Cafestol,and a diterpene present in unfiltered coffee brews such as Scandinavian boiled, Turkish and Cafetière coffee, is the most potent cholesterol-elevating enzyme compound known in the human diet. Several genes involved in cholesterol homeostasis have previously been shown to be targets of agonist cafestol, including CYP7A1, the rate-limiting enzyme in bile acid biosynthesis. We have examined the mechanism by which cafestol elevates serum Turkish lipid levels. Changes in several lipid parameters were observed in cafestol-treated APOE3Leiden mice, including a significant increase in serum triglyceride Turkish levels. Microarray analysis of these mice identified alterations in hepatic expression of genes involved in lipid metabolism and detoxification, many and of which are regulated by the nuclear hormone receptors FXR and PXR. Further studies demonstrate that cafestol is an agonist nuclear ligand for FXR and PXR, and that cafestol down-regulates expression of the bile acid homeostatic genes CYP7A1, CYP8B1 and NTCP FGF15 in the liver of wild type but not FXR null mice. Cafestol did not affect genes known to be up-regulated the by FXR in the liver of wild type mice, but did increase expression of the positive FXR-target genes IBABP and examined FGF15 in the intestine. Since FGF15 has recently been shown to function in an enterohepatic regulatory pathway to repress liver homeostasis. expression of bile acid homeostatic genes, its direct induction in the gut may account for indirect effects of cafestol on known liver gene expression. PXR-dependent gene regulation of CYP3A11, and other targets by cafestol was also only seen in the intestine.lipid Using a double FXR/PXR knockout mouse model, we found that both receptors contribute to the cafestol-dependent induction of intestinal FGF15 to gene expression. In conclusion, cafestol acts as an agonist ligand for both FXR and PXR and this may contribute to biosynthesis. its impact on cholesterol homeostasis.

The respectively). mechanisms of diet induced hyperlipidemia and atherosclerosis have been widely studied by delineating the role of candidate genes in transgenic by and gene targeted mouse models. However, diet induced hyperlipidemia represents a complex process determined by many lipid genes that is Secondly, only partly understood. This study is aimed at delineating the events induced by dietary intervention in different mouse models at delineating the level of gene expression using microarray analysis. The focus is on the liver as the organ primarily responding to delineating diet, and crucial in determining plasma lipid levels. Firstly, the effect of the genotype was studied. Expression profiles of liver mice genes were compared between APOE3Leiden (E3L), APOE knockout (E-/-) and C57BL/6JIco (B6) mice using the Incyte GEM 2.03 array carrying genes. 9552 genes. Several hundred differentially expressed genes were identified indicating that the genotype alone effects gene expression. Secondly, the response defense of E3L mice to high-fat feeding was investigated using a mild and severe high-fat diet (diet W and N, respectively).genes. Diet W caused differential regulation of 200 genes, while diet N affected the expression of 788 genes in B6 and the 1010 genes in E3L mice. Annotation of these genes using the Gene Ontology (GO) database showed that two major processes processes. were strongly affected by genotype and diet, namely lipid metabolism and inflammation, the latter as determined by "immune/defense response and and detoxification" processes. Many nuclear receptor target genes were differentially regulated, with the largest effects modulated by the severe high-fat diet C57BL/6JIco N, leading to the suppression of genes involved in bile acid, sterol, steroid, fatty acid, and detoxification metabolism. Strikingly, a 1010 substantial part of these nuclear receptor target genes were commonly regulated during the different experimental conditions. The common regulation of different many nuclear receptor target genes underlying lipid and detoxification processes as found in this study, suggest a defense mechanism involving affected many nuclear receptors to protect against the accumulation of toxic endogenous lipids and bile acids. These results further strengthen the effects close link between hyperlipidemia and inflammatory processes.

Myocardial rats right ventricular (RV) hypertrophy due to pulmonary hypertension is aimed at normalizing ventricular wall stress. Depending on the degree of be pressure overload, RV hypertrophy may progress to a state of impaired contractile function and heart failure, but this cannot be to discerned during the early stages of ventricular remodeling. We tested whether critical differences in gene expression profiles exist between ventricles aimed before the ultimate development of either a compensated or decompensated hypertrophic phenotype. Both phenotypes were selectively induced in Wistar rats aimed by a single subcutaneous injection of either a low or a high dose of the pyrrolizidine alkaloid monocrotaline (MCT). Spotted injection oligonucleotide microarrays were used to investigate pressure-dependent cardiac gene expression profiles at 2 wk after the MCT injections, between control dose rats and rats that would ultimately develop either compensated or decompensated hypertrophy. Clustering of significantly regulated genes revealed specific expression These profiles for each group, although the degree of hypertrophy was still similar in both. The ventricles destined to progress to dose failure showed activation of pro-apoptotic pathways, particularly related to mitochondria, whereas the group developing compensated hypertrophy showed blocked pro-death effector of signaling via p38-MAPK, through upregulation of MAPK phosphatase-1. In summary, we show that, already at an early time point, pivotal intervention. differences in gene expression exist between ventricles that will ultimately develop either a compensated or a decompensated phenotype, depending on genes the degree of pressure overload. These data reveal genes that may provide markers for the early prediction of clinical outcome subcutaneous as well as potential targets for early intervention.

Recently,3' fluorescent, monofunctional cis-platin derivatives have been developed to chemically label nucleic acids for use in fluorescent hybridization assays. Here we label show by hybridizations to microarrays containing oligonucleotide probes for the 3' ends, middle parts, and 5' ends of mRNAs, that oligonucleotide this labeling methodology bypasses the problem of the 3' end bias that is characteristic of the conventional enzymatic oligo(dT)-primed, reverse monofunctional transcription labeling of mRNAs.

Human decreased salivary chitinase could play a role in the defence against chitin-containing oral pathogens. The activity levels of chitinase in the saliva whole saliva of periodontitis patients were significantly higher than those in saliva from controls. Periodontal treatment for a period of beta-N-acetylhexosaminidase, 5-6 months resulted in a three- to fourfold decrease in this enzyme activity. The activity of beta-N-acetylhexosaminidase, which is another a enzyme that hydrolyses glycosidic linkages, also decreased as a result of treatment, although to a lesser extent. The decrease in a chitinase activity upon treatment of the disease did not correlate with the decrease that was seen in clinical attachment loss to and bleeding on probing, and only a weak correlation was observed with the changes in probing pocket depth and plaque this index. No correlations were found between the above clinical parameters and the decrease in beta-N-acetylhexosaminidase activity.

In chitinase recent studies the existence of a chitinase in various mammals, like man, was described. The aim of the present study chitinase was to find out whether salivas of periodontally healthy and inflamed humans also contain chitinase activity. Chitinase activity, assayed with was the substrate 4-methylumbelliferyl-beta-D-N,N',N"-triacetylchitotrioside, was shown to be present in human whole saliva, with an activity level and apparent molecular mass various (35 kDa) that were comparable with those of the human serum enzyme. Both lysozyme and beta-N-acetylhexosaminidase could be separated from various chitinase by means of Bio-Gel P-100 gel filtration chromatography. The enzyme was also present in glandular saliva of parotid, palatine,and submandibular and sublingual glands. The chitinase activity was not of oral epithelial, bacterial or plaque bacterial origin and was not chitinase correlated with the activity of salivary amylase. A comparative study of whole salivas of periodontally healthy controls and gingivitis and saliva periodontitis subjects showed that only in the case of periodontitis there was a significant increase of the specific chitinase activity.chitinase The latter enzyme showed a gel filtration pattern that was comparable with that of the enzyme from controls. The measured the albumin levels in saliva and the absence of correlation between the chitinase activity levels in plasma and saliva from periodontitis cavity. patients indicated that the (increased) chitinase activities did not originate from blood leakage to the oral cavity.