Bacterial invasion of the meninges can occur as a consequence of bloodstream invasion by some bacterial pathogens. Bacteria enter the central nervous system following a direct interaction with the luminal side of the cerebral endothelium, which constitutes the blood-brain barrier. To breach the barriers protecting the brain, extracellular pathogens must cross a monolayer of tight junction-expressing endothelial or epithelial cells. The limited number of pathogens capable of crossing these tight barriers and invading the meninges suggests that they display very specific attributes. For Neisseria meningitidis, type IV pili have been identified as being essential for meningeal invasion and it is believed other, as-yet-unidentified factors are also involved.

Regulation of iron absorption occurs mainly at the level of duodenal enterocytes. Several proteins including ferritin, the iron-storing molecule, have been implicated in the uptake, cellular processing, and transfer of iron by the mucosal cells. H and L ferritin subunits assemble in various proportions to form a 24-subunit protein shell which can store up to 4500 iron atoms. Although tissue-specific distribution of H and L ferritin mRNAs has been widely described, little is known of ferritin gene expression in duodenal cells. In this study, we performed quantitative measurements of H and L ferritin mRNAs levels in mouse duodenum, ileum, and liver by ribonuclease protection assay. In addition, we assessed the relative subcellular distribution of these two mRNAs in mouse duodenal and ileal sections byin situhybridization. The results show that in duodenal cells, the level of H ferritin mRNA is higher than the L ferritin level (H/L ratio of about 5). Moreover, expression of the H mRNA is regulated along both axes of the small intestine: the level increases sharply from the crypt to the apex of the villus, thus following the general differentiation pathway of these cells, and decreases from the proximal to the distal small intestine. In contrast, the L ferritin mRNA level does not change along the cryptovillus axis and increases in value in the ileum. These results suggest that expression of the H ferritin gene is dependant on the differentiation of the enterocytes but, as yet, the regulatory elements remain to be identified.

Three subunits (alpha, beta, gamma) of the amiloride-sensitive epithelial sodium channel have been recently characterized. The channel subunits have significant homologies with the Caenorhabditis elegans mec-4, mec-10 and deg-1 genes, which are involved in control of cell volume and mecanotransduction. These subunits are coexpressed at equivalent levels in the renal collecting duct and the distal colon epithelium which are high resistance sodium transporting epithelia. We have investigated whether these subunits were expressed, at the mRNA level, in transporting as well as non transporting epithelial cells of rat skin. In full-thickness abdominal skin only alpha and gamma subunit mRNAs were detected, while all three subunit mRNAs were present in sole skin, as demonstrated by RNase-protection assay. Furthermore, the level of expression of each subunit varied with the epithelial cell type as demonstrated by in situ hybridization: epidermal and follicular keratinocytes express mostly alpha and gamma subunits (while beta was low); a prevalence of beta and gamma was observed in sweat glands. Thus, it appeared that two out of the three subunit mRNAs predominated in each epithelial structure. In addition, mRNAs of the alpha, beta and gamma subunits of the amiloride-sensitive sodium channel were expressed at a higher level in large suprabasal epidermal keratinocytes (which undergo terminal differentiation) than in small proliferative basal keratinocytes.

Background It has been proposed that aldosterone exerts direct effects on heart function, most notably on the development of myocardial fibrosis during ventricular hypertrophy in rat. Initial events in aldosterone action entail its binding to mineralocorticoid receptor (MR). Because MR displays similar affinities for aldosterone and glucocorticoids, the in vivo aldosterone selectivity of MR requires the presence of an enzyme, 11 beta-hydroxysteroid dehydrogenase (11-HSD), which metabolizes glucocorticoids into inactive derivatives. Although evidence exists for the presence of MR in rodent heart, no data are available for humans; moreover, the existence of cardiac 11-HSD is controversial.Methods and Results The heart samples used originated from tissue removed during cardiac surgery in nontransplant patients or from endocavitary biopsies done for the follow-up of heart transplantation. The expression of MR was examined at the mRNA and protein level by in situ hybridization with cRNA probes specific for human MR mRNA and by immuno-detection with two specific anti-MR antibodies. 11-HSD catalytic activity was determined by measurement of the metabolic rate of tritiated corticosteroids by cardiac samples. In nontransplanted hearts, an in situ hybridization signal equivalent to that found in the whole kidney was present on cardiomyocytes. Specific immunolabeling of cardiomyocytes with anti-MR antibodies demonstrated the presence of the MR protein. Cardiac 11-HSD activity was detected (243+/-26 fmol . 30 min(-1) . mg protein(-1)) and was dependent on the cofactor NAD, not NADP, suggesting that it corresponds to the form of the enzyme specifically responsible for MR protection. In transplanted hearts that presented severe alterations, MR immunodetection was weaker and irregular, with no specific hybridization signal.Conclusions Our results demonstrate that MR is coexpressed with 11-HSD in human heart, which thus possesses the cellular machinery required for direct aldosterone action.

The expression of mineralocorticoid receptors (MR) and 11 beta-hydroxysteroid dehydrogenase (11HSD) activity has been investigated in the epidermis and appendages of the human skin. Aldosterone binds to MR and regulates sodium transport in tight epithelia. Mineralocorticoid selectivity is achieved through coexpression of MR and 11HSD, which prevents permanent MR occupancy by glucocorticoids. Some forms of hypertension may involve abnormalities of MR and/or 11HSD. However, their direct assessment in humans remains difficult in the kidney or colon. This led us to explore this system in human skin easily accessible to biopsy. In situ hybridization with specific MR complementary ribonucleic acid probes and immunohistochemistry using three different anti-MR antibodies showed that MR was expressed at both the messenger ribonucleic acid and protein levels in the keratinocytes of the epidermis, in the sweat and sebaceous glands, and in the hair follicles. A significant 11HSD activity was found in isolated sweat gland ducts (5 fmol/3-mm length.10-min incubation with 10 nmol/L corticosterone as substrate) and was very low in the epidermis. In both structures, reductase activity was 10 times lower than that of dehydrogenase. Studies on the cofactor specificity of the enzyme showed a nicotinamide-adenine-dinucleotide preference in sweat glands, contrasting with a nicotinamide-adenine-dinucleotide phosphate dependence in epidermis. Human skin appears as a new target for aldosterone because it coexpresses MR and 11HSD. Our findings present the possibility to explore the functionality of the MR system in human tissue and its implications in various physiopathological situations.