No implication of thromboxane prostanoid receptors in reactive hyperemia of skin and skeletal muscle in human forearm.

There is evidence that reactive hyperemia (ie, the transient increase of blood flow above resting level after a short period of ischemia) could be negatively modulated by vasoconstrictor prostanoids. The present study tested whether pharmacological blockade of the thromboxane prostanoid receptors with the specific antagonist S18886 (terutroban) would amplify reactive hyperemia in human skin and skeletal muscle. Twenty healthy young male volunteers were enrolled in a randomized, blinded, crossover trial of oral S18886 30 mg/d for 5 days versus placebo. Reactive hyperemia was evaluated in forearm skin and skeletal muscle, after occlusion of the brachial artery with a pneumatic cuff inflated at suprasystolic pressure. Blood flow was measured with laser Doppler imaging (skin) and strain gauge venous occlusion plethysmography (muscle). On the first and last day of each treatment period, recordings of reactive hyperemia were obtained immediately before and 2 hours after drug intake. Whether in forearm muscle or skin, S18886 had no discernible effect on peak postocclusion blood flow, nor on the global hyperemic response as quantified by the area under curve. These results do not support that thromboxane prostanoid receptor activation could exert a moderating influence on reactive hyperemia in human skin and skeletal muscle, at least in young subjects.

Immunocytochemical and pharmacological characterization of metabotropic glutamate receptors of the vestibular end organs in the frog.

Using immunocytochemistry and multiunit recording of afferent activity of the whole vestibular nerve, we investigated the role of metabotropic glutamate receptors (mGluR) in the afferent neurotransmission in the frog semicircular canals (SCC). Group I (mGluR1alpha) and group II (mGluR2/3) mGluR immunoreactivities were distributed to the vestibular ganglion neurons, and this can be attributed to a postsynaptic locus of metabotropic regulation of rapid excitatory transmission. The effects of group I/II mGluR agonist (1S,3R)-1-aminocyclopentane-trans-1,3-dicarboxylic acid (ACPD) and antagonist (R,S)-alpha-methyl-4-carboxyphenylglycine (MCPG) on resting and chemically induced afferent activity were studied. ACPD (10-100 microM) enhanced the resting discharge frequency. MCPG (5-100 microM) led to a concentration-dependent decrease of both resting activity and ACPD-induced responses. If the discharge frequency had previously been restored by L-glutamate (L-Glu) in high-Mg2+ solution, ACPD elicited a transient increase in the firing rate in the afferent nerve suggesting that ACPD acts on postsynaptic receptors. The L-Glu agonists, alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) and N-methyl-D-aspartate (NMDA), were tested during application of ACPD. AMPA- and NMDA-induced responses were higher in the presence than absence of ACPD, implicating mGluR in the modulation of ionotropic glutamate receptors. These results indicate that activation of mGluR potentiates AMPA and NMDA responses through a postsynaptic interaction. We conclude that ACPD may exert modulating postsynaptic effects on vestibular afferents and that this process is activity-dependent.

Correlation between vasoconstrictor roles and mRNA expression of alpha1-adrenoceptor subtypes in blood vessels of genetically engineered mice.

We examined the contribution of each alpha(1)-adrenoceptor (AR) subtype in noradrenaline (NAd)-evoked contraction in the thoracic aortas and mesenteric arteries of mice. Compared with the concentration-response curves (CRCs) for NAd in the thoracic aortas of wild-type (WT) mice, the CRCs of mutant mice showed a significantly lower sensitivity. The pD(2) value in rank order is as follows: WT mice (8.21) > alpha(1B)-adrenoceptor knockout (alpha(1B)-KO) (7.77) > alpha(1D)-AR knockout (alpha(1D)-KO) (6.44) > alpha(1B)- and alpha(1D)-AR double knockout (alpha(1BD)-KO) (5.15). In the mesenteric artery, CRCs for NAd did not differ significantly between either WT (6.52) and alpha(1B)-KO mice (7.12) or alpha(1D)-KO (6.19) and alpha(1BD)-KO (6.29) mice. However, the CRC maximum responses to NAd in alpha(1D)- and alpha(1BD)-KO mice were significantly lower than those in WT and alpha(1B)-KO mice. Except in the thoracic aortas of alpha(1BD)-KO mice, the competitive antagonist prazosin inhibited the contraction response to NAd with high affinity. However, prazosin produced shallow Schild slopes in the vessels of mice lacking the alpha(1D)-AR gene. In the thoracic aorta, pA(2) values in WT mice for KMD-3213 and BMY7378 were 8.25 and 8.46, respectively, and in alpha(1B)-KO mice they were 8.49 and 9.13, respectively. In the mesenteric artery, pA(2) values in WT mice for KMD-3213 and BMY7378 were 8.34 and 7.47, respectively, and in alpha(1B)-KO mice they were 8.11 and 7.82, respectively. These pharmacological findings were in fairly good agreement with findings from comparison of CRCs, with the exception of the mesenteric arteries of WT and alpha(1B)-KO mice, which showed low affinities to BMY7378. We performed a quantitative analysis of the mRNA expression of each alpha(1)-AR subtype in these vessels in order to examine the correlation between mRNA expression level and the predominance of each alpha(1)-AR subtype in mediating vascular contraction. The rank order of each alpha(1)-AR subtype in terms of its vasoconstrictor role was in fairly good agreement with the level of expression of mRNA of each subtype, that is, alpha(1D)-AR > alpha(1B)-AR > alpha(1A)-AR in the thoracic aorta and alpha(1D)-AR > alpha(1A)-AR > alpha(1B)-AR in the mesenteric artery. No dramatic compensatory change of alpha(1)-AR subtype in mutant mice was observed in pharmacological or quantitative mRNA expression analysis.

Vasopressin dilates the rat carotid artery by stimulating V1 receptors.

The acute effects of various vasopressor agents on the diameter of the common carotid artery were studied in halothane-anesthetized normotensive rats. The animals were infused intravenously for 60 min with equipressor doses of angiotensin II (10 ng/min), the alpha1-stimulant methoxamine (5 microg/min), lysine vasopressin (5 mU/min), or vehicle. The arterial diameter was measured by using a high-resolution ultrasonic echo-tracking device. The three vasoconstrictors increased the carotid artery diameter, but this effect was significantly more pronounced with lysine vasopressin. Even a nonpressor dose of lysine vasopressin (1 mU/min) caused a significant increase in the arterial diameter. The lysine vasopressin-induced vasodilatation could be prevented by the administration of d(CH2)5Tyr(Me)AVP (10 microg, i.v.), a selective V1-vasopressinergic receptor antagonist. These data therefore suggest that a short-term increase in blood pressure induces in rats a distention of the carotid artery. The increase in arterial diameter seems to involve an active mechanism with lysine vasopressin caused by the stimulation of V1-vasopressinergic receptors.

Control of the peroxisomal beta-oxidation pathway by a novel family of nuclear hormone receptors.

Three novel members of the Xenopus nuclear hormone receptor superfamily have been cloned. They are related to each other and similar to the group of receptors that includes those for thyroid hormones, retinoids, and vitamin D3. Their transcriptional activity is regulated by agents causing peroxisome proliferation and carcinogenesis in rodent liver. All three Xenopus receptors activate the promoter of the acyl coenzyme A oxidase gene, which encodes the key enzyme of peroxisomal fatty acid beta-oxidation, via a cognate response element that has been identified. Therefore, peroxisome proliferators may exert their hypolipidemic effects through these receptors, which stimulate the peroxisomal degradation of fatty acids. Finally, the multiplicity of these receptors suggests the existence of hitherto unknown cellular signaling pathways for xenobiotics and putative endogenous ligands.

Influence of sex and age on T3 receptors and T3 concentration in the pituitary gland of the rat: consequences on TSH secretion.

A reduced secretion of thyroid hormones with age has been documented in humans and animals with no substantial increase in TSH secretion, which may be indicative of an age-related impairment of the pituitary sensitivity to the negative control exerted by thyroid hormones. We have evaluated in rats the influence of sex and age on pituitary T3 nuclear receptors--known to be determinant in the regulation of TSH secretion--as well as on T3 concentration in the pituitary gland. As regards sex, the density of T3 receptors and the concentration of T3 in pituitary gland and plasma were greater in females than in males whereas pituitary and plasma TSH concentrations were less. As for age, the density of T3 receptors was greater in old male rats than in young ones with no changes in pituitary T3 and plasma TSH concentrations. In old female rats in contrast, there was no significant increase in T3 receptors but pituitary T3 was less and plasma TSH greater than in young female rats. In both sexes plasma thyroid hormones and pituitary TSH were reduced with age whereas TSH response to TRH was not altered. These results illustrate sex and age differences in pituitary T3 receptors and pituitary T3 concentration as well as in TSH secretion. In young animals of both sexes an inverse correlation is observed between the density of pituitary T3 receptors and plasma TSH. In contrast, in old animals the absence of this correlation is suggestive of an age-related impairment of T3 action on the thyrotrophs or of changes pertaining to other factors modulating TSH secretion.

Characterization of the ligand-binding site of the serotonin 5-HT3 receptor: the role of glutamate residues 97, 224, AND 235.

Ligand-gated ion channels of the Cys loop family are receptors for small amine-containing neurotransmitters. Charged amino acids are strongly conserved in the ligand-binding domain of these receptor proteins. To investigate the role of particular residues in ligand binding of the serotonin 5-HT3AS receptor (5-HT3R), glutamate amino acid residues at three different positions, Glu97, Glu224, and Glu235, in the extracellular N-terminal domain were substituted with aspartate and glutamine using site-directed mutagenesis. Wild type and mutant receptor proteins were expressed in HEK293 cells and analyzed by electrophysiology, radioligand binding, fluorescence measurements, and immunochemistry. A structural model of the ligand-binding domain of the 5-HT3R based on the acetylcholine binding protein revealed the position of the mutated amino acids. Our results demonstrate that mutations of Glu97, distant from the ligand-binding site, had little effect on the receptor, whereas mutations Glu224 and Glu235, close to the predicted binding site, are indeed important for ligand binding. Mutations E224Q, E224D, and E235Q decreased EC50 and Kd values 5-20-fold, whereas E235D was functionally expressed at a low level and had a more than 100-fold increased EC50 value. Comparison of the fluorescence properties of a fluorescein-labeled antagonist upon binding to wild type 5-HT3R and E235Q, allowed us to localize Glu235 within a distance of 1 nm around the ligand-binding site, as proposed by our model.

Dispensable role of myeloid differentiation primary response gene 88 (MyD88) and MyD88-dependent toll-like receptors (TLRs) in a murine model of osteoarthritis.

OBJECTIVES: The aim of our study was to evaluate the role of cell-membrane expressed TLRs and the signaling molecule MyD88 in a murine model of OA induced by knee menisectomy (surgical partial removal of the medial meniscus [MNX]). METHODS: OA was induced in 8-10weeks old C57Bl/6 wild-type (WT) female (n=7) mice and in knockout (KO) TLR-1 (n=7), -2 (n=8), -4 (n=9) -6 (n=5), MyD88 (n=8) mice by medial menisectomy, using the sham-operated contralateral knee as a control. Cartilage destruction and synovial inflammation were evaluated by knee joint histology using the OARSI scoring method. Apoptotic chondrocytes and cartilage metabolism (collagen II synthesis and MMP-mediated aggrecan degradation) were analyzed using immunohistochemistry. RESULTS: Operated knees exhibited OA features at 8weeks post-surgery compared to sham-operated ones. In menisectomized TLR-1, -2, -4, and -6 deficient mice, cartilage lesions, synovial inflammation and cartilage metabolism were similar to that in operated WT mice. Accordingly, using the same approach, we found no significant protection in MyD88-deficient mice in terms of OA progression as compared to WT littermates. CONCLUSIONS: Deficiency of TLRs or their signalling molecule MyD88 did not impact on the severity of experimental OA. Our results demonstrate that MyD88-dependent TLRs are not involved in this murine OA model. Moreover, the dispensable role of MyD88, which is also an adaptor for IL-1 receptor signaling, suggests that IL-1 is not a key mediator in the development of OA. This latter hypothesis is strengthened by the lack of efficiency of IL-1β antagonist in the treatment of OA.

Per què els castellanoparlants no miren TV3? Proposta de recerca sobre gust televisiu: anàlisi de textos i receptors

Amb l’objectiu d’orientar una futura tesi doctoral, aquest treball de recerca planteja una investigació sobre audiència televisiva a Catalunya. A partir de les aportacions fetes pels investigadors del camp, tant a nivell empíric com teòric, es dissenya una metodologia d’anàlisi del text i la recepció per tractar de donar resposta a un fenomen particular: la poca presència de públiccastellanoparlant en l’audiència de TV3, la televisió pública catalana. Sota elparaigües del construccionisme social moderat, la recerca dissenya unes hipòtesis relacionades amb la distància percebuda respecte de la cadena, el gust televisiu i les característiques enunciatives dels missatges televisius, i planteja donar-hi resposta a partir de l’anàlisi de l’enunciació de les principals cadenes generalistes que emeten a Catalunya, per una banda, i amb la realització d’entrevistes i grups de discussió amb una mostra d’espectadors, per l’altra.

Die Töpferin und der Schmied : Basel-Gasfabrik, Grabung 1989/5

RésuméDie Grabung 1989/5 der Archäologischen Bodenforschung Basel-Stadt im Zentrum der Siedlung Basel-Gasfabrik umfasste eine Fläche von etwa 900 m2. Auf etwa 370 m2 konnten intakte latènezeitliche Strukturen festgestellt werden. Der ehemalige Gehhorizont und die darunter liegende Siedlungsschicht waren jedoch nirgends erhalten geblieben.Die in den gewachsenen Boden eingetieften Pfostengrub en erlaubten die Rekonstrulction von zwei rechteckigen Häusern mit je vier Eckpfosten, die als Wohngebäude gedeutet werden.Über das ganze Areal verteilt wurden 14 Gruben unterschiedlicher Form und Grösse ganz oder teilweise ausgegraben. Bei der flachen Grube 261 handelt es sich wohl um den eingetieften Bereich eines kleinen Grubenhauses, das für nicht näher bestimmbare handwerkliche Aktivitäten genutzt wurde. Die beiden mit einem Pfostenbau überdeckten Gruben 258 und 259 können hingegen mit grosser Wahrscheinlichkeit als Schmiedegruben gedeutet werden. In Grube 258 waren gar die Reste von zwei Feuerstellen und der Unterbau eines kaminartigen Abzugs in situ erhalten. Die Gruben 253, 254A, 254B, 255,256 und 257 mit Resten von Lehmausideidungen dienten vermutlich als Getreidesilos. Sie liegen alle im Zentrum des Grabungsareals im Bereich einer anstehenden Lehmschicht. Bei einigen weiteren Gruben könnte es sich um Keller handeln.Im Westen der Grabungsfläche kamen die Heizkanäle von zwei Töpferöfen zum Vorschein. Es handelt sich um einfache Kuppelöfen mit gegenständiger Feuerung. In geringer Entfernung lagen drei kleine Gruben, die vielleicht zur Aufbereitung des Töpferlehms dienten. Aufgrund der klimatischen Verhältnisse ist mit einem saisonalen Betrieb der Töpferöfen während der Sommermonate zu rechnen. Die notwendigen Arbeiten wurden mit grosser Wahrscheinlichkeit von Frauen ausgeführt, da die Männer während dieser Jahreszeit in der Landwirtschaft beschäftigt waren. Der Töpfereibetrieb produzierte grössere Serien scheibengedrehter, reduzierend gebrannter Feinkeramik, von der im Bereich der Öfen zahlreiche Scherben gefunden wurden.Die räumliche Organisation der Bebauung zeugt von einer einheitlichen und wohl kontinuierlichen Nutzung der Grabungsfläche. ImWesten gruppieren sich Haus 1 und die Einrichtungen des Töpfereibetriebs um eine unbebaute Hofzone. Nach Osten zu folgt dann die Zone mit den Getreidesilos. Das östliche Bauensemble besteht aus Haus 2, den Schmiedegruben, dem Grubenhaus sowie einer mutmasslichen Kellergrube. Stratigra-phische Überschneidungen von Befunden sind nur im Bereich der Töpferöfen vorhanden.Die Grubenfüllungen erwiesen sich als sehr fundreich. Die gegen 20000 Keramikscherben repräsentieren das gesamte Spektrum der aus Basel-Gasfabrik bekannten einheimischen Grobund Feinkeramik. Bei den seltenen Scherben von Graphittonkeramik handelt es sich um Importe aus dem ostkeltischen Bereich.Die zahlreichen Amphorenscherben stammen von mindestens 35 verschiedenen Gefässindividuen des Typs Dressel 1A, von denen aber immer nur ein kleiner Prozentsatz vorhanden war. Die Typologie und die Datierung der Befunde zeigen, dass der Amphorenimport erst im Laufe von LT D1 einsetzte.Unter den Kleinfunden verdienen die zahlreichen Silber- und Potinmünzen sowie die Glasfunde besondere Erwähnung. Das Fragment eines möglicherweise latènezeitlichen Glasgefässes stammt leider aus einem unsicheren Fundzusammenhang.Die über 70 Fibeln und Fibelteile sind zu 70% aus Bronze gefertigt. Das Fibelspektrum wird von den Nauheimer Fibeln dominiert. Besonders bei den Eisenfibeln konnten aber auch typologisch ältere Typen identifiziert werden, welche in die Stufen LT C2 und LT D1a gehören. Bei den Glasarmringen machen mittellatènezeitliche Typen gegen 20 % der Fundmenge aus.Von besonderer Bedeutung sind die 23 Menschenknochen von 19 verschiedenen Individuen. Es handelt sich dabei um einen fast vollständigen Schädel, Schädelteile und Fragmente von Langknochen sowie wenige weitere Skelettteile. Die Zusammensetzung des Ensembles und der Zustand der Einzelknochen lässt sich sehr gut mit den Menschenknochen aus den Altgrabungen von Basel-Gasfabrik und aus Manching vergleichen. Eine detaillierte Untersuchung zeigt, dass die Knochen am Ende eines langwierigen Totenrituals, das aufgrund ethno - logischer Vergleiche als mehrstufige Bestattung bezeichnet werden kann, in der Siedlung vergraben wurden. In der Umgebung der Menschenknochen wurden in einigen Fällen auffallend viele Amphorenscherben gefunden, die bezeugen, dass der importierte Wein bei diesen Riten eine bedeutende Rolle spielte. Wahrscheinlich wurden diese Knochen auch als Ahnenrelikte verehrt.Neben den mehrstufigen Bestattungen konnten auch zwei Säuglingsbestattungen identifiziert werden.Die Analyse von Fundmenge und Fundverteilung in den Gruben zeigt, dass die Funde nicht direkt, sondern auf dem Umweg über primäre Deponien in die Gruben gelangten. In den pri-mären Deponien wurden Funde über längere Zeit akkumuliert und mit verschiedenen Erdmaterialien intensiv vermischt.Für den Grossteil der Funde wird eine profane Deutung als Abfälle vorgeschlagen. Daneben können aber auch einige Funde als gezielte Deponierungen angesprochen werden. Ver-schiedene Fibelpaare und wahrscheinlich auch Münzen wurden wohl als Opfergaben in die Gruben gegeben. Ein Zusammenhang dieser Opfergaben mit den Bestattungen von Men-schenknochen ist nicht erkennbar.Die Datierung der Funde zeigt einen Siedlungsbeginn in LT C2 und ein Ende noch vor dem Beginn der Stufe LT D2. Die Kombination dieser Datierungen mit den stratigraphischen Überschneidungen einiger Befunde erlaubt eine Rekonstruktion der Besiedlungsentwicklung auf dem Grabungsareal. Die Gruben 259 und 260 wurden bereits in LT C2 verfüllt, die Schmiedegrube 259 anschliessend durch Grube 258 ersetzt. Die beiden Töpferöfen waren nacheinander in LT D1 in Betrieb. Die beiden Häuser können leider nicht genauer datiert werden.Die Besiedlung des Area's setzte noch vor 150 V. Chr. ein und dauerte maximal 80 Jahre.Die Synthese aller Untersuchungen zeigt, dass auf dem Grabungsareal eine Gruppe von etwa 15 Personen ansässig war, die sich v. a. der Landwirtschaft widmete. Die handwerkliche Tätigkeit (Töpferei, Metallverarbeitung) erreichte keinen vollberuflichen Standard.Die dörfliche Siedlungsgemeinschaft bestand aus bäuerlichen Selbstversorgern, mit anderen Gemeinschaften wurden jedoch Keramik, Handwerksprodukte und Schlachtvieh ausge-tauscht. Von besonderer Bedeutung war offenbar das Getreide, das in grossen Mengen in der Siedlung gelagert wurde und mit grosser Wahrscheinlichkeit für den Export bestimmt war. Die Herkunft dieses Getreides und die sozialen Strukturen hinter diesem Austausch sind nicht ldar, aber mit hoher Wahrscheinlichkeit wurden durch den Getreideexport die Importe wie Salz, Rohstoffe (Metall) und Wein ermöglicht.

Positive regulation of the peroxisomal beta-oxidation pathway by fatty acids through activation of peroxisome proliferator-activated receptors (PPAR).

Peroxisome proliferators regulate the transcription of genes by activating ligand-dependent transcription factors, which, due to their structure and function, can be assigned to the superfamily of nuclear hormone receptors. Three such peroxisome proliferator-activated receptors (PPAR alpha, beta, and gamma) have been cloned in Xenopus laevis. Their mRNAs are expressed differentially; xPPAR alpha and beta but not xPPAR gamma are expressed in oocytes and embryos. In the adult, expression of xPPAR alpha and beta appears to be ubiquitous, and xPPAR gamma is mainly observed in adipose tissue and kidney. Immunocytochemical analysis revealed that PPARs are nuclear proteins, and that their cytoplasmic-nuclear translocation is independent of exogenous activators. A target gene of PPARs is the gene encoding acyl-CoA oxidase (ACO), which catalyzes the rate-limiting step in the peroxisomal beta-oxidation of fatty acids. A peroxisome proliferator response element (PPRE), to which PPARs bind, has been identified within the promoter of the ACO gene. Besides the known xenobiotic activators of PPARs, such as hypolipidemic drugs, natural activators have been identified. Polyunsaturated fatty acids at physiological concentrations are efficient activators of PPARs, and 5,8,11,14-eicosatetraynoic acid (ETYA), which is the alkyne homolog of arachidonic acid, is the most potent activator of xPPAR alpha described to date. Taken together, our data suggest that PPARs have an important role in lipid metabolism.

Genomics of secondary metabolite production by Pseudomonas fluorescens Pf-5

The complete sequence of the 7.07 Mb genome of the biological control agent Pseudomonas fluorescens Pf-5 is now available, providing a new opportunity to advance knowledge of biological control through genomics. P. fluorescens Pf-5 is a rhizosphere bacterium that suppresses seedling emergence diseases and produces a spectrum of antibiotics toxic to plant-pathogenic fungi and oomycetes. In addition to six known secondary metabolites produced by Pf-5, three novel secondary metabolite biosynthesis gene clusters identified in the genome could also contribute to biological control. The genomic sequence provides numerous clues as to mechanisms used by the bacterium to survive in the spermosphere and rhizosphere. These features include broad catabolic and transport capabilities for utilizing seed and root exudates, an expanded collection of efflux systems for defense against environmental stress and microbial competition, and the presence of 45 outer membrane receptors that should allow for the uptake of iron from a wide array of siderophores produced by soil microorganisms. As expected for a bacterium with a large genome that lives in a rapidly changing environment, Pf-5 has an extensive collection of regulatory genes, only some of which have been characterized for their roles in regulation of secondary metabolite production or biological control. Consistent with its commensal lifestyle, Pf-5 appears to lack a number of virulence and pathogenicity factors found in plant pathogen.

Peroxisome proliferator activated receptors: transcriptional regulators of adipogenesis, lipid metabolism and more....

The recent discovery of lipid-activatable transcription factors that regulate the genes controlling lipid metabolism and adipogenesis has provided insight into the way that organisms sense and respond to lipid levels. Identification of the signaling pathways in which these receptors are involved will help us to understand the control of energy balance and the molecular defects underlying its disorders.

DNA binding properties of peroxisome proliferator-activated receptor subtypes on various natural peroxisome proliferator response elements. Importance of the 5'-flanking region.

The three subtypes of the peroxisome proliferator-activated receptors (PPARalpha, beta/delta, and gamma) form heterodimers with the 9-cis-retinoic acid receptor (RXR) and bind to a common consensus response element, which consists of a direct repeat of two hexanucleotides spaced by one nucleotide (DR1). As a first step toward understanding the molecular mechanisms determining PPAR subtype specificity, we evaluated by electrophoretic mobility shift assays the binding properties of the three PPAR subtypes, in association with either RXRalpha or RXRgamma, on 16 natural PPAR response elements (PPREs). The main results are as follows. (i) PPARgamma in combination with either RXRalpha or RXRgamma binds more strongly than PPARalpha or PPARbeta to all natural PPREs tested. (ii) The binding of PPAR to strong elements is reinforced if the heterodimerization partner is RXRgamma. In contrast, weak elements favor RXRalpha as heterodimerization partner. (iii) The ordering of the 16 natural PPREs from strong to weak elements does not depend on the core DR1 sequence, which has a relatively uniform degree of conservation, but correlates with the number of identities of the 5'-flanking nucleotides with respect to a consensus element. This 5'-flanking sequence is essential for PPARalpha binding and thus contributes to subtype specificity. As a demonstration of this, the PPARgamma-specific element ARE6 PPRE is able to bind PPARalpha only if its 5'-flanking region is exchanged with that of the more promiscuous HMG PPRE.