Cytogenetic analysis of human oocytes after subzonal insemination

Subzonal insemination has been proposed to achieve fertilization in cases where standard in vitro fertilization has failed. We present the results of chromosome analysis of oocytes after subzonal insemination. Our data suggest that the main cause (76 per cent) of the absence of cleavage after subzonal insemination is the total absence of sperm nucleus evolution of the injected spermatozoa. Our results also suggest that spermatozoa chromatin development is normal after subzonal insemination. Aneuploidy does not seem to be increased in zygotes after subzonal insemination. However, polyploidy was often more important than predicted by the observation of pronuclei (PN). Pronucleus development might be asynchronous and can appear earlier or later than after standard IVF. The cytogenetic risk after subzonal insemination might therefore be triploidy (if a triploid egg is transferred, because only 2 PN were seen) rather than aneuploidy or structural abnormalities.     

Long-term CO2 enrichment of a forest ecosystem: implications for forest regeneration and succession.

The composition and successional status of a forest affect carbon storage and net ecosystem productivity, yet it remains unclear whether elevated atmospheric carbon dioxide (CO2) will impact rates and trajectories of forest succession. We examined how CO2 enrichment (+200 microL CO2/L air differential) affects forest succession through growth and survivorship of tree seedlings, as part of the Duke Forest free-air CO2 enrichment (FACE) experiment in North Carolina, USA. We planted 2352 seedlings of 14 species in the low light forest understory and determined effects of elevated CO2 on individual plant growth, survival, and total sample biomass accumulation, an integrator of plant growth and survivorship over time, for six years. We used a hierarchical Bayes framework to accommodate the uncertainty associated with the availability of light and the variability in growth among individual plants. We found that most species did not exhibit strong responses to CO2. Ulmus alata (+21%), Quercus alba (+9.5%), and nitrogen-fixing Robinia pseudoacacia (+230%) exhibited greater mean annual relative growth rates under elevated CO2 than under ambient conditions. The effects of CO2 were small relative to variability within populations; however, some species grew better under low light conditions when exposed to elevated CO2 than they did under ambient conditions. These species include shade-intolerant Liriodendron tulipifera and Liquidambar styraciflua, intermediate-tolerant Quercus velutina, and shade-tolerant Acer barbatum, A. rubrum, Prunus serotina, Ulmus alata, and Cercis canadensis. Contrary to our expectation, shade-intolerant trees did not survive better with CO2 enrichment, and population-scale responses to CO2 were influenced by survival probabilities in low light. CO2 enrichment did not increase rates of sample biomass accumulation for most species, but it did stimulate biomass growth of shade-tolerant taxa, particularly Acer barbatum and Ulmus alata. Our data suggest a small CO2 fertilization effect on tree productivity, and the possibility of reduced carbon accumulation rates relative to today's forests due to changes in species composition.     

A comparison of antimony and lead profiles over the past 2500 years in Flanders Moss ombrotrophic peat bog, Scotland

Two cores collected in 2001 and 2004 from Flanders Moss ombrotrophic peat bog in central Scotland were dated (14C, 210Pb) and analysed (ICP-OES, ICP-MS) to derive and compare the historical atmospheric deposition records of Sb and Pb over the past 2500 years. After correction, via Sc, for contributions from soil dust, depositional fluxes of Sb and Pb peaked from ca. 1920-1960 A.D., with >95% of the anthropogenic inventories deposited post-1800 A.D. Over the past two centuries, trends in Sb and Pb deposition have been broadly similar, with fluctuations in the anthropogenic Sb/Pb ratio reflecting temporal variations in the relative input from emission sources such as the mining and smelting of Pb ores (in which Sb is commonly present, as at Leadhills/Wanlockhead in southern Scotland), combustion of coal (for which the Sb/Pb ratio is approximately an order of magnitude greater than in Pb ores) and exhaust emissions (Pb from leaded petrol) and abrasion products from the brake linings (Sb from heat-resistant Sb compounds) of automobiles. The influence of leaded petrol has been most noticeable in recent decades, firstly through the resultant minima in Sb/Pb and 206Pb/207Pb ratios (the latter arising from the use of less radiogenic Australian Pb in alkylPb additives) and then, during its phasing out and the adoption of unleaded petrol, complete by 2000 A.D., the subsequent increase in both Sb/Pb and 206Pb/207Pb ratios. The extent of the 20th century maximum anthropogenic enrichment of Sb and Pb, relative to the natural Sc-normalised levels of the Upper Continental Crust, was similar at approximately 50- to 100-fold. Prior to 1800 A.D., the influence of metallurgical activities on Sb and Pb concentrations in the peat cores during both the Mediaeval and Roman/pre-Roman periods was discernible, small Sb and Pb peaks during the latter appearing attributable, on the basis of Pb isotopic composition, to the mining/smelting of Pb ores indigenous to Britain.     

Effects of environmental concentrations of atrazine on hemocyte density and phagocytic activity in the pond snail Lymnaea stagnalis (Gastropoda, Pulmonata)

Immunotoxicological effects of environmentally relevant concentrations (10, 23, 50, 100 microg/l) of atrazine were studied in Lymnaea stagnalis. Individual hemolymph sampling was performed at 0, 24, 48, 72, 96, 168, 336, 504 and 672 h during exposure. Every atrazine concentration induced a significant increase in the mean number of circulating hemocytes, without any concentration-response relation. A peak (1.6-fold increase) of hemocyte density was observed after 96 h of exposure. After 504 h, the number of hemocytes remained higher only in the snails exposed to the two highest concentrations. Granulocytes contributed most to the increase in hemocyte density in herbicide-exposed snails. Both short- (24 and 96 h) and long-term (504 h) exposures resulted in significant inhibition of hemocyte phagocytic activity upon E. coli. Over the long-term, phagocytosis recovered for the two lowest concentrations. After 504 h of exposure, every herbicide level resulted in a significant reduction of reactive oxygen species production in E. coli-stimulated hemocytes, which was not observed for short-term exposures.     

Effect of photosensitizer riboflavin on the fate of 2,4,6-trinitrotoluene in a freshwater environment

The effect of riboflavin (1 microM) on the fate of TNT (20 mg/l) in a natural water environment was studied. The relative contribution of photolysis, microbial assemblages and freshwater matrix to TNT degradation was examined. The rates, extent and products of TNT and riboflavin transformation were compared under different experimental conditions. It was found that riboflavin significantly enhanced the degradation of TNT in natural water environment. Thus it is a potentially useful photosensitizing agent for the treatment of TNT-contaminated surface water. Furthermore, in the presence of riboflavin, two new intermediates with max. absorption wavelength of 230 nm were found, demonstrating that transformation of TNT in the presence of riboflavin undergoes different pathways.     

Reduction of Ag(I), Au(III), Cu(II), and Hg(II) by Fe(II)/Fe(III) hydroxysulfate green rust

Green rusts are mixed Fe(II)/Fe(III) hydroxides that are found in many suboxic environments where they are believed to play a central role in the biogeochemical cycling of iron. X-ray absorption fine structure analysis of hydroxysulfate green rust suspensions spiked with aqueous solutions of AgCH(3)COO, AuCl(n)(OH)(4-n), CuCl(2), or HgCl(2) showed that Ag(I), Au(III), Cu(II), and Hg(II) were readily reduced to Ag(0), Au(0), Cu(0), and Hg(0). Imaging of the resulting solids from the Ag(I)-, Au(III)-, and Cu(II)-amended green rust suspensions by transmission electron microscopy indicated the formation of submicron-sized particles of Ag(0), Au(0), and Cu(0). The facile reduction of Ag(I), Au(III), Cu(II), and Hg(II) to Ag(0), Au(0), Cu(0), and Hg(0), respectively, by green rust suggests that the presence of green rusts in suboxic soils and sediments can have a significant impact on the biogeochemistry of silver, gold, copper, and mercury, particularly with respect to their mobility.     

Seabird eggs as bioindicators of chemical contamination in Chile

Seabird eggs were used as bioindicators of chemical contamination in Chile. Brown-hooded Gull (Larus maculipennis), Kelp Gull (Larus dominicanus), Trudeau's Tern (Sterna trudeaui), Neotropic Cormorant (Phalacrocorax brasilianus), and Pink-footed Shearwater (Puffinus creatopus) eggs were sampled at different breeding sites during the 1990s. Mercury and organochlorines (PCBs, DDT, HCB, HCH, and PCP) were quantified to reveal the interspecific differences, spatial and temporal trends in contamination levels. Trudeau's Tern displayed the highest levels of mercury (486 ng g(-1) wet weight). The highest sumDDT concentrations were measured in Brown-hooded Gulls (726 ng g(-1)). PCB levels were similar among the species (102-236 ng g(-1)), but the composition of the PCB mixture was different in Pink-footed Shearwaters. With the exception of the Brown-hooded Gull, all species studied presented similar and low levels of organochlorines (sumOHa). Residues of PCB and related compounds were not detected in any of the seabird eggs analyzed in Chile. Geographical variation was low, although levels of industrial chemicals were slightly higher in eggs from Concepción Bay, and agricultural chemicals in eggs from Valdivia. Also interannual variation was low, but some evidence was found of decreasing levels in gull eggs throughout the time of the study. The causes of the low levels and small variability in space and time of environmental chemicals in Chilean seabirds are discussed. We propose the use of seabirds in future monitoring of the development of chemical contamination in Chile.     

Temporal reproductive isolation and gametic compatibility are evolutionary mechanisms in the<Emphasis Type="Italic"> Acropora humilis</Emphasis> species group (Cnidaria; Scleractinia)

Patterns of interbreeding between individuals are fundamental to the structure and maintenance of evolutionary boundaries between species. In corals, both hybridisation and reproductive isolation appear to be important evolutionary mechanisms. In this study, I examine evolutionary boundaries using morphological, molecular and reproductive criteria within the Acropora humilis species group at Lizard Island on the Great Barrier Reef, Australia. Five species and seven morphs are recognised on the basis of morphological appearance of features traditionally used to identify corals of the genus Acropora. In a molecular phylogenetic analysis, I examine relationships for the mitochondrial DNAs putative control region, using maximum-parsimony and maximum-likelihood methods. The reproductive criteria explore whether species and morphs are reproductively isolated on the basis of temporal or fertilisation barriers. Timing of gamete maturity is surveyed for each species and morph, from the month prior to and 3 months after the mass spawning. Time of spawning is documented at the levels of night and hour of spawning, and time taken for egg-sperm bundles to separate. Laboratory fertilisation experiments tested the potential of species and morphs to interbreed. High levels of intraspecific and extremely low or zero fertilisation levels between the five species indicated that they are valid species. Based on the combined assessment of morphological, molecular and reproductive criteria, A. humilis and A. gemmifera appear to be the most closely related species, which are most closely related to the remaining species in the following order: A. samoensis, A. monticulosa and A. digitifera. Evidence derived from one or more of these criteria suggest that the morphs (1) are at various stages of divergence from the species with which they share morphological characters, and (2) may indicate possible zones of speciation and hybridisation. Identification of morphs avoided the possibility of taxonomic error and was essential for accurate interpretation of evolutionary boundaries. Confirmation of morphology as an informative character of evolutionary boundaries is of great significance because most coral research projects rely on morphology as the primary tool for identification of species.Communicated by M.S. Johnson, Crawley