Structural studies of the parainfluenza virus 5 hemagglutinin-neuraminidase tetramer in complex with its receptor, sialyllactose

The paramyxovirus hemagglutinin-neuraminidase (HN) functions in virus attachment to cells, cleavage of sialic acid from oligosaccharides, and stimulating membrane fusion during virus entry into cells. The structural basis for these diverse functions remains to be fully understood. We report the crystal structures of the parainfluenza virus 5 (SV5) HN and its complexes with sialic acid, the inhibitor DANA, and the receptor sialyllactose. SV5 HN shares common structural features with HN of Newcastle disease virus (NDV) and human parainfluenza 3 (HPIV3), but unlike the previously determined HN structures, the SV5 HN forms a tetramer in solution, which is thought to be the physiological oligomer. The sialyllactose complex reveals intact receptor within the active site, but no major conformational changes in the protein. The SV5 HN structures do not support previously proposed models for HN action in membrane fusion and suggest alternative mechanisms by which HN may promote virus entry into cells.     

The pattern of polymorphism in Arabidopsis thaliana

We resequenced 876 short fragments in a sample of 96 individuals of Arabidopsis thaliana that included stock center accessions as well as a hierarchical sample from natural populations. Although A. thaliana is a selfing weed, the pattern of polymorphism in general agrees with what is expected for a widely distributed, sexually reproducing species. Linkage disequilibrium decays rapidly, within 50 kb. Variation is shared worldwide, although population structure and isolation by distance are evident. The data fail to fit standard neutral models in several ways. There is a genome-wide excess of rare alleles, at least partially due to selection. There is too much variation between genomic regions in the level of polymorphism. The local level of polymorphism is negatively correlated with gene density and positively correlated with segmental duplications. Because the data do not fit theoretical null distributions, attempts to infer natural selection from polymorphism data will require genome-wide surveys of polymorphism in order to identify anomalous regions. Despite this, our data support the utility of A. thaliana as a model for evolutionary functional genomics.     

Determination of WR-1065 in human blood by high-performance liquid chromatography following fluorescent derivatization by a maleimide reagent ThioGlo3

In order to improve the sensitivity and stability of human blood samples containing WR-1065 (i.e., active metabolite of the cytoprotective agent amifostine), a high-performance liquid chromatographic method was developed and validated using fluorescent derivatization with ThioGlo3. Using a sample volume of only 100 microl, the method was specific, sensitive (limit of quantitation=10 nM in deproteinized blood or 20 nM in whole blood), accurate (error < or = 3.2%) and reproducible (CV < or = 8.7%). In addition, the stability of WR-1065 in deproteinized and derivatized blood samples was assured for at least four weeks at -20 degrees C. This method should be particularly valuable in translating the kinetic-dynamic relationship of WR-1065 in preclinical models to that in cancer patients.     

Systematics,biogeography and evolution of Asaccus gallagheri (Squamata,Phyllodactylidae) with the description of a new endemic species from Oman

The Hajar Mountains are the highest mountain range in eastern Arabia. Despite being classified as a mountain desert, it is considered one of the top biodiversity hotspots of Arabia. As a result of its relatively old geological origin, complex topography, environmental heterogeneity and geographic isolation from other mountain ranges, its fauna and flora have diversified significantly producing high levels of endemicity, particularly amongst reptiles. Several genetic studies indicate that this diversity may still be underestimated, especially within some groups containing morphologically similar species like the nocturnal geckos of the genus Asaccus. These have radiated extensively on both sides of the Gulf of Oman, in the Hajar Mountains and the Zagros Mountains of south-west Asia, and are a good example of the faunal affinities between these two mountain ranges. In the present work, we analyse A. gallagheri, the smallest species of the Arabian radiation, using an unprecedented sampling across its entire distribution range and an integrative approach combining morphological, macroecological and multilocus molecular data with the objective of clarifying its systematics and phylogeography. The results support the presence of two allopatric species within A. gallagheri that split approximately 6 Ma. The newly discovered species is endemic to the Eastern Hajars and is described herein mainly on the basis of its smaller size and high genetic divergence from A. gallagheri. The molecular analyses also uncovered remarkable levels of genetic diversity within both species. The present study highlights the diversity of the genus Asaccus in south-east Arabia and stresses its relevance from a conservation point of view.

http://www.zoobank.org/urn:lsid:zoobank.org:pub:62EB3146-9F79-4857-8CC6-36FE235D84D4     

The contribution of DNA replication stress marked by high-intensity,pan-nuclear γH2AX staining to chemosensitization by CHK1 and WEE1 inhibitors

Small molecule inhibitors of the checkpoint proteins CHK1 and WEE1 are currently in clinical development in combination with the antimetabolite gemcitabine. It is unclear, however, if there is a therapeutic advantage to CHK1 vs. WEE1 inhibition for chemosensitization. The goals of this study were to directly compare the relative efficacies of the CHK1 inhibitor MK8776 and the WEE1 inhibitor AZD1775 to sensitize pancreatic cancer cell lines to gemcitabine and to identify pharmacodynamic biomarkers predictive of chemosensitization. Cells treated with gemcitabine and either MK8776 or AZD1775 were first assessed for clonogenic survival. With the exception of the homologous recombination-defective Capan1 cells, which were relatively insensitive to MK8776, we found that these cell lines were similarly sensitized to gemcitabine by CHK1 or WEE1 inhibition. The abilities of either the CDK1/2 inhibitor roscovitine or exogenous nucleosides to prevent MK8776 or AZD1775-mediated chemosensitization, however, were both inhibitor-dependent and variable among cell lines. Given the importance of DNA replication stress to gemcitabine chemosensitization, we next assessed high-intensity, pan-nuclear γH2AX staining as a pharmacodynamic marker for sensitization. In contrast to total γH2AX, aberrant mitotic entry or sub-G1 DNA content, high-intensity γH2AX staining correlated with chemosensitization by either MK8776 or AZD1775 (R² 0.83 – 0.53). In summary, we found that MK8776 and AZD1775 sensitize to gemcitabine with similar efficacy. Furthermore, our results suggest that the effects of CHK1 and WEE1 inhibition on gemcitabine-mediated replication stress best predict chemosensitization and support the use of high-intensity or pan-nuclear γH2AX staining as a marker for therapeutic response.     

Free-ranging domestic cats (<Emphasis Type="Italic">Felis catus</Emphasis>) on public lands: estimating density,activity, and diet in the Florida Keys

Feral and free-ranging domestic cats (Felis catus) can have strong negative effects on small mammals and birds, particularly in island ecosystems. We deployed camera traps to study free-ranging cats in national wildlife refuges and state parks on Big Pine Key and Key Largo in the Florida Keys, USA, and used spatial capture–recapture models to estimate cat abundance, movement, and activities. We also used stable isotope analyses to examine the diet of cats captured on public lands. Top population models separated cats based on differences in movement and detection with three and two latent groups on Big Pine Key and Key Largo, respectively. We hypothesize that these latent groups represent feral, semi-feral, and indoor/outdoor house cats based on the estimated movement parameters of each group. Estimated cat densities and activity varied between the two islands, with relatively high densities (~4 cats/km²) exhibiting crepuscular diel patterns on Big Pine Key and lower densities (~1 cat/km²) exhibiting nocturnal diel patterns on Key Largo. These differences are most likely related to the higher proportion of house cats on Big Pine relative to Key Largo. Carbon and nitrogen isotope ratios from hair samples of free-ranging cats (n = 43) provided estimates of the proportion of wild and anthropogenic foods in cat diets. At the population level, cats on both islands consumed mostly anthropogenic foods (>80% of the diet), but eight individuals were effective predators of wildlife (>50% of the diet). We provide evidence that cat groups within a population move different distances, exhibit different activity patterns, and that individuals consume wildlife at different rates, which all have implications for managing this invasive predator.     

Rubella—Some Comments on the 1964-65 Epidemic in California

The rubella epidemic of 1964-1965 resulted in the birth of a group of children with defects of vision, of hearing or of the heart. In this study of cases known to five Los Angeles agencies, it was found that about half of those affected have more than one defect. Findings demonstrate a need for more sensitive communicable disease surveillance and for the development of services for the multiple handicapped child.     

NUMERICAL TAXONOMY OF LIMNANTHACEAE

A numerical taxonomic study of the angiosperm family Limnanthaceae is presented. The 21 OTUs include all taxa recognized in a recent monograph of Limnanthes (Mason, 1952), plus Floerkea proserpinacoides, two undescribed Limnanthes species, and two artificial interspecific F₁ Limnanthes hybrids. One species (L. montana) was treated as two OTUs: measurements for one OTU were taken from field collections and for the other, from garden-grown plants. A maximum of 35 characters was used in the numerical analyses. Raw data for each character were given equal weighting by condensation. Coefficients of resemblance were established using the mean character difference (MCD) method. Tryon's clustering technique (called key communality cluster analysis) was used. Three numerical analyses of Limnanthaceae were made: one with all 35 characters (T analysis), one with 18 floral characters (F analysis), and one with 17 pre- or post-anthesis or vegetative characters (V analysis). Relative positions of OTUs in the T and F analyses are substantially different from those in a conventional taxonomic treatment of the family. The V analysis shows many differences from the T and F analyses. In all analyses the two OTUs representing L. montana were closer to other OTUs than to each other. The two hybrid OTUs showed greater resemblance to one parent than to the other and in two analyses one hybrid was not intermediate between the parents. Similarities between the T and F analyses are explained on the basis of the predominance of floral characters in both. The differences between these two analyses on the one hand and the V analysis and phylogenetic arrangement of the family on the other are attributed to parallelisms in the evolution of floral characters associated with iucreasing autogamy in each of the two phyletic lines in the family. Analyses emphasizing floral characters will place evolutionarily remotely related outcrossing OTUs together and unrelated, autogamous OTUs together. Using characters that are mostly unassociated with the breeding system the positions of OTUs in a numerical analysis are similar to their positions in a phylogenetic arrangement. The V analysis agrees best with suggestions concerning phylogenetic affinities among the taxa in the family, and provides a useful context for the generation of further evolutionary hypotheses.     

Genetic control of basal level of alkaline phosphatase in Escherichia coli

Summary The basis for a difference in basal levels of alkaline phosphatase in two strains of E. coli has been examined. It was found that the enzyme level characteristic of a highlevel strain was not conferred upon a low level strain merely by the transfer of the pho ⁺, phoR⁺ or phoS ⁺ allele from the high-level strain to the low level strain. Such strains may have slightly altered basal levels of enzyme, but these changes are not quantitatively sufficient to account for the strain differences. Approximately 20% of the recombinants which receive the phoS ⁺ gene show a level of phosphatase intermediate between the high and low-level strains. The effect is specific for the basal level of phosphatase, since it does not affect the levels of three other enzymes, or the derepressed level of alkaline phosphatase itself. This latter fact is consistent with the view that the intermediate level strains have an intermediate level of repressor of alkaline phosphatase under repressed conditions. It is suggested that the basal level of enzyme is affected by several genes in addition to those previously described, and that the intermediate level strains have received the allele of one or more of these loci from the high level strain.     

THE ELECTRICAL CONDUCTIVITY OF SODIUM AND POTASSIUM GUAIACOLATES IN GUAIACOL

The data of the author and Uhlig, and new data, on the conductivity of sodium and of potassium guaiacolates in guaiacol at 25° have been computed with an improved conductance equation which is valid to somewhat higher concentrations than the equations formerly used. The new constants are, Λ₀ = 9.0, K = 2.8 x 10^–5 for sodium guaiacolate and Λ₀ = 9.5, K = 3.4 x 10^–5 for potassium guaiacolate.