Reactive oxygen species and Udx1 during early sea urchin development

Sea urchin fertilization is marked by a massive conversion of molecular oxygen to hydrogen peroxide by a sea urchin dual oxidase, Udx1. This enzyme is essential for completing the physical block to polyspermy. Yet, its expression is maintained during development, as indicated by the presence of both Udx1 mRNA and Udx1 protein enriched at the surface of all non-mesenchymal blastomeres. When hydrogen peroxide synthesis by Udx1 is inhibited, either pharmacologically or by specific antibody injection, cleavage is delayed. Application of exogenous hydrogen peroxide, however, partially rescues a fraction of these defective embryos. We also report an unequal distribution of reactive oxygen species between sister blastomeres during early cleavage stages, suggesting a functional role for Udx1 in intracellular signaling.     

Metabolic effects of thia fatty acids

Thia substituted fatty acids are saturated fatty acids which are modified by insertion of a sulfur atom at specific positions in the carbon backbone. During the last few years pleiotropic effects of the 3-thia fatty acid tetradecylthioacetic acid have been revealed. The biological responses to tetradecylthioacetic acid include mitochondrial proliferation, increased catabolism of fatty acids, antiadiposity, improvement in insulin sensitivity, antioxidant properties, reduced proliferation and induction of apoptosis in rapidly proliferating cells, cell differentiation and antiinflammatory action. These biological responses indicate that tetradecylthioacetic acid changes the plasma profile from atherogenic to cardioprotective. As a pan-peroxisome proliferator-activated receptor ligand, tetradecylthioacetic acid regulates the adipose tissue mass and the expression of lipid metabolizing enzymes, particularly those involved in catabolic pathways. In contrast, circumstantial evidences suggest that peroxisome proliferator-activated receptor-independent metabolic pathways may be of importance for the antioxidant, antiproliferative and antiinflammatory action of tetradecylthioacetic acid.     

Number of Lianas per Tree and Number of Trees Climbed by Lianas at Los Tuxtlas,Mexico1

In the tropical rainforest of Los Tuxtlas, Mexico, lianas (woody‐climbing plants) had a clumped distribution on trees, and 63.3 percent of trees ≥10 cm in diameter carried at least one liana. Trees with larger diameters supported more lianas and a higher total basal area of lianas than trees with smaller diameters. There was no relationship between liana diameter and the number of trees a liana climbed, but tendril climbers climbed on more trees than stem‐twiners.     

Membrane trafficking machinery components associated with the mammalian acrosome during spermiogenesis.

Active trafficking from the Golgi apparatus is involved in acrosome formation, both by delivering acrosomal contents to the nascent secretory vesicle and by controlling organelle growth and shaping. During murine spermiogenesis, Golgi antigens (giantin, beta-COP, golgin 97, mannosidase II) are detected in the acrosome until the late cap-phase spermatids, but are not found in testicular spermatozoa (maturation-phase spermatids). This suggests that Golgi-acrosome flow may be relatively unselective, with Golgi residents retrieved before spermiation is complete. Treatment of spermatogenic cells with brefeldin A, a drug that causes the Golgi apparatus to collapse into the endoplasmic reticulum, disrupted the Golgi in both pachytene spermatocytes and round spermatids. However, this treatment did not affect the acrosomal granule, and some beta-COP labeling on the acrosome of elongating spermatids was maintained. Additionally, N-ethylmaleimide sensitive factor, soluble NSF attachment proteins, and homologues of the t-SNARE syntaxin and of the v-SNARE VAMP/synaptobrevin, as well as members of the rab family of small GTPases, are associated with the acrosome (but not the acrosomal granule) in round and elongated spermatids. This suggests that rab proteins and the SNARE machinery for membrane recognition/docking/fusion may be involved in trafficking during mammalian acrosome biogenesis.     

Gastrulation in the sea urchin is accompanied by the accumulation of an endoderm-specific mRNA

Spatial diversification of the endoderm during gastrulation in the sea urchin Lytechinus variegatus was examined with an endoderm-specific cDNA clone. This cDNA clone, LvN1.2, was identified by a differential cDNA screen between the ectoderm and endoderm/mesoderm fractions from prism stage embryos. The LvN 1.2-kb mRNA was first detectable by Northern blots at the mesenchyme blastula stage just prior to gastrulation and then accumulated approximately 15-fold from gastrulation to the pluteus stage. In situ hybridization analysis demonstrated that the mRNA accumulated specifically in endoderm and was restricted to the hindgut-midgut regions. This restricted localization was apparent during gastrulation and predicted the morphological distinction between foregut and midgut eventually seen at prism and pluteus stages. Sequence analysis showed that the 189-amino acid open reading frame represented a novel protein. In vitro translation of synthetically produced LvN1.2 mRNA and Western blot analysis with antibodies to the protein sequence yielded the same 25-kDa polypeptide on SDS-PAGE. The LvN1.2 protein resided within discrete granules of the hindgut and midgut cells. These particles were concentrated to the luminal aspect of the cells, suggesting the LvN1.2 protein participates in the digestive function of this region of the gut.     

Toxicity Studies with the Blue-Green Alga Oscillatoria Agardhii from two Eutrophic Norwegian Lakes

Extracts of the blue-green alga Oscillatoria agardhii were tested for acute toxicity on laboratory mice and rats. Material originating from lake Gjersjøen proved to be toxic to the animals, samples from the nearby lake Årungen did not. Clinical symptoms culminated in the development of a fatal shock due to decrease in circulating blood volume. Pathological examination revealed heavy pooling of blood in the liver and severe damage to the organ. Blood analyses also indicated liver damage. Effects were the same with extracts from a laboratory clone culture as from a natural water bloom, but the toxin content was higher in the bloom material. Toxicity was not affected by heat, acid or alkali treatment.     

Differential In Vivo Regulation of mRNA Encoding the Norepinephrine Transporter and Tyrosine Hydroxylase in Rat Adrenal Medulla and Locus Ceruleus

Abstract: To investigate the regulation of norepinephrine transporter mRNA in vivo, we analyzed the effects of reserpine on its expression in the rat adrenal medulla and locus ceruleus. First, PCR was used to clone a 0.5-kb rat cDNA fragment that exhibits 87% nucleotide identity to the corresponding human norepinephrine transporter cDNA sequence. In situ, the cDNA hybridizes specifically within norepinephrine-secreting cells, but in neither dopamine nor serotonin neurons, suggesting strongly it is a partial rat norepinephrine transporter cDNA. Reserpine, 10 mg/kg administered 24 h premortem, decreased steady-state levels of norepinephrine transporter mRNA in the adrenal medulla by ∼65% and in the locus ceruleus by ∼25%, as determined by quantitative in situ hybridization. Northern analysis confirmed the results of the in situ hybridization analysis in the adrenal medulla but did not detect the smaller changes observed in the locus ceruleus. Both analyses showed that reserpine increased tyrosine hydroxylase expression in the adrenal medulla and locus ceruleus. These results suggest that noradrenergic neurons and adrenal chromaffin cells can coordinate opposing changes in systems mediating catecholamine uptake and synthesis, to compensate for catecholamine depletion.     

Chemical composition of reindeer forage plants in Svalbard and Norway

Several of the most important reindeer forage plants in Svalbard were analysed for content of minerals (Na, K, P, Ca, Mg, S, Cl, Fe, Cu, Mn, Zn, Mo and Co) and for content of ash, crude protein, crude fiber, ether extracts and nitrogen free extracts. Some forage plants were also examined in in vitro digestibility trials. The results were compared with results from similar analyses of alpine mountain plants from southern Norway. Also freshly voided fecal samples were analysed for the same components as the forage plants.
The Svalbard vegetation had a higher content of Na, Ca, Mg, Cl, Fe and Co, than similar plants from Southern Norway. The content of other elements examined seemed to be more equal in Norway and on Svalbard. Within the Svalbard area plants growing in limestone areas had the highest content of calcium and magnesium. Also plants growing on manured soil below bird cliffs appeared to have high nutritional value. It is furthermore concluded that the closeness to the sea as well as the occurrence of several plants with high mineral content, gave the Svalbard reindeer the possibility to further improve their mineral balance compared with continental reindeer. Also the dietary shift from lichens as a major dietary component in Norway, to mosses on Svalbard, may increase mineral intake, but may also have an inferior effect on digestibility.
Although balance experiments and carcass analyses of minerals in Svalbard reindeer are missing, it is concluded that Svalbard reindeer apparently had adequate intake of most mineral elements. In spite of low standing biomass of reindeer forage plants, the quality of the vegetation seemed to compare favourably with continental reindeer pasture.     

Simian virus 40 T-antigen DNA helicase is a hexamer which forms a binary complex during bidirectional unwinding from the viral origin of DNA replication.

The role of simian virus 40 (SV40) large tumor antigen (T antigen) as a DNA helicase at the replication fork was studied. We found that a T-antigen hexamer complex acts during the unidirectional unwinding of appropriate DNA substrates and is localized directly in the center of the fork, contacting the adjacent double strand as well as the emerging single strands. When bidirectional DNA unwinding, initiated at the viral origin of DNA replication, was analyzed, a larger T-antigen complex that is simultaneously active at both branch points of an unwinding bubble was observed. The size and shape of this helicase complex imply that the T-antigen dodecamer complex, assembled at the origin and active in the localized melting of duplex DNA, is subsequently also used to continue DNA unwinding bidirectionally. Then, however, the dodecamer complex does not split into two hexamer subunits that track along the DNA; rather, the DNA is threaded through the intact complex, with the concomitant extrusion of single-stranded loops.