National Primary Care Day \u2797

No abstract available

Repression of GW/P body components and the RNAi microprocessor impacts primary ciliogenesis in human astrocytes

<p>Abstract</p> <p>Background</p> <p>In most cells, the centriolar component of the centrosome can function as a basal body supporting the formation of a primary cilium, a non-motile sensory organelle that monitors information from the extracellular matrix and relays stimuli into the cell via associated signaling pathways. Defects in the formation and function of primary cilia underlie multiple human diseases and are hallmarks of malignancy. The RNA silencing pathway is involved in the post-transcriptional silencing of > 50% of mRNA that occurs within GW/P bodies. GW/P bodies are found throughout the cytoplasm and previously published live cell imaging data suggested that in a malignant cell type (U2OS), two GW/P bodies reside at the centrosome during interphase. This led us to investigate if a similar relationship exists in primary cells and if the inhibition of the miRNA pathway impairs primary cilium formation.</p> <p>Results</p> <p>Two GW/P bodies as marked by GW182 and hAgo2 colocalized to the basal body of primary human astrocytes as well as human synoviocytes during interphase and specifically with the distal end of the basal body in the pericentriolar region. Since it is technically challenging to examine the two centrosomal GW/P bodies in isolation, we investigated the potential relationship between the global population of GW/P bodies and primary ciliogenesis. Astrocytes were transfected with siRNA directed to GW182 and hAgo2 and unlike control astrocytes, a primary cilium was no longer associated with the centrosome as detected in indirect immunofluorescence assays. Ultrastructural analysis of siRNA transfected astrocytes revealed that knock down of GW182, hAgo2, Drosha and DGCR8 mRNA did not affect the appearance of the earliest stage of ciliogenesis but did prevent the formation and elongation of the ciliary axoneme.</p> <p>Conclusions</p> <p>This study confirms and extends a previously published report that GW/P bodies reside at the centrosome in U2OS cells and documents that GW/P bodies are resident at the centrosome in diverse non-malignant cells. Further, our study demonstrates that repression of key effector proteins in the post-transcriptional miRNA pathway impairs primary cilium formation.</p

Retrospective: Adjusting Contaminant Concentrations in Bird Eggs to Account for Moisture and Lipid Loss During Their Incubation

By the 1960s, research and monitoring efforts on chlorinated pesticide residues in tissues of wildlife were well underway in North America and Europe. Conservationists and natural resource managers were attempting to resolve whether pesticide exposure and accumulated residues were related to population declines in several species of predatory and scavenging birds (e.g., bald eagle Haliaeetus leucocephalus, peregrine falcon Falco peregrinus, brown pelican Pelecanus occidentalis and osprey Pandion haliaetus). The avian egg was a favored sampling matrix even before the realization that eggshell thinning was linked to population declines (Ratcliffe 1967; Hickey and Anderson 1968) and that the concentration of p,p’-DDE in an egg was associated with the shell thinning phenomenon (e.g., Blus et al. 1972; Wiemeyer et al. 1988). The necessity for making wet-weight concentration adjustments to account for natural moisture loss during incubation of viable eggs was realized. Correction for the more dramatic moisture loss in non-viable decaying eggs was recognized as being paramount. For example, the ∑DDT residues in osprey eggs were reported to vary by as much as eightfold without accounting for moisture loss adjustments (Stickel et al. 1965). In the absence of adjusting concentrations to the fresh wet-weight that was present at the time of egg laying, the uncorrected values exaggerated contaminant concentrations, yielding artifactual results and ultimately incorrect conclusions. The adjustment to fresh wet-weight concentration is equally important for many other persistent contaminants including PCBs, dioxins, furans, and brominated diphenyl ethers

Drivers of Pb, Sb and As release from spent gunshot in wetlands: Enhancement by organic matter and native microorganisms

In many countries the use of lead-based ammunition is prevalent, and results in exposure and poisoning of waterfowl and other species of birds. In waterfowl hunting areas large quantities of spent shot may be deposited in wetland and terrestrial habitats. These pellets can undergo transformations, which are influenced by various abiotic and biotic factors. In addition to lead (Pb), other elements like antimony (Sb) and arsenic (As) can be leached from Pb shot into the environment. In vitro simulations that included organic matter and microorganisms were utilized to examine elemental leaching from gunshot. We found that leaching efficiency was the greatest in solutions rich in organic matter derived from artificial root exudates (2.69 % for Pb, 1.16 % for Sb, 1.83 % for As), while leaching efficiency was considerably lower in river water (0.04 %). In vitro simulations containing native microorganisms also exhibited greater leaching efficiency (0.49 % for Pb, 0.52 % for Sb, 1.32 % for As) than in ultrapure deionized water and river water. Surface alterations in gunshot included the formation of a weathering crust and secondary phases dominated by carbonates. Spent gunshot is a source of Pb, Sb and As in wetlands that could affect aquatic ecosystems

Mapping the assembly pathways that specify formation of the trilaminar kinetochore plates in human cells

We report the interactions amongst 20 proteins that specify their assembly to the centromere–kinetochore complex in human cells. Centromere protein (CENP)-A is at the top of a hierarchy that directs three major pathways, which are specified by CENP-C, -I, and Aurora B. Each pathway consists of branches that intersect to form nodes that may coordinate the assembly process. Complementary EM studies found that the formation of kinetochore trilaminar plates depends on the CENP-I/NUF2 branch, whereas CENP-C and Aurora B affect the size, shape, and structural integrity of the plates. We found that hMis12 is not constitutively localized at kinetochores, and that it is not essential for recruiting CENP-I. Our studies also revealed that kinetochores in HeLa cells contain an excess of CENP-A, of which ∼10% is sufficient to promote the assembly of normal levels of kinetochore proteins. We elaborate on a previous model that suggested kinetochores are assembled from repetitive modules (Zinkowski, R.P., J. Meyne, and B.R. Brinkley. 1991. J. Cell Biol. 113:1091–110)

Comparative Toxicity of Diphacinone to Northern Bobwhite (\u3ci\u3eColinus virginianus\u3c/i\u3e) and American Kestrels (\u3ci\u3eFalco sparverius\u3c/i\u3e)

The acute oral toxicity of the anticoagulant rodenticide diphacinone was found to be about 20 times greater to American kestrels (LD50=97 mg/kg) than to northern bobwhite (LD50=2,014 mg/kg). Several precise and sensitive clotting assays (prothrombin time, Russell’s Viper venom time, thrombin clotting time) were adapted for use in these species, and this combination of assays is recommended to detect effects of diphacinone and other rodenticides on coagulation. Oral administration of diphacinone over a range of doses (sublethal to the extrapolated LD15) prolonged prothrombin time and Russell’s Viper venom time within 24 to 48 hrs post-exposure. Prolongation of in vitro clotting time reflects impaired coagulation complex activity and was detected before or at the onset of overt signs of toxicity and lethality. These data will assist in the development of a pharmacodynamic model to assess and predict rodenticide toxicity to non-target avian species