Pannexin 1 channels mediate 'find-me' signal release and membrane permeability during apoptosis

Apoptotic cells release 'find-me' signals at the earliest stages of death to recruit phagocytes. The nucleotides ATP and UTP represent one class of find-me signals, but their mechanism of release is not known. Here, we identify the plasma membrane channel pannexin 1 (PANX1) as a mediator of find-me signal/nucleotide release from apoptotic cells. Pharmacological inhibition and siRNA-mediated knockdown of PANX1 led to decreased nucleotide release and monocyte recruitment by apoptotic cells. Conversely, PANX1 overexpression enhanced nucleotide release from apoptotic cells and phagocyte recruitment. Patch-clamp recordings showed that PANX1 was basally inactive, and that induction of PANX1 currents occurred only during apoptosis. Mechanistically, PANX1 itself was a target of effector caspases (caspases 3 and 7), and a specific caspase-cleavage site within PANX1 was essential for PANX1 function during apoptosis. Expression of truncated PANX1 (at the putative caspase cleavage site) resulted in a constitutively open channel. PANX1 was also important for the 'selective' plasma membrane permeability of early apoptotic cells to specific dyes. Collectively, these data identify PANX1 as a plasma membrane channel mediating the regulated release of find-me signals and selective plasma membrane permeability during apoptosis, and a new mechanism of PANX1 activation by caspases.     

Resistance to therapy caused by intragenic deletion in BRCA2

Cells with loss of BRCA2 function are defective in homologous recombination (HR) and are highly sensitive to inhibitors of poly(ADP-ribose) polymerase (PARP), which provides the basis for a new therapeutic approach. Here we show that resistance to PARP inhibition can be acquired by deletion of a mutation in BRCA2. We derived PARP-inhibitor-resistant (PIR) clones from the human CAPAN1 pancreatic cancer cell line, which carries the protein-truncating c.6174delT frameshift mutation. PIR clones could form DNA-damage-induced RAD51 nuclear foci and were able to limit genotoxin-induced genomic instability, both hallmarks of a competent HR pathway. New BRCA2 isoforms were expressed in the resistant lines as a result of intragenic deletion of the c.6174delT mutation and restoration of the open reading frame (ORF). Reconstitution of BRCA2-deficient cells with these revertant BRCA2 alleles rescued PARP inhibitor sensitivity and HR deficiency. Most of the deletions in BRCA2 were associated with small tracts of homology, and possibly arose from error-prone repair caused by BRCA2 deficiency. Similar ORF-restoring mutations were present in carboplatin-resistant ovarian tumours from c.6174delT mutation carriers. These observations have implications for understanding drug resistance in BRCA mutation carriers as well as in defining functionally important domains within BRCA2.     

Event-horizon-scale structure in the supermassive black hole candidate at the Galactic Centre

The cores of most galaxies are thought to harbour supermassive black holes, which power galactic nuclei by converting the gravitational energy of accreting matter into radiation. Sagittarius A* (Sgr A*), the compact source of radio, infrared and X-ray emission at the centre of the Milky Way, is the closest example of this phenomenon, with an estimated black hole mass that is 4,000,000 times that of the Sun. A long-standing astronomical goal is to resolve structures in the innermost accretion flow surrounding Sgr A*, where strong gravitational fields will distort the appearance of radiation emitted near the black hole. Radio observations at wavelengths of 3.5 mm and 7 mm have detected intrinsic structure in Sgr A*, but the spatial resolution of observations at these wavelengths is limited by interstellar scattering. Here we report observations at a wavelength of 1.3 mm that set a size of 37(+16)(-10) microarcseconds on the intrinsic diameter of Sgr A*. This is less than the expected apparent size of the event horizon of the presumed black hole, suggesting that the bulk of Sgr A* emission may not be centred on the black hole, but arises in the surrounding accretion flow.     

Experimentally-derived haplotypes substantially increase the efficiency of linkage disequilibrium studies.

The study of complex genetic traits in humans is limited by the expense and difficulty of ascertaining populations of sufficient sample size to detect subtle genetic contributions to disease. Here we introduce an application of a somatic cell hybrid construction strategy called conversion that maximizes the genotypic information from each sampled individual. The approach permits direct observation of individual haplotypes, thereby eliminating the need for collecting and genotyping DNA from family members for haplotype-based analyses. We describe experimental data that validate the use of conversion as a whole-genome haplotyping tool and evaluate the theoretical efficiency of using conversion-derived haplotypes instead of conventional genotypes in the context of haplotype-frequency estimation. We show that, particularly when phenotyping is expensive, conversion-based haplotyping can be more efficient and cost-effective than standard genotyping.     

PARAMAP vs. Isomap: A Comparison of Two Nonlinear Mapping Algorithms

Dimensionality reduction techniques are used for representing higher dimensional data by a more parsimonious and meaningful lower dimensional structure. In this paper we will study two such approaches, namely Carroll’s Parametric Mapping (abbreviated PARAMAP) (Shepard and Carroll, 1966) and Tenenbaum’s Isometric Mapping (abbreviated Isomap) (Tenenbaum, de Silva, and Langford, 2000). The former relies on iterative minimization of a cost function while the latter applies classical MDS after a preprocessing step involving the use of a shortest path algorithm to define approximate geodesic distances. We will develop a measure of congruence based on preservation of local structure between the input data and the mapped low dimensional embedding, and compare the different approaches on various sets of data, including points located on the surface of a sphere, some data called the "Swiss Roll data", and truncated spheres.     

Least squares algorithms for constructing constrained ultrametric and additive tree representations of symmetric proximity data

A mathematical programming algorithm is developed for fitting ultrametric or additive trees to proximity data where external constraints are imposed on the topology of the tree. The two procedures minimize a least squares loss function. The method is illustrated on both synthetic and real data. A constrained ultrametric tree analysis was performed on similarities between 32 subjects based on preferences for ten odors, while a constrained additive tree analysis was carried out on some proximity data between kinship terms. Finally, some extensions of the methodology to other tree fitting procedures are mentioned.The first author is supported as Bevoegdverklaard Navorser of the Belgian Nationaal Fonds voor Wetenschappelijk Onderzoek.     

Ontogenetic and habitat-related changes in diet of late larval and juvenile suckers (Catostomidae) in Upper Klamath Lake, Oregon

We describe ontogenetic patterns in the diets of shortnose and Lost River suckers (15.8–92.8 mm standard length) from Upper Klamath Lake in summer 1999. Both species made a transition from surface and planktonic prey to benthic prey at about 20–30 mm standard length, corresponding to the approximate size of the juvenile morphological transition. Surface prey was dominated by adult chironomids and undigestable pollen, while benthic prey was dominated by larval chironomids, chydorids, and ostracods. In the 15–20-mm size class, pollen made up >75% of food particles in 68% of specimens, and only 2 specimens in this size class lacked any pollen grains. A better understanding of the prey selection process in larval suckers is needed to determine the importance of this potential source of starvation. The planktonic prey eaten during the surface-to-benthic feeding transition were widely distributed in the lake, suggesting that larvae use emergent vegetation primarily as a refuge from predators. On a numerical basis, crustaceans and larval chironomids contributed most to the diets of larger juveniles. In specimens >40 mm, shortnose suckers ate more Tanytarsini and Lost River suckers ate more chironomid pupae and chydorids. In specimens >40 mm there was a strong habitat-related difference in diet: specimens collected in offshore samples ate more chironomid larvae and pupae, harpacticoid copepods, and chydorids, whereas onshore specimens ate more cyclopoid copepods and invertebrate eggs. The transition from onshore to offshore habitat appears to occur over a broad size range of about 40–90 mm.