Calreticulin-melatonin. An unexpected relationship.

Increasing evidence suggests that melatonin can exert some effect at nuclear level. Previous experiments using binding techniques clearly showed the existence of specific melatonin binding sites in cell nucleus of rat liver. To further identify these sites, nuclear extracts from rat hepatocytes were treated with different percentages of ammonium sulfate and purified by affinity chromatography. Subsequent ligand blot analysis shows the presence of two polypeptides of approximately 60 and approximately 74 kDa that bind specifically to melatonin. N-Terminal sequence analysis showed that the 60 kDa protein shares a high homology with rat calreticulin, whereas the 74 kDa protein shows no homology with any known protein. The binding of melatonin to calreticulin was further characterized incubating 2-[125I]melatonin with recombinant calreticulin. Binding kinetics show a Kd = 1.08 +/- 0.2 nm and Bmax = 290 +/- 34 fmol.mg protein-1, compatible with other binding sites of melatonin in the cell. The presence of calreticulin was further identified by Western blot analysis, and the lack of endoplasmic reticulum contamination in our material was assessed by Western blot and immunostaining with anti-calnexin Ig. The results suggest that calreticulin may represent a new class of high-affinity melatonin binding sites involved in some functions of the indoleamine including genomic regulation.     

An adenovirus agnogene.

The nucleotide sequence of a 550 base pairs long segment, located between map positions 21 and 22.5 in the adenovirus type 2 genome has been determined. S1 nuclease mapping and sequence analysis of cDNA copies of adenovirus mRNA demonstrated that the established sequence includes the i-leader which is spliced to the 5'-end of certain adenovirus mRNAs (1). The i-leader which is 440 nucleotides long contains an open translational reading frame which is preceded by an AUG triplet and which terminates in the third segment of the tripartite leader. A polypeptide with the same molecular weight as predicted from the DNA sequence was identified by in vitro translation of mRNA which had been selected by hybridization to DNA fragments, containing sequences from the i-leader. The results thus suggest that the i-leader, unlike other adenovirus leader segments, is used for translation.     

An adolescent ward.

A ward has been set up for adolescents, who, being neither children nor adults, have special needs. It provides a pleasant and enthusiastic atmosphere that allows the patients to mix together socially alties is important, but not more than 20% should be long-stay patients. Those needing intensive care or specialised investigations and those likely to be a disruptive influence are excluded. No serious sexual problems have been encountered.     

An investigation of non-steady-state algal growth. I. An experimental model ecosystem

In order to test rigorously the transient behaviour of mathematical models of algal growth, detailed laboratory data sets with good temporal resolution are required. A series of algal growth experiments was conducted in transient conditions. Monoculture growth of, and competition for nutrients between, three contrasting species of phytoplankton (the diatom Thalassiosira pseudonana, the harmful flagellate Heterosigma carterae and the toxic dinoflagellate Alexandrium minutum) were investigated in different temperature, light and nutrient regimes. Although growth dynamics were qualitatively similar in batch culture, quantitative differences were evident in the growth response of the different species when grown in single yield-limiting nutrient conditions in identical physical conditions. Quantities such as the carbon:nitrogen (C:N) ratio and C and N per cell varied between species and within species under different growth conditions. Such results have particular significance to the development of mathematical models, which commonly represent algal populations as a single homogeneous group using a single currency such as numbers, C or N. Changes in light and temperature regime influenced algal growth: Alexandrium failed to grow at low temperatures, while specific growth rates of Thalassiosira were more sensitive to changes in temperature than those of Heterosigma. Changes in the dominant organism(s) and/or its size or nutrient status may influence the transfer of nutrients within the food web. Commonly, mathematical models make cell growth a function of a single yield-limiting nutrient. Decreased growth rates and high residual nutrient concentrations in competition experiments indicate that this approach is unlikely to be successful in conditions of limited supply of more than one nutrient, where multiple nutrient stresses will be significant.