The urethral fold of 30 mares was split transversely into dorsal and ventral shelves, and the ventral shelf was used to help create a urethral extension. The dorsal shelf was stretched caudally and sutured to the roof of the extension so that it covered at least the cranial half of the extension. For 20 mares, a relaxing, vaginal incision was created cranial to the external urethral orifice to enable the dorsal shelf to be retracted further caudally. Ten of the 30 mares (33.3 per cent) developed a defect, but none developed a defect in that portion covered by the dorsal shelf of the urethral fold. Two of the 30 mares (6.7 per cent) developed a defect so small that the defect could be detected only by inserting a dye, under pressure, into the tunnel. The total number of mares that developed only a grossly visible and palpable defect was eight of 30 (26.6 per cent). Four of the 10 mares that did not receive the relief incision and six of 20 mares that did receive the relief incision developed a defect in the extension. Modifying the McKinnon technique by transversely splitting the urethral fold and retracting the dorsal half helps prevent a defect from forming in the cranial portion of the extension. The dorsal shelf can be retracted further caudally by creating a relief incision on the floor of the vagina.

The human small nuclear (sn) and small cytoplasmic (sc) RNA gene families encode diverse non-coding RNAs that influence cellular growth and division. Many snRNA and scRNA genes are related via their compact and yet powerful promoters that support RNA polymerase III transcription. We have utilized the human U6 snRNA gene family to examine the mechanism for regulated transcription of these potent transcription units. Analysis of nine U6 family members showed enriched CpG density within the promoters of actively transcribed loci relative to inert genes, implying a relationship between gene potency and DNA methylation. Indeed, both pharmacological inhibition of DNMT activity and the forced diminution of DNA methyltransferases (DNMT)-1, DNMT-3a, and DNMT-3b by siRNA targeting resulted in increased U6 levels in asynchronously growing U2OS osterosarcoma cells. In vitro transcription assays further showed that template methylation impedes U6 transcription by RNA polymerase III. Both DNMT-1 and DNMT-3a were detected at the U6-1 locus by chromatin immunoprecipitation directly linking these factors to RNA polymerase III regulation. Despite this association, the endogenous U6-1 locus was not substantially methylated in actively growing cells. However, both DNMT occupancy and low frequency methylation were correlated with increased Retinoblastoma tumor suppressor (RB) expression, suggesting that the RB status can influence specific epigenetic marks.

Our aim was to compare plastinated sections of the canine heart with corresponding two-dimensional (2D) echocardiographic images. Thirteen dog hearts were fixed by dilation and then processed by the S10 silicon plastination method (Biodur®). Two dogs without evidence of cardiac disease were imaged using 2D echocardiography so as to obtain a complete series of the standard right and left parasternal images, which were compared with corresponding plastinated slices obtained by knife sectioning of the hearts. The plastinated slices revealed the internal anatomy of the heart with great detail and were particularly useful to display the spatial relationship between complex anatomic structures. The plastinated slices corresponded accurately with the echocardiographic images. Because of the dilation of the right heart during the fixation process, it was not possible to obtain plastinated specimens in ventricular systole. This paper may be a reference atlas for assisting 2D echocardiography interpretation.

DNA damage induced by ionizing radiation activates the ataxia telangiectasia mutated pathway resulting in apoptosis or DNA repair. The serine/threonine checkpoint kinase Chk2 is an important transducer of this DNA damage signaling pathway and mediates the ultimate fate of the cell. Chk2 is an advantageous target for the development of adjuvant drugs for cancer therapy, because inhibition of Chk2 allows normal cells to enter cell cycle arrest and DNA repair, whereas many tumors bypass cell cycle checkpoints. Chk2 inhibitors may thus have a radioprotective affect on normal cells. We herein report the synthesis, kinase inhibition and radioprotective abilities of Chk2 inhibitors that are structurally related to the natural product hymenialdisine. Several of the hymenialdisine-derived analogs inhibit Chk2 at nanomolar concentrations, inhibit autophosphorylation of Chk2 at Ser516 in cells and increase the survival of normal cells following ionizing radiation.

With 14 figures SUMMARY: The respiratory tracts of seven grey short-tailed opossums were histologically examined. Six opossums were prepared by perfusion with buffered formalin. Opossum seven was perfused with gluteraldehyde. Samples taken from the respiratory passages and lungs of specimens 1-6 were stained with haematoxylin and eosin. A mixture of methylene and azure blue was used for specimen 7. The trachea and right and left principal bronchi are lined with a pseudostratified ciliated columnar epithelium with occasional goblet cells. The secondary and tertiary bronchi and the primary and secondary bronchioles are lined by a simple ciliated columnar epithelium. The terminal bronchioles and a portion of the respiratory bronchioles are lined by a simple ciliated cuboidal epithelium. The terminal portion of the respiratory bronchioles and the alveolar ducts are lined with simple squamous epithelium. Alveoli are lined by type I and II pneumocytes. Tracheal glands are present in the tela submucosa. The fibromusculocartilaginous tunic of the trachea consists of c-shaped cartilage rings and the trachealis muscle. A lamina muscularis mucosa begins in the intrapulmonary portion of the principal bronchus and continues into the respiratory bronchioles. Bronchial glands are present in the propria submucosa and tela submucosa of the principal bronchi. The musculocartilaginous tunic is localized to the extrapulmonary portion of the principal bronchus. The bronchial cartilages are irregular shaped plates and limited to the extrapulmonary portion of the principal bronchus. The visceral pleura is a simple squamous mesothelium covering the outer surface of the lung.

Here we report the first measurements of polybrominated diphenyl ethers (PBDE-47,-99, and -153) alongside 11 organochlorine pesticides (OCPs) and 28 polychlorinated biphenyls (PCBs) in the plasma of albatross from breeding colonies distributed across a large spatial east-west gradient in the North Pacific Ocean. North Pacific albatross are wide-ranging, top-level consumers that forage in pelagic regions of the North Pacific Ocean, making them an ideal sentinel species for detection and distribution of marine contaminants. Our work on contaminant burdens in albatross tissue provides information on transport of persistent organic pollutants (POPs) to the remote North Pacific and serves as a proxy for regional environmental quality. We sampled Black-footed (Phoebastria nigripes; n = 20) and Laysan albatross (Phoebastria immutabilis; n = 19) nesting on Tern Island, Hawaii, USA, and Laysan albatross (n = 16) nesting on Guadalupe Island, Mexico. Our results indicate that North Pacific albatross are highly exposed to both PCBs and OCPs with levels ranging from 8.8 to 86.9 ng/ml wet weight and 7.4 to 162.3 ng/ml wet weight, respectively. A strong significant gradient between Laysan albatross breeding in the Eastern Pacific having approximately 1.5 fold and 2.5 fold higher levels for PCBs and OCPs, respectively, compared to those from the Central Pacific. Interspecies levels of contaminants within the same breeding site also showed high variation with Tern Black-footed albatross having approximately three fold higher levels of both PCBs and OCPs than Tern Laysan albatross. Surprisingly, while PBDEs are known to travel long distances and bioaccumulate in wildlife of high trophic status, we detected these three PBDE congeners only at trace levels ranging from not detectable (ND) to 0.74 ng/ml wet weight in these albatross. Environ. Toxicol. Chem. © 2011 SETAC.

Thorough dehydration is a key for good plastination and invariably it leads to shrinkage. Shrinkage during plastination has been studied to lesser extent. Shrinkage was studied in 10 pig kidneys including regional shrinkage (cortex, medulla, sinus) and at which stages of the process (dehydration, impregnation, curing) shrinkage occurred. Kidneys were fixation by perfusion of 10% neutral buffered formalin solution via the renal artery. The vessels and ureter were filled with colored silicone (Dow Corning, Silastic E RTV Silicone Rubber) and the kidneys were cut into one centimeter transverse slices. Two slices of each kidney were plastinated via the classic von Hagens' method. Slices were photographed at the same focal length after preparation and at the end of each stage of plastination. Slice surface area was determined by a point-counting planimetry method. Post dehydration shrinkage of the kidney was 10.21% while post impregnation 10.11%. After completion of plastination, total area of kidney slice shrinkage was 19.72%. Cortical area shrunk 12.81% after dehydration and 13.16% after impregnation. After plastination, cortical area had shrunk 24.28%. No significant shrinkage occurred in the medulla and sinus. Results demonstrate that kidney shrinkage during impregnation is as intense as during dehydration. Significant shrinkage occurred in the renal cortex but not in the medulla and sinus. This demonstrates that different tissue types, even in the same specimen, have different rates of shrinkage during dehydration and impregnation. Therefore, plastinated specimens should be used carefully in research where obtaining measures is important.

Pelagic marine predators face unprecedented challenges and uncertain futures. Overexploitation and climate variability impact the abundance and distribution of top predators in ocean ecosystems. Improved understanding of ecological patterns, evolutionary constraints and ecosystem function is critical for preventing extinctions, loss of biodiversity and disruption of ecosystem services. Recent advances in electronic tagging techniques have provided the capacity to observe the movements and long-distance migrations of animals in relation to ocean processes across a range of ecological scales. Tagging of Pacific Predators, a field programme of the Census of Marine Life, deployed 4,306 tags on 23 species in the North Pacific Ocean, resulting in a tracking data set of unprecedented scale and species diversity that covers 265,386 tracking days from 2000 to 2009. Here we report migration pathways, link ocean features to multispecies hotspots and illustrate niche partitioning within and among congener guilds. Our results indicate that the California Current large marine ecosystem and the North Pacific transition zone attract and retain a diverse assemblage of marine vertebrates. Within the California Current large marine ecosystem, several predator guilds seasonally undertake north-south migrations that may be driven by oceanic processes, species-specific thermal tolerances and shifts in prey distributions. We identify critical habitats across multinational boundaries and show that top predators exploit their environment in predictable ways, providing the foundation for spatial management of large marine ecosystems.

During blastocyst formation the segregation of the inner cell mass (ICM) and trophectoderm is governed by the mutually antagonistic effects of the transcription factors Oct4 and Cdx2. Evidence indicates that suppression of Oct4 expression in the trophectoderm is mediated by Cdx2. Nonetheless, the underlying epigenetic modifiers required for Cdx2-dependent repression of Oct4 are largely unknown. Here we show that the chromatin remodeling protein Brg1 is required for Cdx2-mediated repression of Oct4 expression in mouse blastocysts. By employing a combination of RNA interference (RNAi) and gene expression analysis we found that both Brg1 Knockdown (KD) and Cdx2 KD blastocysts exhibit widespread expression of Oct4 in the trophectoderm. Interestingly, in Brg1 KD blastocysts and Cdx2 KD blastocysts, the expression of Cdx2 and Brg1 is unchanged, respectively. To address whether Brg1 cooperates with Cdx2 to repress Oct4 transcription in the developing trophectoderm, we utilized preimplantation embryos, trophoblast stem (TS) cells and Cdx2-inducible embryonic stem (ES) cells as model systems. We found that:(1) combined knockdown (KD) of Brg1 and Cdx2 levels in blastocysts resulted in increased levels of Oct4 transcripts compared to KD of Brg1 or Cdx2 alone,(2) endogenous Brg1 co-immunoprecipitated with Cdx2 in TS cell extracts,(3) in blastocysts Brg1 and Cdx2 co-localize in trophectoderm nuclei and (4) in Cdx2-induced ES cells Brg1 and Cdx2 are recruited to the Oct4 promoter. Lastly, to determine how Brg1 may induce epigenetic silencing of the Oct4 gene, we evaluated CpG methylation at the Oct4 promoter in the trophectoderm of Brg1 KD blastocysts. This analysis revealed that Brg1-dependent repression of Oct4 expression is independent of DNA methylation at the blastocyst stage. In toto, these results demonstrate that Brg1 cooperates with Cdx2 to repress Oct4 expression in the developing trophectoderm to ensure normal development.