(Poly)phenolic compounds in pollen and spores of Antarctic plants as indicators of solar UV-B – A new proxy for the reconstruction of past solar UV-B?

The morphology, size and characteristics of the pollen of the plant species Antarctic hairgrass (Deschampsia antarctica, Poaceae) and Antarctic pearlwort (Colobanthus quitensis, Caryophyllaceae) are described by scanning electron microscopy and light microscopy. Based on the number of pores the pollen of Colobanthus quitensis is classified as periporate or polypantorate, while that of Deschampsia antarctica is monoporate.

Pollen of Vicia faba plants, exposed to enhanced UV-B (10.6 kJ m^?2 day^?1 UV-B_BE) in a greenhouse, showed an increased content of UV-B absorbing compounds. There was also an increase of UV-B absorbing compounds in response to exposure to UV-A. By sequential chemical extraction three `compartments' of UV-B absorbance of pollen can be distinguished: a cytoplasmic fraction consisting of, e.g., flavonoids (acid-methanol extractable), a wall-bound fraction, consisting of, e.g., ferulic acid (NaOH extractable) and aromatic groups in the bioresistant polymer sporopollenin possibly consisting of, e.g., para-coumaric acid monomers (fraction remaining after acetolysis). The sporopollenin fraction in the pollen of Helleborus foetidus showed considerable UV-B absorbance (280–320 nm). There is evidence that enhanced solar UV-B induces increased UV-B absorbance (of sporopollenin) in pollen and spores of mosses, which may be preserved in the fossil record. As there are no instrumental records of solar UV-B in the Antarctic before 1970 and no instrumental records of stratospheric ozone over the Antarctic before 1957, the use of UV-B absorbing polyphenolics in pollen (and spores) as bio-indicator, or proxy of solar UV-B, may allow reconstruction of pre-ozone hole and subrecent UV-B and stratospheric ozone levels. Pollen and spores from herbarium specimens and from frozen moss banks (about 5000–10?000 years old) in the Antarctic may, therefore, represent a valuable archive of historical UV-B levels.

     

Stimulated platelets release equivalent amounts of arachidonate from phosphatidylcholine, phosphatidylethanolamine, and inositides

Thrombin-induced changes in arachidonate content of platelet phospholipids were quantitated to establish the ultimate origins of this eicosanoid precursor. Fifteen seconds following thrombin addition (15 U/5 X 10(9) platelets), phosphatidylcholine lost 11.8 nmol of arachidonate and phosphatidylethanolamine lost 10.5 nmol. Arachidonate in phosphatidate, phosphatidylinositol, and phosphatidylinositol-4,5-bisphosphate combined decreased by 11.0 nmol. Increases in free and oxygenated arachidonate (41 nmol) exceeded decreases in inositides. Thus phospholipase A2 released at least twice as much arachidonate as phospholipase C-diglyceride lipase. Phosphatidylinositol-4-phosphate levels remained unchanged upon stimulation. Therefore, increases in phosphatidylinositol-4,5-bisphosphate indicated the minimum rate of phosphorylation of phosphatidylinositol to resynthesize phosphatidylinositol-4,5-bisphosphate, following stimulus-induced breakdown by phospholipase C. Phosphatidylinositol-4, 5-bisphosphate increased 1.4 nmol between 10 and 15 sec following thrombin, markedly less than phosphatidylinositol decreased (2.1 nmol). This could be due to phospholipase A2, in addition to phospholipase C, acting directly on phosphatidylinositol to a greater extent than estimated by accumulation of lysophosphatidylinositol, degraded rapidly by lysophospholipase. Thus, upon high-dose thrombin stimulation of human platelets inositide metabolism via phospholipase C directs initial formation of intracellular second messengers, and sequentially, or in parallel, arachidonate release by phospholipase A2 supplies the larger proportion of arachidonate for syntheses of eicosanoids involved in intercellular communication.     

Cytochemical demonstration of hydrogen peroxide in polymorphonuclear leukocyte phagosomes

Phagocytosis by polymorphonuclear leukocytes (PMN) is accompanied by specific morphological and metabolic events which may result in the killing of internalized micro-organism. Hydrogen peroxide is produced in increased amounts during phagocytosis (17) and in combination with myeloperoxidase and halide ions constitute a potent, microbicidal mechanism (8,9,11). There can be direct iodination of micro-organisms (10), or alternatively, other intermediate reaction products, i.e. chloramines and aldehydes (21), can exert a microbicidal effect. The H2O2-peroxidase-halide system is presumed to operate within the phagocytic vacuole (12,18). Myeloperoxidase, present in the primary granules of PMN, enters the phagocytic vacuole during degranulation (1,4,7), and halide ions are probably derived from the extracellular medium or are present in the PMN (see 11, 18). For the operation of this system in intact cells, the presence of H2O2 in the phagocytic vacuole is necessary, and indeed this has been suggested by the work of several investigators (12, 18, 21). In the present investigation, the diaminobenzidine reaction of Graham and Karnovsky (5), modified to utilize endogenous myeloperoxidase and hydrogen peroxide, has been applied to actively phagocytizing PMN to demonstrate cytochemically the presence of H2O2 in the phagocytic vacuole.     

Molecular evolution of the period gene in Drosophila athabasca

We measured nucleotide variability within and between the three semispecies of the Drosophila athabasca complex, at the period (per) gene by using a polymerase chain reaction-based four-cutter restriction- enzyme analysis. The levels of polymorphism varied considerably between the three semispecies. Our results for per, combined with previous data for X-linked allozymes, suggest that the X chromosome in the western- northern semispecies is less variable than expected under an equilibrium-neutral model. Both the pattern of divergence between the semispecies and a cladistic clustering of per haplotypes support the previously hypothesized grouping of eastern A and eastern B as the two most recently diverged semispecies. A 21-bp in-frame segment in the region of per which shares sequence similarity with the neuronal development gene single minded is deleted in all eastern A and eastern B flies examined but is present in all of the western-northern flies and all other published per sequences. Despite these hints that there may be significant differences at the per gene between the semispecies, especially the western-northern group versus the two eastern groups, there is no compelling evidence that per is involved in the mating song differences between the semispecies.      

Mapping of fabC, a locus for the biosynthesis of unsaturated fatty acids in Escherichia coli K12

Summary A mutation affecting the biosynthesis of unsaturated fatty acids in Escherichia coli K12 has been mapped. This mutation called fabC, which is cotransducible with nalA, lies between the nalA and his loci.This paper is taken from a thesis to be submitted by J. H. F. F. Broekman to the faculty of Science of the State University in Utrecht in partial fulfilment of the requirements for the Ph. D. degree.     

Using eggshell membranes as a DNA source for population genetic research

In the context of population genetic research, a faster and less invasive method of DNA sampling would allow large-scale assessments of genetic diversity and genetic differentiation with the help of volunteer observers. The aim of this study was to investigate the usefulness of eggshell membranes as a DNA source for population genetic research, by addressing eggshell membrane DNA quality, degeneration and cross-contamination. To this end, a comparison was made with blood-derived DNA samples. We have demonstrated 100% successful DNA extraction from post-hatched Black-tailed Godwit (Limosa limosa) eggshell membranes as well as from blood samples. Using 11 microsatellite loci, DNA amplification success was 99.1% for eggshell membranes and 97.7% for blood samples. Genetic information within eggshell membrane DNA in comparison to blood DNA was not affected (F _ST = −0.01735, P = 0.999) by degeneration or possible cross-contamination. Furthermore, neither degeneration nor cross-contamination was apparent in total genotypic comparison of eggshell membrane DNA and blood sample DNA. Our research clearly illustrates that eggshell membranes can be used for population genetic research.