Sphingomyelinase treatment of rat hepatocytes inhibits cell-swelling-stimulated glycogen synthesis by causing cell shrinkage.

Breakdown of plasma-membrane sphingomyelin caused by TNF-alpha is known to inhibit glucose metabolism and insulin signalling in muscle and fat cells. In hepatocytes, conversion of glucose to glycogen is strongly activated by amino acid-induced cell swelling. In order to find out whether breakdown of plasma-membrane sphingomyelin also inhibits this insulin-independent process, the effect of addition of sphingomyelinase was studied in rat hepatocytes. Sphingomyelinase (but not ceramide) inhibited glycogen synthesis, caused cell shrinkage, decreased the activity of glycogen synthase a, but had no effect on phosphorylase a. Cell integrity was not affected by sphingomyelinase addition as gluconeogenesis and the intracellular concentration of ATP were unchanged. As a control, glycogen synthesis was studied in HepG2 cells. In these cells, the basal rate of glycogen production was high, could not be stimulated by amino acids, nor be inhibited by sphingomyelinase. Regarding the mechanism responsible for the inhibition of glycogen synthase a, sphingomyelinase did not affect amino acid-induced, PtdIns 3-kinase-dependent, phosphorylation of p70S6 kinase, but caused an increase in intracellular chloride, which is known to inhibit glycogen synthase phosphatase. It is concluded that the decrease in cell volume, following the breakdown of sphingomyelin in the plasma membrane of the hepatocyte, may contribute to the abnormal metabolism of glucose when TNF-alpha levels are high.     

The antifungal plant defensin RsAFP2 from radish induces apoptosis in a metacaspase independent way in Candida albicans

We show that the antifungal plant defensin Raphanus sativus antifungal protein 2 (RsAFP2) from radish induces apoptosis and concomitantly triggers activation of caspases or caspase-like proteases in the human pathogen Candida albicans. Furthermore, we demonstrate that deletion of C. albicans metacaspase 1, encoding the only reported (putative) caspase in C. albicans, significantly affects caspase activation by the apoptotic stimulus acetic acid, but not by RsAFP2. To our knowledge, this is the first report on the induction of apoptosis with concomitant caspase activation by a defensin in this pathogen. Moreover, our data point to the existence of at least two different types of caspases or caspase-like proteases in C. albicans.     

Changes in cytoplasmic pH are involved in the cell type regulation of Dictyostelium

The cytoplasmic pH (pHi) of populations of developing Dictyostelium discoideum cells was determined by means of two independent pH null-point methods. Both methods reveal in populations containing 75-80% prespore cells a pHi value of about 0.2 pH units higher than in populations containing 50% prespore cells. During the process of cell type regulation, decreases and increases in the percentage of prespore cells of about 15-20% are accompanied by decreases and increases in pHi of about 0.2 pH units. Abolition of these changes in pHi by means of a weak base or acid also prevents the regulation process. It is concluded that changes in pHi are involved in the prespore cell type regulation in D. discoideum.     

External nutrient inputs into terrestrial ecosystems of the Falkland Islands and the Maritime Antarctic region

Antarctic terrestrial ecosystems are nutrient-poor and depend for their functioning in part on external nutrients. However, little is known about the relative importance of various sources. We measured external mineral nutrient sources (wind blown material, precipitation and guano) at three locations, the cold temperate oceanic Falkland Islands (51°76′S), and the Maritime Antarctic Signy (60°71′S) and Anchorage Islands (67°61′S). These islands differ in the level of vegetation development through different environmental constraints and historical factors. Total mineral nitrogen input differed considerably between the islands. During the 3 month summer period it amounted to 18 mg N m⁻² on the Falkland Islands and 6 and 102 mg N m⁻² at Signy and Anchorage Islands, respectively. The high value for Anchorage was a result of guano deposition. By measuring stable isotopic composition (δ¹⁵N) of the different nitrogen sources and the dominant plant species, we investigated the relative utilisation of each source by the vegetation at each island. We conclude that external mineral nitrogen inputs to Antarctic terrestrial ecosystems show great spatial variability, with the local presence of bird (or other vertebrate) colonies being particularly significant.     

Somatic Uniparental Isodisomy Explains Multifocality of Glomuvenous Malformations

Inherited vascular malformations are commonly autosomal dominantly inherited with high, but incomplete, penetrance; they often present as multiple lesions. We hypothesized that Knudson’s two-hit model could explain this multifocality and partial penetrance. We performed a systematic analysis of inherited glomuvenous malformations (GVMs) by using multiple approaches, including a sensitive allele-specific pairwise SNP-chip method. Overall, we identified 16 somatic mutations, most of which were not intragenic but were cases of acquired uniparental isodisomy (aUPID) involving chromosome 1p. The breakpoint of each aUPID is located in an A- and T-rich, high-DNA-flexibility region (1p13.1–1p12). This region corresponds to a possible new fragile site. Occurrences of these mutations render the inherited glomulin variant in 1p22.1 homozygous in the affected tissues without loss of genetic material. This finding demonstrates that a double hit is needed to trigger formation of a GVM. It also suggests that somatic UPID, only detectable by sensitive pairwise analysis in heterogeneous tissues, might be a common phenomenon in human cells. Thus, aUPID might play a role in the pathogenesis of various nonmalignant disorders and might explain local impaired function and/or clinical variability. Furthermore, these data suggest that pairwise analysis of blood and tissue, even on heterogeneous tissue, can be used for localizing double-hit mutations in disease-causing genes.