Cloning of a gene (RIG-G) associated with retinoic acid-induced differentiation of acute promyelocytic leukemia cells and representing a new member of a family of interferon-stimulated genes

In a cell line (NB4) derived from a patient with acute promyelocytic leukemia, all-trans-retinoic acid (ATRA) and interferon (IFN) induce the expression of a novel gene we call RIG-G (for retinoic acid-induced gene G). This gene codes for a 58-kDa protein containing 490 amino acids with several potential sites for post-translational modification. In untreated NB4 cells, the expression of RIG-G is undetectable. ATRA treatment induces the transcriptional expression of RIG-G relatively late (12–24 hr) in a protein synthesis-dependent manner, whereas IFN-α induces its expression early (30 min to 3 hr). Database search has revealed a high-level homology between RIG-G and several IFN-stimulated genes in human (ISG54K, ISG56K, and IFN-inducible and retinoic acid-inducible 58K gene) and some other species, defining a well conserved gene family. The gene is composed of two exons and has been mapped by fluorescence in situ hybridization to chromosome 10q24, where two other human IFN-stimulated gene members are localized. A synergistic induction of RIG-G expression in NB4 cells by combined treatment with ATRA and IFNs suggests that a collaboration exists between their respective signaling pathways.

Defects in embryonic hindbrain development and fetal resorption resulting from vitamin A deficiency in the rat are prevented by feeding pharmacological levels of all-trans-retinoic acid

Vitamin A is required for reproduction and normal embryonic development. We have determined that all-trans-retinoic acid (atRA) can support development of the mammalian embryo to parturition in vitamin A-deficient (VAD) rats. At embryonic day (E) 0.5, VAD dams were fed purified diets containing either 12 μg of atRA per g of diet (230 μg per rat per day) or 250 μg of atRA per g of diet (4.5 mg per rat per day) or were fed the purified diet supplemented with a source of retinol (100 units of retinyl palmitate per day). An additional group was fed both 250 μg of atRA per g of diet in combination with retinyl palmitate. Embryonic survival to E12.5 was similar for all groups. However, embryonic development in the group fed 12 μg of atRA per g of diet was grossly abnormal. The most notable defects were in the region of the hindbrain, which included a loss of posterior cranial nerves (IX, X, XI, and XII) and postotic pharyngeal arches as well as the presence of ectopic otic vesicles and a swollen anterior cardinal vein. All embryonic abnormalities at E12.5 were prevented by feeding pharmacological amounts of atRA (250 μg/g diet) or by supplementation with retinyl palmitate. Embryos from VAD dams receiving 12 μg of atRA per g of diet were resorbed by E18.5, whereas those in the group fed 250 μg of atRA per g of diet survived to parturition but died shortly thereafter. Equivalent results were obtained by using commercial grade atRA or atRA that had been purified to eliminate any potential contamination by neutral retinoids, such as retinol. Thus, 250 μg of atRA per g of diet fed to VAD dams (≈4.5 mg per rat per day) can prevent the death of embryos at midgestation and prevents the early embryonic abnormalities that arise when VAD dams are fed insufficient amounts of atRA.

Distinct leukemia phenotypes in transgenic mice and different corepressor interactions generated by promyelocytic leukemia variant fusion genes PLZF–RARα and NPM–RARα

Acute promyelocytic leukemia (APL) is characterized by a specific chromosome translocation involving RARα and one of four fusion partners: PML, PLZF, NPM, and NuMA genes. To study the leukemogenic potential of the fusion genes in vivo, we generated transgenic mice with PLZF–RARα and NPM–RARα. PLZF–RARα transgenic animals developed chronic myeloid leukemia-like phenotypes at an early stage of life (within 3 months in five of six mice), whereas three NPM–RARα transgenic mice showed a spectrum of phenotypes from typical APL to chronic myeloid leukemia relatively late in life (from 12 to 15 months). In contrast to bone marrow cells from PLZF–RARα transgenic mice, those from NPM–RARα transgenic mice could be induced to differentiate by all-trans-retinoic acid (ATRA). We also studied RARE binding properties and interactions between nuclear corepressor SMRT and various fusion proteins in response to ATRA. Dissociation of SMRT from different receptors was observed at ATRA concentrations of 0.01 μM, 0.1 μM, and 1.0 μM for RARα–RXRα, NPM–RARα, and PML–RARα, respectively, but not observed for PLZF–RARα even in the presence of 10 μM ATRA. We also determined the expression of the tissue factor gene in transgenic mice, which was detected only in bone marrow cells of mice expressing the fusion genes. These data clearly establish the leukemogenic role of PLZF–RARα and NPM–RARα and the importance of fusion receptor/corepressor interactions in the pathogenesis as well as in determining different clinical phenotypes of APL.

Retinoids down-regulate telomerase and telomere length in a pathway distinct from leukemia cell differentiation

Human telomerase, a cellular reverse transcriptase (hTERT), is a nuclear ribonucleoprotein enzyme complex that catalyzes the synthesis and extension of telomeric DNA. This enzyme is specifically activated in most malignant tumors but is usually inactive in normal somatic cells, suggesting that telomerase plays an important role in cellular immortalization and tumorigenesis. Terminal maturation of tumor cells has been associated with the repression of telomerase activity. Using maturation-sensitive and -resistant NB4 cell lines, we analyzed the pattern of telomerase expression during the therapeutic treatment of acute promyelocytic leukemia (APL) by retinoids. Two pathways leading to the down-regulation of hTERT and telomerase activity were identified. The first pathway results in a rapid down-regulation of telomerase that is associated with retinoic acid receptor (RAR)-dependent maturation of NB4 cells. Furthermore, during NB4 cell maturation, obtained independently of RAR by retinoic X receptor (RXR)-specific agonists (rexinoids), no change in telomerase activity was observed, suggesting that hTERT regulation requires a specific signaling and occurs autonomously. A second pathway of hTERT regulation, identified in the RAR-responsive, maturation-resistant NB4-R1 cell line, results in a down-regulation of telomerase that develops slowly during two weeks of all-trans retinoic acid (ATRA) treatment. This pathway leads to telomere shortening, growth arrest, and cell death, all events that are overcome by ectopic expression of hTERT. These findings demonstrate a clear and full dissociation between the process of tumor cell maturation and the regulation of hTERT mRNA expression and telomerase activity by retinoids. We propose telomerase expression as an efficient and selective target of retinoids in the therapy of tumors.

RIG-E, a human homolog of the murine Ly-6 family, is induced by retinoic acid during the differentiation of acute promyelocytic leukemia cell.

In vivo all-trans-retinoic acid (ATRA), a differentiation inducer, is capable of causing clinical remission in about 90% of patients with acute promyelocytic leukemia (APL). The molecular basis for the differentiation of APL cells after treatment with ATRA remains obscure and may involve genes other than the known retinoid nuclear transcription factors. We report here the ATRA-induced gene expression in a cell line (NB4) derived from a patient with APL. By differential display-PCR, we isolated and characterized a novel gene (RIG-E) whose expression is up-regulated by ATRA. The gene is 4.0 kb long, consisting of four exons and three introns, and is localized on human chromosome region 8q24. The deduced amino acid sequence predicts a cell surface protein containing 20 amino acids at the N-terminal end corresponding to a signal peptide and an extracellular sequence containing 111 amino acids. The RIG-E coded protein shares some homology with CD59 and with a number of growth factor receptors. It shares high sequence homology with the murine LY-6 multigene family, whose members are small cysteine-rich proteins differentially expressed in several hematopoietic cell lines and appear to function in signal transduction. It seems that so far RIG-E is the closest human homolog of the LY-6 family. Expression of RIG-E is not restricted to myeloid differentiation, because it is also present in thymocytes and in a number of other tissues at different levels.

Photolabeling reveals the proximity of the alpha-neurotoxin binding site to the M2 helix of the ion channel in the nicotinic acetylcholine receptor.

A photoactivatable derivative of neurotoxin II from Naja naja oxiana containing a 125I-labeled p-azidosalicylamidoethyl-1,3'-dithiopropyl label at Lys-25 forms a photo-induced cross-link with the delta subunit of the membrane-bound Torpedo californica nicotinic acetylcholine receptor (AChR). The cross-linked radioactive receptor peptide was isolated by reverse-phase HPLC after tryptic digestion of the labeled delta subunit. The sequence of this peptide, delta-(260-277), and the position of the label at Ala-268 were established by matrix-assisted laser-desorption-ionization mass spectrometry based on the molecular mass and on post-source decay fragment analysis. With the known dimensions of the AChR molecule, of the photolabel, and of alpha-neurotoxin, finding the cross-link at delta Ala-268 (located in the upper part of the channel-forming transmembrane helix M2) means that the center of the alpha-neurotoxin binding site is situated at least approximately 40 A from the extracellular surface of the AChR, proximal to the channel axis.

(Table 1) Argon dating of ash layers from DSDP Hole 18-178

Ash layers from Deep Sea Drilling Project site 178 in the northeast Pacific Ocean have been dated by the 40Ar-39Ar stepwise heating technique to resolve published discrepancies concerning the length of time explosive volcanism has affected the eastern Aleutian arc and Alaskan Peninsula. The results of the investigation indicate that the record of ash-fall deposition at site 178 extends back at least 6.5 m.y. Assuming that 6.5 m.y. ago marks the onset of renewed calc-alkalic volcanism of the volcanic arc, proposed models of continuous and discontinuous motion between the Pacific and North American lithospheric plates can be evaluated. If appreciable time elapsed between the onset of subduction and the onset of arc volcanism, the 6.5-m.y. record of ash-fall deposition in the north-east Pacific is most compatible with models of continuous plate motion throughout late Cenozoic time.

Radiocarbon dates, grain size measurements and selected foraminifera analysis from sediment cores AvH248260-2 and AvH248260-1, Ameralik Fjord, SW Greenland

Sedimentological and geochemical (XRF) data together with information from diatom and benthic foraminiferal records of a 3.5 m long gravity core from Ameralik Fjord, southern West Greenland, is used for reconstructing late-Holocene environmental changes in this area. The changes are linked to large-scale North Atlantic ocean and climate variability. AMS 14C-dating of benthic foraminifera indicates that the sediment core records the last 4400 years and covers the termination of the Holocene Thermal Maximum (HTM). The late HTM (4.4 3.2 ka BP) is characterized by high accumulation rates of fine (silty) sediments related to strong meltwater discharge from the Inland Ice. The HTM benthic foraminiferal fauna demonstrates the presence of well-ventilated, saline bottom water originating from inflow of subsurface West Greenland Current water of Atlantic (Irminger Sea) origin. The hydrographic conditions were further characterized by limited sea ice probably related to a mild and relatively windy winter climate. After 3.2 ka BP lower fine-grained sedimentation rates, but a larger input from sea-ice rafted or aeolian coarse material prevailed. This can be related to colder atmospheric conditions with a decreased meltwater discharge and more widespread sea-ice cover in the fjord.

Plant species, biomass and environmental characteristics of relevés along the North America Arctic bioclimate gradient

Question: How do interactions between the physical environment and biotic properties of vegetation influence the formation of small patterned-ground features along the Arctic bioclimate gradient? Location: At 68° to 78°N: six locations along the Dalton Highway in arctic Alaska and three in Canada (Banks Island, Prince Patrick Island and Ellef Ringnes Island). Methods: We analysed floristic and structural vegetation, biomass and abiotic data (soil chemical and physical parameters, the n-factor [a soil thermal index] and spectral information [NDVI, LAI]) on 147 microhabitat releves of zonalpatterned-ground features. Using mapping, table analysis (JUICE) and ordination techniques (NMDS). Results: Table analysis using JUICE and the phi-coefficient to identify diagnostic species revealed clear groups of diagnostic plant taxa in four of the five zonal vegetation complexes. Plant communities and zonal complexes were generally well separated in the NMDS ordination. The Alaska and Canada communities were spatially separated in the ordination because of different glacial histories and location in separate floristic provinces, but there was no single controlling environmental gradient. Vegetation structure, particularly that of bryophytes and total biomass, strongly affected thermal properties of the soils. Patterned-ground complexes with the largest thermal differential between the patterned-ground features and the surrounding vegetation exhibited the clearest patterned-ground morphologies.

al-Inshāʾ al-ʻaṣrī lil-kātib wa-al-ḥāsib wa-al-mallāk wa-al-tājir fī al-murāsalāt wa-al-muʻāmalāt wa-al-ḥisāb wa-mask al-dafātir [microform] /

Microfilm.

[Majmūʻat rasāʾil] [microform] /

"Risālah fī ithbāt al-najāh wa-al-imān" by ʻAlī al-Dāghistānī: 8 p. at end.

MARKETING DE RELACIONAMENTO NAS INSTITUIÇÕES DE ENSINO SUPERIOR: Estudo de caso da Universidade Metodista de São Paulo e da Universidade Metodista de Piracicaba

O marketing de relacionamento como ferramenta para atrair e fidelizar os alunos do ensino superior privado é o tema desta pesquisa. Trata-se de um estudo de caso múltiplo incorporado enfocando a Universidade Metodista de São Paulo (UMESP) e a Universidade Metodista de Piracicaba (UNIMEP) e que tem como objetivos principais apontar fatores que motivam os estudantes a escolher determinadas instituições de ensino superior, identificar a maneira como elas atuam na tarefa de atraí-los com vistas a estabelecer um relacionamento e uma comunicação eficientes com esses públicos de interesse e fidelizálos, bem como analisar a real efetividade das ações desenvolvidas pelas IESs. Propõe-se uma incursão teórica para abordar motivações psicológicas, culturais, econômicas e sociais relacionadas ao ato de consumir e estabelecer uma correlação com o consumidor-aluno . Discutem-se também características do marketing de relacionamento associadas ao contexto educacional e apontam-se dificuldades/desafios vivenciados pelas instituições nos esforços de criar e estreitar os laços com os estudantes com vistas a tornar tais laços duradouros. Foram aplicados 1.400 questionários em ambas as universidades a fim de identificar, entre outros aspectos, atributos desejáveis de uma IES, fatores influenciadores para o abandono dela e características associadas a processos comunicacionais que representam diferenciais. O estudo apontou que, apesar de as dificuldades financeiras serem um componente que interfere na atração e na fidelização dos alunos, outros também são determinantes, como o nível de excelência de ensino, o corpo docente e a qualidade do relacionamento que as instituições de ensino estabelecem com tais públicos. Tal qualidade passa, obrigatoriamente, por atributos como liberdade para expressar ideias, espaço de diálogo e transparência na comunicação, elementos desafiadores ao modelo vigente e estimuladores a um novo fazer educacional.

Legacy and Fate of Mercury and Methylmercury in the Florida Everglades

Mass inventories of total Hg (THg) and methylmercury (MeHg) and mass budgets of Hg newly deposited during the 2005 dry and wet seasons were constructed for the Everglades. As a sink for Hg, the Everglades has accumulated 914, 1138, 4931, and 7602 kg of legacy THg in its 4 management units, namely Water Conservation Area (WCA) 1, 2, 3, and the Everglades National Park (ENP), respectively, with most Hg being stored in soil. The current annual Hg inputs account only for 1−2% of the legacy Hg. Mercury transport across management units during a season amounts to 1% or less of Hg storage, except for WCA 2 where inflow inputs can contribute 4% of total MeHg storage. Mass budget suggests distinct spatiality for cycling of seasonally deposited Hg, with significantly lower THg fluxes entering water and floc in ENP than in the WCAs. Floc in WCAs can retain a considerable fraction (around 16%) of MeHg produced from the newly deposited Hg during the wet season. This work is important for evaluating the magnitude of legacy Hg contamination and for predicting the fate of new Hg in the Everglades, and provides a methodological example for large-scale studies on Hg cycling in wetlands.

Spatial and Temporal Changes in Groundwater Salinity in South Florida

A combination of statistical and interpolation methods and Geographic Information System (GIS) spatial analysis was used to evaluate the spatial and temporal changes in groundwater Cl− concentrations in Collier and Lee Counties (southwestern Florida), and Miami-Dade and Broward Counties (southeastern Florida), since 1985. In southwestern Florida, the average Cl− concentrations in the shallow wells (0–43 m) in Collier and Lee Counties increased from 132 mg L−1 in 1985 to 230 mg L−1 in 2000. The average Cl− concentrations in the deep wells (>43 m) of southwestern Florida increased from 392 mg L−1 in 1985 to 447 mg L−1 in 2000. Results also indicated a positive correlation between the mean sea level and Cl− concentrations and between the mean sea level and groundwater levels for the shallow wells. Concentrations in the Biscayne Aquifer (southeastern Florida) were significantly higher than those of southwestern Florida. The average Cl− concentrations increased from 159 mg L−1 in 1985 to 470 mg L−1 in 2010 for the shallow wells (<33 m) and from 1360 mg L−1 in 1985 to 2050 mg L−1 in 2010 for the deep wells (>33 m). In the Biscayne Aquifer, wells showed a positive or negative correlation between mean sea level and Cl− concentrations according to their location with respect to the saltwater intrusion line. Wells located inland behind canal control structures and west of the saltwater intrusion line showed negative correlation values, whereas wells located east of the saltwater intrusion line showed positive values. Overall, the results indicated that since 1985, there was a potential decline in the available freshwater resources estimated at about 12–17% of the available drinking-quality groundwater of the southeastern study area located in the Biscayne Aquifer.

(Table 2) Collection sites and number of specimens of Usnea subgenus Neuropogon species

Usnea species of the Neuropogon group are amongst the most widespread and abundant macrolichens in Antarctic regions. Four principal species, U. antarctica, U. aurantiaco-atra, U. sphacelata and U. subantarctica, have been described on morphological grounds. However, identification to species level is often difficult and atypical morphologies frequently arise. Over 400 specimens were collected on the Antarctic Peninsula and Falkland Islands. Both morphological and molecular characters (ITS and RPB1) were used to compare samples to clarify taxonomic relationships. Morphological characteristics used included presence of apothecia, apothecial rays, soredia, papillae, fibrils, pigmentation and the diameter of the central axis as a proportion of branch diameter. Results revealed a very close relationship between U. antarctica and U. aurantiaco-atra, suggesting that they might constitute a species pair or be conspecific. Usnea sphacelata was comprised of at least two genetically distinct groups with no clear differences in morphology. One group included the first reported fertile specimen of this species. Usnea subantarctica was phylogenetically distinct from the other main Antarctic Usnea species, but clustered with U. trachycarpa. Genetic variation was evident within all species although there was no clear correlation between geographic origin and genetic relatedness. Phylogenetic analyses indicated that species circumscription in the Neuropogon group needs revision, with the principal species being non-monophyletic. None of the morphological characters, or groups of characters, used in this study proved to be completely unambiguous markers for a single species. However, axis thickness was supported as being informative for the identification of monophyletic lineages within the group.