In Vitro Rescue of the Bile Acid Transport Function of ABCB11 Variants by CFTR Potentiators

ABCB11 is responsible for biliary bile acid secretion at the canalicular membrane of hepatocytes. Variations in the ABCB11 gene cause a spectrum of rare liver diseases. The most severe form is progressive familial intrahepatic cholestasis type 2 (PFIC2). Current medical treatments have limited efficacy. Here, we report the in vitro study of Abcb11 missense variants identified in PFIC2 patients and their functional rescue using cystic fibrosis transmembrane conductance regulator potentiators. Three ABCB11 disease-causing variations identified in PFIC2 patients (i.e., A257V, T463I and G562D) were reproduced in a plasmid encoding an Abcb11-green fluorescent protein. After transfection, the expression and localization of the variants were studied in HepG2 cells. Taurocholate transport activity and the effect of potentiators were studied in Madin–Darby canine kidney (MDCK) clones coexpressing Abcb11 and the sodium taurocholate cotransporting polypeptide (Ntcp/Slc10A1). As predicted using three-dimensional structure analysis, the three variants were expressed at the canalicular membrane but showed a defective function. Ivacaftor, GLP1837, SBC040 and SBC219 potentiators increased the bile acid transport of A257V and T463I and to a lesser extent, of G562D Abcb11 missense variants. In addition, a synergic effect was observed when ivacaftor was combined with SBC040 or SBC219. Such potentiators could represent new pharmacological approaches for improving the condition of patients with ABCB11 deficiency due to missense variations affecting the function of the transporter.     

Nitric Oxide Metabolic Pathway in Drought-Stressed Nodules of Faba Bean (Vicia faba L.)

Drought is an environmental stress that strongly impacts plants. It affects all stages of growth and induces profound disturbances that influence all cellular functions. Legumes can establish a symbiosis with Rhizobium-type bacteria, whose function is to fix atmospheric nitrogen in organs called nodules and to meet plant nitrogen needs. Symbiotic nitrogen fixation (SNF) is particularly sensitive to drought. We raised the hypothesis that, in drought-stressed nodules, SNF inhibition is partly correlated to hypoxia resulting from nodule structure compaction and an increased O₂ diffusion barrier, and that the nodule energy regeneration involves phytoglobin–nitric oxide (Pgb–NO) respiration. To test this hypothesis, we subjected faba bean (Vicia faba L.) plants nodulated with a Rhizobium laguerreae strain to either drought or osmotic stress. We monitored the N₂-fixation activity, the energy state (ATP/ADP ratio), the expression of hypoxia marker genes (alcohol dehydrogenase and alanine aminotransferase), and the functioning of the Pgb–NO respiration in the nodules. The collected data confirmed our hypothesis and showed that (1) drought-stressed nodules were subject to more intense hypoxia than control nodules and (2) NO production increased and contributed via Pgb–NO respiration to the maintenance of the energy state of drought-stressed nodules.     

Circadian variations in the risk of urinary calcium oxalate stone formation

OBJECTIVE: To evaluate the circadian fluctuations in the risk of urinary calcium oxalate stone formation with regard to critical periods of crystallization. PATIENTS AND METHODS: Over a given time period, the Tiselius index depends on urine volume and urinary excretion of oxalate, calcium, citrate and magnesium. This crystallization potential was evaluated during three successive periods spread over 24 h for 25 recurrent stone-formers aged 16-76 years (mean 50) and 25 control subjects aged 27-71 years (mean 44). RESULTS: There was no significant difference in the value of the Tiselius index for all equivalent time periods in both groups of patients. The minimum value was recorded in the afternoon and the circadian pattern of the index illustrated the predominant importance of urinary output in its determination. Morning urinary concentrations and excretions of citrate, and nocturnal levels of magnesium were significantly higher in the stone-formers when compared with the control subjects. CONCLUSION: The lithogenic risk for calcium oxalate stones was maximal at the end of the night or during the early morning, when urinary output was minimal. This circadian study revealed abnormalities that are not apparent from non-fractionated 24 h urine samples, and which were potentially relevant to therapy.     

Behavioral avoidance dynamics in the presence of a virtual spider

Tracking behavior with a virtual spider and a neutral target is compared in fearful and nonfearful subjects. Head-tracking in virtual environments appears to be a scale-free behavior with long-range fractal-like patterns. Moreover, these fractal patterns change according to what the target affords the tracker and the level of behavioral avoidance manifested by the subjects. Results are interpreted in terms of ecological psychology and nonlinear dynamics, and implications for virtual reality (VR) psychology are outlined.     

Capillary zone electrophoresis with laser-induced fluorescence detection for the determination of nanomolar concentrations of noradrenaline and dopamine: application to brain microdialysate analysis

Determination of catecholamines by capillary zone electrophoresis with laser-induced fluorescence detection was performed on low-concentration samples, which were derivatized with naphthalene-2,3-dicarboxaldehyde to give highly fluorescent compounds. When the borate concentration in the derivatization medium was decreased from 130 to 13 mM, sensitivity for noradrenaline (NA) and dopamine (DA) was greatly enhanced while resolution between these two compounds decreased. A 50 mM borate concentration in derivatization medium was chosen since it provided maximal resolution between NA and DA, together with a high separation efficiency (3.1 million theoretical plates per meter for DA). The injection of 2.4 nL of a NA and DA solution derivatized at 10(-9) M produced peaks with signal-to-noise ratio of 8:1 and 3:1, respectively, corresponding to 1.8 amol of each catecholamine. The calibration curves were linear when NA and DA solutions were derivatized at concentrations ranging from 10(-6) to 10(-9) M. This method was used to determine NA in brain extracellular fluid: a peak corresponding to a basal level of 5 x 10(-9) M endogeneous NA was observed in microdialysates from the medial frontal cortex of the rat, and its nature was confirmed by both electrophoretic and pharmacological validations.     

Variations of histamine concentration in sleep and arousal hemodialysates (seasonal, thalamic and reticular influences)

In hemodialysates of cerebral venous blood from rabbits kept asleep by electrical stimulation of the somnogenic intralaminar thalamus, a seasonal correlation was found between the histamine (H) concentration and the external daily temperature, with a minimum in winter and a maximum in summer. By contrast, in dialysates of rabbits aroused by electrical stimulation of the midtrain reticular formation, the H concentration reached in winter the same high level as in summer. During the winter, stimulation of the waking midbrain reticular system increased the H-level of the cerebral blood dialysate, whereas that of the somnogenic intralaminar thalamus did not enhance, sometimes even lowered, the H-concentration. This suggests that the mechanism of H release in blood might be associated with the midbrain activating reticular system and moderated by the intralaminar thalamus.     

X-ray topographic study of dislocations in KH2(1−x)D2xPO4 single crystals

Dislocations in heavily deuterated DKDP single crystals were examined by the Lang technique, and their characteristics were compared with those of the KDP isomorph species. The influence of dislocations on the operation of DKDP targets in large-screen display devices is discussed. The origin of dislocations which takes place during the growth habit formation is briefly discussed.     

Separator materials for use in alkaline water electrolysers

Separator materials suitable for use in industrial alkaline water electrolysers are surveyed, including both inorganic and organic materials. The properties which make asbestos diaphragms unsuitable for use above 100°C are also reviewed.In summary, potassium titanate and polyantimonic acid separator materials perform well in high-temperature alkali. Oxide-coated metallic diaphragms merit further attention. Polysulfone and polyphenylene sulfide polymers are promising, but will need definite improvement in their wettability. Asbestos stabilization by silicate electrolyte additions may be viable.