vitro and in vivo passive permeability as well as P-gp expression levels (as determined by LC-MS/MS), to predict the Kp
  .
 
  A 10-fold difference between in vitro and in vivo passive permeability was observed. Incorporation of the differences between in vitro and in vivo passive permeability and P-gp expression levels resulted in an improved prediction of rat (AAFE 2.17) and human Kp
  (AAFE 2.10).
 
  We have succesfully validated a methodology to use a P-gp overexpressing LLC-PK1 cell line to predict both rat and human Kp
  by correcting for both passive permeability and P-gp expression levels.
 We have succesfully validated a methodology to use a P-gp overexpressing LLC-PK1 cell line to predict both rat and human Kpuu,brain by correcting for both passive permeability and P-gp expression levels.An integrated aflatoxin B1 (AFB1) detection platform with quantum dot (QD)-based electrochemical immunosensor and an automated magneto-controlled pretreatment system was successfully developed. The automated pretreatment system adopts the immunoaffinity magnetic beads (IMB) as the capture probe of AFB1 and QD-labeled AFB1 complete antigen (AFB1-BSA-QDs) as the signal probe. AFB1-BSA-QDs can be easily converted into corresponding metallic cations through acidic treatment, which can be detected electrochemically via anode stripping voltammetry (ASV). Moreover, a disposable screen-printed electrode (SPE) without requiring any further modification is used in the novel electrochemical immunosensor' making routine testing feasible. Under optimal conditions, the detectable concentration range of AFB1 was 0.08-800 μg/kg. The metal ion signal associated linearly with the logarithm of AFB1 concentration within the range of 5-240 μg/kg, with a detection limit of 0.05 μg/kg. The spiked recoveries of three different concentrations in four different matrixes ranged from 83.9 to 118.0%, and inter-day relative standard deviations were below 10%. Furthermore, the methodology was validated by analyzing naturally contaminated samples, and results of the novel immunosensor were in good agreement with those of LC-MS/MS, demonstrating the potentiality of the developed method for the monitor of AFB1 in cereals and oils.The aim of the present investigation was the analysis and identification of antisense oligonucleotide metabolism products after incubation with human liver microsomes regarding four different oligonucleotide modifications. Separation and detection methods based on the use of liquid chromatography coupled with quadrupole time-of-flight mass spectrometry were developed for this purpose. Firstly, the optimization of mass spectrometer parameters was done to select those which ensure the highest possible sensitivity of oligonucleotide analysis. This step was conducted for two chromatographic modes-ion pair chromatography and hydrophilic interaction liquid chromatography-due to their common application in oligonucleotide analysis. Based on sensitivity results, ion pair chromatography coupled with mass spectrometry was selected for the separation of model oligonucleotide mixtures in order to verify its selectivity for N-deleted metabolite separation. Next, the developed method was applied in the examination of oligonucleotides in vitro metabolism. First, wide optimization of incubation parameters was conducted including the concentration of the reaction buffer components. Obtained results indicated that both 3'-exonucleases and 5'-exonucleases contributed to the biotransformation of oligonucleotides. Moreover, it may be concluded that the number of metabolites depends on oligonucleotide modification and consequently its resistance to enzymatic attack. Thus, the number of the oligonucleotide metabolites decreased with the decrease of the resultant polarity of oligonucleotide caused by chemical modification. Graphical abstract.Recently, a crystal structure of V-nitrogenase was presented, showing that one of the µ2 sulphide ions in the active site (S2B) is replaced by a lighter atom, suggested to be NH or NH2, i.e. representing a reaction intermediate. Moreover, a sulphur atom is found 7 Å from the S2B site, suggested to represent a storage site for this ion when it is displaced. We have re-evaluated this structure with quantum refinement, i.e. standard crystallographic refinement in which the empirical restraints (employed to ensure that the final structure makes chemical sense) are replaced by more accurate quantum-mechanical calculations.  https://www.selleckchem.com/products/lenalidomide-s1029.html  This allows us to test various interpretations of the structure, employing quantum-mechanical calculations to predict the ideal structure and to use crystallographic measures like the real-space Z-score and electron-density difference maps to decide which structure fits the crystallographic raw data best. We show that the structure contains an OH--bound state, rather than an N2-derived reaction intermediate. Moreover, the structure shows dual conformations in the active site with ~ 14% undissociated S2B ligand, but the storage site seems to be fully occupied, weakening the suggestion that it represents a storage site for the dissociated ligand.
  The aim of this study was to investigate the detection rate of unruptured intracranial aneurysms (UIAs) and incidence of aneurysmal subarachnoid haemorrhage (SAH) in relation to the rapidly changing smoking rates in Norway between 2008 and 2015.
 
  The registry-based study included all patients (≥ 16 years old) admitted to a hospital in Norway between 2008 and 2015 with a primary diagnosis of aneurysmal SAH or an outpatient diagnosis of UIAs. Age group-specific and total detection rate of UIAs and incidence rate of SAH over the years were calculated. Age group-specific data on smoking habits was retrieved from a national annual survey representative of the whole Norwegian population.
 
  The rate of daily smokers decreased by 48% between 2008 and 2015. The detection rate of UIAs decreased by 47% from 17.3 in 2008 to 9.3 per 100,000 persons in 2015, and the incidence of SAH decreased by 30% from 11.3 in 2008 to 7.9 per 100,000 persons in 2015. The average annual decline in prevalence of daily smoking, UIA detection rate, and SAH incidence was 6.