Effects of deferasirox and deferiprone on cellular iron load in the human hepatoma cell line HepaRG

Two oral chelators, CP20 (deferiprone) and ICL670 (deferasirox), have been synthesized for the purpose of treating iron overload diseases, especially thalassemias. Given their antiproliferative effects resulting from the essential role played by iron in cell processes, such compounds might also be useful as anticancer agents. In the present study, we tested the impact of these two iron chelators on iron metabolism, in the HepaRG cell line which allowed us to study proliferating and differentiated hepatocytes. ICL670 uptake was greater than the CP20 uptake. The iron depletion induced by ICL670 in differentiated cells increased soluble transferrin receptor expression, decreased intracellular ferritin expression, inhibited ⁵⁵Fe (III) uptake, and reduced the hepatocyte concentration of the labile iron pool. In contrast, CP20 induced an unexpected slight increase in intracellular ferritin, which was amplified by iron-treated chelator exposure. CP20 also promoted Fe(III) uptake in differentiated HepaRG cells, thus leading to an increase of both the labile pool and storage forms of iron evaluated by calcein fluorescence and Perls staining, respectively. In acellular conditions, compared to CP20, iron removing ability from the calcein-Fe(III) complex was 40 times higher for ICL670. On the whole, biological responses of HepaRG cells to ICL670 treatment were characteristic of expected iron depletion. In contrast, the effects of CP20 suggest the potential involvement of this compound in the iron uptake from the external medium into the hepatocytes from the HepaRG cell line, therefore acting like a siderophore in this cell model.     

Presence of galactose in precultures induces lacS and leads to short lag phase in lactose-grown Lactococcus lactis cultures

Lactose conversion by lactic acid bacteria is of high industrial relevance and consistent starter culture quality is of outmost importance. We observed that Lactococcus lactis using the high-affinity lactose-phosphotransferase system excreted galactose towards the end of the lactose consumption phase. The excreted galactose was re-consumed after lactose depletion. The lacS gene, known to encode a lactose permease with affinity for galactose, a putative galactose–lactose antiporter, was upregulated under the conditions studied. When transferring cells from anaerobic to respiration-permissive conditions, lactose-assimilating strains exhibited a long and non-reproducible lag phase. Through systematic preculture experiments, the presence of galactose in the precultures was correlated to short and reproducible lag phases in respiration-permissive main cultivations. For starter culture production, the presence of galactose during propagation of dairy strains can provide a physiological marker for short culture lag phase in lactose-grown cultures.

     

Cladistic analysis of Aradidae (Insecta,Heteroptera) based on morphological and molecular characters

In this study, we present for the first time a cladistic analysis of the subfamilies of Aradidae, using molecular and morphological characters. Eighty‐three taxa, including 79 species representing eight subfamilies, are treated. The analyses were performed using 72 morphological characters and approximately 1650 bp corresponding to three molecular loci (CO1, 16S and 28S). Characters were analysed using parsimony and Bayesian methods. Based on the combined analyses, we find support for the monophyly of Aradidae, including the subfamilies Aradinae, Aneurinae, Calisiinae, Isoderminae, and Mezirinae and the paraphyly of Prosympiestinae and Chinamyersiinae. In all analyses, Prosympiestinae includes Isoderminae. The Termitaphididae were not found sister to the Aradidae in the Aradoidea.     

One-carbon metabolism in synchronized cultures of Euglena gracilis

The levels of folate derivatives in division synchronized cultures of Euglena gracilis Klebs (strain Z) increased rapidly on a per cell basis durin     

Magnesium ions in catecholamine fluorescence histochemistry

Summary The effects of high concentrations of magnesium ions in the cryostat and Vibratome procedures for visualization of catecholamine fluorescence in the central nervous system have been investigated. In cryostat sections, obtained from specimens perfused with a formaldehyde and glyoxylic acid containing buffer, the addition of high concentrations of MgSO₄ to the perfusion solution enhances the fluorescence intensity and reduces the unspecific background fluorescence and the diffusion of the catecholamine fluorophore. This improves the visualization of all portions of the central catecholamine-containing neurons. Similar effects are obtained in the formaldehyde-Vibratome technique by the introduction of an immersion bath containing MgSO₄ after the sectioning procedure. The use of the magnesium perfusion or immersion steps furthermore increases the reproducibility of the Vibratome and cryostat techniques. The paper describes the improved Vibratome and cryostat techniques used in our laboratory.     

An improved histofluorescence procedure for freeze-dried paraffin-embedded tissue based on combined formaldehyde-glyoxylic acid perfusion with high magnesium content and acid pH.]

A technique is described for highly sensitive and precise visualization of central catecholamine systems in paraffin sections of freeze-dried tissue. The procedure is based on perfusion of the animal with a solution containing formaldehyde and/or glyoxylic acid, in the presence of a very high magnesium content (40 g MgSO4/150 ml solution) and acid pH. The perfused tissue is rapidly frozen, freeze-dried, treated with formaldehyde vapours (at +80 degrees C for 1 h), embedded in parffin in vacuo, and finally sectioned. The present technique has a sensitivity for the dopamine- and noradrenaline-containing systems that is comparable with that of the glyoxylic acid-Vibratome technique, which utilizes fresh, glyoxylic acid-perfused tissue. Thus, the preterminal axon pathways become fluorescent throughout their full extent and the several new terminal systems, discovered with the glyoxylic acid-Vibratome method, are well demonstrable. The method is also highly useful for the study of the cell bodies and their dendritic processes. The catacholamine fibre systems are visualized without any signs of diffusion and with a richness in detail. In animals pretreated with L-tryptophan and MAO-inhibitor the technique is also useful for studies on central indolamine-containing systems.     

Gut-type glucagon immunoreactivity in nerves of the rat brain.

Nerve fibers reacting with antisera demonstrating gut-type glucagon were numerous in certain areas of hypothalamus and thalamus but absent from neocortex and hippocampus. They did not react with glucagon antisera specific for pancreatic type glucagon. Immunoreactive cell bodies were not observed.