CAR, the continuously advancing receptor, in drug metabolism and disease.

The detoxification and elimination of potentially toxic foreign and endogenous compounds depends on the concerted action of xenobiotic metabolizing enzymes. Nuclear hormone receptors (NHRs) have emerged as key regulators of the expression of these enzymes and his review focuses on the xenosenor CAR (Constitutive Androstane Receptor, NR1I3). CAR is highly expressed in the liver and the small intestine, two key tissues expressing xenobiotic metabolizing enzymes, and mediates the induction of their expression by the widely used antiepileptic drug, phenobarbital (PB) and the potent synthetic inducer 1, 4-bis-(2-(3, 5, -dichloropyridyloxy)) benzene (TCPOBOP). TCPOBOP is an agonist ligand for CAR. PB induces its nuclear translocation, which results in increased expression of CAR target genes since, unlike the classical, ligand-dependent nuclear receptors, CAR is an apparently constitutive transactivator. This constitutive activity is inhibited by the inverse agonist ligands androstanol and androstenol. The CAR mediated induction of the expression of xenobiotic metabolizing enzymes is generally protective, but can be deleterious if toxic metabolites are produced. CAR also has a protective role in the stress response elicited by hyperbilirubinemia, as well as lithocholic acid induced cholestasis. In addition, recent studies show that CAR activation disrupts thyroid hormone homeostasis. Finally, CAR activation promotes hepatocyte proliferation and blocks apoptosis, and is essential for the tumorigenesis induced by its activators PB and TCPOBOP. The role of CAR in endobiotic and xenobiotics metabolism has clinical implications in disease prevention, drug-drug interactions, and the development of better drug treatments.

Beta-thalassaemia intermedia in Lebanon.

Approximately one third of thalassaemia patients on record in Lebanon have thalassaemia intermedia. We have analysed three factors in a panel of 73 patients with this less severe form of the disease in our population: mild beta-globin gene mutations, deletions in the alpha-globin gene and the presence of a polymorphism for the enzyme Xmn I in the Ggamma-promoter region. The results show that the most important contributing factor is the beta-genotype: 68% of patients have a mild beta+ mutation (IVSI-6, cd29, -88 or -87), while 26% of patients are positive for the Xmn I polymorphism associated with increased production of HbF, which showed strong linkage to particular mutations (IVSII-1, cd8 and cd30). However, the genotype phenotype correlation is difficult, because many patients were initially misdiagnosed as thalassaemia major and were started early on regular blood transfusions, which was stopped later on. This illustrates well the importance of an early accurate diagnosis of thalassaemia intermedia for appropriate clinical management.

Beta-thalassemia mutations and haplotype analysis in Lebanon.

The molecular basis of beta-thalassemia in Lebanon reflects the heterogeneity of the Lebanese population. Eighteen different mutations were identified among a total of 277 chromosomes. There is evidence of clustering of some mutations in particular geographic regions or among specific religious groups. Haplotype analysis, using seven restriction sites was performed on a total of 110 samples and 11 different haplotypes were identified. The five most common mutations were each found on two different haplotypes, and most linkages were as previously reported in other Mediterranean populations, with a few exceptions, also showing some clustering.

Emergence of Salmonella enteritidis outbreaks in broiler chickens in the Lebanon: epidemiological markers and competitive exclusion control.

This study investigates the first emergence of Salmonella Enteritidis outbreaks among chickens in the Lebanon and identifies the epidemiological markers of selected recovered Enteritidis strains. In addition, the authors evaluate a competitive exclusion approach to control infection in broiler chickens by Enteritidis organisms which possess the prevalent identified markers. The basic procedure in this investigation involved recording signs and lesions in eleven broiler chicken flocks on eleven farms, and culturing livers, spleens, and caeca of ten randomly selected birds per flock for Salmonella isolation and serotyping. Furthermore, culturing for Salmonella and serotyping was attempted from the livers, spleens, caeca and oviduct swabs of ten hens in four broiler breeder flocks which provided hatching eggs for the broilers under study. The identification of epidemiological markers in recovered S. Enteritidis included the determination of drug-resistance patterns and plasmid profiling. The competitive exclusion was evaluated by spraying the microflora on day-old broilers in the hatchery, followed by a controlled oral challenge at three days of age, with 2.85 x 10(5) colony-forming units of S. Enteritidis organisms per bird. Exclusion was evaluated by culturing for S. Enteritidis in anal swabs, spleens, livers, and caeca of individual challenged birds treated with the microflora and in untreated challenged birds. A total of 112 invasive S. Enteritidis strains were recovered on eleven farms from individual organs of broiler chickens with typical signs and lesions of salmonellosis. The prevalent resistance to drugs in such strains was to furaltadone and gentamycin, a marker identified in 93 strains (83%), recovered from nine out of eleven farms. The same resistance pattern was present in S. Enteritidis strains recovered from breeders on one out of four farms. The prevalent plasmid profile in nine S. Enteritidis organisms selected randomly from a pool of 93 strains (one per each of the nine broiler farms) was 14.1 kilobases (kb) and approximately 50.0 kb, a typical pattern to that identified in S. Enteritidis organisms recovered from oviducts of breeders on one out of four breeder farms. The exclusion significantly reduced cumulative mortality in birds of up to 45 days of age by 3.93%, in comparison to that observed in untreated challenged birds (P < 0.05). At 45 days of age, exclusion resulted in a 15.6% reduction in the percentage infection rate by S. Enteritidis in spleens or livers and a 34.4% reduction in the percentage infection rate of the caeca (P < 0.05).

Analysis of epitope-tagged forms of the dyskeratosis congenital protein (dyskerin): identification of a nuclear localization signal.

The X-linked form of the bone marrow failure syndrome Dyskeratosis congenital is caused by mutations in dyskerin, a 514 amino acid protein that is presumed to play a role in ribosome biogenesis. Here we report that dyskerin tagged with the human immunoglobulin epitope localizes to nuclei of transfected HeLa and COS-1 cells. A carboxyl-terminal domain consisting of amino acids 467-475 and encoding KKEKKKSKK is both necessary and sufficient to mediate nuclear entry. Immunoglobulin-tagged dyskerin did not interact with the Fanconi anemia group A protein, FANCA. These results suggest a nuclear role for dyskerin. Moreover, hematopoietic failure observed in both Dyskeratosis congenital and the most common type of Fanconi anemia is unlikely to have a common mechanism resulting from abnormal physical interactions between the respective gene products of these disorders.