Serotonin localization in Phallusia mammillata larvae and effects of 5-HT antagonists during larval development

The neurotransmitter 5-hydroxytryptamine (5-HT, serotonin) plays an important role in a wide range of non-neural processes. Using immunofluorescence with an antiserotonin antibody, 5-HT was localized in the brain and in some neurons of the larval tail of Phallusia mammillata. To test the effect of 5-HT on development, we treated embryos with two different 5-HT receptor subtype antagonists. Treatment at the gastrula stage with 10 microM ondansetron, an antagonist of the 5-HT(3) receptor, induced anterior truncation and a short tail. At 10 microM, ritanserin, a 5-HT(2B) receptor antagonist, induced larval phenotypes characterized by a roundish trunk region with flat papillae. The juveniles developed from these larvae had an abnormal cardiocirculatory system: their heart contractions were ineffective and their blood cells accumulated in the heart cavity. We conclude that an appropriate level of 5-HT is necessary for correct development and morphogenesis. Moreover, a different key role for multiple receptors in modulating the morphogenetic effects of 5-HT is suggested.     

Distribution of class I, III and IV alcohol dehydrogenase mRNAs in the adult rat, mouse and human brain.

The localization of different classes of alcohol dehydrogenases (ADH) in the brain is of great interest because of their role in both ethanol and retinoic acid metabolism. Conflicting data have been reported in the literature. By Northern blot and enzyme activity analyses only class III ADH has been detected in adult brain specimens, while results from riboprobe in situ hybridization indicate class I as well as class IV ADH expression in different regions of the rat brain. Here we have studied the expression patterns of three ADH classes in adult rat, mouse and human tissues using radioactive oligonucleotide in situ hybridization. Specificity of probes was tested on liver and stomach control tissue, as well as tissue from class IV ADH knock-out mice. Only class III ADH mRNA was found to be expressed in brain tissue of all three investigated species. Particularly high expression levels were found in neurons of the red nucleus in human tissue, while cortical neurons, pyramidal and granule cells of the hippocampus and dopamine neurons of substantia nigra showed moderate expression levels. Purkinje cells of cerebellum were positive for class III ADH mRNA in all species investigated, whereas granular layer neurons were positive only in rodents. The choroid plexus was highly positive for class III ADH, while no specific signal for class I or class IV ADH was detected. Our results thus support the notion that the only ADH expressed in adult mouse, rat and human brain is class III ADH.     

Ericoid mycorrhizal fungi: some new perspectives on old acquaintances

Many ericaceous species colonize as pioneer plants substrates ranging from arid sandy soils to moist mor humus, in association with their mycorrhizal fungi. Thanks to the symbiosis with ericoid mycorrhizal fungi, ericaceous plants are also able to grow in highly polluted environments, where metal ions can reach toxic levels in the soil substrate. For a long time this mycorrhizal type has been regarded as an example of a highly specific interaction between plants and fungi. More recent studies have been challenging this view because some ericoid mycorrhizal endophytes seem also able to colonise plants from very distant taxa. A molecular approach has allowed the investigation of genetic diversity and molecular ecology of ericoid mycorrhizal fungi, and has revealed that ericaceous plants can be very promiscuous, with multiple occupancy of their thin roots. The molecular analysis of sterile morphotypes involved in this symbiosis has also led to deeper understanding of the species diversity of ericoid fungi. Genetic polymorphism of ericoid fungi is wider than previously thought, and often increased by the presence of Group I introns in the nuclear small subunit rDNA.     

Alternative pathway respiration is associated with ammonium ion sensitivity in spinach and pea plants

Spinach and pea plants were grown in hydroponic culture with nitrate orammonium salts as the nitrogen source. Dry matter accumulation andphotosynthetic rate declined in spinach plants fed with ammonium salts, whereasthey did not change in pea plants compared with nitrate-fed plants. Measurementof organic nitrogen and free amino acid content showed that ammonium ions wereassimilated in shoots in spinach plants and in roots in pea plants. Ammoniumionnutrition led to a decline in starch content in both species. Organic acidsincreased in roots of pea plants fed with ammonium ions whereas they declinedinspinach plants. In both species ammonium ions increased root respiration ratebut the contribution of both routes (cytochromic and alternative pathway) tothis increase was different depending on the species. In spinach plants,ammonium ions increased the cytochromic path and decreased the alternativepathway, whereas in pea plants both routes were stimulated mainly through thealternative pathway. The differences in the sensitivity to ammonium ionsbetweenboth species are discussed in terms of differences in the availability of Cskeletons and energy, which could be due in part to differences in the capacityto stimulate the alternative pathway.     

Self Organized Terminode Routing

We consider the problem of routing in a wide area mobile ad hoc network called Terminode Network. Routing in this network is designed with the following objectives. First, it should scale well in terms of the number of nodes and geographical coverage; second, routing should have scalable mechanisms that cope with the dynamicity in the network due to mobility; and third, nodes need to be highly collaborative and redundant, but, most of all, cannot use complex algorithms or protocols. Our routing scheme is a combination of two protocols called Terminode Local Routing (TLR) and Terminode Remote Routing (TRR). TLR is used to route packets to close destinations. TRR is used to route to remote destinations. The combination of TLR and TRR has the following features: (1) it is highly scalable because every node relies only on itself and a small number of other nodes for packet forwarding; (2) it acts and reacts well to the dynamicity of the network because as a rule multipath routing is considered; and (3) it can be implemented and run in very simple devices because the algorithms and protocols are very simple and based on high collaboration. We performed simulations of the TLR and TRR protocols using the GloMoSim simulator. The simulation results for a large, highly mobile ad hoc environment demonstrate benefits of the combination of TLR and TRR over an existing protocol that uses geographical information for packet forwarding.     

High-yield method for isolation and culture of endothelial cells from rat coronary blood vessels suitable for analysis of intracellular calcium and nitric oxide biosynthetic pathways

We describe here a method for isolating endothelial cells from rat heart blood vessels by means of coronary microperfusion with collagenase. This method makes it possible to obtain high amounts of endothelial cells in culture which retain the functional properties of their in vivo counterparts, including the ability to uptake fluorescently-labeled acetylated low-density lipoproteins and to respond to vasoactive agents by modulating intracellular calcium and by upregulating intrinsic nitric oxide generation. The main advantages of our technique are: (i) good reproducibility, (ii) accurate sterility that can be maintained throughout the isolation procedure and (iii) high yield of pure endothelial cells, mainly due to microperfusion and temperature-controlled incubation with collagenase which allow an optimal distribution of this enzyme within the coronary vascular bed.     

Enzymatic conversions of polar lipids. Principles, problems and solutions

This text provides a brief overview of the principles of enzymatic lipid conversion and some recent advances in the enzymatic conversion of glycerophospholipids and galactolipids. Lipases and phospholipases are used to exchange fatty acids or the polar group in the lipids. The reactions can be carried out either as hydrolysis–esterification sequences or as one-step transferase reactions. The scope and limitations of the different methods are discussed.     

Statistical modelling of insect behavioural responses in relation to the chemical composition of test extracts

Abstract. The use of generalized linear models (GLM) for relating changes in insect behaviour to changes in the chemical composition of a plant extract is presented and applied to data from an experimental study of the olfactory response of Cydia pomonella L. (Lepidoptera: Tortricidae) to apple volatiles. The volatiles were collected from healthy apples, artificially damaged apples or apples infested with C. pomonella larvae (either instar I, IV or V). These treatments produced a blend of 23 major components and the chemical composition of the blends differed substantially amongst the treatments.
A statistically significant relationship was found between the concentration of hexyl hexanoate and 2-methylbutyl acetate in each extract and the number of moths moving upwind. Statistically significant models were developed which suggested that a relationship exists between the concentration of Z , E -α-farnesene, hexyl hexanoate and 2-methylbutyl acetate and the number and duration of movements made by the moths.
Subsequently Y-tube assays were carried out to validate the predictions made with respect to the orientation of mated female C. pomonella . The results of these assays confirm hexyl hexanoate as an attractant. There were indications that 2-methylbutyl acetate acted as a repellent although differences were not statistically significant. Previous bioassays have shown that C. pomonella displays a statistically significant negative linear dose–response to α-farnesene ( Hern & Dorn, 1999 ).
The statistical methods employed are very flexible and fairly easy to implement, offering the potential to screen plant extracts for bioactive compounds with a minimum of biological constraints. Their general applicability has yet to be demonstrated and as such these analyses only offer evidence of statistical relationships; the results must be validated by additional bioassays before conclusions can be drawn.