Flagellar membranes are rich in raft-forming phospholipids.

The observation that the membranes of flagella are enriched in sterols and sphingolipids has led to the hypothesis that flagella might be enriched in raft-forming lipids. However, a detailed lipidomic analysis of flagellar membranes is not available. Novel protocols to detach and isolate intact flagella from Trypanosoma brucei procyclic forms in combination with reverse-phase liquid chromatography high-resolution tandem mass spectrometry allowed us to determine the phospholipid composition of flagellar membranes relative to whole cells. Our analyses revealed that phosphatidylethanolamine, phosphatidylserine, ceramide and the sphingolipids inositol phosphorylceramide and sphingomyelin are enriched in flagella relative to whole cells. In contrast, phosphatidylcholine and phosphatidylinositol are strongly depleted in flagella. Within individual glycerophospholipid classes, we observed a preference for ether-type over diacyl-type molecular species in membranes of flagella. Our study provides direct evidence for a preferential presence of raft-forming phospholipids in flagellar membranes of T. brucei.

Casimir effect with quantized charged spinor matter in background magnetic field

We study the influence of a background uniform magnetic field and boundary conditions on the vacuum of a quantized charged spinor matter field confined between two parallel neutral plates; the magnetic field is directed orthogonally to the plates. The admissible set of boundary conditions at the plates is determined by the requirement that the Dirac Hamiltonian operator be self-adjoint. It is shown that, in the case of a sufficiently strong magnetic field and a sufficiently large separation of the plates, the generalized Casimir force is repulsive, being independent of the choice of a boundary condition, as well as of the distance between the plates. The detection of this effect seems to be feasible in the foreseeable future.

Function and regulation of anionic phospholipids in mitochondria of Saccharomyces cerevisiae

As the major anionic phospholipids predominantly found in the mitochondrial inner membrane of eukaryotic cells, cardiolipin (CL) and its precursor phosphatidylglycerol (PG) are of great importance in many critical mitochondrial processes. Pgs1Δ cells of Saccharomyces cerevisiae lacking both PG and CL display severe mitochondrial defects. Translation of several proteins including products of four mitochondrial DNA (mtDNA) encoded genes (COX1, COX2, COX3, and COB ) and one nuclear-encoded gene (COX4) is inhibited. The molecular basis of this phenotype was analyzed using a combined biochemical, molecular and genetic approach. ^ Using a mitochondrial targeted green fluorescence protein (mtGFP) fused to the COX4 promoter and its 5′ and 3′ untranslated regions (UTRs), lack of mtGFP expression independent of carbon source and strain background was confirmed to be at the translational level. The translational defect was not due to deficiency of mitochondrial respiratory function but rather caused directly by the lack of PG/CL in the mitochondrial membrane. Re-introduction of a functional PGS1 gene restored PG synthesis and expression of the above mtGFP. Deletional analysis of the 5′ UTR of COX4 mRNA revealed the presence of a 50 nt sequence as a cis-acting element inhibiting COX4 translation. Using similar constructs with HIS3 and lacZ as reporter genes, extragenic spontaneous mutations that allowed expression of His3p and β-galactosidase were isolated, which appeared to be recessive and derived from loss-of-function mutations as determined by mating analysis. Using a tetracycline repressible plasmid-borne PGS1 expression system and an in vivo mitochondrial protein translation method, the translation of mtDNA encoded COX1 and COX3 mRNAs was shown to be significantly inhibited in parallel with reduced levels of PG/CL content. Therefore, the cytoplasmic translation machinery appears to be able to sense the level of PG/CL in mitochondria and regulate COX4 translation coordinately with the mtDNA encoded subunits. ^ The essential requirement of PG and CL in mitochondrial function was further demonstrated in the study of CL synthesis by factors affecting mitochondrial biogenesis such as carbon source, growth phase or mitochondrial mutations at the level of transcription. We have also demonstrated that CL synthesis is dependent on the level of PG and INO2/INO4 regulatory genes. ^

Phospholipids in sediments of the deep Arabian Sea

Eight different sites from 2300 to 4420 m water depth in the Arabian Sea were sampled for a biochemical quantification of phospholipid concentrations in the sediments. This method serves as a measure of microbial biomass in marine sediments comprising all small-sized organisms, including bacteria, fungi, protozoa and metazoa. Phospholipid concentrations can be converted to carbon units as an estimate of total microbial biomass in the sediments. The average phospholipid concentrations in the surface sediments (0-1 cm) of the 4 abyssal sites ranged from 7 nmol cm?3 at the southern site (SAST, 10°N 65°E, 4425 m) to 29 nmol/cm**3 at the western site (WAST, 16°N 60°E, 4045 m). The high values detected at the abyssal station WAST exceeded those in the literature for other abyssal sites and were comparable to values from the upper continental slope of the NE-Atlantic and the Arctic. At the four continental slope sites in the Arabian Sea, average phospholipid concentrations ranged from 9 to 53 nmol/cm**3 with the maximum values at stations A (2314 m) and D (3142 m) close to the Omani coast. Records of particulate organic carbon flux to the deep sea are available for four of the investigated locations, allowing a test of the hypothesis that the standing stock of benthic microorganisms in the deep sea is controlled by substrate availability, i.e. particle sedimentation. Total microbial biomass in the surface sediments of the Arabian Sea was positively correlated with sedimentation rates, consistent with previous studies of other oceans. The use of the measurement of phospholipid concentrations as a proxy for input of particulate organic matter is discussed.

Percentage of cultured cell-populations that stained positively and/or negatively for apoptotic markers cleaved caspase-3 and cleaved PARP, following DNA damage treatments induced by doxorubicin and TNF-alpha co-treatments

The present dataset contains the source data for Figure 2B of Tentner et al. (2012). The data shows the percentage of cultured cell-populations that stained positively and/or negatively for apoptotic markers cleaved caspase-3 and cleaved PARP, following DNA damage treatments induced by various doses of doxorubicin (0, 2 and 10 µmole/L) in the presence (100 ng/mL) or absence (0 ng/mL) of TNF-alpha co-treatment. For the six treatment conditions investigated, cell counts were made by flow cytometry at times 6, 12, 24, and 48 h following treatment; CULTURE DETAILS: U2OS cells were obtained from ATCC were maintained at 21% oxygen and 5% CO2 in Dulbecco's modified Eagle medium supplemented with 10% fetal bovine serum, penicillin, streptomycin, 2mM L-glutamine, and used within 15-20 passages. The first thymidine block was released by washing the plates three times with PBS, and incubating them in fresh thymidine-free media for 12 h. A second thymidine block was then performed by re-addition of thymidine to 2.5 mM followed by incubation for an additional 18 h. Media was aspirated, plates were washed 3 with PBS, and replaced with fresh media in the presence or absence of 10 mM aphidicolin; ANALYSIS DETAILS: See supplementary journal publication; RESULT: The authors of the supplementary journal publication conclude that TNF enhances dose-dependent cell death following doxorubicin-induced DNA damage with minimal affect on dose-dependent cell-cycle arrest.