Transformation of mammalian cells by overexpressing H2O2-generating peroxisomal fatty acyl-CoA oxidase.

Peroxisome proliferators induce qualitatively predictable pleiotropic responses, including development of hepatocellular carcinomas in rats and mice despite the inability of these compounds to interact with and damage DNA directly. In view of the nongenotoxic nature of peroxisome proliferators, it has been postulated that hepatocarcinogenesis by this class of chemicals is due to a receptor-mediated process leading to transcriptional activation of H2O2-generating peroxisomal fatty acyl-CoA oxidase (ACOX) in liver. To test this hypothesis, we overexpressed rat ACOX in African green monkey kidney cells (CV-1 cells) under control of the cytomegalovirus promoter. A stably transfected CV-1 cell line overexpressing rat ACOX, designated CV-ACOX4, when exposed to a fatty acid substrate (150 microM linoleic acid) for 2-6 weeks, formed transformed foci, grew efficiently in soft agar, and developed adenocarcinomas when transplanted into nude mice. These findings indicate that sustained overexpression of H2O2-generating ACOX causes cell transformation and provide further support for the role of peroxisome proliferation in hepatocarcinogenesis induced by peroxisome proliferators.

Endogenous mutagenesis by an insertion sequence element identifies Aeromonas salmonicida AbcA as an ATP-binding cassette transport protein required for biogenesis of smooth lipopolysaccharide.

Analysis of an Aeromonas salmonicida A layer-deficient/O polysaccharide-deficient mutant carrying a Tn5 insertion in the structural gene for A protein (vapA) showed that the abcA gene immediately downstream of vapA had been interrupted by the endogenous insertion sequence element ISAS1. Immunoelectron microscopy showed that O polysaccharides did not accumulate at the inner membrane-cytoplasm interface of this mutant. abcA encodes an unusual protein; it carries both an amino-terminal ATP-binding cassette (ABC) domain showing high sequence similarity to ABC proteins implicated in the transport of certain capsular and O polysaccharides and a carboxyl-terminal potential DNA-binding domain, which distinguishes AbcA from other polysaccharide transport proteins in structural and evolutionary terms. The smooth lipopolysaccharide phenotype was restored by complementation with abcA but not by abcA carrying site-directed mutations in the sequence encoding the ATP-binding site of the protein. The genetic organization of the A. salmonicida ABC polysaccharide system differs from other bacteria. abcA also differs in apparently being required for both O-polysaccharide synthesis and in energizing the transport of O polysaccharides to the cell surface.

Immortalization of distinct human mammary epithelial cell types by human papilloma virus 16 E6 or E7.

Multiple mammary epithelial cell (MEC) types are observed both in mammary ducts in vivo and in primary cultures in vitro; however, the oncogenic potential of different cell types remains unknown. Here, we used human papilloma virus 16 E6 and E7 oncogenes, which target p53 and Rb tumor suppressor proteins, respectively, to immortalize MECs present in early or late passages of human mammary tissue-derived cultures or in milk. One MEC subtype was exclusively immortalized by E6; such cells predominated in late-passage cultures but were rare at early passages and apparently absent in milk. Surprisingly, a second cell type, present only in early-passage tissue-derived cultures, was fully immortalized by E7 alone. A third cell type, observed in tissue-derived cultures and in milk, showed a substantial extension of life span with E7 but eventually senesced. Finally, both E6 and E7 were required to fully immortalize milk-derived MECs and a large proportion of MECs in early-passage tissue-derived cultures, suggesting the presence of another discrete subpopulation. Identification of MECs with distinct susceptibilities to p53- and Rb-targeting human papillomavirus oncogenes raises the possibility that these cells may serve as precursors for different forms of breast cancer.

The Wsh and Ph mutations affect the c-kit expression profile: c-kit misexpression in embryogenesis impairs melanogenesis in Wsh and Ph mutant mice.

The receptor tyrosine kinases (RTKs) c-kit and platelet-derived growth factor receptor alpha chain (PDG-FRa) are encoded at the white spotting (W) and patch (Ph) loci on mouse chromosome 5. While W mutations affect melanogenesis, gametogenesis, and hematopoiesis, the Ph mutation affects melanogenesis and causes early lethality in homozygotes. W-sash (Wsh) is an expression mutation and blocks c-kit expression in certain cell types and enhances c-kit expression in others, including at sites important for early melanogenesis. We have determined the effect of Ph on c-kit expression during embryogenesis in Ph heterozygotes. Immunohistochemical analysis revealed enhanced c-kit expression in several cell types, including sites important for early melanogenesis. We propose that in both Wsh and Ph mutant mice c-kit misexpression affects early melanogenesis and is responsible for the pigment deficiency. Moreover, we have defined the organization of the RTKs in the W/Ph region on chromosome 5 and characterized the Wsh mutation by using pulsed-field gel electrophoresis. Whereas the order of the RTK genes was determined as Pdgfra-c-kit-flk1, analysis of the Wsh mutation revealed that the c-kit and Pdgfra genes are unlinked in Wsh, presumably because of an inversion of a small segment of chromosome 5. The Ph mutation consists of a deletion including Pdgfra and the 3' deletion endpoint of Ph lies between Pdgfra and c-kit. Therefore, positive 5' upstream elements controlling c-kit expression in mast cells and some other cell types are affected by the Wsh mutation and negative elements are affected by both the Wsh and the Ph mutation.

Extracellular pH regulation in microdomains of colonic crypts: effects of short-chain fatty acids.

It has been suggested that transepithelial gradients of short-chain fatty acids (SCFAs; the major anions in the colonic lumen) generate pH gradients across the colonic epithelium. Quantitative confocal microscopy was used to study extracellular pH in mouse distal colon with intact epithelial architecture, by superfusing tissue with carboxy SNARF-1 (a pH-sensitive fluorescent dye). Results demonstrate extracellular pH regulation in two separate microdomains surrounding colonic crypts: the crypt lumen and the subepithelial tissue adjacent to crypt colonocytes. Apical superfusion with (i) a poorly metabolized SCFA (isobutyrate), (ii) an avidly metabolized SCFA (n-butyrate), or (iii) a physiologic mixture of acetate/propionate/n-butyrate produced similar results: alkalinization of the crypt lumen and acidification of subepithelial tissue. Effects were (i) dependent on the presence and orientation of a transepithelial SCFA gradient, (ii) not observed with gluconate substitution, and (iii) required activation of sustained vectorial acid/base transport by SCFAs. Results suggest that the crypt lumen functions as a pH microdomain due to slow mixing with bulk superfusates and that crypts contribute significant buffering capacity to the lumen. In conclusion, physiologic SCFA gradients cause polarized extracellular pH regulation because epithelial architecture and vectorial transport synergize to establish regulated microenvironments.

Cuerdas y no conmutatividad

Los conceptos geométricos clásicos que usan las ecuaciones de Einstein para describir el espacio, el tiempo y la gravedad no son compatibles con los principios de la mecánica cuántica. A distancias muy cortas, cercanas o por debajo de la longitud de Planck lp 10−36m, se espera que la estructura del espacio-tiempo se haga difusa, con un principio de incertidumbre asociado a las propias coordenadas espacio temporales. Tal principio de incertidumbre podría derivarse de relaciones de conmutación no triviales [xμ, x ] 6= 0 entre operadores asociados a la posición en el espacio-tiempo. Conmutadores de este tipo aparecen de manera natural en teoría de cuerdas, que, por otra parte, contiene y generaliza las ecuaciones de Einstein. Constituye por ello un escenario idóneo para el estudio de la naturaleza no determinista del espacio-tiempo. En efecto, Chu y Ho [4] demostraron que la cuantización canónica de la cuerda abierta en espacio-tiempo de Minkowski con 2-forma B y dilatón constantes conduce a conmutadores no triviales entre los operadores de posición de los extremos de la cuerda. Este hecho sugiere la interpreción de la D-brana sobre la que pueden moverse dichos extremos como un espacio no conmutativo. Seiberg y Witten [5] dieron un paso más y encontraron un límite de baja energía bien definido en el cual la dinámica de los extremos de la cuerda se desacopla de la de los modos internos y se describe como una teoría de Yang-Mills no conmutativa sobre la D-brana. Es díficil trasladar estos resultados a backgrounds más generales para la cuerda. Los modelos de Wess-Zumino-Witten (WZW) [6] constituyen los backgrounds no triviales mejor conocidos, pero no se conoce una caracterización completa de las D-branas en estos modelos. El objetivo de esta tesis es mejorar la comprensión del origen de no conmutatividad a partir de (i) el estudio de la cuerda abierta en backgrounds no triviales y (ii) la caracterización de las D-branas sobre las que pueden moverse sus extremos. El trabajo de esta tesis ha dado lugar a las publicaciones [I], [II] y [III]...

Paraklētikē : manuscript, 1353

Written by unidentified scribe in Church Slavic of Serbian recension, in semiuncial book hand in brown ink on vellum, headings and initials in red ink; front pages are missing; pagination in pencil starts on f. 1.

(Table 1) Concentrations of the main organochlorine pesticides in liver and plasma samples of ringed seals (Phoca hispida)

The present study investigates the concentrations and patterns of organochlorine pesticides (OCPs) and their metabolites in liver and plasma of two ringed seal populations (Phoca hispida): lower contaminated Svalbard population and more contaminated Baltic Sea population. Among OCPs, p,p'-DDE and sum-chlordanes were the highest in concentration. With increasing hepatic contaminant concentrations and activities of xenobiotic-metabolizing enzymes, the concentrations of 3-methylsulfonyl-p,p'-DDE and the concentration ratios of pentachlorophenol/hexachlorobenzene increased, and the toxaphene pattern shifted more towards persistent Parlar-26 and -50 and less towards more biodegradable Parlar-44. Relative concentrations of the chlordane metabolites, oxychlordane and -heptachlorepoxide, to sum-chlordanes were higher in the seals from Svalbard compared to the seals from the Baltic, while the trend was opposite for cis- and trans-nonachlor. The observed differences in the OCP patterns in the seals from the two populations are probably related to the catalytic activity of xenobiotic-metabolizing enzymes, and also to differences in dietary exposure.

Proglacial river dataset from the Akuliarusiarsuup Kuua River northern tributary, Southwest Greenland, 2008 - 2010, version 1.0

Pressing scientific questions concerning the Greenland ice sheet's climatic sensitivity, hydrology, and contributions to current and future sea level rise require hydrological datasets to resolve. While direct observations of ice sheet meltwater losses can be obtained in terrestrial rivers draining the ice sheet and from lake levels, few such datasets exist. We present a new dataset of meltwater river discharge for the vicinity of Kangerlussuaq, Southwest Greenland. The dataset contains measurements of river stage and discharge for three sites along the Akuliarusiarsuup Kuua (Watson) River's northern tributary, with 30 minute temporal resolution between June 2008 and August 2010. Additional data of water temperature, air pressure, and lake water depth and temperature are also provided. Discharge data were measured at sites with near-ideal properties for such data collection. Regardless, high water bedload and turbulent flow introduce considerable uncertainty. These were constrained and quantified using statistical techniques, thereby providing a high quality dataset from this important site. The greatest data uncertainties are associated with streambed elevation change and measurements. Large portions of stream channels deepened according to statistical tests, but poor precision of streambed depth measurements also added uncertainty. Quality checked data are freely available for scientific use as supplementary online material.

Tu Fu : wanderer and minstrel under moons of Cathay /

Mode of access: Internet.

Beach erosion control study, Homer Spit, Alaska /

"September 1987."

Tongzhi tiao yue /

On double leaves, oriental style, in 2 cases.

Toward 2000: opportunities in transportation evolution.

Mode of access: Internet.