RAIDD aggregation facilitates apoptotic death of PC12 cells and sympathetic neurons

In human cell lines, the caspase 2 adaptor RAIDD interacts selectively with caspase 2 through its caspase recruitment domain (CARD) and leads to caspase 2-dependent death. Whether RAIDD induces such effects in neuronal cells is unknown. We have previously shown that caspase 2 is essential for apoptosis of trophic factor-deprived PC12 cells and rat sympathetic neurons. We report here that rat RAIDD, cloned from PC12 cells, interacts with rat caspase 2 CARD. RAIDD overexpression induced caspase 2 CARD- and caspase 9-dependent apoptosis of PC12 cells and sympathetic neurons. Apoptosis correlated with the formation of discrete perinuclear aggregates. Both death and aggregates required the expression of full-length RAIDD. Such aggregates may enable more effective activation of caspase 2 through close proximity. Following trophic deprivation, RAIDD overexpression increased death and aggregate formation. Therefore, RAIDD aggregation is important for its death-promoting effects and may play a role in trophic factor withdrawal-induced neuronal apoptosis.     

Lethal and sublethal effects of a neem-based insecticide on balsam fir sawfly (Hymenoptera: Diprionidae)

Lethal and sublethal effects of Neemix 4.5 EC, a commercial neem preparation, on balsam fir sawfly, Neodiprion abietis (Harris), were determined in the laboratory. Larval mortality of N. abietis increased in a concentration-dependent manner, and lethal time decreased with increasing Neemix 4.5 EC concentration. Fifty percent of the larvae died after 4.6 d at a concentration of 90 ppm azadirachtin (AZA) and 12.3 d at a concentration of 0.08 ppm. Neemix 4.5 EC showed some deterrent effects to feeding site selection on N. abietis larvae at high concentrations, but not at low concentrations. Strong reduction of food intake by N. abietis larvae after exposure to Neemix 4.5 EC was demonstrated by significant reduction of frass production. Larvae fed on Neemix 4.5 EC-treated foliage at 90 ppm AZA produced only 16% as much frass as that produced by larvae fed on control foliage (0 ppm). Neemix 4.5 EC at a concentration of 0.08 ppm AZA retarded larval and pupal development. Sublethal doses significantly reduced pupal weight and adult emergence, although the sex ratio of N. abietis adults was not affected. Results indicate that sublethal effects of Neemix 4.5 EC on N. abietis may contribute greatly to the overall field efficacy.     

Synthesis and properties of novel psoralen derivatives

We have synthesized a set of new trimethylpsoralen derivatives that are characterized by a chain extending from the 4'-position of the furan ring and linked to this ring by an aminomethylene group. The nature of the side chain can be varied widely. In these derivatives, the chains contain either amino or ethylene oxide units for enhanced water solubility and allow the introduction of a thiol or amine group to nucleic acids. These compounds represent the first set of thiolated psoralen derivatives, and their usefulness is demonstrated in several nucleic acid cross-linking experiments. The reagents can be used to create both intraduplex reversible cross-links between the two single-strand partners in a DNA double helix and interduplex reversible cross-links between two DNA double helices.     

Molecular adaptation of a leaf-eating bird: stomach lysozyme of the hoatzin

This report describes a lysozyme expressed at high levels in the stomach of the hoatzin, the only known foregut-fermenting bird. Evolutionary comparison places it among the calcium-binding lysozymes rather than among the conventional types. Conventional lysozymes were recruited as digestive enzymes twice in the evolution of mammalian foregut fermenters, and these independently recruited lysozymes share convergent structural changes attributed to selective pressures in the stomach. Biochemical convergence and parallel amino acid replacements are observed in the hoatzin stomach lysozyme even though it has a different genetic origin from the mammalian examples and has undergone more than 300 million years of independent evolution.      

Separation of porphyrin isomers by high-performance liquid chromatography.

A reversed-phase gradient elution system is described for the simultaneous separation of the type I and type III isomers of 8-, 7-, 6-, 5- and 4-carboxylated porphyrins and isocoproporphyrins. The method, adaptable for isocratic and stepwise separation of individual groups of isomers, is also suitable for preparative isolation of pure porphyrins. The analyses of porphyrin isomers in the urine and faeces of porphyric patients are examples of applications.     

On the distribution of Na+ pump sites in the frog skin

Exposure of the outside of the isolated frog skin to a Ringer's solution, made hypertonic by the addition of mannitol, causes a rapid and sustained increase in transepithelial permeability through a structural distortion-a focal blistering-of the "tight" junctions of the outermost living cell layer. [(3)H]ouabain, used as an autoradiographic marker for the Na+-pump (Na+-K+-adenosine triphosphatase), is usually unable to penetrate the frog skin from the outside solution, but when added to a hypertonic mannitol- Ringer's solution in the outside bath it readily penetrates the epithelium, presumably through the opened shunt pathway. Radioautographic analysis of [(3)H]ouabain binding sites revealed that most of ouabain enters from the outside solution binds to the sites on the cell membranes of the stratum spinosum, as was the case when it was applied from the inside bath in an earlier study. The outer living cell layer, the first to be exposed to ouabain, does not appear to be the major site for the Na+-pump, and therefore, is not likely to be responsible for most of the active pumping of Na+. This result demonstrates that previous failure to show a high density of Na+-pump sites on the cells of the outermost layer, when [(3)H]ouabain was applied from the inside solution, was not due to the inability of the marker to reach these cells at a sufficient concentration to reveal all pump sites. These results provide further support for a model of Na+-transport across the frog skin which distributes the active pump step on the inward facing membranes of all living cells.     

Molecular phylogeny of the major arthropod groups indicates polyphyly of crustaceans and a new hypothesis for the origin of hexapods

A phylogeny of the arthropods was inferred from analyses of amino acid sequences derived from the nuclear genes encoding elongation factor-1 alpha and the largest subunit of RNA polymerase II using maximum- parsimony, neighbor-joining, and maximum-likelihood methods. Analyses of elongation factor-1 alpha from 17 arthropods and 4 outgroup taxa recovered many arthropod clades supported by previous morphological studies, including Diplopoda, Myriapoda, Insecta, Hexapoda, Branchiopoda (Crustacea), Araneae, Tetrapulmonata, Arachnida, Chelicerata, and Malacostraca (Crustacea). However, counter to previous studies, elongation factor-1 alpha placed Malacostraca as sister group to the other arthropods. Branchiopod crustaceans were found to be more closely related to hexapods and myriapods than to malacostracan crustaceans. Sequences for RNA polymerase II were obtained from 11 arthropod taxa and were analyzed separately and in combination with elongation factor-1 alpha. Results from these analyses were concordant with those derived from elongation factor-1 alpha alone and provided support for a Hexapoda/Branchiopoda clade, thus arguing against the monophyly of the traditionally defined Atelocerata (Hexapoda + Myriapoda).      

Inhibition of herpes simplex virus-induced DNA polymerase activity and viral DNA replication by 9-(2-hydroxyethoxymethyl)guanine and its triphosphate.

The effect of the nucleoside analog 9-(2-hydroxyethoxymethyl)guanine (acycloguanosine) on herpes simplex virus type 1 DNA synthesis was examined. Acycloguanosine inhibited herpesvirus DNA synthesis in virus-infected cells. The synthesis of host cell DNA was only partially inhibited in actively growing cells at acycloguanosine concentrations several hundred-fold greater than the 50% effective dose for herpes simplex virus type 1. Studies using partially purified enzymes revealed that the triphosphate of this compound inhibited the virus-induced DNA polymerases (DNA nucleotidyltransferases) to a greater degree than the DNA polymerase of the host cell, that the inhibition was dependent upon the base composition of the template, and that the triphosphate was a better substrate for the virus-induced polymerases than for the alpha cellular DNA polymerases.     

Major changes in fecundity and the effect on population egg production for three species of north-west Atlantic flatfishes

Temporal and spatial variability in fecundity was investigated for three species of north-west Atlantic flatfishes: American plaice Hippoglossoides platessoides , yellowtail flounder Limanda ferruginea and witch flounder Glyptocephalus cynoglossus . Significant log₁₀ linear relationships existed between fecundity and total length ( L _T) for all three species and for all combinations of area (NAFO regulatory areas) and year. Fecundity differed between areas for yellowtail flounder and witch flounder and changed significantly for all three species since the last reported fecundity studies in the region 30–40 years ago. Three populations experienced decreases in fecundity (3LNO American plaice and yellowtail flounder, 3Ps witch flounder), one experienced an increase (3NO witch flounder) and one did not change (3L witch flounder). Significant yearly variability in fecundity at length and relative fecundity existed for all three species within the 6 year sampling period (1993–1998). Estimates of population egg production based on new and historic fecundity data revealed that using the old, invariant fecundity and L _T relationship resulted in overestimation of total annual egg production by up to 24% for 3LNO American plaice and as high as 41% for 3LNO yellowtail flounder. These results clearly demonstrate the variable nature of fecundity for flatfishes in Newfoundland waters and suggest that the use of an invariant fecundity and L _T relationship may result in inaccurate estimates of stock reproductive potential.