Analysis of PEG 400 and 4000 in urine for gut permeability assessment using solid phase extraction and gel permeation chromatography with refractometric detection.

We developed a treatment of urine samples allowing the analysis of two intestinal permeability markers: polyethylene glycol (PEG) 400 (highly diffusible; basal permeability indicator) and PEG 4000 (poorly diffusible; indicator of an abnormal increase of permeability) by a unique gel permeation chromatography (GPC) with refractometric detection. Urinary PEG were extracted using a mixed-bed resin composed of C2 and C18 layers. Permeability mean values determined in 11 human healthy subjects were 24.20 +/- 9.30% and 0.12 +/- 0.08% for, respectively, PEG 400 and 4000. The percentage of the PEG 4000 permeability value to the one of PEG 400 corresponded to an intestinal permeability index (IPI) of 0.52 +/- 0.35 expressing a low diffusion of this poorly permeability marker.

Pancreatic exocrine secretions as a source of luminal polyamines in pigs.

The goal of the present study was twofold: (1) to detect the possible storage of dietary polyamines (PAs) in various tissues and (2) to investigate the role of dietary PAs in the differentiation of the pig intestinal epithelium. A first experimental series was designed to assess the accumulation of either milk PAs (mostly spermidine) or orally administered spermine (SPM) in piglet red blood cells (RBCs) and plasma, a preliminary stage in their distribution to growing and storage organs. Though PA concentrations of piglet RBCs and plasma were generally significantly higher than their sow counterparts, our experimental conditions failed to demonstrate that this increase could stem from ingested PAs. A second experimental series dealt with the determination of disaccharidase specific activities in proximal and distal parts of piglet gut on the 26th and 29th days after birth (preweaning time). In agreement with observations made previously on rat pups, we observed an increase in maltase specific activity (SA) at the end of the suckling period (the observed increase in sucrase SA was not significant). However, orally administered SPM did not affect this activity. Compared to the constant protein concentrations observed in both parts of the gut, the pancreatic protein content decreased sharply between the 26th and 29th postnatal days. At the same time pancreatic concentrations of spermidine (SPD) also decreased, suggesting that some pancreatic PAs were released as the organ secreted its proteins. In accordance with this hypothesis, we recorded SPM and SPD in pancreatic juice. The increases in PA concentrations seemed to follow the protein secretion pattern (i.e. PA concentrations reached a maximal value when the protein concentration was highest). The presence of PAs in pancreatic juice could be indicative of a control mechanism exerted by the pancreas on PA-induced growth and differentiation of porcine intestinal epithelium.

APH-1, a POU homeobox gene expressed in the salt gland of the crustacean Artemia franciscana.

We characterized the first POU-homeoprotein in a crustacean (designated APH-1 for Artemia POU-Homeoprotein, EMBL Y15070). The amino acid sequence of the APH-1 POU-domain is identical, except for two residues, to that of the two class III POU proteins Cf1-a (Drosophila) and POU-M1 (Bombyx mori). Southern blot analysis suggests that crustaceans have only one class III POU gene. RT-PCR and whole-mount in situ hybridization show that APH-1 mRNA is present in larvae specifically in the salt gland, an organ which is involved in osmoregulation, and disappears in the adult.

[Statement on polyamines]. / Le point sur les polyamines.

Polyamines are ubiquitous substances. Their intracellular concentration is controlled quickly and rigorously by extremely sophisticated systems. It depends on metabolism and cellular permeability. Polyamines act as structural and functional elements in the cell (nucleic acid conformation, cytoskeleton, radioprotection, apoptosis, proliferation and differentiation of cells...). They also play a role in various diseases (origin of food allergy, cancers...). They present a great therapeutic interest (oncology, molecular transfer to cell nucleus, transfer across the blood-brain barrier, parasitosis, effects on NMDA and GABA receptors in the central nervous system...).

Follow-up of protein release from Pseudoplusia includens hemocytes: a first step toward identification of factors mediating encapsulation in insects.

Hemocyte types involved in encapsulation were characterized using monoclonal antibodies (mAbs). This approach revealed that four hemocyte types in Pseudoplusia includens could be classified into two antigenically distinct cell lines. The first line comprised granulocytes (GR) and spherulocytes (SP) and the second line comprised plasmatocytes (PL) and oenocytoids (OE). One of the mAbs labeled a subpopulation of plasmatocytes that spread on culture plate surfaces. This subclass represented approximately 70% of all plasmatocytes. The cytoplasmic punctate staining of granulocytes clearly decreased upon short term culture, suggesting the associated antigens were released into the culture medium during cell spreading. A follow-up of protein secretion into culture medium by Western blotting confirmed this hypothesis for both granulocytes and plasmatocytes. In a few cases, discharged proteins exhibited a short half-life suggesting they may behave as regulatory molecules. Among them, plasmatocyte proteins of +/-25 kDa might be mobilized at an early stage of encapsulation. The same proteins appeared to accumulate at the periphery of the median plasmatocyte multilayer in late capsules. This location coincides with where an outer monolayer of granulocytes attaches and causes termination of capsule growth. These preliminary results raise the possibility that released proteins regulate hemocyte recruitment during encapsulation.

Structure and possible functions of the calcospherite-rich cells (R* cells) in the digestive gland of the shore crab Carcinus maenas.

R*-cells of the digestive gland of Carcinus maenas have been investigated functionally and morphologically. A comparison of the capacity of separated cell suspensions to synthesize glycogen gave support to the hypothesis that R and R* cells belong to the same cell line. The unexpected observation of R* cells in gastric juice suggests that their release could represent a mode of redistribution of carbohydrate stores when the feeding activity of the crab is lower. Under electron microscopy, the calcospherites of R* cells appeared to be surrounded by multiple membranous layers, and displayed tubular and vesicular structures in their core. High glucose-6-phosphatase (G6Pase) activity in the subcellular fraction that is enriched in calcospherites suggests that these membranes are derived from the endoplasmic reticulum, via a process in which the enzyme plays a key role. We propose that this is the way by which the R cell differentiates into R* cell.