Heme regulates cytochrome P-450 gene transcription elongation

Administration of 3-methylcholanthrene (MC) to rats results in a striking increase in the transcription of cytochrome P-450 (c+d) messenger RNA with isolated nuclei, which is blocked by the simultaneous administration of cobalt chloride, an inhibitor of heme biosynthesis. Transcription of cytochrome P-450 (c+d) mRNAs with nuclei isolated from MC treated rats shows a linear increase with time of incubation, whereas it shows a progressive decrease with incubation time in the case of nuclei isolated from MC+CoCl2 treated rats. Addition of heme in vitro (10−6M) to the latter nuclei results in a significant counteraction of the decreased cytochrome P-450 (c+d) mRNA transcription. The inhibition in transcription rates observed in MC+CoCl2 treated rat liver nuclei is more pronounced with the seventh exon probe than with the second exon probe. Once again, in vitro heme addition can counteract the inhibition observed with both the probes. Since run off transcription with isolated nuclei represents essentially elongation of the initiated transcripts, the data obtained can be interpreted on the basis that heme regulates cytochrome P-450 gene transcription elongation.

Toksoplasman esiintyvyys suomalaisissa kissoissa

Toxoplasma gondii on kokkideihin kuuluva alkueläinloinen. Sen pääisäntiä ovat kissaeläimet, joissa tapahtuvan suvullisen lisääntymisen tuloksena tuotetaan ympäristöön ookystia. Väli-isäntiä voivat olla kaikki tasalämpöiset eläimet. Toksoplasma muodostaa isäntien kudoksiin infektiivisiä kudoskystia. Ihminen voi saada tartunnan syömällä kudoskystia tai ookystia, tai sikiö voi infektoitua jo kohdussa istukan kautta. Toxoplasma gondii voi aiheuttaa isäntänsä vakavan sairastumisen ja on siksi maailmanlaajuisesti merkittävä zoonoottinen loinen. Toksoplasman torjunnassa on oleellista tuntea alueen kissojen toksoplasmaseroprevalenssi ja arvioida ympäristön ookystakuormitusta: seropositiivisten kissojen voidaan olettaa joskus erittäneen ookystia. Tässä tutkimuksessa selvitettiin toksoplasman esiintymistä suomalaisissa kissoissa: suoralla agglutinaatiotestillä (ToxoScreen-DA) määritettiin IgG-vasta-aineiden esiintymistä seerumissa sekä vasta-ainetasoja (tiitteri). Flotac® - flotaatiomenetelmällä tutkittiin ookystien esiintymistä ulostenäytteissä. Tutkimuksessa selvitettiin kissojen toksoplasma-vasta-aineiden esiintymiseen vaikuttavia tekijöitä: serologisessa tutkimuksessa oli mukana sekä löytöettä rotukissoja, ja lisäksi tutkittiin kissojen sukupuolen, iän, sekä rotukissoilla lihansyönnin vaikutusta vastaaineiden esiintymiseen seerumissa. Serologisessa tutkimuksessa 398 kissan aineistossa seroprevalenssiksi saatiin 48,2 %. Tutkituista 369 rotukissasta 49,9 % oli seropositiivisia, kun taas 27 tutkitusta löytökissasta seropositiivisia oli 25,9 % (P<0,05). Sukupuolella ei todettu olevan merkitystä kissan seropositiivisuuteen. Aikuiset kissat olivat nuoria kissoja useammin seropositiivisia (53,7 % vs. 23,5 %) (P<0,001), koska kerran tartunnan saaneen kissan seerumista voidaan todeta vasta-aineita, ja vanhemmat kissat ovat nuoria todennäköisemmin ehtineet törmätä loiseen elämänsä aikana ja saada tartunnan. Ruokavalio oli tiedossa 347 rotukissalta, ja näistä 270 (77,8 %) oli joskus saanut raakaa lihaa. Raakaa lihaa saaneista kissoista 151 (55,9 %) oli seropositiivisia; 77 kissasta, jotka eivät olleet saaneet raakaa lihaa sitä vastoin ainoastaan 26 (33,8 %) oli seropositiivisia (P<0,001). Ulostenäytteistä 131 kissan aineistosta 1,5 % eritti toksoplasman kaltaisia ookystia ulosteessaan tutkimushetkellä. Suomessa kissojen ravinnon ja toksoplasmaseropositiivisuuden välistä yhteyttä ei ole aikaisemmin tutkittu. Näiden uusien tulosten valossa olisi toksoplasman torjunnassa erityisen tärkeää kiinnittää huomiota loisen tartuntareitteihin kissan osalta. Kissoja ei tulisi ruokkia raa’alla lihalla: kissalle annettava liha olisi hyvä kuumentaa yli 67ºC:een tai pakastaa alle -12ºC:n lämpötilassa toksoplasmatartunnan ehkäisemiseksi.

Terminal solid solubilities at 900–1000°C in the magnesium oxide-zinc oxide system measured using a magnesium fluoride solid-electrolyte galvanic cell

The terminal solid solubilities of the periclase (MgO-rich) and zincite (ZnO-rich) solid solutions in the MgO---ZnO system have been determined by measuring the activity of MgO using a solid-state galvanic cell of the type 02(g), Pt/MgO, MgF2//MgF2//{χMgO+(1-χ)ZnO}(s, sln), MgF2/Pt, O2(g) in the temperature range 900–1050°C. The ZnO activity was calculated by graphical Gibbs-Duhem integration. The activity-composition plots of both components exhibit a strong positive deviation from ideality and are characterised by a miscibility gap. The terminal solid solubilities of the periclase and zincite solid solutions obtained from the activity-composition plots are found to be in reasonable agreement with those reported in the literature.

Metal-insulator transitions in Cr-doped V2O3 by x-ray and ultraviolet photoelectron spectroscopy

X-ray and ultraviolet photoelectron spectroscopy have been employed to investigate the high temperature metal-insulator transitions in V2O3 and (V0.99Cr0.01)2O3. The high temperature transitions are associated with more gradual changes in the 3d bands compared to the low-temperature transitions

Inhibition of rat liver mevalonate pyrophosphate decarboxylase and mevalonate phosphate kinase by phenyl and phenolic compounds.

1. Mevalonate pyrophosphate decarboxylase of rat liver is inhibited by various phenyl and phenolic acids. 2. Some of the phenyl and phenolic acids also inhibited mevalonate phosphate kinase. 3. Compounds with the phenyl-vinyl structure were more effective. 4. Kinetic studies showed that some of the phenolic acids compete with the substrates, mevalonate 5-phosphate and mevalonate 5-pyrophosphate, whereas others inhibit umcompetitively. 5. Dihydroxyphenyl and trihydroxyphenyl compounds and p-chlorophenoxyisobutyrate, a hypocholesterolaemic drug, had no effect on these enzymes. 6. Of the three mevalonate-metabolizing enzymes, mevalonate pyrophosphate decarboxylase has the lowest specific activity and is probably the rate-determining step in this part of the pathway.

A Priori and the A Posteriori

The distinction between a priori and a posteriori knowledge has been the subject of an enormous amount of discussion, but the literature is biased against recognizing the intimate relationship between these forms of knowledge. For instance, it seems to be almost impossible to find a sample of pure a priori or a posteriori knowledge. In this paper it will be suggested that distinguishing between a priori and a posteriori is more problematic than is often suggested, and that a priori and a posteriori resources are in fact used in parallel. We will define this relationship between a priori and a posteriori knowledge as the bootstrapping relationship. As we will see, this relationship gives us reasons to seek for an altogether novel definition of a priori and a posteriori knowledge. Specifically, we will have to analyse the relationship between a priori knowledge and a priori reasoning, and it will be suggested that the latter serves as a more promising starting point for the analysis of aprioricity. We will also analyse a number of examples from the natural sciences and consider the role of a priori reasoning in these examples. The focus of this paper is the analysis of the concepts of a priori and a posteriori knowledge rather than the epistemic domain of a posteriori and a priori justification.

Information and Decision Technologies

Production scheduling in a flexible manufacturing system (FMS) is a real-time combinatorial optimization problem that has been proved to be NP-complete. Solving this problem needs on-line monitoring of plan execution and requires real-time decision-making in selecting alternative routings, assigning required resources, and rescheduling when failures occur in the system. Expert systems provide a natural framework for solving this kind of NP-complete problems.In this paper an expert system with a novel parallel heuristic approach is implemented for automatic short-term dynamic scheduling of FMS. The principal features of the expert system presented in this paper include easy rescheduling, on-line plan execution, load balancing, an on-line garbage collection process, and the use of advanced knowledge representational schemes. Its effectiveness is demonstrated with two examples.

Antigenic determinants on chicken riboflavin carrier protein. A study with monoclonal antibodies

Monoclonal antibodies raised against chicken egg white riboflavin carrier protein were classified into seven categories each recognizing a distinct epitope. Of these, six were directed against conformation dependent epitopes and one to a sequential epitope. The roles of lysine residues and the post-translationally attached phosphate and oligosaccharide moieties in the antigenicity of riboflavin carrier protein recognized by the monoclonal antibodies were investigated. The binding region of three monoclonal antibodies could be located within the 87–219 amino acid sequence of the protein and one antibody among these recognized a sequence of 182–204 amino acid residues. All the monoclonal antibodies were able to recognize riboflavin carrier proteins present in the sera of pregnant rats, cows and humans indicating that the epitopes to which they are directed are conserved through evolution from chicken to the human.

Drosophila “enhancer-trap” transposants: Gene expression in chemosensory and motor pathways and identification of mutants affected in smell and taste ability

We have isolated about a thousandDrosophila P-element transposants that allow thein situ detection of genomic enhancer elements by a histochemical assay for β-galactosidase activity. We summarize the β-galactosidase staining patterns of over 200 such transposants in the adult. Our aim was to identify genes that are likely to be involved in the chemosensory and motor pathways ofDrosophila. Based on β-galactosidase expression patterns in the tissues of our interest, we have chosen some strains for further analysis. Behavioral tests on a subset of the transposants have, in addition, identified several strains defective in their chemosensory responses.

Characterization of poly- and monoclonal antibodies to physalis mottle (tymo) virus

Polyclonal antibodies were raised against the Physalis mottle virus (PhMV) and its denatured coat protein (PhMV-P). Analysis of the reactivity of the polyclonal antibodies with tryptic peptides of PhMV-P in dot-blot assays revealed that many of the epitopes were common to intact virus and denatured coat protein. Five monoclonal antibodies to the intact virus were obtained using hybridoma technology. These monoclonal antibodies reacted well with the denatured coat protein. Epitope analysis suggested that probably these monoclonal antibodies recognize overlapping epitopes. This was substantiated by epitope mapping using the CNBr digest of PhMV-P in western blots. All the five monoclonals recognized the N-terminal 15 K fragment. Attempts to further delineate the specific region recognized by the monoclonals by various enzymatic cleavages resulted in the loss of reactivity in all the cases. The results indicate that these monoclonals probably recognize epitopes within the N-terminal 15 K fragment of the coat protein.

Identification of tropomyosin as the major shrimp allergen and characterization of its IgE-binding epitopes

The major heat-stable shrimp allergen (designated as Sa-II), capable of provoking IgE-mediated immediate type hypersensitivity reactions after the ingestion of cooked shrimp, has been shown to be a 34-kDa heat- stable protein containing 300 amino acid residues. Here, we report that a comparison of amino acid sequences of different peptides generated by proteolysis of Sa-II revealed an 86% homology with tropomyosin from Drosophila melanogaster, suggesting that Sa-II could be the shrimp muscle protein tropomyosin. To establish that Sa-II is indeed tropomyosin, the latter was isolated from uncooked shrimp (Penaeus indicus) and its physicochemical and immunochemical properties were compared with those of Sa-II. Both tropomyosin and Sa-II had the same molecular mass and focused in the isoelectric pH range of 4.8 to 5.4. In the presence of 6 M urea, the mobility of both Sa-II and shrimp tropomyosin shifted to give an apparent molecular mass of 50 kDa, which is a characteristic property of tropomyosins. Shrimp tropomyosin bound to specific IgE antibodies in the sera of shrimp-sensitive patients as assessed by competitive ELISA inhibition and Western blot analysis. Tryptic maps of both Sa-II and tropomyosin as obtained by reverse phase HPLC were superimposable. Dot-blot and competitive ELISA inhibition using sera of shrimp-sensitive patients revealed that antigenic as well as allergenic activities were associated with two peptide fractions. These IgE-binding tryptic peptides were purified and sequenced. Mouse anti-anti-idiotypic antibodies raised against Sa-II specific human idiotypic antibodies recognized not only tropomyosin but also the two allergenic peptides, thus suggesting that these peptides represent the major IgE binding epitopes of tropomyosin. A comparison of the amino acid sequence of shrimp tropomyosin in the region of IgE binding epitopes (residues 50-66 and 153-161) with the corresponding regions of tropomyosins from different vertebrates confirmed lack of allergenic cross-reactivity between tropomyosins from phylogenetically distinct species.