Nascent histamine induces α-synuclein and caspase-3 on human cells

Histamine (Hia) is the most multifunctional biogenic amine. It is synthetized by histidine decarboxylase (HDC) in a reduced set of mammalian cell types. Mast cells and histaminergic neurons store Hia in specialized organelles until the amine is extruded by exocytosis; however, other immune and cancer cells are able to produce but not store Hia. The intracellular effects of Hia are still not well characterized, in spite of its physiopathological relevance. Multiple functional relationships exist among Hia metabolism/signaling elements and those of other biogenic amines, including growth-related polyamines. Previously, we obtained the first insights for an inhibitory effect of newly synthetized Hia on both growth-related polyamine biosynthesis and cell cycle progression of non-fully differentiated mammalian cells. In this work, we describe progress in this line. HEK293 cells were transfected to express active and inactive versions of GFP-human HDC fusion proteins and, after cell sorting by flow cytometry, the relative expression of a large number of proteins associated with cell signaling were measured using an antibody microarray. Experimental results were analyzed in terms of protein–protein and functional interaction networks. Expression of active HDC induced a cell cycle arrest through the alteration of the levels of several proteins such as cyclin D1, cdk6, cdk7 and cyclin A. Regulation of α-synuclein and caspase-3 was also observed. The analyses provide new clues on the molecular mechanisms underlying the regulatory effects of intracellular newly synthetized Hia on cell proliferation/survival, cell trafficking and protein turnover. This information is especially interesting for emergent and orphan immune and neuroinflammatory diseases.     

Functional Characterization of the Xanthophyllomyces dendrorhous Farnesyl Pyrophosphate Synthase and Geranylgeranyl Pyrophosphate Synthase Encoding Genes That Are Involved in the Synthesis of Isoprenoid Precursors

The yeast Xanthophyllomyces dendrorhous synthesizes the carotenoid astaxanthin, which has applications in biotechnology because of its antioxidant and pigmentation properties. However, wild-type strains produce too low amounts of carotenoids to be industrially competitive. Considering this background, it is indispensable to understand how the synthesis of astaxanthin is controlled and regulated in this yeast. In this work, the steps leading to the synthesis of the carotenoid precursor geranylgeranyl pyrophosphate (GGPP, C₂₀) in X. dendrorhous from isopentenyl pyrophosphate (IPP, C₅) and dimethylallyl pyrophosphate (DMAPP, C₅) was characterized. Two prenyl transferase encoding genes, FPS and crtE, were expressed in E. coli. The enzymatic assays using recombinant E. coli protein extracts demonstrated that FPS and crtE encode a farnesyl pyrophosphate (FPP, C₁₅) synthase and a GGPP-synthase, respectively. X. dendrorhous FPP-synthase produces geranyl pyrophosphate (GPP, C₁₀) from IPP and DMAPP and FPP from IPP and GPP, while the X. dendrorhous GGPP-synthase utilizes only FPP and IPP as substrates to produce GGPP. Additionally, the FPS and crtE genes were over-expressed in X. dendrorhous, resulting in an increase of the total carotenoid production. Because the parental strain is diploid, the deletion of one of the alleles of these genes did not affect the total carotenoid production, but the composition was significantly altered. These results suggest that the over-expression of these genes might provoke a higher carbon flux towards carotenogenesis, most likely involving an earlier formation of a carotenogenic enzyme complex. Conversely, the lower carbon flux towards carotenogenesis in the deletion mutants might delay or lead to a partial formation of a carotenogenic enzyme complex, which could explain the accumulation of astaxanthin carotenoid precursors in these mutants. In conclusion, the FPS and the crtE genes represent good candidates to manipulate to favor carotenoid biosynthesis in X. dendrorhous.     

Redox activities in plasma membrane vesicles isolated from papaya (Carica papaya) leaves

Plasma membrane vesicles were isolated from papaya (Carica papaya) leaf cells by a two-phase partition system. Redox activities of these vesicles were determined by ferricyanide reduction and NADH oxidation. Ferricyanide reductase activity was accompanied by continuous acidification of the medium and was stimulated by fusicoccin. NADH oxidase activity was inhibited by catalase.Abbreviations FC fusicoccin - PCMBS parachloromercuri-benzenesulfonic acid     

Quantitation and characterization of the microtubule associated MAP2 in porcine tissues and its isolation from porcine (PK15) and human (HeLa) cell lines

A radioimmunoassay developed for the microtubule associated protein MAP₂ shows that this protein, or related polypeptides are present in all the porcine tissues studied. Nervous tissues (brain, 11 μg MAP₂/mg protein; cerebellum, 9.7 μg MAP₂/mg protein) contain much higher levels of MAP₂ than non-nervous tissues (kidney, 104 ng MAP₂/mg protein; lung 89 ng MAP₂/mg protein; spleen 66 ng MAP₂/mg protein; thyroid 21 ng MAP₂/mg protein; liver 9.7 ng MAP₂/mg protein). A heat resistant protein doublet of 300,000 with the ability to promote microtubule polymerization has been purified from pig kidney cells by affinity chromatography using MAP₂ antibodies. Using a similar purification method a protein of 200,000 daltons has been isolated from Hela cells.     

Growth-Dependent Stable Carbon Isotope Fractionation by Basidiomycete Fungi: δ13C Pattern and Physiological Process

We grew 11 basidiomycetes in axenic culture to characterize their physiological capacities to fractionate stable C isotopes. Generally, δ¹³C values of the fungal biomass were (i) enriched in ¹³C relative to the growth medium, (ii) variable among the isolates, and (iii) dependent on the growth rate and growth stage of the fungi. We found a multiphasic dynamic of fractionation for Cryptoporus volvatus and Marasmius androsaceus during various growth stages. The first phase, P1, corresponded to the exponential growth stage and was characterized by an increasing enrichment in ¹³C content of the fungal biomass relative to the growth medium ranging between 4.6 and 6.9‰. The second phase, P2, exhibited a continual depletion in ¹³C of the fungal biomass, with the δ¹³C values of the fungal biomass asymptotically returning to the δ¹³C value of the growth medium at inoculation. The expression of the various fractionation phases was dependent on the amount of low-concentration micronutrients and growth factors added to the growth medium. The onset of P2 occurred at reduced concentrations of these elements. All of the sugars in the growth medium (sucrose, maltose, and glucose) were utilized for growth, indicating that the observed fractionation was not an artifact derived from the preferential use of ¹³C-rich maltose, which was found at low concentrations in the growth medium. In this study, we establish a framework with which to explore the impact of physiological fractionations by fungal interfaces on natural distributions of stable C isotopes.     

Traditional farming and key foraging habitats for chough Pyrrhocorax pyrrhocorax conservation in a Spanish pseudosteppe landscape

1. Agricultural land devoted to low-intensity practices supports many declining bird species in Europe. The potential effects of intensification or abandonment of traditional farming practices are assessed from the point of view of the conservation of the chough Pyrrhocorax pyrrhocorax .
2. The study was conducted in Los Monegros (north-east Spain), in a pseudosteppe landscape of special relevance for this species. In this area traditional farming survives in the form of low-intensity arable systems including fallowing and extensive sheep grazing.
3. Foraging choughs showed a strong selection for field margins, avoided any remnant of the original scrubland and halophitic vegetation, and utilized other habitats maintained by traditional crop rotation (e. g. fallow, stubble) at different times of the year.
4. A highly complex pattern of habitat selection was found which was not only related to seasonal changes in habitat availability, but also differed between territorial pairs and the non-breeding population. This emphasizes the importance of taking all the different components of bird populations into account when analysing interactions between habitat selection and the annual farming cycle.
5. Chough habitat selection suggests that both agricultural intensification and land abandonment would have detrimental consequences for this species in Los Monegros. The maintenance of traditional farming is recommended, especially by promoting the use of long fallow rotations grazed extensively by livestock, encouraging the increase of grassland edges around landholdings and sand roads, and maintaining a low input of biocides. These management measurements could also favour most other endangered species of steppe birds, and could be supported by the establishment of Zonal Programmes under the CAP Agri-Environment Regulation (2078/92).     

The relative contributions of sexual reproduction and clonal propagation in Opuntia rastrera from two habitats in the Chihuahuan Desert

1 The clonal cactus Opuntia rastrera shows predominantly sexual reproduction in grasslands (GH) and clonal propagation in nopaleras (NH). We assessed the effects of light, herbivory, water availability and the habitat an offspring came from on the survival and growth of sexual or clonal offspring (i.e. seedlings and cladodes), through 3- and 4-year common garden and short-term greenhouse experiments.
2 Shading by nurse plants increased seedling survival in the field by an order of magnitude, and a small additional advantage due to predator protection by grasses was observed. Strong herbivory transforms a facultative nurse–protégé relationship for seedlings into an obligatory one.
3 In the greenhouse seedlings grew better under shade, but in the field the production of the first cladode was delayed in seedlings in the more shaded GH. Competition for soil resources may be more intense under a dense grass tussock than under a open shrub, thus affecting the nurse–protégé relationship. Seedling survival under nurse plants was similar in GH and NH, but higher plant cover suggests that a larger number of seedlings will establish in GH in the long term.
4 Cladode survival was higher in NH. Cladodes were more successful than seedlings at establishing in intercanopy areas, possibly due to physiological differences as well as their ability to survive partial predation. Cladode survival in intercanopy areas may explain the enhanced clonal propagation in the more open NH scrubland, together with their susceptibility to the flooding which affects GH.
5 The high seedling and cladode survival in the greenhouse experiments contrasted with that observed in the field, indicating that survival is determined by the interaction between herbivores, plants and abiotic conditions rather than the physiological aptitude of the plants.     

Isocitrate dehydrogenases and oxoglutarate dehydrogenase activities of baker's yeast grown in a variety of hypoxic conditions

Molecular and Cellular Biochemistry - The activities of isocitrate dehydrogenase (NAD), isocitrate dehydrogenase (NADP) and oxoglutarate dehydrogenase have been investigated inSaccharomyces...     

Growth and cell wall changes in rice coleoptiles growing under different conditions I. Changes in turgor pressure and cell wall polysaccharides during intact growth

The effect of the oxygen supply on growth, water absorptionof cells and cell wall changes was studied in coleoptiles ofrice seedlings growing under three different conditions: underwater, under water with constant air bubbling and in air. Coleoptilegrowth was larger when they were grown under water than in waterwith air bubbling and in air. Coleoptile growth under waterwas limited by the suction force of their cells rather thanby mechanical properties of the cell wall, while that of thecoleoptiles growing under the other two conditions was limitedby the cell wall rigidity. A decrease in the relative amountof noncellulosic glucose of the cell wall, and an increase inthe noncellulosic xylose during coleoptile growth were foundfor all three culture conditions. ¹ Present address: Departamento de Fisiologia Vegetal, Facultadde Ciencias, Universidad, Salamanca, Spain. (Received May 21, 1979; )