Characterization of the acidic N-linked glycans of the zona pellucida of prepuberal pigs by a mass spectrometric approach

Oocyte maturation is a prerequisite for successful fertilization. Growing evidence suggests that not only the oocyte but also the surrounding zona pellucida has to undergo maturational changes. In the pig, two-dimensional electrophoretic analysis demonstrated an acidic shift of the zona pellucida glycoproteins of about 1.5–2.0 pH units during the maturation process. These findings were corroborated by histological studies that indicated the synthesis of acidic glycoconjugates in the cumulus cells and an increased occurrence of acidic glycans in the zona pellucida after oocyte maturation. In order to provide structural data on prepuberal zona pellucida N-glycosylation, N-glycans were released from prepuberal zona pellucida glycoproteins by N-glycosidase F and studied by mass spectrometry before and after desialylation and treatment with endo-β-galactosidase. Our results verified the presence of high-mannose-type Man₅GlcNAc₂ compounds as well as diantennary N-glycans as major neutral species, whereas sialylated diantennary and triantennary species constituted the dominant non-sulfated acidic sugar chains. The major acidic N-glycans of prepuberal animals, however, represented mono-sulfated diantennary, triantennary and tetraantennary oligosaccharides carrying, in part, N-acetyllactosamine repeating units as well as additional Neu5Ac or Neu5Gc residues. Glycans comprising more than one sulfate residue were not detected. In contrast to the literature data on zona pellucida glycoprotein-N-glycans of cyclic animals, our data thus reveal a lower degree in glycan sulfation of the prepuberal zona pellucida.     

Modeling system states in liver cells: Survival,apoptosis and their modifications in response to viral infection

Background  

The decision pro- or contra apoptosis is complex, involves a number of different inputs, and is central for the homeostasis of an individual cell as well as for the maintenance and regeneration of the complete organism.     

Hemiclonal reproduction slows down the speed of Muller's ratchet in the hybridogenetic frog Rana esculenta

Rare recombination in otherwise asexually reproducing organisms is known to beneficially influence the fitness in small populations. In most of the investigated organisms, asexual and rare sexual generations with recombination follow each other sequentially. Here we present a case where clonal reproduction and rare recombination occur simultaneously in the same population. The hybridogenetic water frog Rana esculenta (E), a hybrid between R. lessonae (L) and R. ridibunda (R) produces gametes that only contain the unaltered maternal R part of their genome. New generations of R. esculenta usually arise from E x L matings. Intraspecific E x E matings produce mostly inviable offspring, but in rare cases, female R. ridibunda arise from such matings which are capable of recombination. In the absence of conspecific males, these R females have to mate with E males, which results in further R females, or with L males, which produces new E lineages. This indirect mechanism reintroduces recombination into the otherwise clonally transmitted R genomes in R. esculenta populations. In this study, we show through Monte Carlo simulations that, in most cases, it is sufficient that only between 1 % and 10 % of mixed water frog populations consist of R females to prevent or significantly reduce the fixation and accumulation of deleterious mutations.     

Human biomonitoring of cadmium and lead exposure of child-mother pairs from Germany living in the vicinity of industrial sources (hot spot study NRW).

Cadmium (Cd) and lead (Pb) exposure of children and their mothers living in the vicinity of industrial sources (metal refining) was assessed by a cross-sectional study performed in 2000. Study areas were the highly industrialized city of Duisburg and a rural area of North Rhine Westphalia, Germany. Exposure to ambient air concentrations of Cd and Pb was calculated from a Lagrange dispersion model using data sets from ambient air quality measurements. Cd in blood and urine and Pb in blood were measured by AAS. Mean age (years) was 6.4 (range 5.5-7.7) for children (n = 238) and 36 (range 23-48) for mothers (n = 213). A total of 49% of the children were males. Factors suspected to influence metal levels in blood or urine were obtained by questionnaire. Individual ambient Cd and Pb levels according to the home address ranged from 0.5 ng/m3 (Cd) and 0.03 microg/m3 (Pb) (rural area) up to 31.2 ng/m3 (Cd) and 0.73 microg/m3 (Pb) (industrialized area). Cd levels (geometric mean) in blood (0.13 and 0.10 microg/L) and urine (both areas 0.09 microg/L) of children did not differ between the two areas. Cd levels in blood and urine of mothers from the industrialized area were higher (blood 0.39 microg/L, urine 0.28 microg/L) than in those from the rural area (blood 0.25 microg/L, urine 0.25 microg/L). Pb levels in the blood of children from the industrialized area were higher (31 microg/L) than in those from the rural area (21 microg/L). Pb levels in the blood of mothers did not differ between the two areas (both 24 microg/L). Pb levels in blood showed a significant association between child and mother (n = 192; r = 0.26, p < 0.001). This did not apply for Cd in blood or urine. Regression analysis clearly revealed that Pb levels in ambient air were associated with Pb in the blood of children. Minor associations were also found between Cd in air and Cd in the blood of mothers and between Cd in air and urine of mothers.