Metabolism of (±) 2-[14C]-abscisic acid in Lactuca sativa achenes

Achenes ofLactuca sativa L. cv. Grand Rapids were treated with (±) 2-[¹⁴C]-abscisic acid (ABA) at 10⁵ - or 2-10⁶ M for 6, 12, 24, 48 or 96 h in darkness at 24°C. They were then extracted in 80% ethanol. Two acidic diethyl ether phases which contained the free acids and the acids released after mild alkaline hydrolysis respectively, were analyzed as well as the radioactivity which remained in the final aqueous phase. For treatment durations between 6 and 96 h, the major part of the radioactivity was found in the free phase, in the form of ABA. For treatment durations up to 48 h, no radioactivity was detected at the Rf of phaseic acid or dihydrophaseic acid (free and hydrolysed phases). After 96 h culture on 10⁵ M ABA, dihydrophaseic acid was present, but only in very small quantities. Two ABA metabolites were detected. One was characterized as β-d -glucopyranosyl abscisate since its Rf was the same as that of an authentic sample in three different solvent systems and also since it released ABA on mild alkaline hydrolysis. It increased steadily with time and represented the main metabolite. The other metabolite found in the aqueous phase after mild alkaline hydrolysis and extraction with ether at pH 3 was a very polar compound, resistant to alkaline hydrolysis in the presence of concentrated ammonia and to methylation. It was, however, metabolized by apple embryo, yielding essentially dihydrophaseic acid and an ester which released dihydrophaseic acid on mild alkaline hydrolysis. These results indicate that under the conditions tried, the metabolism of [¹⁴C]-ABA by lettuce achenes leads almost exclusively to the formation of conjugates, oxidative metabolism of ABA being almost non-existent. Separate analysis of the integuments and of the endosperm plus embryo after culture of whole achenes for 48 h in the presence of 10⁵ M [¹⁴C]-ABA showed that ABA metabolism occurred only in the endospermembryo tissue.     

Synthesis and dopaminergic properties of the two enantiomers of 3-(3,4-dimethylphenyl)-1-propylpiperidine, a potent and selective dopamine D4 receptor ligand

The synthesis of the two enantiomers of 3-(3,4-dimethylphenyl)-1-propylpiperidine 1, a potent and selective D4 dopaminergic ligand, was performed. The 3-(3,4-dimethylphenyl)- 1-propylpiperidine with the R configuration showed an affinity for the D4 receptors 6-fold higher than the corresponding enantiomer with the S configuration. Furthermore, the (R)-1 enantiomer proved to be highly selective for D4 receptors with respect to D2-D3 receptors, with a Ki ratio higher than 25,000, while the (S)-1 enantiomer was about 100-fold less selective than the (R)-1 one.     

Histochemistry of mucosubstances in the gallbladder epithelium of the chick embryo

Mucosubstances of epithelial cells of the chick embryo gallbladder were investigated by histochemical methods from days 12-21 of incubation (stages 38-46). On incubation day 14, only neutral mucins were detected; on day 15, neutral and sulfated mucosubstances were observed in the epithelial cells that invaginate the underlying mesenchyme. In the same sites, at day 16 of incubation, neutral, carboxylated and sulfated mucins were seen. From the 17th day of incubation until hatching, neutral, carboxylated and sulfated mucosubstances were present in the surface cells and in the cells lining the epithelial invaginations. During this period, the chemical characteristics of the secretory material are similar to those observed by Yamada and Hoshino (1972) in the fowl.     

Sphingosine 1-phosphate induces Ca2+ transients and cytoskeletal rearrangement in C2C12 myoblastic cells

In many cell systems, sphingosine 1-phosphate (SPP) increases cytosolic Ca2+ concentration ([Ca2+]i) by acting as intracellular mediator and extracellular ligand. We recently demonstrated (Meacci E, Cencetti F, Formigli L, Squecco R, Donati C, Tiribilli B, Quercioli F, Zecchi-Orlandini S, Francini F, and Bruni P. Biochem J 362: 349-357, 2002) involvement of endothelial differentiation gene (Edg) receptors (Rs) specific for SPP in agonist-mediated Ca2+ response of a mouse skeletal myoblastic (C2C12) cell line. Here, we investigated the Ca2+ sources of SPP-mediated Ca2+ transients in C2C12 cells and the possible correlation of ion response to cytoskeletal rearrangement. Confocal fluorescence imaging of C2C12 cells preloaded with Ca2+ dye fluo 3 revealed that SPP elicited a transient Ca2+ increase propagating as a wave throughout the cell. This response required extracellular and intracellular Ca2+ pool mobilization. Indeed, it was significantly reduced by removal of external Ca2+, pretreatment with nifedipine (blocker of L-type plasma membrane Ca2+ channels), and inositol 1,4,5-trisphosphate [Ins(1,4,5)P3]-mediated Ca2+ pathway inhibitors. Involvement of EdgRs was tested with suramin (specific inhibitor of Edg-3). Fluorescence associated with Ins(1,4,5)P3Rs and L-type Ca2+ channels was evident in C2C12 cells. SPP also induced C2C12 cell contraction. This event, however, was unrelated to [Ca2+]i increase, because the two phenomena were temporally shifted. We propose that SPP may promote C2C12 cell contraction through Ca2+-independent mechanisms.     

Row orientation effect on UV-B,UV-A and PAR solar irradiation components in vineyards at Tuscany,Italy

Besides playing an essential role in plant photosynthesis, solar radiation is also involved in many other important biological processes. In particular, it has been demonstrated that ultraviolet (UV) solar radiation plays a relevant role in grapevines (Vitis vinifera) in the production of certain important chemical compounds directly responsible for yield and wine quality. Moreover, the exposure to UV-B radiation (280–320 nm) can affect plant–disease interaction by influencing the behaviour of both pathogen and host. The main objective of this research was to characterise the solar radiative regime of a vineyard, in terms of photosynthetically active radiation (PAR) and UV components. In this analysis, solar spectral UV irradiance components, broadband UV (280–400 nm), spectral UV-B and UV-A (320–400 nm), the biological effective UVBE, as well as the PAR (400–700 nm) component, were all considered. The diurnal patterns of these quantities and the UV-B/PAR and UV-B/UV-A ratios were analysed to investigate the effect of row orientation of the vineyard in combination with solar azimuth and elevation angles. The distribution of PAR and UV irradiance at various heights of the vertical sides of the rows was also studied. The results showed that the highest portion of plants received higher levels of daily radiation, especially the UV-B component. Row orientation of the vines had a pronounced effect on the global PAR received by the two sides of the rows and, to a lesser extent, UV-A and UV-B. When only the diffused component was considered, this geometrical effect was greatly attenuated. UV-B/PAR and UV-A/PAR ratios were also affected, with potential consequences on physiological processes. Because of the high diffusive capacity of the UV-B radiation, the UV-B/PAR ratio was significantly lower on the plant portions exposed to full sunlight than on those in the shade.     

Sphingosine 1-phosphate induces cell contraction via calcium-independent/Rho-dependent pathways in undifferentiated skeletal muscle cells

We have previously shown that sphingosine 1-phosphate (S1P) can induce intracellular Ca(2+) mobilization and cell contraction in C2C12 myoblasts and that the two phenomena are temporally unrelated. Although Ca(2+)-independent mechanisms of cell contraction have been the focus of numerous studies on Ca(2+) sensitization of smooth muscle, comparatively less studies have focused on the role that these mechanisms play in the regulation of skeletal muscle contractility. Phosphorylation and activation of myosin by Rho-dependent kinase mediate most of Ca(2+)-independent contractile responses. In the present study, we examined the potential role of Rho/Rho-kinase cascade activation in S1P-induced C2C12 cell contraction. First, we showed that depletion of Ca(2+), by pre-treatment with BAPTA, did not affect S1P-induced myoblastic contractility, whereas it abolished S1P-induced Ca(2+) transients. These results correlated with the absence of troponin C and with the immature cytoskeletal organization of these cells. Experimental evidence demonstrating the involvement of Rho pathway in S1P-stimulated myoblast contraction included: the activation/translocation of RhoA to the membrane in response to agonist-stimulation in cells depleted of Ca(2+) and the inhibition of dynamic changes of the actin cytoskeleton in cells where Rho functions had been inhibited either by overexpression of RhoGDI, a physiological inhibitor of GDP dissociation from Rho proteins, or by pretreatment with Y-27632, a specific Rho kinase inhibitor. Contribution of protein kinase C in this cytoskeletal rearrangement was also evaluated. However, the pretreatment with G?6976 or rottlerin, specific inhibitors of PKC alpha and PKC delta, respectively, failed to inhibit the agonist-induced myoblastic contraction. Single particle tracking of G-actin fluorescent probe was performed to statistically evaluate actin cytoskeletal dynamics in response to S1P. Stimulation with S1P was also able to increase the phosphorylation level of myosin light chain II. In conclusion, our results strongly suggest that Ca(2+)-independent/Rho-Rho kinase-dependent pathways may exert an important role in S1P-induced myoblastic cell contraction.     

Impact of 2003 heat waves on aerobiological indices of allergenic herbaceous family pollen season in Tuscany (Italy)

The aim of this study was to assess the current aerobiological situation and to investigate the influence of the hot and dry summer 2003 on pollen season (onset, end and duration of the pollen season, peak pollen day and value, total seasonal amount of pollen grains) of herbaceous family as Poaceae, Urticaceae and Compositeae. Heat wave of 2003 influenced the phenology of the main pollen families in Tuscany: the high temperatures occurred during 2003 affected pollen season of different family with different responses. This study confirms the role that the climate has on the flora species and in particular on herbaceous species phenology and the high variability of the pollination among different places, during extreme events. In general, high spring temperature induces an advance of the flowering period and a release of higher pollen quantity of Urticaceae and Poaceae; however, exceptional weather conditions (i.e., summer 2003) could exert an opposite effect, resulting in an impairment of flowering of Urticaceae during autumn. Compositeae species produced a low amount of pollen in 2003, even if the peak value was higher than the average in some stations.