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Sample records for rodentia cricetidae k1

  1. [Molecular characterization of Sigmodon hirsutus (Rodentia: Cricetidae) populations in Venezuela].

    PubMed

    Lessmann, Janeth; Arrivillaga, Jazzmín; Aguilera, Marisol

    2011-06-01

    Recent phylogenetic studies based on cytochrome b gene sequence, have determined that the species historically known as Sigmodon hispidus (Rodentia) from South America comprises a species S. hirsutus of paraphyletic origin. The aim of this study was to test the hypothesis that populations from Venezuela, represent the sensu strict form, ancestral haplotypes, and monophyletic subspecieS. For this, 12 individual sequences from three localities of different biogeographic regions in Venezuela were evaluated and sequenced based on cyto b. Additionally, the sequences were used to develop a cladistic analysis and genetic distance calculations, and to compare this information with two individual sequences of Sigmodon specimens available in Genbank. Phylogenetic analyses show that the three populations of S. hirsutus of Venezuela form an ancestral and monophyletic subclade supported by high bootstrap values and significant genetic distance between subclade within the S. hirsutus. Besides, the existence of two lineages suggests two subspecies, S. hirsutus hirsutus from Venezuela, and S. hirsutus mexicanus from Mexico-Central America, but, both species need formal description.

  2. New species and records of the mite genus Prolistrophorus (Acariformes: Listrophoridae) from rodents of the subfamily Sigmodontinae (Rodentia: Cricetidae).

    PubMed

    Bochkov, Andre V; Lareschi, Marcela; Barreto, Mauricio

    2014-10-01

    Six fur-mite species of the genus Prolistrophorus Fain, 1970 (Acariformes: Listrophoridae) were recorded from Central and South American rodents of the subfamily Sigmodontinae (Rodentia: Cricetidae). Among them, Prolistrophorus (Aprolistrophorus) parabidentatus sp. nov. from Akodon azarae from Argentina and Prolistrophorus (Aprolistrophorus) tylomys sp. nov. from Tylomys nudicaudus from Guatemala are described as new for science. New hosts are recorded for the following species: Prolistrophorus (Prolistrophorus) grassii (Radford, 1954) from Zygodontomys brevicauda from Colombia, P. (P.) frontalis (Hirst, 1921) from Oligoryzomys sp. from Argentina, P. (P.) argentinus (Hirst, 1921) from Melanomys caliginosus, Akodon affinis from Colombia and Scapteromys aquaticus from Argentina, Prolistrophorus (Beprolistrophorus) hirstianus Fain, 1973 from Scapteromys aquaticus from Argentina.

  3. The tubular compartment and the spermatogenic dynamics of the wild rodent Oxymycterus nasutus (Rodentia: Cricetidae).

    PubMed

    Morais, Ana Carolina Torre; Balarini, Maytê Koch; Lopes, Elizabeth Oliveira; Menezes, Tatiana Prata; Quintela, Fernando Marques; Morais, Danielle Barbosa; Gomes, Marcos de Lucca M; Matta, Sérgio Luis P da

    2014-10-01

    Despite the order Rodentia present worldwide distribution and large number of species in the Brazilian fauna, detailed studies on testicular morphophysiology are still scarce. Therefore, this study aimed to analyze the dynamics of the spermatogenic process of Oxymycterus nasutus using morphometrical and stereological tools. Testicles from ten sexually mature males were used, showing a gonadosomatic index of 0.89%. The testicular parenchyma showed one of the highest tubulesomatic indexes reported among wild rodents - 0.82% - from which 65.12% was allocated into seminiferous epithelium. The average tubular diameter was 249.89 μm, whereas the epithelium height was 62.47 μm and the total length was 18.62 m per gram of testis. Eight different stages of the seminiferous epithelium cycle were described. Stage 1 was used for counting the germ cell population as well as the Sertoli cells. On average, 3.47 type-A spermatogonia, 24.39 primary spermatocytes in preleptotene/leptotene, 24.13 primary spermatocytes in pachytene, 68.38 round spermatids and 7.33 Sertoli cells were found per tubular cross section. There were 91.02 × 10(6) Sertoli cells per gram of testis and each cell was able to support 9.33 spermatids and 16.43 germ cells. The coefficient of spermatogonial mitosis was 7.02, while 2.83 spermatids were produced for each primary spermatocyte in pachytene. The overall efficiency of spermatogenesis was 19.70 cells, whereas the sperm reserve per gram of testis totalized 849.63 × 10(6) spermatids. Therefore, the presented data showed that O. nasutus shows a high energetic investment in reproduction, corroborating the findings for other species of the Cricetidae family.

  4. Histological and serological evidence of experimental paracoccidioidomycosis in Calomys callosus (Rodentia: Cricetidae)

    PubMed Central

    Berbert, Alceu LCV; Faria, Gabriele G; Gennari-Cardoso, Margareth L; Silva, Maria MMD; Mineo, José R; Loyola, Adriano M

    2007-01-01

    The responses of animal experimental models related to the infectivity, virulence and pathogenicity of Paracoccidioides brasiliensis is constantly used to develop new perspectives of investigation. The rodent Calomys callosus, Rengger 1830 (Rodentia: Cricetidae) is an indigenous inhabitant of the savannah environment found in the central regions of Brazil. The aim of the present work was to evaluate the histopathological and serological features of C. callosus after inoculation with the Pb18 strain of P. brasiliensis. Furthermore, A/Sn and B10.A mice strains were also tested to compare the results obtained in C. callosus to these well-established experimental models of resistance and susceptibility respectively. In every instance, survival analysis was performed, and histopathological study of the lungs, liver and spleen was employed to investigate tissue involvement, degree of inflammation and fungal presence. Levels of antibodies to P. brasiliensis were measured by using an enzyme-linked immunosorbent assay after 4 weeks and at the advanced stage of infection. The mortality rate was proportional to inoculation dose in all groups, but overall it was much superior in C. callosus than in the B10.A-susceptible mice. Macroscopical and microscopical pathological alterations were also more extensive and remarkable for C. callosus, once again proportional to inoculation dose, but more noticeable differences among the studied groups were found with 0.6 × 105 inoculum. In addition, the serological profile of C. callosus was similar to that found for B10.A-susceptible mice. Infection of C. callosus with 0.6 × 108 Pb18 inoculum resulted in more serious illness, and it decreased in severity in proportion to the inoculum dose. This difference was more pronounced in C. callosus, and the clinical, serological and pathological findings in this animal were more intense and precocious compared with the B10.A-susceptible mice. The present results suggest that C. callosus is a

  5. Histological and serological evidence of experimental paracoccidioidomycosis in Calomys callosus (Rodentia: Cricetidae).

    PubMed

    Berbert, Alceu L C V; Faria, Gabriele G; Gennari-Cardoso, Margareth L; Silva, Maria M M D; Mineo, José R; Loyola, Adriano M

    2007-02-01

    The responses of animal experimental models related to the infectivity, virulence and pathogenicity of Paracoccidioides brasiliensis is constantly used to develop new perspectives of investigation. The rodent Calomys callosus, Rengger 1830 (Rodentia: Cricetidae) is an indigenous inhabitant of the savannah environment found in the central regions of Brazil. The aim of the present work was to evaluate the histopathological and serological features of C. callosus after inoculation with the Pb18 strain of P. brasiliensis. Furthermore, A/Sn and B10.A mice strains were also tested to compare the results obtained in C. callosus to these well-established experimental models of resistance and susceptibility respectively. In every instance, survival analysis was performed, and histopathological study of the lungs, liver and spleen was employed to investigate tissue involvement, degree of inflammation and fungal presence. Levels of antibodies to P. brasiliensis were measured by using an enzyme-linked immunosorbent assay after 4 weeks and at the advanced stage of infection. The mortality rate was proportional to inoculation dose in all groups, but overall it was much superior in C. callosus than in the B10.A-susceptible mice. Macroscopical and microscopical pathological alterations were also more extensive and remarkable for C. callosus, once again proportional to inoculation dose, but more noticeable differences among the studied groups were found with 0.6x10(5) inoculum. In addition, the serological profile of C. callosus was similar to that found for B10.A-susceptible mice. Infection of C. callosus with 0.6x10(8) Pb18 inoculum resulted in more serious illness, and it decreased in severity in proportion to the inoculum dose. This difference was more pronounced in C. callosus, and the clinical, serological and pathological findings in this animal were more intense and precocious compared with the B10.A-susceptible mice. The present results suggest that C. callosus is a

  6. DNA barcoding of Murinae (Rodentia: Muridae) and Arvicolinae (Rodentia: Cricetidae) distributed in China.

    PubMed

    Li, Jing; Zheng, Xin; Cai, Yansen; Zhang, Xiuyue; Yang, Min; Yue, Bisong; Li, Jing

    2015-01-01

    Identification of rodents is very difficult mainly due to high similarities in morphology and controversial taxonomy. In this study, mitochondrial cytochrome oxidase subunit I (COI) was used as DNA barcode to identify the Murinae and Arvicolinae species distributed in China and to facilitate the systematics studies of Rodentia. In total, 242 sequences (31 species, 11 genera) from Murinae and 130 sequences (23 species, 6 genera) from Arvicolinae were investigated, of which 90 individuals were novel. Genetic distance, threshold method, tree-based method, online BLAST and BLOG were employed to analyse the data sets. There was no obvious barcode gap. The average K2P distance within species and genera was 2.10% and 12.61% in Murinae, and 2.86% and 11.80% in Arvicolinae, respectively. The optimal threshold was 5.62% for Murinae and 3.34% for Arvicolinae. All phylogenetic trees exhibited similar topology and could distinguish 90.32% of surveyed species in Murinae and 82.60% in Arvicolinae with high support values. BLAST analyses yielded similar results with identification success rates of 92.15% and 93.85% for Murinae and Arvicolinae, respectively. BLOG successfully authenticated 100% of detected species except Leopoldamys edwardsi based on the latest taxonomic revision. Our results support the species status of recently recognized Micromys erythrotis, Eothenomys tarquinius and E. hintoni and confirm the important roles of comprehensive taxonomy and accurate morphological identification in DNA barcoding studies. We believe that, when proper analytic methods are applied or combined, DNA barcoding could serve as an accurate and effective species identification approach for Murinae and Arvicolinae based on a proper taxonomic framework.

  7. Evolutionary story of a satellite DNA from Phodopus sungorus (Rodentia, Cricetidae).

    PubMed

    Paço, Ana; Adega, Filomena; Meštrović, Nevenka; Plohl, Miroslav; Chaves, Raquel

    2014-10-21

    With the goal to contribute for the understanding of satellite DNA evolution and its genomic involvement, in this work it was isolated and characterized the first satellite DNA (PSUcentSat) from Phodopus sungorus (Cricetidae). Physical mapping of this sequence in P. sungorus showed large PSUcentSat arrays located at the heterochromatic (peri)centromeric region of five autosomal pairs and Y-chromosome. The presence of orthologous PSUcentSat sequences in the genomes of other Cricetidae and Muridae rodents was also verified, presenting however, an interspersed chromosomal distribution. This distribution pattern suggests a PSUcentSat-scattered location in an ancestor of Muridae/Cricetidae families, that assumed afterwards, in the descendant genome of P. sungorus a restricted localization to few chromosomes in the (peri)centromeric region. We believe that after the divergence of the studied species, PSUcentSat was most probably highly amplified in the (peri)centromeric region of some chromosome pairs of this hamster by recombinational mechanisms. The bouquet chromosome configuration (prophase I) possibly displays an important role in this selective amplification, providing physical proximity of centromeric regions between chromosomes with similar size and/or morphology. This seems particularly evident for the acrocentric chromosomes of P. sungorus (including the Y-chromosome), all presenting large PSUcentSat arrays at the (peri)centromeric region. The conservation of this sequence in the studied genomes and its (peri)centromeric amplification in P. sungorus strongly suggests functional significance, possibly displaying this satellite family different functions in the different genomes. The verification of PSUcentSat transcriptional activity in normal proliferative cells suggests that its transcription is not stage-limited, as described for some other satellites.

  8. Chromosomal evolution of Arvicolinae (Cricetidae, Rodentia). III. Karyotype relationships of ten Microtus species.

    PubMed

    Lemskaya, Natalia A; Romanenko, Svetlana A; Golenishchev, Feodor N; Rubtsova, Nadezhda V; Sablina, Olga V; Serdukova, Natalya A; O'Brien, Patricia C M; Fu, Beiyuan; Yiğit, Nuri; Ferguson-Smith, Malcolm A; Yang, Fengtang; Graphodatsky, Alexander S

    2010-06-01

    The genus Microtus consists of 65 extant species, making it one of the rodentia genera with the highest number of species. The extreme karyotype diversification in Microtus has made them an ideal species group for comparative cytogenetics and cytotaxonomy. Conventional comparative cytogenetic studies in Microtus have been based mainly on chromosomal banding patterns; the number of Microtus species examined by molecular cytogenetics-cross-species chromosome painting-is limited. In this study, we used whole chromosome painting probes of the field vole Microtus agrestis to detect regions of homology in the karyotypes of eight Microtus species. For almost all investigated species, species-specific associations of conserved chromosomal segments were revealed. Analysis of data obtained here and previously published data allowed us to propose that the ancestral Microtus species had a 2n = 54 karyotype, including two associations of field vole chromosomal segments (MAG 1/17 and 2/8). Further mapping of the chromosome rearrangements onto a molecular phylogenetic tree allows the reconstruction of a karyotype evolution pathway in the Microtus genus.

  9. Rediscovery and New Morphological Data on Two Hassalstrongylus (Nematoda: Heligmonellidae) Coparasitic in the Marsh Rat Holochilus chacarius (Rodentia: Cricetidae) from Argentina.

    PubMed

    Digiani, María Celina; Notarnicola, Juliana; Navone, Graciela T

    2015-10-01

    Two species of Hassalstrongylus Durette-Desset, 1971, coparasitic in Holochilus chacarius Thomas (Rodentia, Cricetidae) and not recorded since their original description in 1937, were newly found in their type host and locality. Hassalstrongylus mazzai (Freitas, Lent and Almeida, 1937) and Hassalstrongylus argentinus (Freitas, Lent and Almeida, 1937) were obtained from Ho. chacarius from 2 different populations: one from Salta Province (northwest Argentina) and another from Chaco Province (northeast Argentina). The species described as Heligmonoides mazzai Freitas, Lent and Almeida, 1937 had been transferred to Hassalstrongylus even though its synlophe had never been studied. We provide the first descriptions and illustrations of the synlophe of males and females of Hassalstrongylus mazzai and the female of H. argentinus and account for morphological and metrical variability. We confirm, through the study of the synlophe, the placement of Hassalstrongylus mazzai in the genus Hassalstrongylus and designate neotypes for the species because the type material deposited by the authors could not be found. Females of both species were morphologically very similar, and a principal components analysis (PCA) performed on some morphometrical characters showed that the body length, uterus length, and an unexpected character as the number of eggs were useful characters in the discrimination of both species.

  10. A new species of Syphacia (Seuratoxyuris) (Nematoda: Oxyuridae) from Sooretamys angouya Fischer, 1814 (Rodentia: Cricetidae) in Argentina.

    PubMed

    Robles, María del Rosario; Panisse, Guillermo; Navone, Graciela Teresa

    2014-11-01

    Syphacia (Seuratoxyuris) hugoti n. sp. (Nematoda: Oxyuridae) is described from the cecum of Sooretamys angouya (Cricetidae: Sigmodontinae: Oryzomyini) captured in Formosa Province, Argentina. The diagnosis of the subgenus is emended, and the new species is separated from eight congeners by the distribution of submedian papillae and amphids, shape of the cephalic plate, presence of deirids, absence of cervical and lateral alae, length of the spicule, structure of the accessory hook of the gubernaculum and distance of excretory pore and vulva from the anterior extremity. The analysis suggests that S. (Se.) oryzomyos should be removed from Seuratoxyuris and redesignated as S. (Syphacia) oryzomyos n. comb. To date, of the species of Syphacia found in South and North American, 7 parasitize Oryzomyini rodents, of which two are distributed in Argentina. The present study constitutes the first record of the subgenus Seuratoxyuris from Argentina and the third record of a Syphacia species from rodents of the tribe Oryzomyini.

  11. Cytogenetics and karyosystematics of Oryzomys albigularis (Rodentia, Cricetidae) from Venezuela.

    PubMed

    Aguilera, M; Pérez-Zapata, A; Martino, A

    1995-01-01

    Several authors have proposed that Oryzomys albigularis constitutes a supraspecific complex and that chromosomal pericentromeric inversions have played a fundamental role in the diversifying process. With the purpose of clarifying the unclear taxonomic situation of the nominal forms of O. albigularis living in Venezuela, a cytogenetic study was carried out on individuals from five different localities along the Andean range and the Cordillera de la Costa. All of the individuals examined showed a diploid number (2n) of 66 chromosomes, but there were differences in the number of autosomal arms (FN = 90, 92, and 104) and in the morphology of the X chromosome (metacentric or acrocentric). The C-banding pattern was similar in all populations, autosomal heterochromatin was restricted to the centromere, and the Y chromosome was the only one that had completely heterochromatic arms. G-banding was useful in making arm-to-arm comparison between the FN = 90 and FN = 104 karyomorphs; 23 shared pairs were found, 7 pairs differed due to pericentric inversions, and 3 pairs had no correspondence. We postulate that these karyomorphs probably correspond to allospecies, and that the specific denominations must correspond to two previously recognized populations of sigmodontine rodents: O. caracolus Thomas 1914 (2n = 66, FN = 90), for the populations from the Cordillera de la Costa, and O. meridensis Thomas 1894 (2n = 66, FN = 104), for the populations distributed between the middle and extreme north of the Andean range. The specific denomination for the populations of animals from the southern portion of the Andean range (Oryzomys sp., 2n = 66, FN = 92) is still to be determined.

  12. Chromosomal evolution in Rodentia.

    PubMed

    Romanenko, S A; Perelman, P L; Trifonov, V A; Graphodatsky, A S

    2012-01-01

    Rodentia is the most species-rich mammalian order and includes several important laboratory model species. The amount of new information on karyotypic and phylogenetic relations within and among rodent taxa is rapidly increasing, but a synthesis of these data is currently lacking. Here, we have integrated information drawn from conventional banding studies, recent comparative painting investigations and molecular phylogenetic reconstructions of different rodent taxa. This permitted a revision of several ancestral karyotypic reconstructions, and a more accurate depiction of rodent chromosomal evolution.

  13. Chromosomal variation in Argentine populations of Akodon montensis Thomas, 1913 (Rodentia, Cricetidae, Sigmodontinae)

    PubMed Central

    Malleret, Matías Maximiliano; Labaroni, Carolina Alicia; García, Gabriela Verónica; Ferro, Juan Martín; Martí, Dardo Andrea; Lanzone, Cecilia

    2016-01-01

    Abstract The genus Akodon Meyen, 1833 is one of the most species-rich among sigmodontine rodents and has great chromosome variability. Akodon montensis has a relatively broad distribution in South America, and Argentine populations are located in the southernmost region of its range. Brazilian populations have important chromosomal variability, but cytogenetic data from Argentina are scarce. We performed a chromosome characterization of natural populations of Akodon montensis using conventional staining, C-banding, Ag-NORs and base-specific fluorochromes. A total of 31 specimens from five localities of Misiones Province, in Argentina, were analyzed. The 2n=24 chromosomes was the most frequently observed karyotype. However, five individuals presented 25 chromosomes due to a supernumerary B-chromosome; and one individual had 2n=26 due to one B plus a trisomy for chromosome 11. Additionally, two XY females and two variants of the X chromosomes were found. C-positive centromeric bands occurred in all chromosomes; additional C-bands were observed in some autosomes, the X, Y and B chromosomes. Ag-NORs were observed in five autosomes, and the B chromosome was frequently marked. Fluorochrome banding was similar among karyotypes of the analyzed populations. Comparisons of cytogenetic data among populations of Argentina and Brazil showed the presence of high intraspecific variability in Akodon montensis and some differences among regions. PMID:27186343

  14. [Supraspecies relationships in the subfamily (Rodentia, Cricetidae, Arvicolinae): unexpexted result of nuclear genes analysis].

    PubMed

    Abramson, N I; Lebedev, V S; Tesakov, A S; Bannikova, A A

    2009-01-01

    Phylogenetic analysis of supraspecies relationships in one of the most young and species rich group of myomorph rodents - subfamily Arvicolinae was carried out on the base of two nuclear genes. Results have shown that mole-voles - Ellobiusini, steppe voles - (Lagurini) and grey voles (Arvicolini) are sister groups. This divergence is the most late, third wave of radiation within the family. The sister clade to this group is the tribe of red-back voles - Myodini (=Clethrionomini) - "second radiation". The order of divergence for earliest radiation remains still unresolved (Ondatrini, Prometheomyini, Dicrostonychini, Lemmini). New data on the close relationships of mole voles, grey voles and steppe voles are unexpected one and contradict to the conventional views. The latter ideas on the significant ancientry and separation of Ellobiusini from all other voles is based on extreme simplicity of their rooted molars and very peculiar structure of the skull and postcranial skeleton. However, many of these characters most likely indicate on significant degree of adaptation to the subterranean life and have no phylogenetic signal.

  15. No differences in the Sry gene between males and XY females in Akodon (Rodentia, Cricetidae).

    PubMed

    Sánchez, A; Marchal, J A; Romero-Fernández, I; Pinna-Senn, E; Ortiz, M I; Bella, José L; Lisanti, J A

    2010-01-01

    Several species of the South American genus Akodon present fully fertile XY females besides XX ones. To analyze the possibility of a Sry mutation as the cause of sex reversal in A. azarae and A. boliviensis, we determined the sequence of the Sry gene in 2 males and 3 XY females from each of these species. The Sry gene sequence was also studied in A. dolores, a species that does not have XY females. In inter-specific comparisons, the percentage identities with respect to the region analyzed varied between 96.8% and 97.9%. An ORF of 543 nucleotides was identified, and the predicted Sry proteins comprised 180 amino acids, with an HMG domain of 83 amino acids. Our results indicate that female sex reversal in A. azarae and A. boliviensis cannot be explained by sequence differences in the Sry region analyzed here, which includes the complete ORF and, together with previous results concerning the inheritance of the XY condition, show that Sry mutation is not the basis of sex reversal.

  16. Karyotypic variation in the Andean rodent Phyllotisxanthopygus (Waterhouse, 1837) (Rodentia, Cricetidae, Sigmodontinae).

    PubMed

    Labaroni, Carolina Alicia; Malleret, Matías Maximiliano; Novillo, Agustina; Ojeda, Agustina; Rodriguez, Daniela; Cuello, Pablo; Ojeda, Ricardo; Dardo Martí; Lanzone, Cecilia

    2014-01-01

    Phyllotisxanthopygus (Waterhouse, 1837) is an Andean rodent endemic to South America. Despite its wide geographical distribution in Argentina, few individuals have been studied on the cytogenetic level and only through conventional staining. In this work, chromosome characterization of Argentine samples of this species was performed using solid staining, C-banding and base-specific fluorochromes. Twenty two specimens were analyzed, collected in the provinces of Jujuy, Catamarca, and the north and south of Mendoza. All studied specimens showed 2n=38, having mostly the bi-armed autosomes, metacentric or submetacentric. Fundamental Number varied between 70 and 72. These changes were due to the presence of chromosome heteromorphisms in individuals from southern Mendoza and Jujuy. C-banding revealed pericentromeric blocks of constitutive heterochromatin in most chromosomes. Acrocentric chromosomes involved in heteromorphisms showed high variation in the amount of heterochromatin within and among populations. Additionally, banding with fluorochromes (DAPI and chromomycin A3) revealed homologous localization of AT and GC rich regions among chromosomes of the different populations analyzed. Comparisons among heteromorphic pairs suggested, however, that the variation might be the result of complex chromosome rearrangements, involving possibly amplifications and/or deletions of heterochromatic segments. These results are in accordance with molecular studies that indicate genetic variability within and among the populations of this taxon.

  17. First cytogenetic information for Drymoreomys albimaculatus (Rodentia, Cricetidae), a recently described genus from Brazilian Atlantic Forest

    PubMed Central

    Suárez-Villota, Elkin Y.; Di-Nizo, Camilla B.; Neves, Carolina L.; Silva, Maria José de Jesus

    2013-01-01

    Abstract The recently described taxon Drymoreomys albimaculatus is endemic to the Brazilian Atlantic Forest and its biology and genetics are still poorly known. Herein, we present, for the first time, the karyotype of the species using classical and molecular cytogenetics, which showed 2n=62, FN=62, and interstitial telomeric signals at the sex chromosomes. Nuclear and mitochondrial DNA sequences from the two karyotyped individuals verify the taxonomic identity as the recently described Drymoreomys albimaculatus and confirm the relationship of the species with other Oryzomyini. Additionally, external morphological information is provided. PMID:23794904

  18. Chromosomal variation in Argentine populations of Akodon montensis Thomas, 1913 (Rodentia, Cricetidae, Sigmodontinae).

    PubMed

    Malleret, Matías Maximiliano; Labaroni, Carolina Alicia; García, Gabriela Verónica; Ferro, Juan Martín; Martí, Dardo Andrea; Lanzone, Cecilia

    2016-01-01

    The genus Akodon Meyen, 1833 is one of the most species-rich among sigmodontine rodents and has great chromosome variability. Akodon montensis has a relatively broad distribution in South America, and Argentine populations are located in the southernmost region of its range. Brazilian populations have important chromosomal variability, but cytogenetic data from Argentina are scarce. We performed a chromosome characterization of natural populations of Akodon montensis using conventional staining, C-banding, Ag-NORs and base-specific fluorochromes. A total of 31 specimens from five localities of Misiones Province, in Argentina, were analyzed. The 2n=24 chromosomes was the most frequently observed karyotype. However, five individuals presented 25 chromosomes due to a supernumerary B-chromosome; and one individual had 2n=26 due to one B plus a trisomy for chromosome 11. Additionally, two XY females and two variants of the X chromosomes were found. C-positive centromeric bands occurred in all chromosomes; additional C-bands were observed in some autosomes, the X, Y and B chromosomes. Ag-NORs were observed in five autosomes, and the B chromosome was frequently marked. Fluorochrome banding was similar among karyotypes of the analyzed populations. Comparisons of cytogenetic data among populations of Argentina and Brazil showed the presence of high intraspecific variability in Akodon montensis and some differences among regions.

  19. Reservoir competence of Microtus pennsylvanicus (Rodentia: Cricetidae) for the Lyme disease spirochete, Borrelia burgdorferi

    USGS Publications Warehouse

    Markowski, D.; Ginsberg, H.S.; Hyland, K.E.; Hu, R.

    1998-01-01

    The reservoir competence of the meadow vole, Microtus pennsylvanicus Ord, for the Lyme disease spirochete, Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner was established on Patience Island, RI. Meadow voles were collected from 5 locations throughout Rhode Island. At 4 of the field sites, M. pennsylvanicus represented only 4.0% (n = 141) of the animals captured. However, on Patience Island, M. pennsylvanicus was the sole small mammal collected (n = 48). Of the larval Ixodes scapularis Say obtained from the meadow voles on Patience Island, 62% (n = 78) was infected with B. burgdorferi. Meadow voles from all 5 locations were successfully infected with B. burgdorferi in the laboratory and were capable of passing the infection to xenodiagnostic I. scapularis larvae for 9 wk. We concluded that M. pennsylvanicus was physiologically capable of maintaining B. burgdorferi infection. However, in locations where Peromyscus leucopus (Rafinesque) is abundant, the role of M. pennsylvanicus as a primary reservoir for B. burgdorferi was reduced.

  20. Nitric oxide production by Peromyscus yucatanicus (Rodentia) infected with Leishmania (Leishmania) mexicana

    PubMed Central

    Loría-Cervera, Elsy Nalleli; Sosa-Bibiano, Erika Ivett; Villanueva-Lizama, Liliana Estefanía; Van Wynsberghe, Nicole Raymonde; Canto-Lara, Silvia Beatriz; Batún-Cutz, José Luis; Andrade-Narváez, Fernando José

    2013-01-01

    Peromyscus yucatanicus (Rodentia: Cricetidae) is a primary reservoir of Leishmania (Leishmania) mexicana (Kinetoplastida: Trypanosomatidae). Nitric oxide (NO) generally plays a crucial role in the containment and elimination of Leishmania. The aim of this study was to determine the amount of NO produced by P. yucatanicus infected with L. (L.) mexicana. Subclinical and clinical infections were established in P. yucatanicus through inoculation with 1 x 102 and 2.5 x 106 promastigotes, respectively. Peritoneal macrophages were cultured alone or co-cultured with lymphocytes with or without soluble Leishmania antigen. The level of NO production was determined using the Griess reaction. The amount of NO produced was significantly higher (p ≤ 0.0001) in co-cultured macrophages and lymphocytes than in macrophages cultured alone. No differences in NO production were found between P. yucatanicus with subclinical L. (L.) mexicana infections and animals with clinical infections. These results support the hypothesis that the immunological mechanisms of NO production in P. yucatanicus are similar to those described in mouse models of leishmaniasis and, despite NO production, P. yucatanicus is unable to clear the parasite infection. PMID:23579796

  1. SSCA1-K1

    SciTech Connect

    Baker, Matthew

    2015-11-06

    SSCA1-K1 is a parallel implementation of kernel 1 of the SSCA1 benchmark suite released by the DARPA HPCS program. This kernel is able to run in parallel on a distributed shared memory system at extreme scales using OpenSHMEM.

  2. A new species of Argyromys (Rodentia, Mammalia) from the Oligocene of the Valley of Lakes (Mongolia): Its importance for palaeobiogeographical homogeneity across Mongolia, China and Kazakhstan

    PubMed Central

    Maridet, Olivier; Zhang, Zhaoqun; Daxner-Höck, Gudrun

    2017-01-01

    We describe a new species of Rodentia (Mammalia), Argyromys cicigei sp. nov. from Toglorhoi (fossil bed TGW-A/2a) in Mongolia and Ulantatal (fossil beds UTL 1 and UTL 7) in China. Its tooth morphology differs from the type species Argyromys aralensis from Akespe in Kazakhstan by smaller size and simpler structures. Argyromys has been assigned in different families of Muroidea, such as Tachyoryctoididae and Spalacidae. However, the presence of common characters indicates a closer relationship of Argyromys with the genera of Cricetidae s.l. (subfamilies Eucricetodontinae; Cricetopinae; Cricetodontinae and Gobicricetodontinae among others) from Asia than with the earliest representatives of Spalacidae or the endemic Tachyoryctoididae. Argyromys cicigei sp. nov. possesses a simple anterocone and anteroconid in the upper and lower first molars, respectively, which is characteristic for Cricetidae s.l. It has a flat occlusal surface in worn specimens; weakly-developed posterolophs; an oblique protolophule and metaloph on the upper molars and it lacks a labial anterolophid on the m1. These traits are also typical of the Oligocene genera Aralocricetodon and Plesiodipus, included in the subfamilies Cricetodontinae and Gobicricetodontinae respectively. The cladistic analysis performed here supports this hypothesis. The clade formed by Argyromys species is grouped with other cricetid taxa (s.l). Spalacids, however, form a different clade, as do the tachyoryctoids. Previous authors state that the Aral Formation (Kazakhstan) should be dated to the Oligocene instead of the Miocene, based on the presence of several taxa. The finds of Argyromys in both regions supports the statement that they are closer in age than previously thought. The occurrence of Argyromys in Kazakhstan, Mongolia and China evidences the biogeographic unity of the Central Asian bioprovince during the Oligocene. PMID:28328975

  3. A new species of Argyromys (Rodentia, Mammalia) from the Oligocene of the Valley of Lakes (Mongolia): Its importance for palaeobiogeographical homogeneity across Mongolia, China and Kazakhstan.

    PubMed

    López-Guerrero, Paloma; Maridet, Olivier; Zhang, Zhaoqun; Daxner-Höck, Gudrun

    2017-01-01

    We describe a new species of Rodentia (Mammalia), Argyromys cicigei sp. nov. from Toglorhoi (fossil bed TGW-A/2a) in Mongolia and Ulantatal (fossil beds UTL 1 and UTL 7) in China. Its tooth morphology differs from the type species Argyromys aralensis from Akespe in Kazakhstan by smaller size and simpler structures. Argyromys has been assigned in different families of Muroidea, such as Tachyoryctoididae and Spalacidae. However, the presence of common characters indicates a closer relationship of Argyromys with the genera of Cricetidae s.l. (subfamilies Eucricetodontinae; Cricetopinae; Cricetodontinae and Gobicricetodontinae among others) from Asia than with the earliest representatives of Spalacidae or the endemic Tachyoryctoididae. Argyromys cicigei sp. nov. possesses a simple anterocone and anteroconid in the upper and lower first molars, respectively, which is characteristic for Cricetidae s.l. It has a flat occlusal surface in worn specimens; weakly-developed posterolophs; an oblique protolophule and metaloph on the upper molars and it lacks a labial anterolophid on the m1. These traits are also typical of the Oligocene genera Aralocricetodon and Plesiodipus, included in the subfamilies Cricetodontinae and Gobicricetodontinae respectively. The cladistic analysis performed here supports this hypothesis. The clade formed by Argyromys species is grouped with other cricetid taxa (s.l). Spalacids, however, form a different clade, as do the tachyoryctoids. Previous authors state that the Aral Formation (Kazakhstan) should be dated to the Oligocene instead of the Miocene, based on the presence of several taxa. The finds of Argyromys in both regions supports the statement that they are closer in age than previously thought. The occurrence of Argyromys in Kazakhstan, Mongolia and China evidences the biogeographic unity of the Central Asian bioprovince during the Oligocene.

  4. Characterization of the satellite DNA Msat-160 from species of Terricola (Microtus) and Arvicola (Rodentia, Arvicolinae).

    PubMed

    Acosta, Manuel J; Marchal, Juan A; Fernández-Espartero, Cecilia; Romero-Fernández, Ismael; Rovatsos, Michail T; Giagia-Athanasopoulou, Eva B; Gornung, Ekaterina; Castiglia, Riccardo; Sánchez, Antonio

    2010-10-01

    In the subfamily Arvicolinae (Cricetidae, Rodentia) the satellite DNA Msat-160 has been so far described in only some species from the genus Microtus and in one species from another genus, Chionomys nivalis. Here we cloned and characterized this satellite in two new arvicoline species, Microtus (Terricola) savii and Arvicola amphibius (terrestris). We have also demonstrated, by PCR and FISH, its existence in the genomes of several other species from both genera. These results suggest that Msat-160 already occurred in the common ancestor of the four genera/subgenera of Arvicolinae (Microtus, Chionomys, Arvicola, and Terricola). In Arvicola and Terricola, Msat-160 showed the basic monomer length of 160 bp, although a higher-order repeat (HORs) of 640 bp could have been probably replacing the original monomeric unit in A. a. terrestris. Msat-160 was localized by FISH mostly on the pericentromeric regions of the chromosomes, but the signal intensity and the number of carrier chromosomes varied extremely even between closely related species, resulting in a species-specific pattern of chromosomal distribution of this satellite. Such a variable pattern most likely is a consequence of a rapid amplification and contraction of particular repeats in the pericentromeric regions of chromosomes. In addition, we proposed that the rapid variation of pericentromeric repeats is strictly related to the prolific species radiation and diversification of karyotypes that characterize Arvicolinae lineage. Finally, we performed phylogenetic analysis in this group of related species based on Msat-160 that results to be in agreement with previously reported phylogenies, derived from other molecular markers.

  5. Ultrastructure of the spermatozoon of the trematode Notocotylus noyeri (Digenea: Notocotylidae), a parasite of Microtus arvalis (Rodentia: Cricetidae).

    PubMed

    Ibnou Ndiaye, Papa; Torres, Jordi; Eira, Catarina; Shimalov, Vladimir V; Miquel, Jordi

    2015-01-01

    In the present paper, we describe the ultrastructure of the spermatozoon of the notocotylid Notocotylus noyeri (Joyeux, 1922) by means of transmission electron microscopy. The mature spermatozoon of N. noyeri exhibits the general pattern described in the majority of digeneans: two axonemes of the 9 + "1" pattern of the Trepaxonemata, nucleus, mitochondria, parallel cortical microtubules, spine-like bodies and ornamentation of the plasma membrane. The glycogenic nature of the electron-dense granules was evidenced applying the test of Thiéry. The ultrastructural features of the spermatozoon of N. noyeri present some differences in relation to those of the Pronocephalidea described until now, but confirm a general pattern for the Notocotylidae, namely a spermatozoon with two mitochondria and an anterior region with ornamentation of the plasma membrane associated with spine-like bodies. The posterior extremity of the spermatozoon exhibits only some microtubules after the disorganisation of the second axoneme. The present study confirms that some ultrastructural characters of the sperm cell such as the presence or absence of lateral expansions, the number of mitochondria and the morphology of both anterior and posterior spermatozoon extremities are useful for phylogenetic purposes within the Pronocephaloidea. Thus, unlike notocotylids, pronocephalids exhibit external ornamentation and a lateral expansion in the anterior spermatozoon region. Moreover, notocotylid spermatozoa present two mitochondria, whereas pronocephalid spermatozoa exhibit a single mitochondrion. Finally, pronocephalids are characterised by a type 2 posterior spermatozoon extremity, whereas notocotylids exhibit a type 3 posterior spermatozoon extremity.

  6. Analysis of meiotic chromosome structure and behavior in Robertsonian heterozygotes of Ellobius tancrei (Rodentia, Cricetidae): a case of monobrachial homology

    PubMed Central

    Matveevsky, Sergey; Bakloushinskaya, Irina; Tambovtseva, Valentina; Romanenko, Svetlana; Kolomiets, Oxana

    2015-01-01

    Abstract Synaptonemal complex (SC) chains were revealed in semisterile intraspecific F1 hybrids of Ellobius tancrei Blasius, 1884 (2n = 49, NF=56 and 2n=50, NF=56), heterozygous for Robertsonian (Rb) translocations. Chains were formed by Rb submetacentrics with monobrachial homology. Chromosome synapsis in spermatocytes of these hybrids was disturbed, apparently because of the problematic release of the chromosomes from the SC chains. These hybrids suffer from low fertility, and our data support the opinion that this is because a formation of Rb metacentrics with monobrachial homology within different races of the same species might be an initial event for the divergence of chromosomal forms. PMID:26752380

  7. Descriptive morphometry and stereology of the tubular compartment in the wild rodent Hylaeamys megacephalus (Rodentia: Cricetidae) from Central Brazil.

    PubMed

    de Melo, Fabiana Cristina Silveira Alves; de Sousa, Tatiane Pires; Costa, Kyvia Lugate C; da Matta, Sérgio Luis P; de Melo, Fabiano Rodrigues; Santa-Rita, Ricardo de Mattos

    2013-04-01

    Information on reproductive characteristics of wild rodents is scarce in the literature. This study aimed to assess the testis morphometry and stereology of Hylaeamys megacephalus. We used five animals in the study, captured in forest fragments in southwestern Goias State, between April and August 2009. The testes were fixed in Karnovsky solution, dehydrated, and embedded in methacrylate. Two-micrometer-thick sections from each sample were stained with toluidine blue/sodium borate 1%. Images of the testicular parenchyma were obtained from photomicroscope and morphometric and stereological analyses were carried out using the Image Pro-Plus software. The average body weight observed in the specimens of H. megacephalus in the study was 47.84 g, of which, 0.40% is allocated to the gonads (GSI) and 0.36% to the seminiferous tubules (TSI). These parameters suggest promiscuous reproductive behavior, of the polyandrous type, favoring males with higher sperm production and consequently, larger testes. The volume density of the seminiferous tubules was 94.46%, which represented a volume of 0.18 mL. The volume density and volume of the interstitium were 5.54% and 0.011 mL, respectively. The diameter of the seminiferous tubules was 206.5 μm and the height of seminiferous epithelium was 71.27 μm. H. megacephalus presents 5.06 m of seminiferous tubules and an average of 27.96 m of seminiferous tubules per gram of testis. The mitotic and meiotic indexes showed losses of 85 and 42%, respectively and an overall loss of 90% over the full spermatogenic process. The number of Sertoli cells per testis and per gram of testis was 7.8×10(6) and 95.28×10(6), respectively. Most of the morphometric parameters evaluated in H. megacephalus in this study are within the range of values described for most mammals.

  8. Shippingport, Kentucky, is the type locality for the white-footed mouse, Peromyscus leucopus (Rafinesque, 1818) (Mammalia: Rodentia: Cricetidae)

    USGS Publications Warehouse

    Woodman, Neal

    2015-01-01

    The white-footed mouse, Musculus leucopus Rafinesque, 1818 (= Peromyscus leucopus), is a common small mammal that is widespread in the eastern and central United States. Its abundance in many habitats renders it ecologically important, and its status as a reservoir for hantavirus and Lyme disease gives the species medical and economic significance. The recognition of two cytotypes and up to 17 morphological subspecies of P. leucopus indicates considerable variation in the species, and to understand this variation, it is important that the nominate subspecies be adequately defined so as to act as a standard for comparison. Relevant to this standard for the white-footed mouse is its type locality, which has generally been accepted to be either the vague "pine barrens of Kentucky" or the mouth of the Ohio River. Newly assembled information regarding the life and travels of Constantine S. Rafinesque, the North American naturalist who described P. leucopus, establishes that Rafinesque observed this species in July 1818 while visiting Shippingport, Kentucky, which is now within the city limits of Louisville, Jefferson Co., Kentucky. Shippingport is therefore the actual type locality for this species.

  9. Encephalitozoon infections in Rodentia and Soricomorpha in Japan.

    PubMed

    Tsukada, Ryusuke; Tsuchiyama, Atsuko; Sasaki, Mizuki; Park, Chun-Ho; Fujii, Yoshito; Takesue, Masataka; Hatai, Hitoshi; Kudo, Noboru; Ikadai, Hiromi

    2013-11-15

    Encephalitozoon is an obligate intracellular microsporidian parasite that infects a wide range of mammalian hosts. In this study, we used nested PCR to investigate the presence of Encephalitozoon infection in Rodentia and Soricomorpha in Japan. We attempted to amplify and sequence Encephalitozoon-specific DNA from brain and viscera samples of 180 animals collected between 2008 and 2010. Forty-three samples (23.9%) from the orders Rodentia and Soricomorpha were positive for Encephalitozoon. This study is the first report of Encephalitozoon infection in Rodentia and Soricomorpha in Japan, and our findings suggest that these hosts may play a role in the spread of microsporidian spores in the environment.

  10. Pterygodermatites (Paucipectines) baiomydis n. sp. (Nematoda: Rictulariidae), a parasite of Baiomys taylori (Cricetidae)

    PubMed Central

    Lynggaard, Christina; García-Prieto, Luis; Guzmán-Cornejo, Carmen; Osorio-Sarabia, David

    2014-01-01

    Pterygodermatites (Paucipectines) baiomydis n. sp., an intestinal parasite of the northern pygmy mouse, Baiomys taylori (Cricetidae), collected in La Yerbabuena, Colima, Mexico, is described herein. Specimens were studied using light and scanning electronic microscopy. This is the 19th species of the subgenus Paucipectines described worldwide and the fourth collected in Mexico. It is differentiated from the remaining species in the subgenus by having 25 perioral denticles, arranged in a triangle (seven on each lateroventral margin, and eleven on the dorsal margin), and 10 pairs of caudal papillae. PMID:25375029

  11. Nematofauna of Rodents of the Families Heteromyidae and Cricetidae from the Mexican Plateau.

    PubMed

    Iturbe-Morgado, José Carlos; Falcón-Ordaz, Jorge; Lira-Guerrero, Georgina; Fernández, Jesús A; Acosta, Roxana

    2017-02-01

    As a part of an ongoing project to inventory the helminth parasites of rodents in Mexico, 85 specimens of 2 families of rodents were collected from the Mexican Plateau: Cricetidae ( Neotoma sp., Neotoma leucodon , Onychomys arenicola , Peromyscus sp., Peromyscus eremicus , and Reithrodontomys sp.) and Heteromyidae ( Chaetodipus sp., Chaetodipus eremicus , Chaetodipus hispidus , Dipodomys merriami , Dipodomys ordii , Dipodomys ornatus, Dipodomys spectabilis , Liomys irroratus , Perognathus sp., and Perognathus flavus ). A total of 13 taxa of helminths were found: Heteromyoxyuris longejector, Heteromyoxyuris otomii, Heteromyoxyuris sp., Onchocercidae gen. sp. 1 and sp. 2, Physalopteridae gen. sp., Protospirura dipodomis, Pterygodermatites dipodomis, Subulura sp., Syphacia sp., Trichuris dipodomis, Vexillata liomyos, and Vexillata armande. The highest species richness was recorded in D. merriami (7 taxa). This study is the first report of nematodes from O. arenicola (Physalopteridae gen. sp.) and C. eremicus (H. longejector) and for V. liomyos from D. merriami . All reports of these species of nematodes represent new collection localities in Mexico.

  12. Chromosomal evolution of Arvicolinae (Cricetidae, Rodentia). II. The genome homology of two mole voles (genus Ellobius), the field vole and golden hamster revealed by comparative chromosome painting.

    PubMed

    Romanenko, Svetlana A; Sitnikova, Natalia A; Serdukova, Natalya A; Perelman, Polina L; Rubtsova, Nadezhda V; Bakloushinskaya, Irina Yu; Lyapunova, Elena A; Just, Walter; Ferguson-Smith, Malcolm A; Yang, Fengtang; Graphodatsky, Alexander S

    2007-01-01

    Using cross-species chromosome painting, we have carried out a comprehensive comparison of the karyotypes of two Ellobius species with unusual sex determination systems: the Transcaucasian mole vole, Ellobius lutescens (2n = 17, X in both sexes), and the northern mole vole, Ellobius talpinus (2n = 54, XX in both sexes). Both Ellobius species have highly rearranged karyotypes. The chromosomal paints from the field vole (Microtus agrestis) detected, in total, 34 and 32 homologous autosomal regions in E. lutescens and E. talpinus karyotypes, respectively. No difference in hybridization pattern of the X paint (as well as Y paint) probes on male and female chromosomes was discovered. The set of golden hamster (Mesocricetus auratus) chromosomal painting probes revealed 44 and 43 homologous autosomal regions in E. lutescens and E. talpinus karyotypes, respectively. A comparative chromosome map was established based on the results of cross-species chromosome painting and a hypothetical ancestral Ellobius karyotype was reconstructed. A considerable number of rearrangements were detected; 31 and 7 fusion/fission rearrangements differentiated the karyotypes of E. lutescens and E. talpinus from the ancestral Ellobius karyotype. It seems that inversions have played a minor role in the genome evolution of these Ellobius species.

  13. Chromosomal evolution of Arvicolinae (Cricetidae, Rodentia). I. The genome homology of tundra vole, field vole, mouse and golden hamster revealed by comparative chromosome painting.

    PubMed

    Sitnikova, Natalia A; Romanenko, Svetlana A; O'Brien, Patricia C M; Perelman, Polina L; Fu, Beiyuan; Rubtsova, Nadezhda V; Serdukova, Natalya A; Golenishchev, Feodor N; Trifonov, Vladimir A; Ferguson-Smith, Malcolm A; Yang, Fengtang; Graphodatsky, Alexander S

    2007-01-01

    Cross-species chromosome painting has become the mainstay of comparative cytogenetic and chromosome evolution studies. Here we have made a set of chromosomal painting probes for the field vole (Microtus agrestis) by DOP-PCR amplification of flow-sorted chromosomes. Together with painting probes of golden hamster (Mesocricetus auratus) and mouse (Mus musculus), the field vole probes have been hybridized onto the metaphases of the tundra vole (Microtus oeconomus). A comparative chromosome map between these two voles, golden hamster and mouse has been established based on the results of cross-species chromosome painting and G-banding comparisons. The sets of paints from the field vole, golden hamster and mouse identified a total of 27, 40 and 47 homologous autosomal regions, respectively, in the genome of tundra vole; 16, 41 and 51 fusion/fission rearrangements differentiate the karyotype of the tundra vole from the karyotypes of the field vole, golden hamster and mouse, respectively.

  14. Taxonomic revision of the Andean leaf-eared mouse, Phyllotis andium Thomas 1912 (Rodentia: Cricetidae), with the description of a new species.

    PubMed

    Rengifo, Edgardo M; Pacheco, Víctor

    2015-09-16

    The Andean Leaf-eared mouse, Phyllotis andium Thomas 1912, has been considered a widespread medium-size sigmodontine rodent (230 mm of total length and 35 grams approximately) that occurs from Tungurahua, Ecuador, through the Andes, to Lima, Peru. Previous studies performed on Phyllotis noted evidence of morphological geographical variation within the species, which is likely because of the several potential geographical barriers that exist within the distribution range of P. andium. We carried out a taxonomic revision of this species based on qualitative and quantitative morphological analyses of 330 specimens from 92 localities. This included appropriate comparisons with other species of the andium/amicus group and performed molecular analysis based on cytochrome b sequences. As a result, morphologic qualitative analysis suggested the recognition of three different taxa, which are supported by morphologic quantitative and molecular analyses. The three taxa here identified have allopatric distributional ranges separated by important geographic barriers. Following these identification criteria, P. andium is now recognized for the samples from Tungurahua, Ecuador to Huánuco, Peru, and includes melanius and fruticicolus as synonymous; the southern populations from the Ancash and Lima departments, in the western Peruvian Andes, are proposed to represent a new species; and we recognize P. stenops as a valid species with tamborum as a synonym. Finally, we postulate that the diversification of these three species is related to key events in the Andean orogeny.

  15. An integrative appraisal of the diversification in the Atlantic forest genus Delomys (Rodentia: Cricetidae: Sigmodontinae) with the description of a new species.

    PubMed

    Gonçalves, Pablo Rodrigues; de Oliveira, João Alves

    2014-01-30

    Recent taxonomic studies on Neotropical mammals have benefited from the use of genetic data to unravel and recognize species diversity in a number of genera, including the Atlantic forest endemic genus Delomys. However, the success of this approach depends on ability to link genetically identified lineages to species names based on voucher specimens that lack genetic data. Cytogenetic studies in the Atlantic forest endemic rodent genus Delomys have revealed two widespread karyotypes, 2n=72/FN=90 and 2n=82/FN=80, which have been respectively ascribed to Delomys sublineatus (Thomas, 1903) and D. dorsalis (Hensel, 1872). More recently, a third karyotype, 2n=82/FN=86, reported from specimens collected on two montaintops in southeastern Brazil, was interpreted as evidence for a third species, D. collinus Thomas, 1917. This nominal form had originally been described as a subspecies of D. dorsalis from Itatiaia, one of the mountain ranges where the third karyotype was later detected. The detection of two sympatric karyotypes at the type locality of D. collinus in the Itatiaia mountain range, Southeastern Brazil, prompted a reevaluation of the association of karyomorphs and species names. In this paper, we assessed the congruence of molecular (cytochrome b), cytogenetic and morphological characters, to diagnose the species in the genus, including data from recently collected series and type specimens. Our results indicate that the genetic and morphological patterns are largely congruent with the recognition of three species, each of which is karyotypically and morphologically diagnosable. Our morphological analyses of sympatric samples from Itatiaia refute the former association of the 2n=82/FN=86 karyotype with the holotype of D. dorsalis collinus (which is more similar to D. dorsalis with 2n=82/FN=80). Instead, we recognize and describe a new species for the 2n=82/FN=86 populations from the highest altitudinal zones of the Itatiaia and Caparaó mountains. The geographical variation in D. dorsalis is also explored and the status of D. d. collinus is discussed in the light of the molecular and morphological evidence. Finally, we discuss biogeographic hypotheses concerning the disjunct distributions of D. dorsalis and the new species.

  16. Differences in richness and composition of gastrointestinal parasites of small rodents (Cricetidae, Rodentia) in a continental and insular area of the Atlantic Forest in Santa Catarina state, Brazil.

    PubMed

    Kuhnen, V V; Graipel, M E; Pinto, C J C

    2012-08-01

    The first and only study on gastrointestinal parasites of wild rodents in the Island of Santa Catarina was done in 1987. The aim of this study was to identify intestinal parasites from wild rodents in Santo Amaro da Imperatriz and Santa Catariana Island, and to compare the richness and composition of the gastrointestinal parasite community of both areas. Rodents were captured with live traps, and feces were screened using the sedimentation method and optical microscopy. The following species of rodents were captured in the two areas: Akodon montensis, Euryoryzomys russatus, Oligoryzomys nigripes and Nectomys squamipes. In Santo Amaro da Impetratriz, prevalent parasites were: A. montensis (51%), E. russatus (62%), O. nigripes (53%) and N. squamipes (20%). From the Island of Santa Catarina the rodent prevalence rates were: A. montensis (43%), E. russatus (59%), O. nigripes (30%) and N. squamipes (33%) and the collected parasites were: Hymenolepis sp., Longistriata sp., Strongyloides sp., Hassalstrongylus sp., Syphacia sp., Trichomonas sp., Ancylostomidae, Trichuridae, Oxyuridae and Eucoccidiorida. The species richness (10.6 ± 0.7) of the endoparasite comunity in the area located on the continent was higher (p < 0.01) and different (p = 0.001) from that of the area located on the island (6.9 ± 0.5).

  17. Chromosome homologies of the highly rearranged karyotypes of four Akodon species (Rodentia, Cricetidae) resolved by reciprocal chromosome painting: the evolution of the lowest diploid number in rodents.

    PubMed

    Ventura, Karen; O'Brien, Patricia C M; Yonenaga-Yassuda, Yatiyo; Ferguson-Smith, Malcolm A

    2009-01-01

    Traditionally comparative cytogenetic studies are based mainly on banding patterns. Nevertheless, when dealing with species with highly rearranged genomes, as in Akodon species, or with other highly divergent species, cytogenetic comparisons of banding patterns prove inadequate. Hence, comparative chromosome painting has become the method of choice for genome comparisons at the cytogenetic level since it allows complete chromosome probes of a species to be hybridized in situ onto chromosomes of other species, detecting homologous genomic regions between them. In the present study, we have explored the highly rearranged complements of the Akodon species using reciprocal chromosome painting through species-specific chromosome probes obtained by chromosome sorting. The results revealed complete homology among the complements of Akodon sp. n. (ASP), 2n = 10; Akodon cursor (ACU), 2n = 15; Akodon montensis (AMO), 2n = 24; and Akodon paranaensis (APA), 2n = 44, and extensive chromosome rearrangements have been detected within the species with high precision. Robertsonian and tandem rearrangements, pericentric inversions and/or centromere repositioning, paracentric inversion, translocations, insertions, and breakpoints, where chromosomal rearrangements, seen to be favorable, were observed. Chromosome painting using the APA set of 21 autosomes plus X and Y revealed eight syntenic segments that are shared with A. montensis, A. cursor, and ASP, and one syntenic segment shared by A. montensis and A. cursor plus five exclusive chromosome associations for A. cursor and six for ASP chromosome X, except for the heterochromatin region of ASP X, and even chromosome Y shared complete homology among the species. These data indicate that all those closely related species have experienced a recent extensive process of autosomal rearrangement in which, except for ASP, there is still complete conservation of sex chromosomes homologies.

  18. Laelapinae mites (Acari: Parasitiformes: Laelapidae) parasitic of sigmodontine rodents from northern Peru, with the description of a new species from Akodon aerosus (Rodentia: Cricetidae: Sigmodontinae).

    PubMed

    Lareschi, Marcela; Velazco, Paúl M

    2013-04-01

    Laelapine mites are common parasites of sigmodontine rodents in the Neotropics. However, few species are reported from Peru as a result of the low number of mammal surveys that include ectoparasite collections. Herein we report 12 species of mites from northern Peru. From these, 8 are reported for the first time for the country, and 1 is new to science , Androlaepaps aerosus sp. nov., the latter associated exclusively with the sigmodontine Akodon aerosus . Most of the laelapine species were host specific. The new species, included in the Androlaelaps rotundus species group, resembles An. rotundus "sensu stricto" and An. ulysespardinasi in general appearance but is unique in the length of the hypostomal seta h3 (>58 μm), which is 3 times as long as the gnathosomal seta, and its tip reaching or over-reaching the gnathosomal setal bases; dorsal seta j2 is very long (>70 μm), almost reaching the point of j3.

  19. Ectoparasite occurrence associated with males and females of wild rodents Oligoryzomys flavescens (Waterhouse) and Akodon azarae (Fischer) (Rodentia: Cricetidae: Sigmodontinae) in the Punta Lara wetlands, Argentina.

    PubMed

    Lareschi, Marcela

    2010-01-01

    The objective of this research was to study infestation parameters and indexes of ectoparasites associated with each sex of the wild rodents Oligoryzomys flavescens (Waterhouse) andAkodon azarae (Fischer) in the Punta Lara wetlands, Argentina. A trend towards higher mean abundance (MA) and ectoparasite specific richness was observed in males of O. flavescens whereas those values were similar for both A. azarae sexes. The prevalence of the following ectoparasites was significantly higher on males (P<0.05): Mysolaelaps microspinosus Fonseca (65.2%) and Hoplopleura travassosi Werneck (73.9%) on O. flavescens, and Ixodes loricatus Neumann (71.4%) on A. azarae. Only H. travassosi mean abundance was significantly higher on males (MA=44.1). Since I. loricatus and Hoplopleura spp. are involved in the transmission of pathogens that cause diseases in animals and humans, and whose reservoirs are rodent hosts, these results are epidemiologically important.

  20. Morphometric analysis of the placenta in the New World mouse Necromys lasiurus (Rodentia, Cricetidae): a comparison of placental development in cricetids and murids

    PubMed Central

    2013-01-01

    Background Stereology is an established method to extrapolate three-dimensional quantities from two-dimensional images. It was applied to placentation in the mouse, but not yet for other rodents. Herein, we provide the first study on quantitative placental development in a sigmodontine rodent species with relatively similar gestational time. Placental structure was also compared to the mouse, in order to evaluate similarities and differences in developmental patterns at the end of gestation. Methods Fetal and placental tissues of Necromys lasiurus were collected and weighed at 3 different stages of gestation (early, mid and late gestation) for placental stereology. The total and relative volumes of placenta and of its main layers were investigated. Volume fractions of labyrinth components were quantified by the One Stop method in 31 placentae collected from different individuals, using the Mercator® software. Data generated at the end of gestation from N. lasiurus placentae were compared to those of Mus musculus domesticus obtained at the same stage. Results A significant increase in the total absolute volumes of the placenta and its main layers occurred from early to mid-gestation, followed by a reduction near term, with the labyrinth layer becoming the most prominent area. Moreover, at the end of gestation, the total volume of the mouse placenta was significantly increased compared to that of N. lasiurus although the proportions of the labyrinth layer and junctional zones were similar. Analysis of the volume fractions of the components in the labyrinth indicated a significant increase in fetal vessels and sinusoidal giant cells, a decrease in labyrinthine trophoblast whereas the proportion of maternal blood space remained stable in the course of gestation. On the other hand, in the mouse, volume fractions of fetal vessels and sinusoidal giant cells decreased whereas the volume fraction of labyrinthine trophoblast increased compared to N. lasiurus placenta. Conclusions Placental development differed between N. lasiurus and M. musculus domesticus. In particular, the low placental efficiency in N. lasiurus seemed to induce morphological optimization of fetomaternal exchanges. In conclusion, despite similar structural aspects of placentation in these species, the quantitative dynamics showed important differences. PMID:23433040

  1. Climatic niche conservatism and ecological opportunity in the explosive radiation of arvicoline rodents (Arvicolinae, Cricetidae).

    PubMed

    Lv, Xue; Xia, Lin; Ge, Deyan; Wu, Yongjie; Yang, Qisen

    2016-05-01

    Climatic niche conservatism shapes patterns of diversity in many taxonomic groups, while ecological opportunity (EO) can trigger rapid speciation that is less constrained by the amount of time a lineage has occupied a given habitat. These two processes are well studied, but limited research has considered their joint and relative roles in shaping diversity patterns. We characterized climatic and biogeographic variables for 102 species of arvicoline rodents (Arvicolinae, Cricetidae), testing the effects of climatic niche conservatism and EO on arvicoline diversification as lineages transitioned between biogeographic regions. We found that the amount of time a lineage has occupied a precipitation niche is positively correlated with diversity along a precipitation gradient, suggesting climatic niche conservatism. In contrast, shift in diversification rate explained diversity patterns along a temperature gradient. Our results suggest that an indirect relationship exists between temperature and diversification that is associated with EO as arvicoline rodents colonized warm Palearctic environments. Climatic niche conservatism alone did not fully explain diversity patterns under density-dependence, highlighting the additional importance of EO-related processes in promoting the explosive radiation in arvicoline rodents and shaping diversity pattern among biogeographic regions and along climatic gradients.

  2. Epizootiology of Tacaribe Serocomplex Viruses (Arenaviridae) Associated with Neotomine Rodents (Cricetidae, Neotominae) in Southern California

    PubMed Central

    Milazzo, Mary Louise; Cajimat, Maria N. B.; Mauldin, Matthew R.; Bennett, Stephen G.; Hess, Barry D.; Rood, Michael P.; Conlan, Christopher A.; Nguyen, Kiet; Wekesa, J. Wakoli; Ramos, Ronald D.; Bradley, Robert D.

    2015-01-01

    Abstract The objective of this study was to advance our knowledge of the epizootiology of Bear Canyon virus and other Tacaribe serocomplex viruses (Arenaviridae) associated with wild rodents in California. Antibody (immunoglobulin G [IgG]) to a Tacaribe serocomplex virus was found in 145 (3.6%) of 3977 neotomine rodents (Cricetidae: Neotominae) captured in six counties in southern California. The majority (122 or 84.1%) of the 145 antibody-positive rodents were big-eared woodrats (Neotoma macrotis) or California mice (Peromyscus californicus). The 23 other antibody-positive rodents included a white-throated woodrat (N. albigula), desert woodrat (N. lepida), Bryant's woodrats (N. bryanti), brush mice (P. boylii), cactus mice (P. eremicus), and deer mice (P. maniculatus). Analyses of viral nucleocapsid protein gene sequence data indicated that Bear Canyon virus is associated with N. macrotis and/or P. californicus in Santa Barbara County, Los Angeles County, Orange County, and western Riverside County. Together, analyses of field data and antibody prevalence data indicated that N. macrotis is the principal host of Bear Canyon virus. Last, the analyses of viral nucleocapsid protein gene sequence data suggested that the Tacaribe serocomplex virus associated with N. albigula and N. lepida in eastern Riverside County represents a novel species (tentatively named “Palo Verde virus”) in the genus Arenavirus. PMID:25700047

  3. Complete mitochondrial genome sequence of Marmota himalayana (Rodentia: Sciuridae) and phylogenetic analysis within Rodentia.

    PubMed

    Chao, Q J; Li, Y D; Geng, X X; Zhang, L; Dai, X; Zhang, X; Li, J; Zhang, H J

    2014-04-14

    This is the first report of a complete mitochondrial genome sequence from Himalayan marmot (Marmota himalayana, class Marmota). We determined the M. himalayana mitochondrial (mt) genome sequence by using long-PCR methods and a primer-walking sequencing strategy with genus-specific primers. The complete mt genome of M. himalayana was 16,443 bp in length and comprised 13 protein-coding genes, 2 ribosomal RNA (rRNA) genes, 22 transfer RNA (tRNA) genes, and a typical control region (CR). Gene order and orientation were identical to those in mt genomes of most vertebrates. The heavy strand showed an overall A+T content of 63.49%. AT and GC skews for the mt genome of the M. himalayana were 0.012 and -0.300, respectively, indicating a nucleotide bias against T and G. The control region was 997 bp in size and displayed some unusual features, including absence of repeated motifs and two conserved sequence blocks (CSB2 and CSB3), which is consistent with observations from two other rodent species, Sciurus vulgaris and Myoxus glis. Phylogenetic analysis of complete mt DNA sequences without the control region including 30 taxa of Rodentia was performed with Maximum-Likelihood (ML) and Bayesian Inference (BI) methods and provided strong support for Sciurognathi polyphyly and Hystricognathi monophyly. This analysis also provided evidence that M. himalayana mt DNA was closely related to that from Sciurus vulgaris (Sciuridae) and was similar to mt DNA from Myoxus glis.

  4. Acariform mites (Acariformes) - permanent symbionts of Hapalomysdelacouri Thomas (Rodentia, Muridae) in Vietnam.

    PubMed

    Bochkov, Andre V; Abramov, Alexei V

    2014-01-01

    Two new species of parasitic acariform mites (Acariformes) are described from the Delacour's marmoset rat Hapalomysdelacouri Thomas (Rodentia: Muridae) in Vietnam: Afrolistrophorus (Afrolistrophorus) hapalomyssp. n. (Listrophoridae) and Radfordia (Radfordia) mirabilissp. n. (Myobiidae). Based on morphological evidences, we show that species of both mite genera associated with Hapalomys Blyth do not demonstrate clear phylogenetic links with respective congeners from rodents of the closest genus Chiropodomys Peters (Rodentia: Muridae).

  5. Arthropod symbiotes of Laonastes aenigmamus (Rodentia:Diatomyidae).

    PubMed

    Bochkov, A V; Abramov, A V; Durden, L A; Apanaskevich, D A; Stekolnikov, A A; Stanyukovich, M K; Gnophanxay, S; Tikhonov, A N

    2011-04-01

    Arthropod symbiotes of the Laotian rock-rat, Laonastes aenigmamus (Rodentia:Diatomyidae), from Laos are examined. This host is a member of Diatomyidae previously thought to have gone extinct >10 million yr ago. Permanent symbiotes are represented by 2 species, a new species of sucking louse, Polyplax sp., near rhizomydis (Phthiraptera:Polyplacidae), and a new species of fur mite, Afrolistrophorus sp., near maculatus (Acariformes:Listrophoridae). The temporary parasites are represented by 18 species, i.e., 1 mesostigmatan species, i.e., a new species of Androlaelaps near casalis (Parasitiformes:Laelapidae); immature stages of 2 tick species, Ixodes granulatus and Haemaphysalis sp. (Parasitiformes:Ixodidae); and a rich fauna of chiggers (Acariformes:Trombiculidae) comprising 8 genera and 15 species. It is hypothesized that this host completely lost its initial fauna of ectosymbiotes and that ancestors of the recorded symbiotes switched to this host from rodents of the superfamily Muroidea.

  6. Taxonomic position of Eothenomys wardi (Arvicolinae: Cricetidae) based on morphological and molecular analyses with a detailed description of the species.

    PubMed

    Zeng, Tao; Jin, Wei; Sun, Zhi Yu; Liu, Yang; Murphy, Robert W; Fu, Jian Rong; Wang, Xin; Hou, Quan Fen; Tu, Fei Yun; Liao, Rui; Liu, Shao Ying; Yue, Bi Song

    2013-01-01

    Ward's Red-backed Vole (Eothenomys wardi) is a rodent from the family Cricetidae. This endemic species occurs only in extreme northwestern Yunnan province, China in the Mekong and Salween river divide. It occupies steep cliffs at 2,800 to 4,250 m above sea level on the remote Qinghai-Tibetan plateau. The validity of E. wardi is controversial and no specimens exist apart from the nominal series. In 2010, we collected 38 topotypes of E. wardi from Meri Snow Mountain. The results of our phylogenetic analyses based on nucleotide sequences of the mitochondrial genes cytochrome b (cytb) and cytochrome c oxidase subunit one (COI) suggest that E. wardi is the sister group of E. custos, against its previously presumed sister species or conspecific species E. chinensis. In addition, seven out of 34 morphological characters differentiate E. wardi from other members of the genus Eothenomys. Therefore, we consider E. wardi to be a valid species and we provide its detailed morphological description.

  7. Environmental Compliance Assessment and Management System Program (ECAMP). U.S. Air Force European Economic Community (EEC)

    DTIC Science & Technology

    1993-09-01

    pipistrellus RODENTIA Sciuridae Citellus citellus Cricetidae Cricetus cricetus Hystricidae Hystrix crisata CARNIVORA Canidae Canis lupus Alopex lagopus...cabrerae Microtus oeconomus arenicola Zapodidae Sicista betulina Hystricidae Hystrix cristata CARNIVORA Canidae Canis lupus except Spanish populations

  8. PPIP5K1 Suppresses Etoposide-triggered Apoptosis

    PubMed Central

    Machkalyan, Gayane

    2016-01-01

    Inositol hexakisphosphate kinase 2 (IP6K2) potentiates pro-apoptotic signalling and increases the sensitivity of mammalian cells to cytotoxic agents. Diphosphoinositol pentakisphosphate kinase (PPIP5K) generates inositol pyrophosphates (InsPPs) that are structurally distinct from those produced by IP6K2 and their possible roles in affecting cell viability remain unclear. In the present study, we tested the impact of PPIP5K1 on cellular sensitivity to various genotoxic agents to determine if PPIP5K1 and IP6K2 contribute similarly to apoptosis. We observed that PPIP5K1 overexpression decreased sensitivity of cells toward several cytotoxic agents, including etoposide, cisplatin, and sulindac. We further tested the impact of PPIP5K1 overexpression on an array of apoptosis markers and observed that PPIP5K1 decreased p53 phosphorylation on key residues, including Ser-15, -46, and -392. Overexpression of a kinase-impaired PPIP5K1 mutant failed to protect cells from apoptosis, indicating this protection is a consequence PPIP5K1 catalytic activity, in contrast with the sensitivity conferred by IP6K2, which is dependent on both catalytic and non-catalytic functions. These observations reveal distinct roles for PPIP5K1 and IP6K2 and the InsPPs they produce in controlling cell death.

  9. New karyotypes of Atlantic tree rats, genus Phyllomys (Rodentia: Echimyidae).

    PubMed

    Araújo, Naiara Pereira; Loss, Ana Carolina; Cordeiro-Junior, Dirceu A; da Silva, Kátia Regina; Leite, Yuri L R; Svartman, Marta

    2014-01-01

    Phyllomys (Echimyidae, Rodentia) is a genus of Neotropical rodents with available cytogenetic data restricted to six out of 13 species, mainly based on simple staining methods, without detailed analyses. In this work, we present new karyotypes for Phyllomys lamarum (diploid number 2n = 56, fundamental number or number of autosomal arms FN = 102) and Phyllomys sp. (2n = 74, FN = 140) from the state of Minas Gerais, southeastern Brazil. We provide the first GTG- and CBG-banding patterns, silver-staining of the nucleolar organizer regions (Ag-NORs), and fluorescence in situ hybridization (FISH) with telomeric and 45S rDNA probes of Phyllomys. In addition to examining their chromosomes and phenotypic characters, we sequenced mitochondrial DNA from the specimens analyzed to confirm their taxonomic identification. The comparison of the distinctive chromosome complements of our specimens with those of other species of Phyllomys already published allowed us to conclude that chromosome data may be very useful for the taxonomy of the genus, as no two species analyzed presented the same diploid and fundamental numbers (2n and FN).

  10. A Transitional Gundi (Rodentia: Ctenodactylidae) from the Miocene of Israel.

    PubMed

    López-Antoñanzas, Raquel; Gutkin, Vitaly; Rabinovich, Rivka; Calvo, Ran; Grossman, Aryeh

    2016-01-01

    We describe a new species of gundi (Rodentia: Ctenodactylidae: Ctenodactylinae), Sayimys negevensis, on the basis of cheek teeth from the Early Miocene of the Rotem Basin, southern Israel. The Rotem ctenodactylid differs from all known ctenodactylid species, including Sayimys intermedius, which was first described from the Middle Miocene of Saudi Arabia. Instead, it most resembles Sayimys baskini from the Early Miocene of Pakistan in characters of the m1-2 (e.g., the mesoflexid shorter than the metaflexid, the obliquely orientated hypolophid, and the presence of a strong posterolabial ledge) and the upper molars (e.g., the paraflexus that is longer than the metaflexus). However, morphological (e.g., presence of a well-developed paraflexus on unworn upper molars) and dimensional (regarding, in particular, the DP4 and M1 or M2) differences between the Rotem gundi and Sayimys baskini distinguish them and testify to the novelty and endemicity of the former. In its dental morphology, Sayimys negevensis sp. nov. shows a combination of both the ultimate apparition of key-characters and incipient features that would be maintained and strengthened in latter ctenodactylines. Thus, it is a pivotal species that bridges the gap between an array of primitive ctenodactylines and the most derived, Early Miocene and later, gundis.

  11. A Transitional Gundi (Rodentia: Ctenodactylidae) from the Miocene of Israel

    PubMed Central

    López-Antoñanzas, Raquel; Gutkin, Vitaly; Rabinovich, Rivka; Calvo, Ran; Grossman, Aryeh

    2016-01-01

    We describe a new species of gundi (Rodentia: Ctenodactylidae: Ctenodactylinae), Sayimys negevensis, on the basis of cheek teeth from the Early Miocene of the Rotem Basin, southern Israel. The Rotem ctenodactylid differs from all known ctenodactylid species, including Sayimys intermedius, which was first described from the Middle Miocene of Saudi Arabia. Instead, it most resembles Sayimys baskini from the Early Miocene of Pakistan in characters of the m1-2 (e.g., the mesoflexid shorter than the metaflexid, the obliquely orientated hypolophid, and the presence of a strong posterolabial ledge) and the upper molars (e.g., the paraflexus that is longer than the metaflexus). However, morphological (e.g., presence of a well-developed paraflexus on unworn upper molars) and dimensional (regarding, in particular, the DP4 and M1 or M2) differences between the Rotem gundi and Sayimys baskini distinguish them and testify to the novelty and endemicity of the former. In its dental morphology, Sayimys negevensis sp. nov. shows a combination of both the ultimate apparition of key-characters and incipient features that would be maintained and strengthened in latter ctenodactylines. Thus, it is a pivotal species that bridges the gap between an array of primitive ctenodactylines and the most derived, Early Miocene and later, gundis. PMID:27049960

  12. Hymenolepis folkertsi n. sp. (Eucestoda: Hymenolepididae) in the oldfield mouse Peromyscus polionotus (Wagner) (Rodentia: Cricetidae: Neotominae) from the southeastern Nearctic with comments on tapeworm faunal diversity among deer mice.

    PubMed

    Makarikov, Arseny A; Nims, Todd N; Galbreath, Kurt E; Hoberg, Eric P

    2015-06-01

    A previously unrecognized species of hymenolepidid cestode attributable to Hymenolepis is described based on specimens in Peromyscus polionotus, oldfield mouse, from Georgia near the southeastern coast of continental North America. Specimens of Hymenolepis folkertsi n. sp. differ from those attributed to most other species in the genus by having testes arranged in a triangle and a scolex with a prominent rostrum-like protrusion. The newly recognized species is further distinguished by the relative position and length of the cirrus sac, shape of seminal receptacle, and relative size of external seminal vesicle and seminal receptacle. Hymenolepidid cestodes have sporadically been reported among the highly diverse assemblage of Peromyscus which includes 56 distinct species in the Nearctic. Although the host genus has a great temporal duration and is endemic to the Nearctic, current evidence suggests that tapeworm faunal diversity reflects relatively recent assembly through bouts of host switching among other cricetid, murid, and geomyid rodents in sympatry.

  13. Mazzanema n. gen. and Mazzanema fortuita n. comb. for Longistriata fortuita Freitas, Lent, and Almeida, 1937 (Nematoda: Heligmonellidae), a parasite of the marsh rat Holochilus chacarius (Rodentia: Cricetidae) from northern Argentina.

    PubMed

    Digiani, María Celina; Notarnicola, Juliana; Paulos, María Soledad

    2013-10-01

    The species described as Longistriata fortuita Freitas, Lent, and Almeida, 1937 is here redescribed from new material collected from the type host, Holochilus chacarius balnearum Thomas, and the type locality, San Martín del Tabacal, Salta, Argentina. Neotypes are designed for the species since the type material deposited by the authors is lost. The original description did not include the synlophe or the female and both are here described. Several characters of the synlophe as the number of ridges (14-19), the ridges continuous and all around body, and the presence of a gradient of size of the ridges allow us to place the species within the Heligmonellidae, Nippostrongylinae. The species possesses a unique combination of characters as the synlophe having a carene together with characters of the caudal bursa as the pattern 1-3-1 and the strong development of the dorsal lobe and ray, which precludes its inclusion in any known genus of Nippostrongylinae. A new genus Mazzanema n. gen. is proposed for it, resulting in the new combination Mazzzanema fortuita n. comb.

  14. Karyotypes of two rare rodents, Hapalomys delacouri and Typhlomys cinereus (Mammalia, Rodentia), from Vietnam.

    PubMed

    Abramov, Alexei V; Aniskin, Vladimir M; Rozhnov, Viatcheslav V

    2012-01-01

    Karyotypes of Hapalomys delacouri (Rodentia, Muridae) and Typhlomys cinereus (Rodentia, Platacanthomyidae) from Vietnam are described for the first time. The diploid karyotype of Hapalomys delacouri is 38 (NFa=48), consisting of six pairs of bi-armed and 12 pairs of acrocentric autosomes decreasing in size; plus a large metacentric X chromosome and Y chromosome, also metacentric, that is equal in size to the largest pair of acrocentric autosomes. The newly described karyotype differs significantly from that reported for Hapalomys delacouri from northern Thailand. The latter record very likely represents a different species of Hapalomys, possibly the taxon Hapalomys pasquieri described from north-central Laos.The diploid karyotype of Typhlomys cinereus is 38 (NF=48), consisting of five pairs of meta- to submetacentric and 14 pairs of acrocentric chromosomes varying in size from large to small; sex chromosomes were not defined.

  15. MAP2K1 — EDRN Public Portal

    Cancer.gov

    MAP2K1, or mitogen-activated protein kinase kinase 1, is a dual specificity protein kinase which acts as an essential component of the MAP kinase signal transduction pathway. MAP kinases, also known as extracellular signal-regulated kinases (ERKs), act as an integration point for multiple biochemical signals. MAP2K1 is located upstream of MAP kinases and stimulates the enzymatic activity of MAP kinases upon wide variety of extra- and intracellular signals. MAP2K1 is thought to be involved in binding of extracellular ligands such as growth factors, cytokines and hormones to their cell-surface receptors, as well as cellular processes such as proliferation, differentiation, transcription regulation and development.

  16. Characterizing the Severe Reactions of Parenteral Vitamin K1.

    PubMed

    Britt, Rachel B; Brown, Jamie N

    2016-01-01

    Parenteral vitamin K1 (phytonadione) is used for anticoagulant reversal, and a boxed warning exists with intravenous and intramuscular administration due to the possibility of severe reactions, including fatalities. These reactions resemble hypersensitivity or anaphylaxis, including anaphylactoid reaction, and have led to shock and cardiac and/or respiratory arrest. The objective of this review is to summarize the available literature detailing the anaphylactic/anaphylactoid reactions with parenteral vitamin K1 in order to better characterize the reaction and provide a more in-depth understanding of its importance. A comprehensive literature search of MEDLINE (1946 to June 2016) and EMBASE (1947 to June 2016) was conducted using the terms vitamin K1, phytonadione, phytomenadione, vitamin K group, anaphylaxis, polyoxyethylated castor oil, and cremophor. A total of 2 retrospective surveillance studies, 2 retrospective cohort studies, and 17 case reports were identified for inclusion and assessment. Based on a review of the literature, use of parenteral vitamin K1 may result in severe hypotension, bradycardia or tachycardia, dyspnea, bronchospasm, cardiac arrest, and death. These reactions are most consistent with a nonimmune-mediated anaphylactoid mechanism. It appears that intravenous administration is more frequently associated with these reactions and occurs at an incidence of 3 per 10 000 doses of intravenous vitamin K1. The solubilizer may also increase the risk of adverse reactions, which occurred in patients with and without previous exposure to vitamin K1. Although there are known factors that increase the risk of an adverse drug event occurring, reactions have been reported despite all precautions being properly followed.

  17. Intestinal scavenger receptors are involved in vitamin K1 absorption.

    PubMed

    Goncalves, Aurélie; Margier, Marielle; Roi, Stéphanie; Collet, Xavier; Niot, Isabelle; Goupy, Pascale; Caris-Veyrat, Catherine; Reboul, Emmanuelle

    2014-10-31

    Vitamin K1 (phylloquinone) intestinal absorption is thought to be mediated by a carrier protein that still remains to be identified. Apical transport of vitamin K1 was examined using Caco-2 TC-7 cell monolayers as a model of human intestinal epithelium and in transfected HEK cells. Phylloquinone uptake was then measured ex vivo using mouse intestinal explants. Finally, vitamin K1 absorption was compared between wild-type mice and mice overexpressing scavenger receptor class B type I (SR-BI) in the intestine and mice deficient in cluster determinant 36 (CD36). Phylloquinone uptake by Caco-2 cells was saturable and was significantly impaired by co-incubation with α-tocopherol (and vice versa). Anti-human SR-BI antibodies and BLT1 (a chemical inhibitor of lipid transport via SR-BI) blocked up to 85% of vitamin K1 uptake. BLT1 also decreased phylloquinone apical efflux by ∼80%. Transfection of HEK cells with SR-BI and CD36 significantly enhanced vitamin K1 uptake, which was subsequently decreased by the addition of BLT1 or sulfo-N-succinimidyl oleate (CD36 inhibitor), respectively. Similar results were obtained in mouse intestinal explants. In vivo, the phylloquinone postprandial response was significantly higher, and the proximal intestine mucosa phylloquinone content 4 h after gavage was increased in mice overexpressing SR-BI compared with controls. Phylloquinone postprandial response was also significantly increased in CD36-deficient mice compared with wild-type mice, but their vitamin K1 intestinal content remained unchanged. Overall, the present data demonstrate for the first time that intestinal scavenger receptors participate in the absorption of dietary phylloquinone.

  18. Intestinal Scavenger Receptors Are Involved in Vitamin K1 Absorption*

    PubMed Central

    Goncalves, Aurélie; Margier, Marielle; Roi, Stéphanie; Collet, Xavier; Niot, Isabelle; Goupy, Pascale; Caris-Veyrat, Catherine; Reboul, Emmanuelle

    2014-01-01

    Vitamin K1 (phylloquinone) intestinal absorption is thought to be mediated by a carrier protein that still remains to be identified. Apical transport of vitamin K1 was examined using Caco-2 TC-7 cell monolayers as a model of human intestinal epithelium and in transfected HEK cells. Phylloquinone uptake was then measured ex vivo using mouse intestinal explants. Finally, vitamin K1 absorption was compared between wild-type mice and mice overexpressing scavenger receptor class B type I (SR-BI) in the intestine and mice deficient in cluster determinant 36 (CD36). Phylloquinone uptake by Caco-2 cells was saturable and was significantly impaired by co-incubation with α-tocopherol (and vice versa). Anti-human SR-BI antibodies and BLT1 (a chemical inhibitor of lipid transport via SR-BI) blocked up to 85% of vitamin K1 uptake. BLT1 also decreased phylloquinone apical efflux by ∼80%. Transfection of HEK cells with SR-BI and CD36 significantly enhanced vitamin K1 uptake, which was subsequently decreased by the addition of BLT1 or sulfo-N-succinimidyl oleate (CD36 inhibitor), respectively. Similar results were obtained in mouse intestinal explants. In vivo, the phylloquinone postprandial response was significantly higher, and the proximal intestine mucosa phylloquinone content 4 h after gavage was increased in mice overexpressing SR-BI compared with controls. Phylloquinone postprandial response was also significantly increased in CD36-deficient mice compared with wild-type mice, but their vitamin K1 intestinal content remained unchanged. Overall, the present data demonstrate for the first time that intestinal scavenger receptors participate in the absorption of dietary phylloquinone. PMID:25228690

  19. Extension of practical k1 limit in EUV lithography

    NASA Astrophysics Data System (ADS)

    Park, Sarohan; Lee, Inwhan; Koo, Sunyoung; Lee, Junghyung; Lim, Chang-Moon

    2016-03-01

    Sub 0.3k1 regime has been widely adopted for high volume manufacturing (HVM) of optical lithography due to various resolution enhancement technologies (RETs). It is not certain when such low k1 is feasible in EUV, though most technologies are available in EUV also. In this paper, experimental results on patterning performance of line space (L/S) and contact hole (C/H) in EUV lithography will be presented. First, practical k1 value with 0.33NA EUV lithography was investigated through experiment using NXE3300 EUV tool. Patterning limit, as defined by local critical dimension uniformity (LCDU) for C/H array pattern were measured with respect to various design rules. It was evaluated that the effect of off axis illumination (OAI) mode with various illumination conditions to improve the patterning performance and to reduce k1 limit. Then the experimental results of LCDU were compared with normalized image log slope (NILS) values from simulation. EUV source mask optimization (SMO) technologies to increase NILS with FlexPupil option of EUV scanner were evaluated and possibility of further improvement was also discussed.

  20. Space Access for Small Satellites on the K-1

    NASA Astrophysics Data System (ADS)

    Faktor, L.

    Affordable access to space remains a major obstacle to realizing the increasing potential of small satellites systems. On a per kilogram basis, small launch vehicles are simply too expensive for the budgets of many small satellite programs. Opportunities for rideshare with larger payloads on larger launch vehicles are still rare, given the complications associated with coordinating delivery schedules and deployment orbits. Existing contractual mechanisms are also often inadequate to facilitate the launch of multiple payload customers on the same flight. Kistler Aerospace Corporation is committed to lowering the price and enhancing the availability of space access for small satellite programs through the fully-reusable K-1 launch vehicle. Kistler has been working with a number of entities, including Astrium Ltd., AeroAstro, and NASA, to develop innovative approaches to small satellite missions. The K-1 has been selected by NASA as a Flight Demonstration Vehicle for the Space Launch Initiative. NASA has purchased the flight results during the first four K-1 launches on the performance of 13 advanced launch vehicle technologies embedded in the K-1 vehicle. On K-1 flights #2-#4, opportunities exist for small satellites to rideshare to low-earth orbit for a low-launch price. Kistler's flight demonstration contract with NASA also includes options to fly Add-on Technology Experiment flights. Opportunities exist for rideshare payloads on these flights as well. Both commercial and government customers may take advantage of the rideshare pricing. Kistler is investigating the feasibility of flying dedicated, multiple small payload missions. Such a mission would launch multiple small payloads from a single customer or small payloads from different customers. The orbit would be selected to be compatible with the requirements of as many small payload customers as possible, and make use of reusable hardware, standard interfaces (such as the existing MPAS) and verification plans

  1. Prevalence of genes encoding virulence factors among Escherichia coli with K1 antigen and non-K1 E. coli strains.

    PubMed

    Kaczmarek, Agnieszka; Budzynska, Anna; Gospodarek, Eugenia

    2012-10-01

    Multiplex PCR was used to detect genes encoding selected virulence determinants associated with strains of Escherichia coli with K1 antigen (K1(+)) and non-K1 E. coli (K1(-)). The prevalence of the fimA, fimH, sfa/foc, ibeA, iutA and hlyF genes was studied for 134 (67 K1(+) and 67 K1(-)) E. coli strains isolated from pregnant women and neonates. The fimA gene was present in 83.6 % of E. coli K1(+) and in 86.6 % of E. coli K1(-) strains. The fimH gene was present in all tested E. coli K1(+) strains and in 97.0 % of non-K1 strains. E. coli K1(+) strains were significantly more likely to possess the following genes than E. coli K1(-) strains: sfa/foc (37.3 vs 16.4 %, P = 0.006), ibeA (35.8 vs 4.5 %, P<0.001), iutA (82.1 vs 35.8 %, P<0.001) and hlyF (28.4 vs 6.0 %, P<0.001). In conclusion, E. coli K1(+) seems to be more virulent than E. coli K1(-) strains in developing severe infections, thereby increasing possible sepsis or neonatal bacterial meningitis.

  2. The K1 beta-lactamase of Klebsiella pneumoniae.

    PubMed Central

    Joris, B; De Meester, F; Galleni, M; Frère, J M; Van Beeumen, J

    1987-01-01

    beta-Lactamase K1 was purified from Klebsiella pneumoniae SC10436. It is very similar to the enzyme produced by Klebsiella aerogenes 1082E and described by Emanuel, Gagnon & Waley [Biochem. J. (1986) 234, 343-347]. An active-site peptide was isolated after labelling of the enzyme with tritiated beta-iodopenicillanate. A cysteine residue was found just before the active-site serine residue. This result could explain the properties of the enzyme after modification by thiol-blocking reagents. The sequence of the active-site peptide clearly established the enzyme as a class A beta-lactamase. PMID:3307765

  3. Identification of lymphocytic choriomeningitis mammarenavirus in house mouse (Mus musculus, Rodentia) in French Guiana.

    PubMed

    Lavergne, Anne; de Thoisy, Benoît; Tirera, Sourakhata; Donato, Damien; Bouchier, Christiane; Catzeflis, François; Lacoste, Vincent

    2016-01-01

    Thirty-seven house mice (Mus musculus, Rodentia) caught in different localities in French Guiana were screened to investigate the presence of lymphocytic choriomeningitis mammarenavirus (LCMV). Two animals trapped in an urban area were found positive, hosting a new strain of LCMV, that we tentatively named LCMV "Comou". The complete sequence was determined using a metagenomic approach. Phylogenetic analyses revealed that this strain is related to genetic lineage I composed of strains inducing severe disease in humans. These results emphasize the need for active surveillance in humans as well as in house mouse populations, which is a rather common rodent in French Guianese cities and settlements.

  4. Loop quantum cosmology of k=1 FRW models

    SciTech Connect

    Ashtekar, Abhay; Pawlowski, Tomasz; Singh, Parampreet; Vandersloot, Kevin

    2007-01-15

    The closed, k=1, FRW model coupled to a massless scalar field is investigated in the framework of loop quantum cosmology using analytical and numerical methods. As in the k=0 case, the scalar field can be again used as emergent time to construct the physical Hilbert space and introduce Dirac observables. The resulting framework is then used to address a major challenge of quantum cosmology: resolving the big-bang singularity while retaining agreement with general relativity at large scales. It is shown that the framework fulfills this task. In particular, for states which are semiclassical at some late time, the big bang is replaced by a quantum bounce and a recollapse occurs at the value of the scale factor predicted by classical general relativity. Thus, the 'difficulties' pointed out by Green and Unruh in the k=1 case do not arise in a more systematic treatment. As in k=0 models, quantum dynamics is deterministic across the deep Planck regime. However, because it also retains the classical recollapse, in contrast to the k=0 case one is now led to a cyclic model. Finally, we clarify some issues raised by Laguna's recent work addressed to computational physicists.

  5. A new species of Reithrodontomys, subgenus Aporodon (Cricetidae: Neotominae), from the highlands of Costa Rica, with comments on Costa Rican and Panamanian Reithrodontomys

    USGS Publications Warehouse

    Gardner, Alfred L.; Carleton, Michael D.

    2009-01-01

    A new species of the rodent genus Reithrodontomys (Cricetidae: Neotominae) is described from Cerro Asuncion in the western Cordillera de Talamanca, Costa Rica. The long tail, elongate rostrum, bulbous braincase, and complex molars of the new species associate it with members of the subgenus Aporodon, tenuirostris species group. In its diminutive size and aspects of cranial shape, the new species (Reithrodontomys musseri, sp. nov.) most closely resembles R. microdon, a form known from highlands in Guatemala and Chiapas, Mexico. In the course of differentially diagnosing the new species, we necessarily reviewed the Costa Rican and Panamanian subspecies of R. mexicanus based on morphological comparisons, study of paratypes and vouchers used in recent molecular studies, and morphometric analyses. We recognize Reithrodontomys cherrii (Allen, 1891) and R. garichensis finders and Pearson, 1940, as valid species, and allocate R. mexicanus potrerograndei Goodwin, 1945, as a subjective synonym of R. brevirostris Goodwin, 1943. Critical review of museum specimens collected subsequent to Hooper's (1952) revision is needed and would do much to improve understanding of Reithrodontomys taxonomy and distribution in Middle America.

  6. Phylloquinone (Vitamin K1): Occurrence, Biosynthesis and Functions.

    PubMed

    Basset, Gilles J; Latimer, Scott; Fatihi, Abdelhak; Soubeyrand, Eric; Block, Anna

    2016-06-22

    Phylloquinone is a prenylated naphthoquinone that is synthesized exclusively by plants, green algae, and some species of cyanobacteria, where it serves as a vital electron carrier in photosystem I and as an electron acceptor for the formation of protein disulfide bonds. In humans and other vertebrates, phylloquinone plays the role of a vitamin (vitamin K1) that is required for blood coagulation and bone and vascular metabolism. Phylloquinone from green leafy vegetables and vegetable oil represents the major dietary source of vitamin K for humans. In recent years, reverse genetics and biochemical approaches using the model plant Arabidopsis thaliana have shown that phylloquinone biosynthesis in plants involves paralogous and multifunctional enzymes, a compartmentation of the corresponding pathway in plastids and peroxisomes, and trafficking of some biosynthetic intermediates within plastids themselves. Furthermore, phylloquinone biosynthetic intermediates create crucial metabolic branch-points with other plastid-synthesized metabolites such as chlorophylls, tocopherols and salicylate. This review presents an update on recent studies of the central role of plastids in the biosynthesis of phylloquinone, in particular on the discovery of novel enzymatic steps that are likely paradigms for phylloquinone and menaquinone (vitamin K2)-synthesizing organisms alike.

  7. Measurement of Branching Fractions of B0 Decays to K1(1270)+ pi- and K1(1400)+ pi-

    SciTech Connect

    Aubert, Bernard; Bona, M.; Karyotakis, Y.; Lees, J.P.; Poireau, V.; Prencipe, E.; Prudent, X.; Tisserand, V.; Garra Tico, J.; Grauges, E.; Lopez, L.; Palano, Antimo; Pappagallo, M.; Eigen, G.; Stugu, Bjarne; Sun, L.; Abrams, G.S.; Battaglia, M.; Brown, D.N.; Cahn, Robert N.; Jacobsen, R.G.; /LBL, Berkeley /Birmingham U. /Ruhr U., Bochum /Bristol U. /British Columbia U. /Brunel U. /Novosibirsk, IYF /UC, Irvine /UCLA /UC, Riverside /UC, San Diego /UC, Santa Barbara /UC, Santa Cruz /Caltech /Cincinnati U. /Colorado U. /Colorado State U. /Dortmund U. /Dresden, Tech. U. /Ecole Polytechnique /Edinburgh U. /Ferrara U. /INFN, Ferrara /Frascati /Genoa U. /INFN, Genoa /Harvard U. /Heidelberg U. /Humboldt U., Berlin /Imperial Coll., London /Iowa U. /Iowa State U. /Johns Hopkins U. /Orsay, LAL /LLNL, Livermore /Liverpool U. /Queen Mary, U. of London /Royal Holloway, U. of London /Louisville U. /Mainz U., Inst. Kernphys. /Manchester U. /Maryland U. /Massachusetts U., Amherst /MIT /McGill U. /Consorzio Milano Ricerche /INFN, Milan /Mississippi U. /Montreal U. /Mt. Holyoke Coll. /Napoli Seconda U. /INFN, Naples /NIKHEF, Amsterdam /Notre Dame U. /Ohio State U. /Oregon U. /Padua U. /INFN, Padua /Paris U., VI-VII /Pennsylvania U. /Perugia U. /INFN, Perugia /INFN, Pisa /Princeton U. /Banca di Roma /Frascati /Rostock U. /Rutherford /DAPNIA, Saclay /South Carolina U. /SLAC /Stanford U., Phys. Dept. /SUNY, Albany /Tennessee U. /Texas U. /Texas U., Dallas /Turin U. /INFN, Turin /Trieste U. /INFN, Trieste /Valencia U., IFIC /Victoria U. /Warwick U. /Wisconsin U., Madison

    2008-08-04

    We present a measurement of the branching fraction of neutral B meson decaying to final states containing a K1 meson, i.e. K{sub 1}(1270) and K{sub 1}(1400), and a charged pion. The data, collected with the BABAR detector at the Stanford Linear Accelerator Center, represent 454 million B{bar B} pairs produced in e{sup +}e{sup -} annihilation. We measure the branching fraction {Beta}(B{sup 0} {yields} K{sub 1}{sup +}{pi}{sup -}) = (31.0 {+-} 2.7 {+-} 6.9) x 10{sup -6}, where the first error quoted is statistical and the second is systematic. In the framework of the K-matrix formalism used to describe these decays, we also set limits on the ratio of the production constants for the K{sub 1}(1270){sup +} and K{sub 1}(1400){sup +} mesons in B{sup 0} decays.

  8. The KSHV K1 Protein Modulates AMPK Function to Enhance Cell Survival

    PubMed Central

    Anders, Penny M.; Zhang, Zhigang; Bhende, Prasana M.; Giffin, Louise

    2016-01-01

    Kaposi’s sarcoma herpesvirus (KSHV) is the etiologic agent of Kaposi’s sarcoma (KS) as well as two lymphoproliferative diseases, primary effusion lymphoma and multicentric Castleman’s disease. KSHV encodes viral proteins, such as K1, that alter signaling pathways involved in cell survival. Expression of K1 has been reported to transform rodent fibroblasts, and K1 transgenic mice develop multiple tumors, suggesting that K1 has an important role in KSHV pathogenesis. We found that cells infected with a KSHV virus containing a WT K1 gene had a survival advantage under conditions of nutrient deprivation compared to cells infected with KSHV K1 mutant viruses. 5’ adenosine monophosphate-activated protein kinase (AMPK) responds to nutrient deprivation by maintaining energy homeostasis, and AMPK signaling has been shown to promote cell survival in various types of cancers. Under conditions of AMPK inhibition, we also observed that cells infected with KSHV containing a WT K1 gene had a survival advantage compared to KSHV K1 mutant virus infected cells. To explore the underpinnings of this phenotype, we identified K1-associated cellular proteins by tandem affinity purification and mass spectrometry. We found that the KSHV K1 protein associates with the gamma subunit of AMPK (AMPKγ1). We corroborated this finding by independently confirming that K1 co-immunoprecipitates with AMPKγ1. Co-immunoprecipitations of wild-type K1 (K1WT) or K1 domain mutants and AMPKγ1, revealed that the K1 N-terminus is important for the association between K1 and AMPKγ1. We propose that the KSHV K1 protein promotes cell survival via its association with AMPKγ1 following exposure to stress. PMID:27829024

  9. Penguin-dominated B →ϕ K1(1270) and ϕ K1(1400 ) decays in the perturbative QCD approach

    NASA Astrophysics Data System (ADS)

    Liu, Xin; Zou, Zhi-Tian; Xiao, Zhen-Jun

    2014-11-01

    We investigate the C P -averaged branching ratios, the polarization fractions, the relative phases, and the C P -violating asymmetries of the penguin-dominated B →ϕ K1(1270 ) and ϕ K1(1400 ) decays in the perturbative QCD (pQCD) approach, where K1(1270 ) and K1(1400 ) are believed to be the mixtures of two distinct types of axial-vector K1 A(P1 3 ) and K1 B(P1 1 ) states with different behavior, however, their mixing angle θK1is still a hot and controversial topic presently. By numerical evaluations with two different mixing angles θK 1˜3 3∘ and 58° and phenomenological analysis, we find that: (i) the pQCD predictions for the branching ratio, the longitudinal polarization fraction and the direct C P violation of B±→ϕ K1(1270 )±decay with the smaller angle 33° are in good agreement with the currently available data; (ii) though the central values significantly exceed the available upper limit, both pQCD predictions of Br (B±→ϕ K1(1400 )±) with two different mixing angles are consistent with that obtained in QCD factorization and with the preliminary data in 2 σ errors. These results and other relevant predictions for the considered decays will be further tested by the LHCb and the forthcoming Super-B experiments; (iii) the weak annihilation contributions can play an important role in B →ϕ K1(1270 ) and ϕ K1(1400 ) decays; (d) these pQCD predictions combined with the future precision measurements can examine the reliability of the factorization approach employed here, but also explore the complicated QCD dynamics and mixing angle θK 1 of the axial-vector K1(1270 ) and K1(1400 ) system.

  10. 26 CFR 1.162(k)-1 - Disallowance of deduction for reacquisition payments.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... payments. 1.162(k)-1 Section 1.162(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE... and Corporations § 1.162(k)-1 Disallowance of deduction for reacquisition payments. (a) In general... corporation to reacquire its stock from an ESOP that are used in a manner described in section...

  11. 26 CFR 1.501(k)-1 - Communist-controlled organizations.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 7 2010-04-01 2010-04-01 true Communist-controlled organizations. 1.501(k)-1 Section 1.501(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Exempt Organizations § 1.501(k)-1...

  12. 26 CFR 31.3402(k)-1 - Special rule for tips.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 15 2010-04-01 2010-04-01 false Special rule for tips. 31.3402(k)-1 Section 31.3402(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) EMPLOYMENT... Collection of Income Tax at Source § 31.3402(k)-1 Special rule for tips. (a) Withholding of income tax...

  13. 17 CFR 201.550 - Summary suspensions pursuant to Exchange Act Section 12(k)(1)(A).

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... to Exchange Act Section 12(k)(1)(A). 201.550 Section 201.550 Commodity and Securities Exchanges... Suspensions § 201.550 Summary suspensions pursuant to Exchange Act Section 12(k)(1)(A). (a) Petition for termination of suspension. Any person adversely affected by a suspension pursuant to Section 12(k)(1)(A)...

  14. 26 CFR 1.168(k)-1 - Additional first year depreciation deduction.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 26 Internal Revenue 2 2013-04-01 2013-04-01 false Additional first year depreciation deduction. 1.168(k)-1 Section 1.168(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Itemized Deductions for Individuals and Corporations § 1.168(k)-1 Additional first...

  15. 26 CFR 1.168(k)-1 - Additional first year depreciation deduction.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 Internal Revenue 2 2014-04-01 2014-04-01 false Additional first year depreciation deduction. 1.168(k)-1 Section 1.168(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Itemized Deductions for Individuals and Corporations § 1.168(k)-1 Additional first...

  16. 26 CFR 1.168(k)-1 - Additional first year depreciation deduction.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 26 Internal Revenue 2 2011-04-01 2011-04-01 false Additional first year depreciation deduction. 1.168(k)-1 Section 1.168(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Itemized Deductions for Individuals and Corporations § 1.168(k)-1 Additional first...

  17. 26 CFR 1.168(k)-1 - Additional first year depreciation deduction.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 26 Internal Revenue 2 2012-04-01 2012-04-01 false Additional first year depreciation deduction. 1.168(k)-1 Section 1.168(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Itemized Deductions for Individuals and Corporations § 1.168(k)-1 Additional first...

  18. 26 CFR 31.3121(k)-1 - Waiver of exemption from taxes.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 26 Internal Revenue 15 2011-04-01 2011-04-01 false Waiver of exemption from taxes. 31.3121(k)-1 Section 31.3121(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED... § 31.3121(k)-1 Waiver of exemption from taxes. (a) Who may file a waiver certificate—(1) In general....

  19. 26 CFR 31.3121(k)-1 - Waiver of exemption from taxes.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 26 Internal Revenue 15 2013-04-01 2013-04-01 false Waiver of exemption from taxes. 31.3121(k)-1 Section 31.3121(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED... § 31.3121(k)-1 Waiver of exemption from taxes. (a) Who may file a waiver certificate—(1) In general....

  20. 26 CFR 31.3121(k)-1 - Waiver of exemption from taxes.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 15 2010-04-01 2010-04-01 false Waiver of exemption from taxes. 31.3121(k)-1 Section 31.3121(k)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED... § 31.3121(k)-1 Waiver of exemption from taxes. (a) Who may file a waiver certificate—(1) In general....

  1. Pharmacologic targeting of S6K1 in PTEN-deficient neoplasia

    PubMed Central

    Liu, Hongqi; Feng, Xizhi; Ennis, Kelli N.; Behrmann, Catherine A.; Sarma, Pranjal; Jiang, Tony T.; Kofuji, Satoshi; Niu, Liang; Stratton, Yiwen; Thomas, Hala Elnakat; Yoon, Sang-Oh; Sasaki, Atsuo T.; Plas, David R.

    2017-01-01

    SUMMARY Genetic S6K1 inactivation can induce apoptosis in PTEN-deficient cells. We analyzed the therapeutic potential of S6K1 inhibitors in PTEN-deficient T cell leukemia and glioblastoma. Results revealed that the S6K1 inhibitor LY-2779964 was relatively ineffective as a single agent, while S6K1-targeting AD80 induced cytotoxicity selectively in PTEN-deficient cells. In vivo, AD80 rescued 50% of mice transplanted with PTEN-deficient leukemia cells. Cells surviving LY-2779964 treatment exhibited inhibitor-induced S6K1 phosphorylation due to increased mTOR-S6K1 co-association, which primed rapid recovery of S6K1 signaling. In contrast, AD80 avoided S6K1 phosphorylation and mTOR co-association, resulting in durable suppression of S6K1-induced signaling and protein synthesis. Kinome analysis revealed that AD80 coordinately inhibits S6K1 together with the TAM family tyrosine kinase AXL. TAM suppression by BMS-777607 or genetic knockdown potentiated cytotoxic responses to LY-2779964 in PTEN-deficient glioblastoma cells. These results reveal that combination targeting of S6K1 and TAMs is a potential strategy for treatment of PTEN-deficient malignancy. PMID:28249155

  2. P70S6K 1 regulation of angiogenesis through VEGF and HIF-1{alpha} expression

    SciTech Connect

    Bian, Chuan-Xiu; Shi, Zhumei; Meng, Qiao; Jiang, Yue; Liu, Ling-Zhi; Jiang, Bing-Hua

    2010-07-30

    Research highlights: {yields} P70S6K1 regulates VEGF expression; {yields} P70S6K1 induces transcriptional activation through HIF-1{alpha} binding site; {yields} P70S6K1 regulates HIF-1{alpha}, but not HIF-1{beta} protein expression; {yields} P70S6K1 mediates tumor growth and angiogenesis through HIF-1{alpha} and VEGF expression. -- Abstract: The 70 kDa ribosomal S6 kinase 1 (p70S6K1), a downstream target of phosphoinositide 3-kinase (PI3K) and ERK mitogen-activated protein kinase (MAPK), is an important regulator of cell cycle progression, and cell proliferation. Recent studies indicated an important role of p70S6K1 in PTEN-negative and AKT-overexpressing tumors. However, the mechanism of p70S6K1 in tumor angiogenesis remains to be elucidated. In this study, we specifically inhibited p70S6K1 activity in ovarian cancer cells using vector-based small interfering RNA (siRNA) against p70S6K1. We found that knockdown of p70S6K1 significantly decreased VEGF protein expression and VEGF transcriptional activation through the HIF-1{alpha} binding site at its enhancer region. The expression of p70S6K1 siRNA specifically inhibited HIF-1{alpha}, but not HIF-1{beta} protein expression. We also found that p70S6K1 down-regulation inhibited ovarian tumor growth and angiogenesis, and decreased cell proliferation and levels of VEGF and HIF-1{alpha} expression in tumor tissues. Our results suggest that p70S6K1 is required for tumor growth and angiogenesis through HIF-1{alpha} and VEGF expression, providing a molecular mechanism of human ovarian cancer mediated by p70S6K1 signaling.

  3. Angular analysis of B → J/ψK1: Towards a model independent determination of the photon polarization with B → K1γ

    NASA Astrophysics Data System (ADS)

    Kou, E.; Le Yaouanc, A.; Tayduganov, A.

    2016-12-01

    We propose a model independent extraction of the hadronic information needed to determine the photon polarization of the b → sγ process by the method utilizing the B →K1 γ → Kππγ angular distribution. We show that exactly the same hadronic information can be obtained by using the B → J / ψK1 → J / ψKππ channel, which leads to a much higher precision.

  4. S6K1-mediated disassembly of mitochondrial URI/PP1gamma complexes activates a negative feedback program that counters S6K1 survival signaling.

    PubMed

    Djouder, Nabil; Metzler, Stefan Christian; Schmidt, Alexander; Wirbelauer, Christiane; Gstaiger, Matthias; Aebersold, Ruedi; Hess, Daniel; Krek, Wilhelm

    2007-10-12

    S6 kinase 1 (S6K1) acts to integrate nutrient and growth factor signals to promote cell growth but also cell survival as a mitochondria-tethered protein kinase that phosphorylates and inactivates the proapoptotic molecule BAD. Here we report that the prefoldin chaperone URI represents a mitochondrial substrate of S6K1. In growth factor-deprived or rapamycin-treated cells, URI forms stable complexes with protein phosphatase (PP)1gamma at mitochondria, thereby inhibiting the activity of the bound enzyme. Growth factor stimulation induces disassembly of URI/PP1gamma complexes through S6K1-mediated phosphorylation of URI at serine 371. This activates a PP1gamma-dependent negative feedback program that decreases S6K1 activity and BAD phosphorylation, thereby altering the threshold for apoptosis. These findings establish URI and PP1gamma as integral components of an S6K1-regulated mitochondrial pathway dedicated, in part, to oppose sustained S6K1 survival signaling and to ensure that the mitochondrial threshold for apoptosis is set in accord with nutrient and growth factor availability.

  5. Breast milk lymphocyte response to K1 antigen of Escherichia coli.

    PubMed Central

    Keller, M A; Turner, J L; Stratton, J A; Miller, M E

    1980-01-01

    Comparison milk and blood lymphocyte blastogenic responses to the K1 antigen of Escherichia coli and lipopolysaccharide (LPS) from E. coli O127,B8 were examined in 16 postpartum women by [3H]thymidine uptake. Rabbit hemolysincoated sheep erythrocyte monolayers were used to deplete macrophages from milk lymphocyte preparations and to enrich for T lymphocytes in order to make milk preparations more comparable to blood preparations. Response was defined as a stimulation index of greater than or equal to 2.0. There was no evidence of selective response to K1 antigen by milk lymphocytes, since both blood and milk lymphocytes responded in four women and neither blood nor milk lymphocytes responded in nine. Milk lymphocytes alone responded to K1 in one woman, whereas blood lymphocytes alone responded in two women. Additional nonpaired milk or blood cultures were available from three women. None of these responded to K1 antigen. Corresponding lymphocyte cultures were stimulated with LPS. A positive K1 response was always accompanied by an LPS response, and the LPS response correlated with the K1 response in 17 of 19 women. Stool cultures examined with an antiserum agar showed no correlation between the presence of K1 E. coli in the stool and milk or blood lymphocyte response to K1 antigen. In the system used here, no selectivity of response of breast milk lymphocytes to K1 antigen was noted. PMID:6991433

  6. The size of sebaceous glands in relation to the size of hair follicles on the heads of some small mammals (Insectivora, Chiroptera, Rodentia).

    PubMed

    Haffner, M

    1998-04-01

    Many large sebaceous glands have been described, and functions such as scent production suggested, but gland size has seldom been studied in relation to the surface area of their hair follicles. However, investigating this relationship is essential for establishing whether glands produce more sebum than would be required for lubricating associated hair alone. Here, the relationship between sebaceous gland size and the surface area of the associated hair follicles has been studied. Glands on the heads of 20 species from three orders were compared with those associated with fur hairs. In all species, the fur hairs had small hair follicles with small sebaceous glands, as had the mystacial and submandibular hairs of Rodentia. Medium-sized glands and hairs were found in the mystacial and submandibular region in Insectivora, and in the circumoral region in Chiroptera and Rodentia. Large glands and hairs were found on a pad in the corner of the mouth in Rodentia. Although the size of glands was not directly proportional to hair size, some gross trends were noted. Vibrissae had either no or very small sebaceous glands. It is likely that sebum has to be provided from elsewhere to lubricate their surfaces. The glands on the snout of Insectivora and Chiroptera are clearly enlarged and could probably produce more sebum than would be required for grooming vibrissae alone. In Rodentia, vibrissae were surrounded by small hair and glands, and the nearest glands large enough to provide sebum were the glands on the pad in the mouth corner.

  7. Broadening diversity in the Arostrilepis horrida complex: Arostrilepis kontrimavichusi n. sp. (Cyclophyllidea: Hymenolepididae) in the western red-backed vole Myodes californicus (Merriam) (Cricetidae: Arvicolinae) from temperate latitudes of the Pacific Northwest, North America.

    PubMed

    Makarikov, Arseny A; Hoberg, Eric P

    2016-06-01

    Specimens originally identified provisionally as Hymenolepis horrida (Linstow, 1901) [later Arostrilepis horrida (Linstow, 1901)] in Myodes californicus (Merriam) from near the Pacific coastal zone of southern Oregon are revised. Specimens in western red-backed voles represent an undescribed species of Arostrilepis Mas Coma & Tenora, 1997, contributing to recognition and resolution of a broadening complex encompassing cryptic diversity for these hymenolepidid tapeworms distributed across the Holarctic region. Consistent with recent studies defining diversity in the genus, the form, dimensions, and spination (pattern, shape and size) of the cirrus are diagnostic. Among 12 nominal congeners, specimens of A. kontrimavichusi n. sp. are further distinguished by the relative position and length of the cirrus-sac, arrangement of the testes and relative size of the external seminal vesicle and seminal receptacle. Specimens from Oregon voles represent the fifth endemic hymenolepidid in this genus from the Nearctic. Host range for the North American assemblage of species includes Cricetidae (Arvicolinae and Neotominae), Heteromyidae, Geomyidae, and rarely Sciuridae.

  8. A new species, Litomosoides odilae n. sp (Nematoda: Onchocercidae) from Oligoryzomys nigripes (Rodentia: Muridae) in the rainforest of Misiones, Argentina.

    PubMed

    Notarnicola, Juliana; Navone, Graciela

    2002-10-01

    A new species of Litomosoides was collected from the abdominal cavity of Oligoryzomys nigripes (Rodentia: Muridae) in a semideciduous secondary rainforest of Misiones, Argentina. Litomosoides odilae n. sp. belongs to the carinii group and is characterized by the amphids displaced dorsally; buccal capsule with an anterior segment transparent and an annular asymmetrical thickening; esophagus divided, with the posterior glandular portion slightly wider than the muscular; male cloacal aperture strongly protruded; and microfilaria sheathed with an attenuated tail. The morphology of the new species, which is similar to that of L petteri, a parasite of marsupials in Brazil, suggests that host-switching events may have occurred in the diversification of this genus.

  9. A new genus and species of Heligmonellidae (Nematoda: Trichostrongylina) parasitic in Delomys dorsalis (Rodentia: Sigmodontinae) from Misiones, Argentina.

    PubMed

    Digiani, María Celina; Kinsella, John M

    2014-10-01

    Alippistrongylus bicaudatus gen. et sp. n. (Nematoda: Heligmonellidae) is described from the striped Atlantic forest rat, Delomys dorsalis (Hensel) (Rodentia: Sigmodontinae), from the province of Misiones in Argentina. The new genus and species is characterised by a synlophe of 21 unequal ridges in both sexes without a gradient in size, with two ridges weakly sclerotised and oriented perpendicularly in the dorsal left quadrant; males with a highly dissymmetrical bursa with a hypertrophied right lobe, and females with a dorsal conical appendage just posterior to the vulva, conferring a two-tailed appearance to the female worms.

  10. The role of S6K1 in ER-positive breast cancer.

    PubMed

    Holz, Marina K

    2012-09-01

    The 40S ribosomal S6 kinase 1 (S6K1) is a conserved serine/threonine protein kinase that belongs to the AGC family of protein kinases, which also includes Akt and many others. S6K1 is the principal kinase effector downstream of the mammalian target of rapamycin complex 1 (mTORC1). S6K1 is sensitive to a wide range of signaling inputs, including growth factors, amino acids, energy levels and hypoxia. S6K1 relays these signals to regulate a growing list of substrates and interacting proteins in control of oncogenic processes, such as cell growth and proliferation, cell survival and apoptosis and cell migration and invasion. Several lines of evidence suggest an important role for S6K1 in estrogen receptor (ER)-positive breast cancer. S6K1 directly phosphorylates and activates ERα. Furthermore, S6K1 expression is estrogenically regulated. Therefore, hyperactivation of mTORC1/S6K1 signaling may be closely related to ER-positive status in breast cancer and may be utilized as a marker for prognosis and a therapeutic target.

  11. 26 CFR 1.501(k)-1 - Communist-controlled organizations.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 26 Internal Revenue 7 2011-04-01 2009-04-01 true Communist-controlled organizations. 1.501(k)-1...) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Exempt Organizations § 1.501(k)-1 Communist-controlled organizations. Under section 11(b) of the Internal Security Act of 1950 (50 U.S.C. 790(b)), as amended, which...

  12. 26 CFR 1.501(k)-1 - Communist-controlled organizations.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 26 Internal Revenue 7 2013-04-01 2013-04-01 false Communist-controlled organizations. 1.501(k)-1...) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Exempt Organizations § 1.501(k)-1 Communist-controlled organizations. Under section 11(b) of the Internal Security Act of 1950 (50 U.S.C. 790(b)), as amended, which...

  13. An Analysis of the Brigance K & 1 Screen with a Disadvantaged Pre-School Sample.

    ERIC Educational Resources Information Center

    Mantzicopoulos, Panayota; Jarvinen, Denis W.

    The Brigance K&1 test is a battery of 12 subtests widely used to identify entering kindergarten children in need of more comprehensive evaluation. To gather data on the reliability of the test, an analysis was conducted of results for 134 children from low socioeconomic backgrounds. The children were administered both the Brigance K&1

  14. Involvement of S6K1 in mitochondria function and structure in HeLa cells.

    PubMed

    Park, Jisoo; Tran, Quangdon; Mun, Kisun; Masuda, Kouhei; Kwon, So Hee; Kim, Seon-Hwan; Kim, Dong-Hoon; Thomas, George; Park, Jongsun

    2016-12-01

    The major biological function of mitochondria is to generate cellular energy through oxidative phosphorylation. Apart from cellular respiration, mitochondria also play a key role in signaling processes, including aging and cancer metabolism. It has been shown that S6K1-knockout mice are resistant to obesity due to enhanced beta-oxidation, with an increased number of large mitochondria. Therefore, in this report, the possible involvement of S6K1 in regulating mitochondria dynamics and function has been investigated in stable lenti-shS6K1-HeLa cells. Interestingly, S6K1-stably depleted HeLa cells showed phenotypical changes in mitochondria morphology. This observation was further confirmed by detailed image analysis of mitochondria shape. Corresponding molecular changes were also observed in these cells, such as the induction of mitochondrial fission proteins (Drp1 and Fis1). Oxygen consumption is elevated in S6K1-depeleted HeLa cells and FL5.12 cells. In addition, S6K1 depletion leads to enhancement of ATP production in cytoplasm and mitochondria. However, the relative ratio of mitochondrial ATP to cytoplasmic ATP is actually decreased in lenti-shS6K1-HeLa cells compared to control cells. Lastly, induction of mitophagy was found in lenti-shS6K1-HeLa cells with corresponding changes of mitochondria shape on electron microscope analysis. Taken together, our results indicate that S6K1 is involved in the regulation of mitochondria morphology and function in HeLa cells. This study will provide novel insights into S6K1 function in mitochondria-mediated cellular signaling.

  15. The complete mitochondrial genome of Microtus fortis calamorum (Arvicolinae, Rodentia) and its phylogenetic analysis.

    PubMed

    Jiang, Xianhuan; Gao, Jun; Ni, Liju; Hu, Jianhua; Li, Kai; Sun, Fengping; Xie, Jianyun; Bo, Xiong; Gao, Chen; Xiao, Junhua; Zhou, Yuxun

    2012-05-01

    Microtus fortis is a special resource of rodent in China. It is a promising experimental animal model for the study on the mechanism of Schistosome japonicum resistance. The first complete mitochondrial genome sequence for Microtus fortis calamorum, a subspecies of M. fortis (Arvicolinae, Rodentia), was reported in this study. The mitochondrial genome sequence of M. f. calamorum (Genbank: JF261175) showed a typical vertebrate pattern with 13 protein coding genes, 2 ribosomal RNAs, 22 transfer RNAs and one major noncoding region (CR region).The extended termination associated sequences (ETAS-1 and ETAS-2) and conserved sequence block 1 (CSB-1) were found in the CR region. The putative origin of replication for the light strand (O(L)) of M. f. calamorum was 35bp long and showed high conservation in stem and adjacent sequences, but the difference existed in the loop region among three species of genus Microtus. In order to investigate the phylogenetic position of M. f. calamorum, the phylogenetic trees (Maximum likelihood and Bayesian methods) were constructed based on 12 protein-coding genes (except for ND6 gene) on H strand from 16 rodent species. M. f. calamorum was classified into genus Microtus, Arvcicolinae for the highly phylogenetic relationship with Microtus kikuchii (Taiwan vole). Further phylogenetic analysis results based on the cytochrome b gene ranged from M. f. calamorum to one of the subspecies of M. fortis, which formed a sister group of Microtus middendorfii in the genus Microtus.

  16. Paternal behavior and testosterone plasma levels in the Volcano Mouse Neotomodon alstoni (Rodentia: Muridae).

    PubMed

    Luis, Juana; Ramírez, Lorena; Carmona, Agustín; Ortiz, Guadalupe; Delgado, Jesús; Cárdenas, René

    2009-01-01

    Paternal behavior and testosterone plasma levels in the Volcano Mouse Neotomodon alstoni (Rodentia: Muridae). Although initially it was thought that testosterone inhibited the display of paternal behavior in males of rodents, it has been shown that in some species high testosterone levels are needed for exhibition of paternal care. In captivity, males of Volcano Mouse (Neotomodon alstoni) provide pups the same care provided by the mother, with the exception of suckling. Here we measured plasmatic testosterone concentrations 10 days after mating, five and 20 days postpartum, and 10 days after males were isolated from their families in order to determine possible changes in this hormone, associated to the presence and age of pups. Males of Volcano Mouse exhibited paternal behavior when their testosterone levels were relatively high. Although levels of this hormone did not change with the presence or pups age, males that invested more time in huddling showed higher testosterone levels. It is possible that in the Volcano Mouse testosterone modulates paternal behavior indirectly, as in the California mouse.

  17. Description of the karyotype of Rhagomys rufescens Thomas, 1886 (Rodentia, Sigmodontinae) from Southern Brazil Atlantic forest

    PubMed Central

    2010-01-01

    Rhagomys rufescens (Rodentia: Sigmodontinae) is an endemic species of the Atlantic forest from Southern and Southeastern Brazil. Some authors consider Rhagomys as part of the tribe Thomasomyini; but its phylogenetic relationships remain unclear. Chromosomal studies on eight specimens of Rhagomys rufescens revealed a diploid number of 2n = 36 and a number of autosome arms FN = 50. GTG, CBG and Ag-NOR banding and CMA3 /DAPI staining were performed on metaphase chromosomes. Eight biarmed and nine acrocentric pairs were found in the karyotype of this species. The X and Y chromosomes were both acrocentric. Most of the autosomes and the sex chromosomes showed positive C-bands in the pericentromeric region. The X chromosome showed an additional heterochromatic block in the proximal region of the long arm. Nucleolus organizer regions (NORs) were located in the pericentromeric region of three biarmed autosomes (pairs 4, 6 and 8) and in the telomeric region of the short arm of three acrocentrics (pairs 10, 12 and 17). CMA 3 /DAPI staining produced fluorescent signals in many autosomes, especially in pairs 4, 6, and 8. This study presents cytogenetic data of Rhagomys rufescens for the first time. PMID:21637420

  18. Codivergence in heteromyid rodents (Rodentia: heteromyidae) and their sucking lice of the genus Fahrenholzia (Phthiraptera: anoplura).

    PubMed

    Light, Jessica E; Hafner, Mark S

    2008-06-01

    Although most studies of codivergence rely primarily on topological comparisons of host and parasite phylogenies, temporal assessments are necessary to determine if divergence events in host and parasite trees occurred contemporaneously. A combination of cophylogenetic analyses and comparisons of branch lengths are used in this study to understand the host-parasite association between heteromyid rodents (Rodentia: Heteromyidae) and their sucking lice of the genus Fahrenholzia (Phthiraptera: Anoplura). Cophylogenetic comparisons based on nucleotide substitutions in the mitochondrial COI gene reveal a significant, but not perfect, pattern of cophylogeny between heteromyids and their sucking lice. Regression analyses show a significant functional relationship between the lengths of analogous branches in the host and parasite trees, indicating that divergence events in hosts and parasites were approximately contemporaneous. Thus, the topological similarity observed between heteromyids and their lice is the result of codivergence. These analyses also show that the COI gene in lice is evolving two to three times faster than the same gene in their hosts (similar to the results of studies of other lice and their vertebrate hosts) and that divergence events in lice occurred shortly after host divergence. We recommend that future studies of codivergence include temporal comparisons and, when possible, use the same molecular marker(s) in hosts and parasites to achieve the greatest insight into the history of the host-parasite relationship.

  19. Helminth fauna of the Siberian chipmunk, Tamias sibiricus Laxmann (Rodentia, Sciuridae) introduced in suburban French forests.

    PubMed

    Pisanu, Benoît; Jerusalem, Christelle; Huchery, Cindy; Marmet, Julie; Chapuis, Jean-Louis

    2007-05-01

    The spread of an immigrant host species can be influenced both by its specific helminth parasites that come along with it and by newly acquired infections from native fauna. The Siberian chipmunk, Tamias sibiricus Laxmann (Rodentia, Sciuridae), a northeastern Eurasiatic ground nesting Sciurid, has been introduced in France for less than three decades. Thirty individuals were collected from three suburban forests in the Ile-de-France Region between 2002 and 2006. Two intestinal nematode species dominated the helminth fauna: Brevistriata skrjabini [Prevalence, P, 99% C.I., 87% (64-97%); mean intensity, M.I., 99% C.I., 43 (28-78)] and Aonchotheca annulosa [P, 47% (25-69%); M.I., 35 (3-157)]. B. skrjabini is a direct life cycle nematode species of North Eurasiatic origin, with a restricted spectrum of phylogenetically related suitable hosts. This result indicates that B. skrjabini successfully settled and spread with founder pet chipmunks maintained in captivity and released in natura. Chipmunks acquired A. annulosa, a nematode species with a large spectrum of phylogenetically unrelated suitable host species, from local Muroid rodent species with similar behavior, life-history traits and habitats. Quantitative studies are needed to evaluate the potential for both B. skrjabini and A. annulosa to impede the spread of Tamias and for B. skrjabini to favor chipmunk colonization through detrimental effects upon native co-inhabiting host species.

  20. A new species of porcupine, genus Coendou (Rodentia: Erethizontidae) from the Atlantic forest of northeastern Brazil.

    PubMed

    Pontes, Antonio Rossano Mendes; Gadelha, José Ramon; Melo, Éverton R A; de Sá, Fabrício Bezerra; Loss, Ana Carolina; Caldara Junior, Vilacio; Costa, Leonora Pires; Leite, Yuri L R

    2013-01-01

    We report the discovery of a new species of Coendou (Rodentia, Erethizontidae), here designated Coendou speratus sp. nov. This small porcupine, locally known as coandumirim, is found in the Pernambuco Endemism Centre in the Atlantic coast of northeastern Brazil north of the São Francisco river, one of the most important known biodiversity hotspots. The geographic range of C. speratus overlaps with that of the larger, widespread C. prehensilis, but not with that of C. insidiosus from the southeastern Atlantic forest, nor with that of C. nycthemera, an eastern Amazonian species. Coendou speratus is a small-bodied, long-tailed species that appears to be completely spiny because it lacks long dorsal fur. The dorsal quills have conspicuously brownish red tips that contrast with the blackish dorsal background color. The new species is overall similar to C. nycthemera, but the dorsal body quills are typically tricolored in the former and bicolored in the latter. The new species is externally very distinct from C. insidiosus, especially because the latter has bicolored dorsal quills that are almost completely hidden beneath longer and homogeneous pale or dark hairs.

  1. DNA extraction from bristles and quills of Chaetomys subspinosus (Rodentia: Erethizontidae) using a novel protocol.

    PubMed

    Oliveira, C G; Martinez, R A; Gaiotto, F A

    2007-09-30

    DNA extraction protocols are as varied as DNA sources. When it comes to endangered species, it is especially important to pay attention to all details that ensure the completion of the study goals and effectiveness in attaining useful data for conservation. Chaetomys subspinosus (Rodentia: Erethizontidae) is a secretive arboreal porcupine endemic to certain ecosystems of the Brazilian Atlantic Forest. A multidisciplinary study (including genetic data) was performed to create a management plan for the conservation of this species. Individuals from natural populations of the states of Bahia, Espírito Santo and Sergipe were sampled. To obtain a reliable and abundant amount of starting material, non-destructive methods were tested, extracting DNA from the bristles and quills that comprise most of this animal's hide. This method has also been innovative in adapting a DNA extraction protocol traditionally used for plants. Digestion using proteinase K was followed by protein precipitation with CTAB, a chloroform-isoamyl alcohol cleaning and DNA precipitation with isopropyl alcohol. This protocol supplies good-quality DNA for genetic analysis with molecular markers based on PCR.

  2. Conservation of plasmids among Escherichia coli K1 isolates of diverse origins.

    PubMed

    Mercer, A A; Morelli, G; Heuzenroeder, M; Kamke, M; Achtman, M

    1984-12-01

    Escherichia coli K1 isolates of various O types were previously assigned to different clonal groups. Members of the two clones defined by membrane pattern 9 (MP9) and serotypes O18:K1 and O1:K1 had been found to be very similar to each other. The plasmid contents of these bacteria confirmed this conclusion. Both groups carried a self-transmissible plasmid of the FI incompatibility group that coded for colicin production and a major outer membrane protein called the plasmid-coded protein (PCP). The size of this plasmid varied from 76 to 96 megadaltons, but restriction endonuclease digestion and DNA heteroduplex analysis revealed that these plasmids were highly related. O18:K1 bacteria of MP6 had previously been determined to represent a subclone, related to but different from O18:K1 MP9 bacteria. These MP6 bacteria carried a different, smaller IncFI plasmid which did not code for colicin production or the PCP protein. This smaller plasmid was primarily related to the larger plasmid within the regions of DNA encoding incompatibility, replication, and conjugation. O1:K1 bacteria of MP5 contained other unrelated plasmids in agreement with the previous conclusion that they are unrelated to O1:K1 bacteria of MP9. The bacteria examined had been isolated from two continents over a time span of 38 years, and the results attest to conservative inheritance of plasmids within bacteria of common descent.

  3. B→K1π(K) decays in the perturbative QCD approach

    NASA Astrophysics Data System (ADS)

    Zhang, Zhi-Qing; Hou, Zhi-Wei; Yang, Yueling; Sun, Junfeng

    2014-10-01

    Within the framework of the perturbative QCD approach, we study the two-body charmless decays B→K1(1270)(K1(1400))π(K). We find the following results: (i) The decays B¯0→K1(1270)+π-, K1(1400)+π- are incompatible with the present experimental data. There exists a similar situation for the decays B¯0→a1(1260)+K-, b1(1235)+K-, which are usually considered that the nonperturbative contributions are needed to explain the data. But the difference is that the nonperturbative contributions seem to play opposite roles in these two groups of decays. (ii) The pure annihilation type decays B¯0→K1±(1270)K∓, K1±(1400)K∓ are good channels to test whether an approach can be used to calculate correctly the strength of the penguin-annihilation amplitudes. Their branching ratios are predicted at 10-7 order, which are larger than the QCDF results. (iii) The dependence of the direct CP-violating asymmetries of these decays on the mixing angle θK_1 are also considered.

  4. Contribution of vitamin K1 to the electron spin polarization in spinach photosystem I

    SciTech Connect

    Rustandi, R.R.; Snyder, S.W.; Feezel, L.L.; Michalski, T.J.; Norris, J.R.; Thurnauer, M.C.; Biggins, J. )

    1990-09-04

    The electron spin polarized (ESP) electron paramagnetic resonance (EPR) signal observed in spinach photosystem I (PSI) particles was examined in preparations depleted of vitamin K1 by solvent extraction and following biological reconstitution by the quinone. The ESP EPR signal was not detected in the solvent-extracted PSI sample but was restored upon reconstitution with either protonated or deuterated vitamin K1 under conditions that also restored electron transfer to the terminal PSI acceptors. Reconstitution using deuterated vitamin K1 resulted in a line narrowing of the ESP EPR signal, supporting the conclusion that the ESP EPR signals in the reconstituted samples arise from a radical pair consisting of the oxidized PSI primary donor, P700+, and reduced vitamin K1.

  5. Lectin Complement Protein Collectin 11 (CL-K1) and Susceptibility to Urinary Schistosomiasis

    PubMed Central

    Antony, Justin S.; Ojurongbe, Olusola; Kremsner, Peter G.; Velavan, Thirumalaisamy P.

    2015-01-01

    Background Urinary Schistosomiasis is a neglected tropical disease endemic in many sub Saharan -African countries. Collectin Kidney 1 (CL-K1, encoded by COLEC11 on chromosome 2p25.3), a member of the vertebrate C-type lectin super family, has recently been identified as pattern-recognition molecule (PRR) of the lectin complement pathway. CL-K1 is preferentially expressed in the kidneys, but also in other organs and it is considered to play a role in host defense to some infectious agents. Schistosome teguments are fucosylated and CL-K1 has, through its collagen-like domain, a high binding affinity to fucose. Methodology/Principal Findings We utilized a Nigerian study group consisting of 167 Schistosoma haematobium infected individuals and 186 matched healthy subjects, and investigated the contribution of CL-K1 deficiency and of COLEC11 polymorphisms to infection phenotype. Higher CL-K1 serum levels were associated with decreased risk of schistosome infection (Pcorr = 0.0004). CL-K1 serum levels were differentially distributed between the COLEC11 genotypes and haplotypes observed. The non-synonymous variant p.R216H was associated with the occurrence of schistosomiasis (OR = 0.44, 95%CI = 0.22–0.72, Pcorr = 0.0004). The reconstructed COLEC11*TCCA haplotypes were associated with higher CL-K1 serum levels (P = 0.002) and with decreased schistosomiasis (OR = 0.38, 95%CI = 0.23–0.63, Pcorr = 0.0001). Conclusions In agreement with findings from our earlier published study, our findings support the observation that CL-K1 and their functional variants may be host factors associated with protection in schistosomiasis and may be a useful marker for further investigations. PMID:25807310

  6. Prediction of 3-hydroxypyridin-4-one (HPO) log K1 values for Fe(III).

    PubMed

    Chen, Yu-Lin; Barlow, Dave J; Kong, Xiao-Le; Ma, Yong-Min; Hider, Robert C

    2012-09-21

    As a means to aid in the design of 3-hydroxypyridin-4-ones (HPOs) intended for use as therapeutic Fe(3+) chelating agents, a novel methodology has been developed using quantum mechanical (QM) calculations for predicting the iron binding affinities of the compounds (more specifically, their log K(1) values). The reported/measured HPO log K(1) values were verified through their correlation with the corresponding sum of the compounds' ligating group pK(a) values. Using a training set of eleven HPOs with known log K(1) values, reliable predictions are shown to be obtained with QM calculations using the B3LYP/6-31+G(d)/CPCM model chemistry (with Bondi radii, and water as solvent). With this methodology, the observed log K(1) values for the training set compounds are closely matched by the predicted values, with the correlation between the observed and predicted values giving r(2) = 0.9. Predictions subsequently made by this method for a test set of 42 HPOs of known log K(1) values gave predicted values accurate to within ±0.32 log units. In order to further investigate the predictive power of the method, four novel HPOs were synthesised and their log K(1) values were determined experimentally. Comparison of these predicted log K(1) values against the measured values gave absolute deviations of 0.22 (13.87 vs. 14.09), 0.02 (14.31 vs. 14.29), 0.12 (14.62 vs. 14.50), and 0.13 (15.04 vs. 15.17). The prediction methodology reported here is the first to be provided for predicting the absolute log K(1) values of iron-chelating agents in the absence of pK(a) values.

  7. The lipid kinase PIP5K1C regulates pain signaling and sensitization

    PubMed Central

    Wright, Brittany D.; Loo, Lipin; Street, Sarah E.; Ma, Anqi; Taylor-Blake, Bonnie; Stashko, Michael A.; Jin, Jian; Janzen, William P.; Frye, Stephen V.; Zylka, Mark J.

    2014-01-01

    SUMMARY Numerous pain-producing (pronociceptive) receptors signal via phosphatidylinositol 4,5- bisphosphate (PIP2) hydrolysis. However, it is currently unknown which lipid kinases generate PIP2 in nociceptive dorsal root ganglia (DRG) neurons and if these kinases regulate pronociceptive receptor signaling. Here, we found that phosphatidylinositol 4-phosphate 5 kinase type 1C (PIP5K1C) is expressed at higher levels than any other PIP5K and, based on experiments with Pip5k1c+/− mice, generates at least half of all PIP2 in DRG neurons. Additionally, Pip5k1c haploinsufficiency reduces pronociceptive receptor signaling and TRPV1 sensitization in DRG neurons as well as thermal and mechanical hypersensitivity in mouse models of chronic pain. We identified a novel small molecule inhibitor of PIP5K1C (UNC3230) in a high-throughput screen. UNC3230 lowered PIP2 levels in DRG neurons and attenuated hypersensitivity when administered intrathecally or into the hindpaw. Our studies reveal that PIP5K1C regulates PIP2- dependent nociceptive signaling and suggest that PIP5K1C is a novel therapeutic target for chronic pain. PMID:24853942

  8. Autonomous safety and reliability features of the K-1 avionics system

    NASA Astrophysics Data System (ADS)

    Mueller, George E.; Kohrs, Dick; Bailey, Richard; Lai, Gary

    2004-03-01

    Kistler Aerospace Corporation is developing the K-1, a fully reusable, two-stage-to-orbit launch vehicle. Both stages return to the launch site using parachutes and airbags. Initial flight operations will occur from Woomera, Australia. K-1 guidance is performed autonomously. Each stage of the K-1 employs a triplex, fault tolerant avionics architecture, including three fault tolerant computers and three radiation hardened Embedded GPS/INS units with a hardware voter. The K-1 has an Integrated Vehicle Health Management (IVHM) system on each stage residing in the three vehicle computers based on similar systems in commercial aircraft. During first-stage ascent, the IVHM system performs an Instantaneous Impact Prediction (IIP) calculation 25 times per second, initiating an abort in the event the vehicle is outside a predetermined safety corridor for at least 3 consecutive calculations. In this event, commands are issued to terminate thrust, separate the stages, dump all propellant in the first-stage, and initiate a normal landing sequence. The second-stage flight computer calculates its ability to reach orbit along its state vector, initiating an abort sequence similar to the first stage if it cannot. On a nominal mission, following separation, the second-stage also performs calculations to assure its impact point is within a safety corridor. The K-1's guidance and control design is being tested through simulation with hardware-in-the-loop at Draper Laboratory. Kistler's verification strategy assures reliable and safe operation of the K-1.

  9. A RING to rule them all? Insights into the Map3k1 PHD motif provide a new mechanistic understanding into the diverse roles of Map3k1.

    PubMed

    Suddason, T; Gallagher, E

    2015-04-01

    Despite the sizable number of components that comprise Mapk cascades, Map3k1 is the only element that contains both a kinase domain and a plant homeodomain (PHD) motif, allowing Map3k1 to regulate the protein phosphorylation and ubiquitin proteasome systems. As such, Map3k1 has complex roles in the regulation of cell death, survival, migration and differentiation. Numerous mouse and human genetic analyses have demonstrated that Map3k1 is of critical importance for the immune system, cardiac tissue, testis, wound healing, tumorigenesis and cancer. Recent gene knockin of Map3k1 to mutate the E2 binding site within the Map3k1 PHD motif and high throughput ubiquitin protein array screening for Map3k1 PHD motif substrates provide critical novel insights into Map3k1 PHD motif signal transduction and bring a brand-new understanding to Map3k1 signaling in mammalian biology.

  10. KEY COMPARISON: Force key comparison CCM.F-K1.a and CCM.F-K1.b: 5 kN and 10 kN

    NASA Astrophysics Data System (ADS)

    Pusa, Aimo

    2009-01-01

    This report describes the key comparisons named CCM.F-K1.a and CCM.F-K1.b, for force with loads of 5 kN and 10 kN. The Draft A Report, reporting the measurement results of the key comparisons, has been accepted at the force expert group meeting in Pretoria on 23 March 2004. This became Part 1 of the preliminary Draft B Report. Then, there have been several discussions to find the best way for the determination of the reference value for force values 5 kN and 10 kN in the key comparisons CCM.F-K1.a and CCM.F-K1.b. Following the meeting held in Queretaro, Mexico, by CENAM, from 3 to 5 December 2007, the reference values have been calculated for each single transducer (see chapter 2), and as one reference value for 5 kN and a second reference value for 10 kN (see chapter 3). To get a better consistency a linear model for the drift of transducers has been applied. The results have been evaluated according to the paper from M G Cox, 'The evaluation of key comparison data' (2002 Metrologia 39 589-595). Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCM, according to the provisions of the CIPM Mutual Recognition Arrangement (MRA).

  11. Postnatal ontogeny of limb proportions and functional indices in the subterranean rodent Ctenomys talarum (Rodentia: Ctenomyidae).

    PubMed

    Echeverría, Alejandra Isabel; Becerra, Federico; Vassallo, Aldo Iván

    2014-08-01

    Burrow construction in the subterranean Ctenomys talarum (Rodentia: Ctenomyidae) primarily occurs by scratch-digging. In this study, we compared the limbs of an ontogenetic series of C. talarum to identify variation in bony elements related to fossorial habits using a morphometrical and biomechanical approach. Diameters and functional lengths of long bones were measured and 10 functional indices were constructed. We found that limb proportions of C. talarum undergo significant changes throughout postnatal ontogeny, and no significant differences between sexes were observed. Five of six forelimb indices and two of four hindlimb indices showed differences between ages. According to discriminant analysis, the indices that contributed most to discrimination among age groups were robustness of the humerus and ulna, relative epicondylar width, crural and brachial indices, and index of fossorial ability (IFA). Particularly, pups could be differentiated from juveniles and adults by more robust humeri and ulnae, wider epicondyles, longer middle limb elements, and a proportionally shorter olecranon. Greater robustness indicated a possible compensation for lower bone stiffness while wider epicondyles may be associated to improved effective forces in those muscles that originate onto them, compensating the lower muscular development. The gradual increase in the IFA suggested a gradual enhancement in the scratch-digging performance due to an improvement in the mechanical advantage of forearm extensors. Middle limb indices were higher in pups than in juveniles-adults, reflecting relatively more gracile limbs in their middle segments, which is in accordance with their incipient fossorial ability. In sum, our results show that in C. talarum some scratch-digging adaptations are already present during early postnatal ontogeny, which suggests that they are prenatally shaped, and other traits develop progressively. The role of early digging behavior as a factor influencing on

  12. Skeletal myocyte hypertrophy requires mTOR kinase activity and S6K1

    SciTech Connect

    Park, In-Hyun . E-mail: ihpark@uiuc.edu; Erbay, Ebru; Nuzzi, Paul; Chen Jie

    2005-09-10

    The protein kinase mammalian target of rapamycin (mTOR) is a central regulator of cell proliferation and growth, with the ribosomal subunit S6 kinase 1 (S6K1) as one of the key downstream signaling effectors. A critical role of mTOR signaling in skeletal muscle differentiation has been identified recently, and an unusual regulatory mechanism independent of mTOR kinase activity and S6K1 is revealed. An mTOR pathway has also been reported to regulate skeletal muscle hypertrophy, but the regulatory mechanism is not completely understood. Here, we report the investigation of mTOR's function in insulin growth factor I (IGF-I)-induced C2C12 myotube hypertrophy. Added at a later stage when rapamycin no longer had any effect on normal myocyte differentiation, rapamycin completely blocked myocyte hypertrophy as measured by myotube diameter. Importantly, a concerted increase of average myonuclei per myotube was observed in IGF-I-stimulated myotubes, which was also inhibited by rapamycin added at a time when it no longer affected normal differentiation. The mTOR protein level, its catalytic activity, its phosphorylation on Ser2448, and the activity of S6K1 were all found increased in IGF-I-stimulated myotubes compared to unstimulated myotubes. Using C2C12 cells stably expressing rapamycin-resistant forms of mTOR and S6K1, we provide genetic evidence for the requirement of mTOR and its downstream effector S6K1 in the regulation of myotube hypertrophy. Our results suggest distinct mTOR signaling mechanisms in different stages of skeletal muscle development: While mTOR regulates the initial myoblast differentiation in a kinase-independent and S6K1-independent manner, the hypertrophic function of mTOR requires its kinase activity and employs S6K1 as a downstream effector.

  13. Metaphase chromosome and nucleoid differences between CHO-K1 and its radiosensitive derivative xrs-5

    SciTech Connect

    Schwartz, J.L. |; Cowan, J.M.; Moan, E.; Sedita, B.A.; Stephens, J.; Vaughan, A.T.M.

    1992-05-01

    The Chinese hamster ovary (CHO) cell line xrs-5 is a radiation-sensitive mutant isolated from CHO-K1 cells. The radiation sensitivity is associated with a defect in DNA double-strand break rejoining. Chromatin structure also appears altered in xrs-5 cells compared to the parental CHO-K1 cells. Metaphase chromosomes from xrs-5 are more condensed in appearance than CHO-K1 chromosomes. The overcondensed look is not the result of colcemid sensitivity. Electron microscopy studies suggest that xrs-5 metaphase chromosomes have larger loops of chromatin extending out from the chromosome core. There are also differences between CHO-K1 and xrs-5 cells in the size and fluorescence pattern of ethidium bromide-stained nucleoid preparations. These results suggest that there is a fundamental difference between CHO-K1 and xrs-5 in either the organization of the supercoiled loops of DNA attached to the nuclear matrix or in the nature of the proteins that attach the DNA to the matrix. These alterations in chromosome structure may underlie, in part, the radiation sensitivity of xrs-5 cells.

  14. PHLPP-mediated dephosphorylation of S6K1 inhibits protein translation and cell growth.

    PubMed

    Liu, Jianyu; Stevens, Payton D; Li, Xin; Schmidt, Micheal D; Gao, Tianyan

    2011-12-01

    PHLPP is a family of Ser/Thr protein phosphatases that contains PHLPP1 and PHLPP2 isoforms. We have shown previously that PHLPP functions as a tumor suppressor by negatively regulating Akt signaling in cancer cells. Here we report the identification of ribosomal protein S6 kinase 1 (S6K1) as a novel substrate of PHLPP. Overexpression of both PHLPP isoforms resulted in a decrease in S6K1 phosphorylation in cells, and this PHLPP-mediated dephosphorylation of S6K1 was independent of its ability to dephosphorylate Akt. Conversely, S6K1 phosphorylation was increased in cells depleted of PHLPP expression. Furthermore, we showed that the insulin receptor substrate 1 (IRS-1) expression and insulin-induced Akt phosphorylation were significantly decreased as the result of activation of the S6K-dependent negative feedback loop in PHLPP knockdown cells. Functionally, the phosphorylation of ribosomal protein S6 (rpS6) and the amount of phosphorylated rpS6 bound to the translation initiation complex were increased in PHLPP-knockdown cells. This correlated with increased cell size, protein content, and rate of cap-dependent translation. Taken together, our results demonstrate that loss of PHLPP expression activates the S6K-dependent negative feedback loop and that PHLPP is a novel player involved in regulating protein translation initiation and cell size via direct dephosphorylation of S6K1.

  15. Determination of ultimate carbonaceous BOD and the specific rate constant (K1)

    USGS Publications Warehouse

    Stamer, J.K.; Bennett, J.P.; McKenzie, Stuart W.

    1982-01-01

    Ultimate carbonaceous biochemical oxygen demand (BODu) and the specific rate constant (K1) at which the demand is exerted are important parameters in designing biological wastewater treatment plants and in assessing the impact of wastewater on receiving streams. An analytical method is presented which uses time-series concentrations of BOD, defined as the calculated sum of dissolved oxygen (DO) losses at each time of measurement, for determining BODu and K1. Time-series DO measurements are obtained from a water sample that is incubated in darkness at 20 degrees Celsius in the presence of nitrapyrin, a chemical nitrification inhibitor. Time-series concentrations of BOD that approximate first order kinetics can be analyzed graphically or mathematically to compute BODu and K1.

  16. Novel assist feature design to improve depth of focus in low k1 EUV lithography

    NASA Astrophysics Data System (ADS)

    Kang, Hoyoung

    2009-12-01

    With the expected continual progress of micro-electronics scaling, low k1 techniques may be required even with EUV lithography. One of important techniques of low k1, the off axis illumination (OAI) in combination with sub-resolution assist features (SRAF) on reticles, has been used extensively in optical lithography. Use of assist features combined with off axis illumination typically requires extremely small pattern sizes. The assist pattern enables printing dense and isolated lines simultaneous. In a low k1 region of around 0.4, assist features will increase depth of focus (DOF) of isolated and semi-isolated lines even in EUV. Since EUVL process operates at a relatively higher k1 value than that for the optical lithography, the assist feature size needed is relatively smaller. In addition, with the mask shadowing effect of EUVL, all horizontal lines should be biased thinner by a couple of nanometers, and horizontal assist features will need to do the same. Fabricating such narrow features on masks is challenging, and could potentially limit the application of SRAF in EUVL in the low k1 regime. A novel approach is proposed to create assist features with similar width as the main critical dimension features. The proposed technique creates assist patterns using thinner absorber which would have higher reflectance than normal absorber. Thinner absorber assist pattern can perform similarly with narrower assist pattern and easier to fabricate. With off axis illumination in EUVL and assist patterns, process margin of semi-isolated and isolated lines can be increased for k1 lower than 0.4.

  17. Characterization of S6K2, a novel kinase homologous to S6K1.

    PubMed

    Lee-Fruman, K K; Kuo, C J; Lippincott, J; Terada, N; Blenis, J

    1999-09-09

    Rapamycin is an immunosuppressant which antagonizes cellular proliferation by inhibiting the function of mTOR. The mTOR:FKBP12: rapamycin complex blocks G1/S transition by inhibiting downstream targets essential for cell cycle progression. One such target is p70S6k1 (S6K1), a serine/threonine kinase which is inactivated by the mTOR : FKBP12 : rapamycin complex, and which has been linked to translational control by virtue of its ability to phosphorylate the ribosomal protein S6. In the current work, we describe cloning and characterization of a novel S6K1 homolog, p54 S6 kinase 2 (p54S6k2/S6K2). Similar to S6K1, S6K2 is activated by mitogens and by constitutively active PI3K, and is inhibited by rapamycin as well as wortmannin. Differences between activation of S6K1 and S6K2 by PDK1 were observed, suggesting potential differences in the regulation of these homologs. Strikingly, S6K2 activity and S6 phosphorylation were both intact in S6K1-/-ES cell, indicating a possible role for S6K2 in in vivo S6 phosphorylation. Interestingly, we found two isoforms of S6K2 which are localized to distinct cellular compartments; the smaller form resides in the detergent-soluble fraction, whereas the larger form is found in the particulate fraction. Our findings demonstrate the existence of a family of rapamycin-sensitive protein kinases potentially involved in S6 phosphorylation, translational control, and transduction of mTOR signals.

  18. Characterization of the mouse ClC-K1/Barttin chloride channel.

    PubMed

    L'Hoste, Sébastien; Diakov, Alexei; Andrini, Olga; Genete, Mathieu; Pinelli, Laurent; Grand, Teddy; Keck, Mathilde; Paulais, Marc; Beck, Laurent; Korbmacher, Christoph; Teulon, Jacques; Lourdel, Stéphane

    2013-11-01

    Several Cl(-) channels have been described in the native renal tubule, but their correspondence with ClC-K1 and ClC-K2 channels (orthologs of human ClC-Ka and ClC-Kb), which play a major role in transcellular Cl(-) absorption in the kidney, has yet to be established. This is partly because investigation of heterologous expression has involved rat or human ClC-K models, whereas characterization of the native renal tubule has been done in mice. Here, we investigate the electrophysiological properties of mouse ClC-K1 channels heterologously expressed in Xenopus laevis oocytes and in HEK293 cells with or without their accessory Barttin subunit. Current amplitudes and plasma membrane insertion of mouse ClC-K1 were enhanced by Barttin. External basic pH or elevated calcium stimulated currents followed the anion permeability sequence Cl(-)>Br(-)>NO3(-)>I(-). Single-channel recordings revealed a unit conductance of ~40pS. Channel activity in cell-attached patches increased with membrane depolarization (voltage for half-maximal activation: ~-65mV). Insertion of the V166E mutation, which introduces a glutamate in mouse ClC-K1, which is crucial for channel gating, reduced the unit conductance to ~20pS. This mutation shifted the depolarizing voltage for half-maximal channel activation to ~+25mV. The unit conductance and voltage dependence of wild-type and V166E ClC-K1 were not affected by Barttin. Owing to their strikingly similar properties, we propose that the ClC-K1/Barttin complex is the molecular substrate of a chloride channel previously detected in the mouse thick ascending limb (Paulais et al., J Membr. Biol, 1990, 113:253-260).

  19. Emergence of Carbapenem-Resistant Serotype K1 Hypervirulent Klebsiella pneumoniae Strains in China.

    PubMed

    Zhang, Rong; Lin, Dachuan; Chan, Edward Wai-Chi; Gu, Danxia; Chen, Gong-Xiang; Chen, Sheng

    2015-11-16

    We report the emergence of five carbapenem-resistant K1 hypervirulent Klebsiella pneumoniae (hvKP) strains which caused fatal infections in hospital patients in Zhejiang Province, China, upon entry through surgical wounds. Genotyping results revealed the existence of three genetically related strains which exhibited a new sequence type, ST1797, and revealed that all strains harbored the magA and wcaG virulence genes and a plasmid-borne bla(KPC-2) gene. These findings indicate that K1 hvKP is simultaneously hypervirulent, multidrug resistant, and transmissible.

  20. Emergence of Carbapenem-Resistant Serotype K1 Hypervirulent Klebsiella pneumoniae Strains in China

    PubMed Central

    Zhang, Rong; Lin, Dachuan; Chan, Edward Wai-chi; Gu, Danxia

    2015-01-01

    We report the emergence of five carbapenem-resistant K1 hypervirulent Klebsiella pneumoniae (hvKP) strains which caused fatal infections in hospital patients in Zhejiang Province, China, upon entry through surgical wounds. Genotyping results revealed the existence of three genetically related strains which exhibited a new sequence type, ST1797, and revealed that all strains harbored the magA and wcaG virulence genes and a plasmid-borne blaKPC-2 gene. These findings indicate that K1 hvKP is simultaneously hypervirulent, multidrug resistant, and transmissible. PMID:26574010

  1. Cyclic sulfoxides-garlicnins K1, K2, and H1-extracted from Allium sativum.

    PubMed

    Nohara, Toshihiro; Fujiwara, Yukio; Komota, Yusuke; Kondo, Yoshihiko; Saku, Taiki; Yamaguchi, Koki; Komohara, Yoshihiro; Takeya, Motohiro

    2015-01-01

    Newly identified cyclic sulfoxides-garlicnins K1 (1), K2 (2), and H1 (3)-were isolated from the acetone extracts of the bulbs of garlic, Allium sativum. Garlicnin H1 (3) demonstrated potential to suppress tumor cell proliferation by regulating macrophage activation. The structures of garlicnins K1 and K2, 3,4-dimethyl-5-allyl-tetrahydrothiophen-2-one-S-oxides, and the structure of garlicnin H1, 3-carboxy-3-hydroxy-4-methyl-5-allylsulfoxide-tetrahydrothiophen-2-(ethane-1,2-diol)-S-oxide were characterized by spectroscopic analysis.

  2. Redescription of Trichuris pampeana (Nematoda: Trichuridae) from the South American subterranean rodent Ctenomys talarum Thomas, 1898 (Rodentia: Octodontidae).

    PubMed

    Rossin, M Alejandra; Malizia, Ana I

    2005-02-01

    Trichuris pampeana Suriano and Navone, 1994 (Nematoda: Trichuridae) is redescribed from voucher specimens from the type host Ctenomys azarae Thomas, 1903 (Rodentia: Octodontidae) and from parasites collected from 2 populations of the subterranean rodent C. talarum Thomas, 1898 from Mar de Cobo and Necochea, Buenos Aires Province, Argentina. After a revision of these nematodes, it was confirmed that the following characters were not considered in the original description: bacillary band, cells from the esophagointestinal junction, ejaculatory duct, vas deferens, adanal papillae, vagina, oviduct, and rectum. Additional information about the spicular sheath, vulva, uteri, and ovary is provided. The morphological features given in this redescription allow to confirm the identity of T. pampeana as a valid species and also to distinguish it more clearly from other species of the genus.

  3. Squamasnema amazonica n. gen. n. sp. (Heligmonellinae): A new parasite of Proechimys roberti (Rodentia: Echimyidae) in the Brazilian Amazon.

    PubMed

    Cordeiro, Helrik da Costa; Melo, Francisco Tiago de Vasconcelos; Furtado, Adriano Penha; Giese, Elane Guerreiro; Maldonado, Arnaldo; dos Santos, Jeannie Nascimento

    2015-08-01

    A new species of nematode, Squamasnema amazonica n. gen. n. sp., is described based on specimens found parasitizing the small intestine of Proechimys roberti (Rodentia: Echimyidae) collected during a survey of the fauna of Tapirapé-Aquirí National Forest (Brazil, Eastern Brazilian Amazon). The nematodes were fixed and processed for light microscopy and scanning electron microscopy (SEM). These nematodes were classified under the family Heligmonellidae and the subfamily Heligmonellinae. Although several species in the family Heligmonellidae exhibit discontinuous ridges, Squamasnema n. gen. and Trichotravassosia are the only genera with columns of scales along their entire body, as an apomorphy of the synlophe. Squamasnema n. gen. has columns of cuticular cells along its body, except for on the left flank, and exhibits a synlophe with no size gradient or inclination and does not present chitinized structures supporting the synlophe. Therefore, due to these morphological differences of Squamasnema n. gen., the creation of a new genus was necessary.

  4. 77 FR 64848 - Proposed Collection; Comment Request for Form 1120S, Schedule D, Schedule K-1, and Schedule M-3

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-10-23

    ... Internal Revenue Service Proposed Collection; Comment Request for Form 1120S, Schedule D, Schedule K-1, and... With Total Assets of $10 Million or More, and Schedule K-1 (Form 1120S), Shareholder's Share of Income... Losses and Built-in Gains, Schedule K-1 (Form 1120S), Shareholder's Share of Income, Credits,...

  5. 26 CFR 1.404(k)-1T - Questions and answers relating to the deductibility of certain dividend distributions. (Temporary)

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... deductibility of certain dividend distributions. (Temporary) 1.404(k)-1T Section 1.404(k)-1T Internal Revenue... Pension, Profit-Sharing, Stock Bonus Plans, Etc. § 1.404(k)-1T Questions and answers relating to the deductibility of certain dividend distributions. (Temporary) Q-1: What does section 404(k) provide? A-1:...

  6. Crystal structures of the S6K1 kinase domain in complexes with inhibitors.

    PubMed

    Niwa, Hideaki; Mikuni, Junko; Sasaki, Shunta; Tomabechi, Yuri; Honda, Keiko; Ikeda, Mariko; Ohsawa, Noboru; Wakiyama, Motoaki; Handa, Noriko; Shirouzu, Mikako; Honma, Teruki; Tanaka, Akiko; Yokoyama, Shigeyuki

    2014-09-01

    Ribosomal protein S6 kinase 1 (S6K1) is a serine/threonine protein kinase that plays an important role in the PIK3/mTOR signaling pathway, and is implicated in diseases including diabetes, obesity, and cancer. The crystal structures of the S6K1 kinase domain in complexes with staurosporine and the S6K1-specific inhibitor PF-4708671 have been reported. In the present study, five compounds (F108, F109, F176, F177, and F179) were newly identified by in silico screening of a chemical library and kinase assay. The crystal structures of the five inhibitors in complexes with the S6K1 kinase domain were determined at resolutions between 1.85 and 2.10 Å. All of the inhibitors bound to the ATP binding site, lying along the P-loop, while the activation loop stayed in the inactive form. Compound F179, with a carbonyl group in the middle of the molecule, altered the αC helix conformation by interacting with the invariant Lys123. Compounds F176 and F177 bound slightly distant from the hinge region, and their sulfoamide groups formed polar interactions with the protein. The structural features required for the specific binding of inhibitors are discussed.

  7. Structural and kinetic studies on beta-lactamase K1 from Klebsiella aerogenes.

    PubMed Central

    Emanuel, E L; Gagnon, J; Waley, S G

    1986-01-01

    beta-Lactamase K1 from Klebsiella aerogenes 1082E hydrolyses both penicillins and cephalosporins comparably and is inhibited by mercurials but not by cloxacillin. These properties distinguish it from those other beta-lactamases that have been allotted to classes on the basis of their amino sequences. beta-Lactamase K1 has been isolated by affinity chromatography; its composition shows resemblances to class A beta-lactamases. Moreover, the N-terminal sequence is similar to those of class A beta-lactamases: there is about 30% identity over the first 32 residues. Furthermore, a putative active-site octapeptide has been isolated and its sequence is similar to the region around the active-site serine residue in class A beta-lactamases. There is one thiol group in beta-lactamase K1; it is not essential for activity. The pH-dependence of kcat. and kcat./Km for the hydrolysis of benzylpenicillin by beta-lactamase K1 were closely similar, suggesting that the rate-determining step is cleavage of the beta-lactam ring. PMID:3521585

  8. The Greatest Educational Change America Has Ever Seen, 2002: Lesson Plans for Grades K-1.

    ERIC Educational Resources Information Center

    United States Mint (Dept. of Treasury), Washington, DC.

    This teacher's guide on the 50 state quarters emitted by the United States Mint includes 6 "teacher-friendly" lesson plans that fit easily into the curriculum of grades K-1; reproducible student worksheets that coincide with each lesson; "fun" state facts and information on the new quarter designs; and USA map template with…

  9. The Greatest Educational Change America Has Ever Seen. [Teaching Guide]. Grades K-1.

    ERIC Educational Resources Information Center

    United States Mint (Dept. of Treasury), Washington, DC.

    This teaching guide for grades K-1 focuses on the 1999-2000 United States Mint 50 State Quarters Program, which includes new quarter designs for the following states: Delaware, Pennsylvania, New Jersey, Georgia, Connecticut, Massachusetts, Maryland, South Carolina, New Hampshire, and Virginia. The guide includes six lesson plans that fit easily…

  10. The Greatest Educational Change America Has Ever Seen, 2001: Lesson Plans for Grades K-1.

    ERIC Educational Resources Information Center

    United States Mint (Dept. of Treasury), Washington, DC.

    This teacher's guide about the 50 state quarters produced by the United States Mint includes 6 lesson plans that fit easily into the social studies curriculum for grades K-1. The lesson plans include reproducible student work pages that coincide with each lesson; state facts and information about the new 2001 state quarter designs (New York, North…

  11. K-1 Teachers' Visual Arts Beliefs and Their Role in the Early Childhood Classroom

    ERIC Educational Resources Information Center

    Goodman-Schanz, Blythe Annette

    2012-01-01

    The purpose of this qualitative study was to explore and describe the visual arts beliefs and practices of eight K-1 teachers in four schools and in two different school districts in a southern state. Using a phenomenological framework (Creswell, 2007; Leedy & Ormrod, 2005), the research revealed the teachers' understandings of beliefs and how…

  12. easyCBM® Reading Criterion Related Validity Evidence: Grades K-1. Technical Report #1309

    ERIC Educational Resources Information Center

    Lai, Cheng-Fei; Alonzo, Julie; Tindal, Gerald

    2013-01-01

    In this technical report, we present the results of a study to gather criterion-related evidence for Grade K-1 easyCBM® reading measures. We used correlations to examine the relation between the easyCBM® measures and other published measures with known reliability and validity evidence, including the Dynamic Indicators of Basic Early Literacy…

  13. Analysis of synonymous codon usage in Aeropyrum pernix K1 and other Crenarchaeota microorganisms.

    PubMed

    Jiang, Peng; Sun, Xiao; Lu, Zuhong

    2007-03-01

    In this study, a comparative analysis of the codon usage bias was performed in Aeropyrum pernix K1 and two other phylogenetically related Crenarchaeota microorganisms (i.e., Pyrobaculum aerophilum str. IM2 and Sulfolobus acidocaldarius DSM 639). The results indicated that the synonymous codon usage in A. pernix K1 was less biased, which was highly correlated with the GC(3S) value. The codon usage patterns were phylogenetically conserved among these Crenarchaeota microorganisms. Comparatively, it is the species function rather than the gene function that determines their gene codon usage patterns. A. pernix K1, P. aerophilum str. IM2, and S. acidocaldarius DSM 639 live in differently extreme conditions. It is presumed that the living environment played an important role in determining the codon usage pattern of these microorganisms. Besides, there was no strain-specific codon usage among these microorganisms. The extent of codon bias in A. pernix K1 and S. acidocaldarius DSM 639 were highly correlated with the gene expression level, but no such association was detected in P. aerophilum str. IM2 genomes.

  14. Absence of micronucleus formation in CHO-K1 cells cultivated in platelet lysate enriched medium.

    PubMed

    Bernardi, Martina; Adami, Valentina; Albiero, Elena; Madeo, Domenico; Rodeghiero, Francesco; Astori, Giuseppe

    2014-03-01

    Human platelet lysate (PL) represents an effective substitute of fetal bovine serum (FBS) for mesenchymal stromal cell (MSC) cultivation. Compared to FBS, PL favors MSC proliferation significantly shortening the population doubling time and avoiding the risks related to the use of animal derivatives. Growth factors contained in the platelets are released upon platelet disruption following freezing/thawing cycles or as we have recently described by using ultrasound. We have investigated whether the increased cell proliferation achieved by using PL could induce mitotic stress and whether the potential formation of free radicals during PL production by ultrasound could cause chromosomal instability in mammalian cells. We have applied an image analysis assisted high content screening (HCS) in vitro micronucleus assay in the Chinese Hamster Ovarian K1 (CHO-K1) rodent mammalian cell line. PL was produced by sonication; for the micronucleus assay, CHO-K1 cells were exposed to increasing concentrations of PL. Cytokinesis was blocked by cytochalasin B, nuclei were stained with bisbenzimide and images were acquired and analyzed automatically using an HCS system, both with a 20× and a 10× objective. Our results suggest that growth stimulus induced by the use of PL did not significantly increase micronucleus formation in CHO-K1 cells compared to negative control. Micronucleus testing in conjunction with HCS could represent a valid tool to evaluate the safety of ancillary materials used in the production of cell-based medicinal products.

  15. Structure of an extended-spectrum class A beta-lactamase from Proteus vulgaris K1.

    PubMed

    Nukaga, Michiyoshi; Mayama, Kayoko; Crichlow, Gregg V; Knox, James R

    2002-03-15

    The structure of a chromosomal extended-spectrum beta-lactamase (ESBL) having the ability to hydrolyze cephalosporins including cefuroxime and ceftazidime has been determined by X-ray crystallography to 1.75 A resolution. The species-specific class A beta-lactamase from Proteus vulgaris K1 was crystallized at pH 6.25 and its structure solved by molecular replacement. Refinement of the model resulted in crystallographic R and R(free) of 16.9 % and 19.3 %, respectively. The folding of the K1 enzyme is broadly similar to that of non-ESBL TEM-type beta-lactamases (2 A rmsd for C(alpha)) and differs by only 0.35 A for all atoms of six conserved residues in the catalytic site. Other residues promoting extended-spectrum activity in K1 include the side-chains of atypical residues Ser237 and Lys276. These side-chains are linked by two water molecules, one of which lies in the position normally filled by the guanidinium group of Arg244, present in most non-ESBL enzymes but absent from K1. The ammonium group of Lys276, ca 3.5 A from the virtual Arg244 guanidinium position, may interact with polar R2 substitutents on the dihydrothiazene ring of cephalosporins.

  16. Neuropathogenic Escherichia coli K1 does not exhibit proteolytic activities to exert its pathogenicity

    PubMed Central

    2013-01-01

    Background Proteases are well-known virulence factors that promote survival, pathogenesis and immune evasion of many pathogens. Several lines of evidence suggest that the blood–brain barrier permeability is a prerequisite in microbial invasion of the central nervous system. Because proteases are frequently associated with vascular permeability by targeting junctional proteins, here it is hypothesized that neuropathogenic Escherichia coli K1 exhibit proteolytic activities to exert its pathogenicity. Methods Zymographic assays were performed using collagen and gelatin as substrates. The lysates of whole E. coli K1 strain E44, or E. coli K-12 strain HB101 were tested for proteolytic activities. The conditioned media were prepared by incubating bacteria in RPMI-1640 in the presence or absence of serum. The cell-free supernatants were collected and tested for proteases in zymography as mentioned above. Additionally, proteolytic degradation of host immune factors was determined by co-incubating conditioned media with albumin/immunoglobulins using protease assays. Results When collagen or gelatin were used as substrates in zymographic assays, neither whole bacteria nor conditioned media exhibited proteolytic activities. The conditioned media of neuropathogenic E. coli K1 strain E44, or E. coli K-12 strain HB101 did not affect degradation of albumin and immunoglobulins using protease assays. Conclusions Neither zymographic assays nor protease assays detected proteolytic activities in either the whole bacteria or conditioned media of E. coli K1 strain E44 and E. coli K-12 strain HB101. These findings suggest that host cell monolayer disruptions and immune evasion strategies are likely independent of proteolytic activities of neuropathogenic E. coli K1. PMID:23634997

  17. KEY COMPARISON: Report on acceleration comparison SIM.AUV.V-K1

    NASA Astrophysics Data System (ADS)

    Evans, David J.; Hornikova, Adriana; Leigh, Stefan D.; Rukhin, Andrew L.; Strawderman, William

    2009-01-01

    The first regional key comparison in vibration conducted under the auspices of the Inter American System of Metrology (SIM) was begun in 1996. The comparison was registered with the International Bureau of Weights and Measures (BIPM) through the Consultative Committee for Acoustics, Ultrasound and Vibration (CCAUV) under the key comparison designation of SIM.AUV.V K1. The national measurement institutes (NMIs) of five countries participating in SIM performed and reported calibrations of one back-to-back and two single-ended accelerometers for this comparison during the time period of 1997 to 1999. The participants in the comparison were: Argentina (INTI), Brazil (INMETRO), Canada (NRC), Mexico (CENAM) and the United States (NIST). Reference Values (RVs) were established for the SIM.AUV.V-K1 comparison using maximum likelihood consensus estimation applied to the summary values of charge sensitivity with the nominal numbers of repeat measurements reported by the participating NMIs. Degrees of equivalence between the results reported by the NMIs and the RVs as well as pairwise degrees of equivalence between the results reported by the NMIs were computed using the uncertainties associated with the results reported by the NMIs and those of the RVs as appropriate. Transformation values were found in order to link the results of SIM.AUV.V-K1 with those of the first international key comparison in vibration conducted under the auspices of the CCAUV during 1999 to 2001 (CCAUV.V-K1). The degrees of equivalence between the NMIs participating in SIM.AUV.V-K1 and those participating in CCAUV.V-K1 were established using linkage values computed from the transformation values. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCAUV, according to the provisions of the CIPM Mutual

  18. Going for baroque at the Escherichia coli K1 cell surface.

    PubMed

    King, Michael R; Steenbergen, Susan M; Vimr, Eric R

    2007-05-01

    Phase variation is usually thought of as the stochastic switching between alternatively expressed ('on') and unexpressed ('off') phenotypic states. However, coupling synthesis of a monotonous homopolysaccharide to a mechanism of random but incomplete chemical modification produces almost infinite structural variation. Potentially limitless variability implies that evolution can produce highly ornate or extravagant flourishes reminiscent of the baroque style. Here, we describe an analysis of capsular polysialic acid form variation in Escherichia coli K1, demonstrating that the large number of variant structures is controlled by a single contingency locus. The mechanism for generating maximum structural diversity from maximal genetic parsimony is conferred by a simple translational switch carried on a K1-specific prophage.

  19. Determination Of Stress Intensity Factors (K1, K2, K3) By Optical Methods

    NASA Astrophysics Data System (ADS)

    Smith, C. W.

    1987-02-01

    An early description of the photoelastic method was provided by Coker and Filon in 1931. Then, in 1937, Oppel introduced the concept of frozen stress photoelasticity. However, one of the first applications of photoelasticity to the field of fracture mechanics did not come until 1958, when Wells and Post conducted their photoelastic study of a running crack. In a discussion of this latter paper, Irwin described a method for extracting the Mode I stress intensity factor (SIP) hereafter referred to as K1, from the photoelastic stress fringe signatures. Subsequently, Fessler and Mansell were among the first to apply Oppel's frozen stress method to the extraction of K1 distributions from three dimensional cracked body problems.

  20. The Circadian Protein BMAL1 Regulates Translation in Response to S6K1-Mediated Phosphorylation.

    PubMed

    Lipton, Jonathan O; Yuan, Elizabeth D; Boyle, Lara M; Ebrahimi-Fakhari, Darius; Kwiatkowski, Erica; Nathan, Ashwin; Güttler, Thomas; Davis, Fred; Asara, John M; Sahin, Mustafa

    2015-05-21

    The circadian timing system synchronizes cellular function by coordinating rhythmic transcription via a transcription-translational feedback loop. How the circadian system regulates gene expression at the translational level remains a mystery. Here, we show that the key circadian transcription factor BMAL1 associates with the translational machinery in the cytosol and promotes protein synthesis. The mTOR-effector kinase, ribosomal S6 protein kinase 1 (S6K1), an important regulator of translation, rhythmically phosphorylates BMAL1 at an evolutionarily conserved site. S6K1-mediated phosphorylation is critical for BMAL1 to both associate with the translational machinery and stimulate protein synthesis. Protein synthesis rates demonstrate circadian oscillations dependent on BMAL1. Thus, in addition to its critical role in circadian transcription, BMAL1 is a translation factor that links circadian timing and the mTOR signaling pathway. More broadly, these results expand the role of the circadian clock to the regulation of protein synthesis.

  1. Palmitoylethanolamide stimulates phagocytosis of Escherichia coli K1 and Streptococcus pneumoniae R6 by microglial cells.

    PubMed

    Redlich, Sandra; Ribes, Sandra; Schütze, Sandra; Czesnik, Dirk; Nau, Roland

    2012-03-01

    The ability of microglial cells to phagocytose bacteria after stimulation with the endocannabinoid palmitoylethanolamide (PEA) was studied in vitro. PEA increased the phagocytosis of unencapsulated Streptococcus pneumoniae R6 and encapsulated Escherichia coli K1 by murine microglial cells significantly after 30 min of microglial stimulation. This suggested that stimulation of microglial cells by PEA can increase the resistance of the brain against CNS infections.

  2. Final report on RMO Vickers key comparison COOMET M.H-K1

    NASA Astrophysics Data System (ADS)

    Aslanyan, E.; Menelao, F.; Herrmann, K.; Aslanyan, A.; Pivovarov, V.; Galat, E.; Dovzhenko, Y.; Zhamanbalin, M.

    2013-01-01

    This report describes a COOMET key comparison on Vickers hardness scales involving five National Metrology Institutes: PTB (Germany), BelGIM (Belarus), NSC IM (Ukraine), KazInMetr (Kazakhstan) and VNIIFTRI (Russia). The pilot laboratory was VNIIFTRI, and PTB acted as the linking institute to key comparisons CCM.H-K1.b and CCM.H-K1.c conducted for the Vickers hardness scales HV1 and HV30, respectively. The comparison was also conducted for the HV5 Vickers hardness scale, since this scale is most frequently used in practice in Russia and CIS countries that work according to GOST standards. In the key comparison, two sets of hardness reference blocks for the Vickers hardness scales HV1, HV5 and HV30 consisting each of three hardness reference blocks with hardness levels of 450 HV and 750 HV were used. The measurement results and uncertainty assessments for HV1 and HV30 hardness scales, as announced by BelGIM, NSC IM, KazInMetr and VNIIFTRI, are in good agreement with the key comparison reference values of CCM.H-K1.b and CCM.H-K1.c. The comparison results for the HV5 hardness scale are viewed as additional information, since up to today no CCM key comparisons on this scale have yet been carried out. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

  3. K1.33Mn8O16 as an electrocatalyst and a cathode

    NASA Astrophysics Data System (ADS)

    Jalili, Seifollah; Moharramzadeh Goliaei, Elham; Schofield, Jeremy

    2017-02-01

    Density functional theory (DFT) calculations are carried out to investigate the electronic, magnetic and thermoelectric properties of bulk and nanosheet K1.33Mn8O16 materials. The catalytic activity and cathodic performance of bulk and nanosheet structures are examined using the Tran-Blaha modified Becke-Johnson (TB-mBJ) exchange potential. Electronic structure calculations reveal an anti-ferromagnetic ground state, with a TB-mMBJ band gap in bulk K1.33Mn8O16 that is in agreement with experimental results. Density of state plots indicate a partial reduction of Mn4+ ions to Mn3+, without any obvious sign of Jahn-Teller distortion. Moreover, use of the O p-band center as a descriptor of catalytic activity suggests that the nanosheet has enhanced catalytic activity compared to the bulk structure. Thermoelectric parameters such as the Seebeck coefficient, electrical conductivity, and thermal conductivity are also calculated, and it is found that the Seebeck coefficients decrease with increasing temperature. High Seebeck coefficients for both spin-up and spin-down states are found in the nanosheet relative to their value in the bulk K1.33Mn8O16 structure, whereas the electrical and thermal conductivity are reduced relative to the bulk. In addition, figures of merit values are calculated as a function of the chemical potential and it is found that the nanosheet has a figure of merit of 1 at room temperature, compared to 0.5 for the bulk material. All results suggest that K1.33Mn8O16 nanosheets can be used both as a material in waste heat recovery and as an electrocatalyst in fuel cells and batteries.

  4. Curcumin inhibits hypoxia-induced migration in K1 papillary thyroid cancer cells

    PubMed Central

    Tan, Cheng; Zhang, Li; Cheng, Xian; Lin, Xiu-Feng; Lu, Rong-Rong; Bao, Jian-Dong

    2014-01-01

    Curcumin, traditionally used as food and medicinal purposes, has recently been reported to have protective efficacy against hypoxia. Hypoxia is one of the important reactive factors in tumor metastasis, which is a key problem in clinical thyroid cancer therapy. In present study, we investigate the anti-metastatic effect of curcumin on the K1 papillary thyroid cancer cells as well as its potential mechanisms. The results show that curcumin effectively inhibits hypoxia-induced reactive oxygen species (ROS) upregulation and significantly decreases the mRNA and protein expression levels of hypoxia-inducible factor-1α (HIF-1α) in K1 cells. Curcumin also decreases the DNA binding ability of HIF-1α to hypoxia response element (HRE). Furthermore, curcumin enhances E-cadherin expression, inhibits metalloproteinase-9 (MMP-9) enzyme activity, and weakens K1 cells migration under hypoxic conditions. In summary, these results indicate that curcumin possesses a potent anti-metastatic effect and might be an effective tumoristatic agent for the treatment of aggressive papillary thyroid cancers. PMID:25349216

  5. Volatile Composition of Comet C/2012 K1 (PanSTARRS)

    NASA Astrophysics Data System (ADS)

    Roth, Nathan; Gibb, Erika; Bonev, Boncho P.; DiSanti, Michael A.; Villanueva, Geronimo L.; Paganini, Lucas; Mumma, Michael J.

    2015-11-01

    On 2014 May 22 and 24 we characterized the volatile composition of the dynamically new Oort Cloud comet C/2012 K1 (PanSTARRS) using the long-slit, high resolution (λ/Δλ ≈ 25,000) infared echelle spectrograph (NIRSPEC) at the 10m Keck 2 telescope on Maunakea, HI. We detected fluorescent emission from six primary species (H2O, HCN, CH4, C2H6, CH3OH, and CO) and prompt emission from one product species (OH* - a directory proxy for H­2O). Upper limits were derived for C2H2 and H2CO. We report rotational temperatures, production rates, and mixing ratios (relative to water). Based on the inventory of comets characterized to date, mixing ratios of trace gases in C/2012 K1 (PanSTARRS) are about normal - CH3OH and C2H6 are slightly enriched, CO, CH4, HCN, and H2CO are average, and C­2H2 is depleted. I will discuss C/2012 K1 (PanSTARRS) in the context of an emerging taxonomy for comets based on volatile composition.This work is supported through the NASA Missouri Space Grant Consortium and the National Science Foundation (NSF 1211362), the NASA Astrobiology Institute through a grant to the Goddard Center for Astrobiology (811073.02.12.03.91), and the NASA Planetary Astronomy Program (811073.02.03.03.43).

  6. [Role of mitochondrial alternative oxidase (AOX) pathway in photoprotection in Rumex K-1 leaves].

    PubMed

    Meng, Xiang-Long; Zhang, Li-Tao; Zhang, Zi-Shan; Gao, Hui-Yuan; Meng, Qing-Wei

    2012-07-01

    Taking Rumex K-1 leaves as test materials, this paper studied the role of mitochondrial alternative oxidase (AOX) pathway in photoprotection under different light intensities. Under low light intensity (200 micromol x m(-2) x s(-1)), and after treated with salicylhydroxamic acid to inhibit the AOX pathway, the leaf actual photochemical efficiency of PS II, linear electron transport rate of photosynthesis, and photosynthetic O2 evolution rate all decreased significantly while the non-Q(B) reducing reaction center had a significant increase, indicating that under low light, the photoinhibition was aggravated while the scavenging enzymes of reactive oxygen species (ROS) increased, which avoided the over-accumulation of ROS and partially alleviated the photoinhibition of Rumex K-1 leaves. Under high light intensity (800 micromol x m(-2) x s(-1)), the inhibition of AOX pathway caused more severe photoinhibition, and the increased activities of ROS scavenging enzymes were insufficient to prevent the over-accumulation of ROS. This study demonstrated that AOX pathway played an important role in the photoprotection in Rumex K-1 leaves under both high and low light intensities, and the role of AOX pathway in photoprotection under high light could be irreplaceable by the other photoprotection pathways in chloroplast.

  7. Precision process calibration and CD predictions for low-k1 lithography

    NASA Astrophysics Data System (ADS)

    Chen, Ting; Park, Sangbong; Berger, Gabriel; Coskun, Tamer H.; de Vocht, Joep; Chen, Fung; Yu, Linda; Hsu, Stephen; van den Broeke, Doug; Socha, Robert; Park, Jungchul; Gronlund, Keith; Davis, Todd; Plachecki, Vince; Harris, Tom; Hansen, Steve; Lambson, Chuck

    2005-06-01

    Leading resist calibration for sub-0.3 k1 lithography demands accuracy <2nm for CD through pitch. An accurately calibrated resist process is the prerequisite for establishing production-worthy manufacturing under extreme low k1. From an integrated imaging point of view, the following key components must be simultaneously considered during the calibration - high numerical aperture (NA>0.8) imaging characteristics, customized illuminations (measured vs. modeled pupil profiles), resolution enhancement technology (RET) mask with OPC, reticle metrology, and resist thin film substrate. For imaging at NA approaching unity, polarized illumination can impact significantly the contrast formation in the resist film stack, and therefore it is an important factor to consider in the CD-based resist calibration. For aggressive DRAM memory core designs at k1<0.3, pattern-specific illumination optimization has proven to be critical for achieving the required imaging performance. Various optimization techniques from source profile optimization with fixed mask design to the combined source and mask optimization have been considered for customer designs and available imaging capabilities. For successful low-k1 process development, verification of the optimization results can only be made with a sufficiently tunable resist model that can predicate the wafer printing accurately under various optimized process settings. We have developed, for resist patterning under aggressive low-k1 conditions, a novel 3D diffusion model equipped with double-Gaussian convolution in each dimension. Resist calibration with the new diffusion model has demonstrated a fitness and CD predication accuracy that rival or outperform the traditional 3D physical resist models. In this work, we describe our empirical approach to achieving the nm-scale precision for advanced lithography process calibrations, using either measured 1D CD through-pitch or 2D memory core patterns. We show that for ArF imaging, the

  8. Compilation of a provisional UK database for the phylloquinone (vitamin K1) content of foods.

    PubMed

    Bolton-Smith, C; Price, R J; Fenton, S T; Harrington, D J; Shearer, M J

    2000-04-01

    This paper reports the compilation of a food composition database for phylloquinone (vitamin K1) derived from the direct analysis of foods, recipe calculation and the assignment of values based on food similarities. All the basic and other food items used in these calculations had been analysed by HPLC and about 170 of the items had been obtained and assayed in the UK. Recipe calculations took account of the cooking method and changes in water and fat content. Currently, approximately 1501 food items with Royal Society of Chemistry/Ministry of Agriculture, Fisheries and Food food codes have been allocated a vitamin K1 value, and a further 282 new recipe codes are included in the database. Representative values from each food group are reported together with an indication of the potential variation. Detailed examples of some recipe calculations are included, and also the impact of changing the type of fat in recipes. Vitamin K1 is associated with, and most abundant in, photosynthetic tissues of plants. Accordingly, the highest concentrations (3000-6000 micrograms/kg) are found in dark-green leafy vegetables and herbs, such as kale, parsley, spinach and green cabbage. Intermediate concentrations (1000-2000 micrograms/kg) are found in plants with paler leaves such as white cabbage and lettuce or in green, non-leafy vegetables such as broccoli and brussel sprouts. Fats and oils contain variable amounts of vitamin K1 with the highest concentrations (300-1300 micrograms/kg) in soyabean, rapeseed and olive oils and the margarines based on them. Other foods such as dairy products, meat dishes and cereal-based foods (bread, biscuits, cakes, desserts etc.), although not in themselves particularly rich in vitamin K1 (< 200 micrograms/kg), may contribute significantly to intakes when consumption of green vegetables is poor. Within the scope of this present study, it has not been possible to address issues such as inter-sample variability, losses during storage or the

  9. PDGF induces SphK1 expression via Egr-1 to promote pulmonary artery smooth muscle cell proliferation.

    PubMed

    Sysol, Justin R; Natarajan, Viswanathan; Machado, Roberto F

    2016-06-01

    Pulmonary arterial hypertension (PAH) is a progressive, life-threatening disease for which there is currently no curative treatment available. Pathologic changes in this disease involve remodeling of the pulmonary vasculature, including marked proliferation of pulmonary artery smooth muscle cells (PASMCs). Recently, the bioactive lipid sphingosine-1-phosphate (S1P) and its activating kinase, sphingosine kinase 1 (SphK1), have been shown to be upregulated in PAH and promote PASMC proliferation. The mechanisms regulating the transcriptional upregulation of SphK1 in PASMCs are unknown. In this study, we investigated the role of platelet-derived growth factor (PDGF), a PAH-relevant stimuli associated with enhanced PASMC proliferation, on SphK1 expression regulation. In human PASMCs (hPASMCs), PDGF significantly increased SphK1 mRNA and protein expression and induced cell proliferation. Selective inhibition of SphK1 attenuated PDGF-induced hPASMC proliferation. In silico promoter analysis for SphK1 identified several binding sites for early growth response protein 1 (Egr-1), a PDGF-associated transcription factor. Luciferase assays demonstrated that PDGF activates the SphK1 promoter in hPASMCs, and truncation of the 5'-promoter reduced PDGF-induced SphK1 expression. Stimulation of hPASMCs with PDGF induced Egr-1 protein expression, and direct binding of Egr-1 to the SphK1 promoter was confirmed by chromatin immunoprecipitation analysis. Inhibition of ERK signaling prevented induction of Egr-1 by PDGF. Silencing of Egr-1 attenuated PDGF-induced SphK1 expression and hPASMC proliferation. These studies demonstrate that SphK1 is regulated by PDGF in hPASMCs via the transcription factor Egr-1, promoting cell proliferation. This novel mechanism of SphK1 regulation may be a therapeutic target in pulmonary vascular remodeling in PAH.

  10. 17 CFR 240.13k-1 - Foreign bank exemption from the insider lending prohibition under section 13(k).

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... the insider lending prohibition under section 13(k). 240.13k-1 Section 240.13k-1 Commodity and....13k-1 Foreign bank exemption from the insider lending prohibition under section 13(k). (a) For the...(k) of the Act (15 U.S.C. 78m(k)) with respect to any such loan made by the foreign bank as long...

  11. Vaccinia virus K1 ankyrin repeat protein inhibits NF-κB activation by preventing RelA acetylation.

    PubMed

    Bravo Cruz, Ariana G; Shisler, Joanna L

    2016-10-01

    The vaccinia virus (VACV) K1 protein inhibits dsRNA-dependent protein kinase (PKR) activation. A consequence of this function is that K1 inhibits PKR-induced NF-κB activation during VACV infection. However, transient expression of K1 also inhibits Toll-like receptor (TLR)-induced NF-κB activation. This suggests that K1 has a second NF-κB inhibitory mechanism that is PKR-independent. This possibility was explored by expressing K1 independently of infection and stimulating NF-κB under conditions that minimized or excluded PKR activation. K1 inhibited both TNF- and phorbol 12-myristate 13-acetate (PMA)-induced NF-κB activation, as detected by transcription of synthetic (e.g. luciferase) and natural (e.g. CXCL8) genes controlled by NF-κB. K1 also inhibited NF-κB activity in PKRkd cells, cells that have greatly decreased amounts of PKR. K1 no longer prevented IκBα degradation or NF-κB nuclear translocation in the absence of PKR, suggesting that K1 acted on a nuclear event. Indeed, K1 was present in the nucleus and cytoplasm of stimulated and unstimulated cells. K1 inhibited acetylation of the RelA (p65) subunit of NF-κB, a nuclear event known to be required for NF-κB activation. Moreover, p65-CBP (CREB-binding protein) interactions were blocked in the presence of K1. However, K1 did not preclude NF-κB binding to oligonucleotides containing κB-binding sites. The current interpretation of these data is that NF-κB-promoter interactions still occur in the presence of K1, but NF-κB cannot properly trigger transcriptional activation because K1 antagonizes acetylation of RelA. Thus, in comparison to all known VACV NF-κB inhibitory proteins, K1 acts at one of the most downstream events of NF-κB activation.

  12. Delay in Human Neutrophil Constitutive Apoptosis after Infection with Klebsiella pneumoniae Serotype K1

    PubMed Central

    Lee, Chen-Hsiang; Chuah, Seng-Kee; Tai, Wei-Chen; Chang, Chia-Chi; Chen, Fang-Ju

    2017-01-01

    Klebsiella pneumoniae serotype K1 is a major cause of invasive syndrome defined by liver abscess with metastatic infections at other body sites. This culprit is known to be resistant to neutrophil phagocytosis and bactericidal activity. We hypothesized that K. pneumoniae serotype K1 might regulate neutrophil apoptosis and enhance the survival of the infected neutrophils that might serve as a vector for dissemination of the bacteria. Two serotypes of K. pneumoniae, KP-M1 isolated from a patient with liver abscess and DT-X (an acapsular mutant strain of KP-M1), were used to infect human neutrophils. The infected neutrophils were examined for their cytotoxicity, annexin V staining, proteins, DNA fragmentation, cytokine production, and viability that are involved in apoptosis. We found that KP-M1 was not destroyed and the ingested bacteria survived within neutrophils. While the uninfected neutrophils became apoptotic within 10 h, the neutrophils infected with KP-M1 could survive up to 24 h post infection. Constitutive apoptosis of KP-M1-infected neutrophils was significantly delayed compared to that of DT-X-infected or uninfected neutrophils (p < 0.01). KP-M1 modulated the anti-apoptotic effects by down-regulating the ratio of Bax to Bcl-2 and Mcl-1, and then delayed caspase-3 activation in the neutrophils, which was accompanied by inducing the anti-apoptotic cytokine, IL-8. These data suggest that K. pneumoniae serotype K1 can prolong the lifespan of infected neutrophils by delaying constitutive apoptosis within the first several hours of infection.

  13. Pathoadaptive Mutations of Escherichia coli K1 in Experimental Neonatal Systemic Infection

    PubMed Central

    McCarthy, Alex J.; Negus, David; Martin, Patricia; Pechincha, Catarina; Oswald, Eric; Stabler, Richard A.; Taylor, Peter W.

    2016-01-01

    Although Escherichia coli K1 strains are benign commensals in adults, their acquisition at birth by the newborn may result in life-threatening systemic infections, most commonly sepsis and meningitis. Key features of these infections, including stable gastrointestinal (GI) colonization and age-dependent invasion of the bloodstream, can be replicated in the neonatal rat. We previously increased the capacity of a septicemia isolate of E. coli K1 to elicit systemic infection following colonization of the small intestine by serial passage through two-day-old (P2) rat pups. The passaged strain, A192PP (belonging to sequence type 95), induces lethal infection in all pups fed 2–6 x 106 CFU. Here we use whole-genome sequencing to identify mutations responsible for the threefold increase in lethality between the initial clinical isolate and the passaged derivative. Only four single nucleotide polymorphisms (SNPs), in genes (gloB, yjgV, tdcE) or promoters (thrA) involved in metabolic functions, were found: no changes were detected in genes encoding virulence determinants associated with the invasive potential of E. coli K1. The passaged strain differed in carbon source utilization in comparison to the clinical isolate, most notably its inability to metabolize glucose for growth. Deletion of each of the four genes from the E. coli A192PP chromosome altered the proteome, reduced the number of colonizing bacteria in the small intestine and increased the number of P2 survivors. This work indicates that changes in metabolic potential lead to increased colonization of the neonatal GI tract, increasing the potential for translocation across the GI epithelium into the systemic circulation. PMID:27861552

  14. Pathoadaptive Mutations of Escherichia coli K1 in Experimental Neonatal Systemic Infection.

    PubMed

    McCarthy, Alex J; Negus, David; Martin, Patricia; Pechincha, Catarina; Oswald, Eric; Stabler, Richard A; Taylor, Peter W

    2016-01-01

    Although Escherichia coli K1 strains are benign commensals in adults, their acquisition at birth by the newborn may result in life-threatening systemic infections, most commonly sepsis and meningitis. Key features of these infections, including stable gastrointestinal (GI) colonization and age-dependent invasion of the bloodstream, can be replicated in the neonatal rat. We previously increased the capacity of a septicemia isolate of E. coli K1 to elicit systemic infection following colonization of the small intestine by serial passage through two-day-old (P2) rat pups. The passaged strain, A192PP (belonging to sequence type 95), induces lethal infection in all pups fed 2-6 x 106 CFU. Here we use whole-genome sequencing to identify mutations responsible for the threefold increase in lethality between the initial clinical isolate and the passaged derivative. Only four single nucleotide polymorphisms (SNPs), in genes (gloB, yjgV, tdcE) or promoters (thrA) involved in metabolic functions, were found: no changes were detected in genes encoding virulence determinants associated with the invasive potential of E. coli K1. The passaged strain differed in carbon source utilization in comparison to the clinical isolate, most notably its inability to metabolize glucose for growth. Deletion of each of the four genes from the E. coli A192PP chromosome altered the proteome, reduced the number of colonizing bacteria in the small intestine and increased the number of P2 survivors. This work indicates that changes in metabolic potential lead to increased colonization of the neonatal GI tract, increasing the potential for translocation across the GI epithelium into the systemic circulation.

  15. EUV resolution enhancement techniques (RETs) for k1 0.4 and below

    NASA Astrophysics Data System (ADS)

    Hsu, Stephen; Howell, Rafael; Jia, Jianjun; Liu, Hua-Yu; Gronlund, Keith; Hansen, Steve; Zimmermann, Jörg

    2015-03-01

    Due to the exponential growth of mobile wireless devices, low-power logic chips continue to drive device scaling. To enable sub-10 nm device scaling at an affordable cost, there is a strong need for single exposure advanced lithography. Extreme ultraviolet lithography (EUVL) is one of the most promising candidates to support the design rules for sub-10 nm. The aggressive mobile device design rules continue to push the critical dimension (CD) and pitch and put very stringent demands on the lithography performance such as pattern placement control, image contrast, critical dimension uniformity (CDU), and line width roughness (LWR). In this paper we report the latest advances in resolution enhancement techniques to address low k1 challenges in EUV lithography, specifically: minimizing the pattern placement error, enhancing the through-focus contrast, and reducing the impact of stochastic effects. We have developed an innovative source-mask optimization (SMO) method to significantly reduce edge placement errors (EPE) [1] [2]. Aggressive design rules using the state-of-the-art NA of 0.33 of the NXE:3300B and its successor tools can have imaging below k1 = 0.4, which can extend the current process capabilities for single exposure high volume manufacturing (HVM). Burkhardt et al. reported in a previous study that inserting a sub-resolution assist feature (SRAF) within semi-isolated features introduces strong Bossung tilts and best focus shifts, and a general solution for random pitches is not apparent [3]. Kang observed the same issues and proposed to introduce spherical aberrations to correct these effects while having a global impact on the full-chip [4]. In this work we introduce a new methodology to apply SRAFs to improve contrast, reduce best focus shift, and improve process window. Finally, the lower number of photons of EUV and the small feature size brings serious issue of the stochastic effect that causes the line-edge-roughness (LER) and local CD uniformity

  16. Mixed Symmetry-Tipe (k,1) Massless Tensor Fields. Consistent Interactions Of Dual Linearized Gravity

    NASA Astrophysics Data System (ADS)

    Bizdadea, C.; Saliu, S. O.; Toma, M.

    2012-12-01

    A particular case of interactions of a single massless tensor field with the mixed symmetry corresponding to a two-column Young diagram (k,1) with k=4, dual to linearized gravity in D=7, is considered in the context of: self-couplings, cross-interactions with a Pauli-Fierz field, cross-couplings to purely matter theories, and interactions with an Abelian 1-form. The general approach relies on the deformation of the solution to the master equation from the antifield-BRST formalism by means of the local cohomology of the BRST differential.

  17. A new genus and two new species of chigger mites (Acari: Trombiculidae) from the Laotian rock-rat Laonastes aenigmamus Jenkins, Kilpatrick, Robinson & Timmins (Rodentia: Diatomyidae).

    PubMed

    Stekolnikov, Alexandr A

    2014-01-01

    A new chigger mite genus Laotrombicula n. g. and two new species, Laotrombicula khunboromi n. sp. (type-species) and L. fangumi n. sp., are described from the Laotian rock-rat Laonastes aenigmamus Jenkins, Kilpatrick, Robinson & Timmins (Rodentia: Diatomyidae). The new genus is most similar to Trombiculindus Radford, 1948 and Leptotrombidium Nagayo, Miyagawa, Mitamura & Imamura, 1916 and differs from these genera by having the scutum of subhexagonal or semicircular shape vs widely rectangular; pinnatifid dorsocentral idiosomal setae vs foliaceous in Trombiculindus and unexpanded in Leptotrombidium; and by the presence of serrated longitudinal crests in the middle part of scutum.

  18. Neto auxiliary proteins control both the trafficking and biophysical properties of the kainate receptor GluK1

    PubMed Central

    Sheng, Nengyin; Shi, Yun S; Lomash, Richa Madan; Roche, Katherine W; Nicoll, Roger A

    2015-01-01

    Kainate receptors (KARs) are a subfamily of glutamate receptors mediating excitatory synaptic transmission and Neto proteins are recently identified auxiliary subunits for KARs. However, the roles of Neto proteins in the synaptic trafficking of KAR GluK1 are poorly understood. Here, using the hippocampal CA1 pyramidal neuron as a null background system we find that surface expression of GluK1 receptor itself is very limited and is not targeted to excitatory synapses. Both Neto1 and Neto2 profoundly increase GluK1 surface expression and also drive GluK1 to synapses. However, the regulation GluK1 synaptic targeting by Neto proteins is independent of their role in promoting surface trafficking. Interestingly, GluK1 is excluded from synapses expressing AMPA receptors and is selectively incorporated into silent synapses. Neto2, but not Neto1, slows GluK1 deactivation, whereas Neto1 speeds GluK1 desensitization and Neto2 slows desensitization. These results establish critical roles for Neto auxiliary subunits controlling KARs properties and synaptic incorporation. DOI: http://dx.doi.org/10.7554/eLife.11682.001 PMID:26720915

  19. Neto auxiliary proteins control both the trafficking and biophysical properties of the kainate receptor GluK1.

    PubMed

    Sheng, Nengyin; Shi, Yun S; Lomash, Richa Madan; Roche, Katherine W; Nicoll, Roger A

    2015-12-31

    Kainate receptors (KARs) are a subfamily of glutamate receptors mediating excitatory synaptic transmission and Neto proteins are recently identified auxiliary subunits for KARs. However, the roles of Neto proteins in the synaptic trafficking of KAR GluK1 are poorly understood. Here, using the hippocampal CA1 pyramidal neuron as a null background system we find that surface expression of GluK1 receptor itself is very limited and is not targeted to excitatory synapses. Both Neto1 and Neto2 profoundly increase GluK1 surface expression and also drive GluK1 to synapses. However, the regulation GluK1 synaptic targeting by Neto proteins is independent of their role in promoting surface trafficking. Interestingly, GluK1 is excluded from synapses expressing AMPA receptors and is selectively incorporated into silent synapses. Neto2, but not Neto1, slows GluK1 deactivation, whereas Neto1 speeds GluK1 desensitization and Neto2 slows desensitization. These results establish critical roles for Neto auxiliary subunits controlling KARs properties and synaptic incorporation.

  20. Draft Genome Sequence of Thermus scotoductus Strain K1, Isolated from a Geothermal Spring in Karvachar, Nagorno Karabakh

    PubMed Central

    Saghatelyan, Ani; Poghosyan, Lianna

    2015-01-01

    The 2,379,636-bp draft genome sequence of Thermus scotoductus strain K1, isolated from geothermal spring outlet located in the Karvachar region in Nagorno Karabakh is presented. Strain K1 shares about 80% genome sequence similarity with T. scotoductus strain SA-01, recovered from a deep gold mine in South Africa. PMID:26564055

  1. 40 CFR Table K-1 to Subpart K of... - Electric Arc Furnace (EAF) CH4 Emission Factors

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 21 2011-07-01 2011-07-01 false Electric Arc Furnace (EAF) CH4.... 98, Subpt. K, Table K-1 Table K-1 to Subpart K of Part 98—Electric Arc Furnace (EAF) CH4 Emission... charging intermittently every minute. b Temperature measured in off-gas channel downstream of the...

  2. 40 CFR Table K-1 to Subpart K of... - Electric Arc Furnace (EAF) CH4 Emission Factors

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 21 2014-07-01 2014-07-01 false Electric Arc Furnace (EAF) CH4.... 98, Subpt. K, Table K-1 Table K-1 to Subpart K of Part 98—Electric Arc Furnace (EAF) CH4 Emission... charging intermittently every minute. b Temperature measured in off-gas channel downstream of the...

  3. 40 CFR Table K-1 to Subpart K of... - Electric Arc Furnace (EAF) CH4 Emission Factors

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 22 2012-07-01 2012-07-01 false Electric Arc Furnace (EAF) CH4.... 98, Subpt. K, Table K-1 Table K-1 to Subpart K of Part 98—Electric Arc Furnace (EAF) CH4 Emission... charging intermittently every minute. b Temperature measured in off-gas channel downstream of the...

  4. 40 CFR Table K-1 to Subpart K of... - Electric Arc Furnace (EAF) CH4 Emission Factors

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 22 2013-07-01 2013-07-01 false Electric Arc Furnace (EAF) CH4.... 98, Subpt. K, Table K-1 Table K-1 to Subpart K of Part 98—Electric Arc Furnace (EAF) CH4 Emission... charging intermittently every minute. b Temperature measured in off-gas channel downstream of the...

  5. Identification of an immunoreceptor tyrosine-based activation motif of K1 transforming protein of Kaposi's sarcoma-associated herpesvirus.

    PubMed

    Lee, H; Guo, J; Li, M; Choi, J K; DeMaria, M; Rosenzweig, M; Jung, J U

    1998-09-01

    Kaposi's sarcoma-associated herpesvirus (KSHV) is consistently identified in Kaposi's sarcoma and body cavity-based lymphoma. KSHV encodes a transforming protein called K1 which is structurally similar to lymphocyte receptors. We have found that a highly conserved region of the cytoplasmic domain of K1 resembles the sequence of immunoreceptor tyrosine-based activation motifs (ITAMs). To demonstrate the signal-transducing activity of K1, we constructed a chimeric protein in which the cytoplasmic tail of the human CD8alpha polypeptide was replaced with that of KSHV K1. Expression of the CD8-K1 chimera in B cells induced cellular tyrosine phosphorylation and intracellular calcium mobilization upon stimulation with an anti-CD8 antibody. Mutational analyses showed that the putative ITAM of K1 was required for its signal-transducing activity. Furthermore, tyrosine residues of the putative ITAM of K1 were phosphorylated upon stimulation, and this allowed subsequent binding of SH2-containing proteins. These results demonstrate that the KSHV transforming protein K1 contains a functional ITAM in its cytoplasmic domain and that it can transduce signals to induce cellular activation.

  6. Draft Genome Sequence of Salmonella enterica subsp. diarizonae Serovar 61:k:1,5,(7) Strain CRJJGF_00165 (Phylum Gammaproteobacteria)

    PubMed Central

    Gupta, Sushim K.; McMillan, Elizabeth A.; Jackson, Charlene R.; Desai, Prerak T.; Porwollik, Steffen; McClelland, Michael; Hiott, Lari M.; Humayoun, Shaheen B.; Barrett, John B.

    2016-01-01

    Here, we report a 4.78-Mb draft genome sequence of the Salmonella enterica subsp. diarizonae serovar 61:k:1,5,(7) strain CRJJGF_00165 [also called S. enterica subsp. IIIb serovar 61:k:1,5,(7) strain CRJJGF_00165], isolated from ground beef in 2007. PMID:27881547

  7. miR-613 inhibits bladder cancer proliferation and migration through targeting SphK1

    PubMed Central

    Yu, Haifeng; Duan, Ping; Zhu, Haibo; Rao, Dapang

    2017-01-01

    Objectives: Increasing evidence has suggested that microRNA (miRNA) dysregulation may contribute to tumor progression and metastasis. However, the role of miR-613 in bladder cancer was still unknown. Materials and methods: qRT-PCR and Western blotting were performed to detect the expression of miR-613 and its direct target gene. CCK-8 analysis, qRT-PCR and cell invasion were performed to measure the cell function. Results: We demonstrated that the expression of miR-613 was downregulated in the bladder cancer cell lines. In addition, miR-613 expression was downregulated in the bladder cancer tissues compared to the adjacent normal tissues. Out of 35 bladder cancer tissues, miR-613 was downregulated in 27 cases compared to the adjacent tissues. Ectopic expression of miR-613 suppressed the bladder cancer cell proliferation and invasion. Moreover, miR-613 overexpression enhanced the expression of epithelial biomarker, Ecadherin, and suppressed the expression of mesenchymal biomarker, Vimentin, Snail and N-cadherin. Furthermore, we identified the Sphingosine kinase 1 (SphK1) as the direct target gene of miR-613 in the bladder cancer cell. Restoration of Sphk1 partially rescued miR-613-inhibited bladder cancer cell proliferation, invasion and EMT. Conclusions: These data suggested that miR-613 acted a tumor suppressive role in bladder cancer through targeting SphK1 in bladder. PMID:28386347

  8. Aerobic expression of Vitreoscilla hemoglobin improves the growth performance of CHO-K1 cells.

    PubMed

    Juárez, Mariana; González-De la Rosa, Claudia H; Memún, Elisa; Sigala, Juan-Carlos; Lara, Alvaro R

    2017-03-01

    Inefficient carbon metabolism is a relevant issue during the culture of mammalian cells for the production of biopharmaceuticals. Therefore, cell engineering strategies to improve the metabolic and growth performance of cell lines are needed. The expression of Vitreoscilla stercoraria hemoglobin (VHb) has been shown to significantly reduce overflow metabolism and improve the aerobic growth of bacteria. However, the effects of VHb on mammalian cells have been rarely studied. Here, the impact of VHb on growth and lactate accumulation during CHO-K1 cell culture was investigated. For this purpose, CHO-K1 cells were transfected with plasmids carrying the vgb or gfp gene to express VHb or green fluorescence protein (GFP), respectively. VHb expression increased the specific growth rate and biomass yields on glucose and glutamine by 60 %, and reduced the amount of lactate produced per cell by 40 %, compared to the GFP-expression controls. Immunofluorescence microscopy showed that VHb is distributed in the cytoplasm and organelles, which support the hypothesis that VHb could serve as an oxygen carrier, enhancing aerobic respiration. These results are useful for the development of better producing cell lines for industrial applications.

  9. OPC model calibration for CPL patterning at extreme low k1

    NASA Astrophysics Data System (ADS)

    Shi, Xuelong; Laidig, Tom; Chen, J. Fung; Van Den Broeke, Douglas; Hsu, Stephen; Hsu, Michael; Wampler, Kurt E.; Hollerbach, Uwe; Park, Jung Chul; Yu, Linda

    2004-12-01

    Model based optical proximity correction (OPC) to enhance image fidelity and process robustness has become one of the most critical components that enable the low k1 optical lithography. To meet the challenges imposed by the previously unthinkable low k1 for manufacturing with most stringent dimension control requirements, a capable OPC model to meet such an aggressive lithography challenges has been urgently called upon. In addition to providing better accuracy for the currently implemented process technologies, the new OPC model must work well with Chromeless Phase Lithography (CPL) in which the topography on the mask is rather significant, and Double Dipole Lithography (DDL) in which two masks and two exposures are needed. It must also be able to intelligently take into account the effect from the more aggressive illuminations, usch as customer designed illuminator and experimental measured illuminator profile from the scanners. The physical and mathematical foundation of the model must be well thought of to meet the requirements for the above-mentioned applications. We have extended our Eigen Decomposition Model (EDM) for model OPC treatment into the high NA regime, in which the vector characteristics of light and thin film stack are taken into account. For CPL calibration, it has been found that 3D mask topography effect cannot be ignored in order to achieve satisfactory model accuracy.

  10. Bending Elasticity Modulus of Giant Vesicles Composed of Aeropyrum Pernix K1 Archaeal Lipid

    PubMed Central

    Genova, Julia; Poklar Ulrih, Nataša; Kralj-Iglič, Veronika; Iglič, Aleš; Bivas, Isak

    2015-01-01

    Thermally induced shape fluctuations were used to study elastic properties of giant vesicles composed of archaeal lipids C25,25-archetidyl (glucosyl) inositol and C25,25-archetidylinositol isolated from lyophilised Aeropyrum pernix K1 cells. Giant vesicles were created by electroformation in pure water environment. Stroboscopic illumination using a xenon flash lamp was implemented to remove the blur effect due to the finite integration time of the camera and to obtain an instant picture of the fluctuating vesicle shape. The mean weighted value of the bending elasticity modulus kc of the archaeal membrane determined from the measurements meeting the entire set of qualification criteria was (1.89 ± 0.18) × 10−19 J, which is similar to the values obtained for a membrane composed of the eukaryotic phospholipids SOPC (1.88 ± 0.17) × 10−19 J and POPC (2.00 ± 0.21) × 10−19 J. We conclude that membranes composed of archaeal lipids isolated from Aeropyrum pernix K1 cells have similar elastic properties as membranes composed of eukaryotic lipids. This fact, together with the importance of the elastic properties for the normal circulation through blood system, provides further evidence in favor of expectations that archaeal lipids could be appropriate for the design of drug delivery systems. PMID:25821933

  11. Observation and polarization measurements of B+/- -->phiK1 +/- and B +/- -->phiK2 *+/-.

    PubMed

    Aubert, B; Bona, M; Karyotakis, Y; Lees, J P; Poireau, V; Prencipe, E; Prudent, X; Tisserand, V; Garra Tico, J; Grauges, E; Lopez, L; Palano, A; Pappagallo, M; Eigen, G; Stugu, B; Sun, L; Abrams, G S; Battaglia, M; Brown, D N; Cahn, R N; Jacobsen, R G; Kerth, L T; Kolomensky, Yu G; Kukartsev, G; Lynch, G; Osipenkov, I L; Ronan, M T; Tackmann, K; Tanabe, T; Hawkes, C M; Soni, N; Watson, A T; Koch, H; Schroeder, T; Walker, D; Asgeirsson, D J; Fulsom, B G; Hearty, C; Mattison, T S; McKenna, J A; Barrett, M; Khan, A; Teodorescu, L; Blinov, V E; Bukin, A D; Buzykaev, A R; Druzhinin, V P; Golubev, V B; Onuchin, A P; Serednyakov, S I; Skovpen, Yu I; Solodov, E P; Todyshev, K Yu; Bondioli, M; Curry, S; Eschrich, I; Kirkby, D; Lankford, A J; Lund, P; Mandelkern, M; Martin, E C; Stoker, D P; Abachi, S; Buchanan, C; Gary, J W; Liu, F; Long, O; Shen, B C; Vitug, G M; Yasin, Z; Zhang, L; Sharma, V; Campagnari, C; Hong, T M; Kovalskyi, D; Mazur, M A; Richman, J D; Beck, T W; Eisner, A M; Flacco, C J; Heusch, C A; Kroseberg, J; Lockman, W S; Schalk, T; Schumm, B A; Seiden, A; Wang, L; Wilson, M G; Winstrom, L O; Cheng, C H; Doll, D A; Echenard, B; Fang, F; Hitlin, D G; Narsky, I; Piatenko, T; Porter, F C; Andreassen, R; Mancinelli, G; Meadows, B T; Mishra, K; Sokoloff, M D; Bloom, P C; Ford, W T; Gaz, A; Hirschauer, J F; Kreisel, A; Nagel, M; Nauenberg, U; Smith, J G; Ulmer, K A; Wagner, S R; Ayad, R; Soffer, A; Toki, W H; Wilson, R J; Altenburg, D D; Feltresi, E; Hauke, A; Jasper, H; Karbach, M; Merkel, J; Petzold, A; Spaan, B; Wacker, K; Kobel, M J; Mader, W F; Nogowski, R; Schubert, K R; Schwierz, R; Sundermann, J E; Volk, A; Bernard, D; Bonneaud, G R; Latour, E; Thiebaux, Ch; Verderi, M; Clark, P J; Gradl, W; Playfer, S; Watson, J E; Andreotti, M; Bettoni, D; Bozzi, C; Calabrese, R; Cecchi, A; Cibinetto, G; Franchini, P; Luppi, E; Negrini, M; Petrella, A; Piemontese, L; Santoro, V; Baldini-Ferroli, R; Calcaterra, A; de Sangro, R; Finocchiaro, G; Pacetti, S; Patteri, P; Peruzzi, I M; Piccolo, M; Rama, M; Zallo, A; Buzzo, A; Contri, R; Lo Vetere, M; Macri, M M; Monge, M R; Passaggio, S; Patrignani, C; Robutti, E; Santroni, A; Tosi, S; Chaisanguanthum, K S; Morii, M; Marks, J; Schenk, S; Uwer, U; Klose, V; Lacker, H M; Bard, D J; Dauncey, P D; Nash, J A; Panduro Vazquez, W; Tibbetts, M; Behera, P K; Chai, X; Charles, M J; Mallik, U; Cochran, J; Crawley, H B; Dong, L; Meyer, W T; Prell, S; Rosenberg, E I; Rubin, A E; Gao, Y Y; Gritsan, A V; Guo, Z J; Lae, C K; Denig, A G; Fritsch, M; Schott, G; Arnaud, N; Béquilleux, J; D'Orazio, A; Davier, M; Firmino da Costa, J; Grosdidier, G; Höcker, A; Lepeltier, V; Le Diberder, F; Lutz, A M; Pruvot, S; Roudeau, P; Schune, M H; Serrano, J; Sordini, V; Stocchi, A; Wormser, G; Lange, D J; Wright, D M; Bingham, I; Burke, J P; Chavez, C A; Fry, J R; Gabathuler, E; Gamet, R; Hutchcroft, D E; Payne, D J; Touramanis, C; Bevan, A J; Clarke, C K; George, K A; Di Lodovico, F; Sacco, R; Sigamani, M; Cowan, G; Flaecher, H U; Hopkins, D A; Paramesvaran, S; Salvatore, F; Wren, A C; Brown, D N; Davis, C L; Alwyn, K E; Bailey, D S; Barlow, R J; Barlow, R J; Chia, Y M; Edgar, C L; Lafferty, G D; West, T J; Yi, J I; Anderson, J; Chen, C; Jawahery, A; Roberts, D A; Simi, G; Tuggle, J M; Dallapiccola, C; Li, X; Salvati, E; Saremi, S; Cowan, R; Dujmic, D; Fisher, P H; Koeneke, K; Sciolla, G; Spitznagel, M; Taylor, F; Yamamoto, R K; Zhao, M; Patel, P M; Robertson, S H; Lazzaro, A; Lombardo, V; Palombo, F; Bauer, J M; Cremaldi, L; Eschenburg, V; Godang, R; Kroeger, R; Sanders, D A; Summers, D J; Zhao, H W; Simard, M; Taras, P; Viaud, F B; Nicholson, H; De Nardo, G; Lista, L; Monorchio, D; Onorato, G; Sciacca, C; Raven, G; Snoek, H L; Jessop, C P; Knoepfel, K J; LoSecco, J M; Wang, W F; Benelli, G; Corwin, L A; Honscheid, K; Kagan, H; Kass, R; Morris, J P; Rahimi, A M; Regensburger, J J; Sekula, S J; Wong, Q K; Blount, N L; Brau, J; Frey, R; Igonkina, O; Kolb, J A; Lu, M; Rahmat, R; Sinev, N B; Strom, D; Strube, J; Torrence, E; Castelli, G; Gagliardi, N; Margoni, M; Morandin, M; Posocco, M; Rotondo, M; Simonetto, F; Stroili, R; Voci, C; del Amo Sanchez, P; Ben-Haim, E; Briand, H; Calderini, G; Chauveau, J; David, P; Del Buono, L; Hamon, O; Leruste, Ph; Ocariz, J; Perez, A; Prendki, J; Gladney, L; Biasini, M; Covarelli, R; Manoni, E; Angelini, C; Batignani, G; Bettarini, S; Carpinelli, M; Cervelli, A; Forti, F; Giorgi, M A; Lusiani, A; Marchiori, G; Morganti, M; Neri, N; Paoloni, E; Rizzo, G; Walsh, J J; Biesiada, J; Lopes Pegna, D; Lu, C; Olsen, J; Smith, A J S; Telnov, A V; Anulli, F; Baracchini, E; Cavoto, G; del Re, D; Di Marco, E; Faccini, R; Ferrarotto, F; Ferroni, F; Gaspero, M; Jackson, P D; Li Gioi, L; Mazzoni, M A; Morganti, S; Piredda, G; Polci, F; Renga, F; Voena, C; Ebert, M; Hartmann, T; Schröder, H; Waldi, R; Adye, T; Franek, B; Olaiya, E O; Roethel, W; Wilson, F F; Emery, S; Escalier, M; Esteve, L; Gaidot, A; Ganzhur, S F; Hamel de Monchenault, G; Kozanecki, W; Vasseur, G; Yèche, Ch; Zito, M; Chen, X R; Liu, H; Park, W; Purohit, M V; White, R M; Wilson, J R; Allen, M T; Aston, D; Bartoldus, R; Bechtle, P; Benitez, J F; Cenci, R; Coleman, J P; Convery, M R; Dingfelder, J C; Dorfan, J; Dubois-Felsmann, G P; Dunwoodie, W; Field, R C; Gabareen, A M; Gowdy, S J; Graham, M T; Grenier, P; Hast, C; Innes, W R; Kaminski, J; Kelsey, M H; Kim, H; Kim, P; Kocian, M L; Leith, D W G S; Li, S; Lindquist, B; Luitz, S; Luth, V; Lynch, H L; MacFarlane, D B; Marsiske, H; Messner, R; Muller, D R; Neal, H; Nelson, S; O'Grady, C P; Ofte, I; Perazzo, A; Perl, M; Ratcliff, B N; Roodman, A; Salnikov, A A; Schindler, R H; Schwiening, J; Snyder, A; Su, D; Sullivan, M K; Suzuki, K; Swain, S K; Thompson, J M; Va'vra, J; Wagner, A P; Weaver, M; West, C A; Wisniewski, W J; Wittgen, M; Wright, D H; Wulsin, H W; Yarritu, A K; Yi, K; Young, C C; Ziegler, V; Burchat, P R; Edwards, A J; Majewski, S A; Miyashita, T S; Petersen, B A; Wilden, L; Ahmed, S; Alam, M S; Ernst, J A; Pan, B; Saeed, M A; Zain, S B; Spanier, S M; Wogsland, B J; Eckmann, R; Ritchie, J L; Ruland, A M; Schilling, C J; Schwitters, R F; Drummond, B W; Izen, J M; Lou, X C; Bianchi, F; Gamba, D; Pelliccioni, M; Bomben, M; Bosisio, L; Cartaro, C; Della Ricca, G; Lanceri, L; Vitale, L; Azzolini, V; Lopez-March, N; Martinez-Vidal, F; Milanes, D A; Oyanguren, A; Albert, J; Banerjee, Sw; Bhuyan, B; Choi, H H F; Hamano, K; Kowalewski, R; Lewczuk, M J; Nugent, I M; Roney, J M; Sobie, R J; Gershon, T J; Harrison, P F; Ilic, J; Latham, T E; Mohanty, G B; Band, H R; Chen, X; Dasu, S; Flood, K T; Pan, Y; Pierini, M; Prepost, R; Vuosalo, C O; Wu, S L

    2008-10-17

    With the full BABAR data sample of 465 x 10(6) B(over)B pairs, we observe the decays B+/- -->phiK_(1)(1270) +/- and B +/- -->phiK*_(2)(1430)+/-. We measure the branching fractions (6.1+/-1.6+/-1.1) x 10(-6) and (8.4+/-1.8+/-1.0) x 10(-6) and the fractions of longitudinal polarization 0.46 (+0.12+0.06) _(-0.13-0.07) and 0.80(+0.09)_(-0.10)+/-0.03, respectively. We also report on the B+/- -->phiK*_(0)(1430)+/- decay branching fraction of (7.0+/-1.3+/-0.9) x 10(-6) and several parameters sensitive to CP violation and interference in the above three decays. Upper limits are placed on the B+/- decay rates to final states with phi and K_1(1400)+/-, K*(1410)+/-, K2(1770)+/-, or K_2(1820)+/-. Understanding the observed polarization pattern requires amplitude contributions from an uncertain source.

  12. Aqueous extract of dioscorea opposita thunb. normalizes the hypertension in 2K1C hypertensive rats

    PubMed Central

    2014-01-01

    Background Dioscorea opposita Thunb. (Huai Shan Yao, DOT), a common staple food in China, has been used for more than 2000 years in traditional Chinese medicine (TCM) to treat different systemic diseases including hypertension. The objective of this study was to investigate the possible antihypertensive effects of the aqueous extract of (DOT) in renovascular hypertensive rats as well as the mechanism in reducing blood pressure. Methods The two-kidney one-clip (2K1C) Goldblatt model of renovascular hypertension was used in Wistar rats. Rats with captopril, low-dose DOT and high-dose DOT treated 2K1C groups for 6 weeks. The blood pressure, cardiac mass index (heart weight/body weight), plasma level of angiotensin-II (Ang-II), endothelin-1(ET-1), superoxide dismutase (SOD) and malondialdehyde (MDA) were evaluated. Results DOT significantly reduced mean systolic and diastolic blood pressure after treatment. DOT also significantly increased plasma SOD activity but decreased plasma MDA concentration. Renal function was improved with captopril and DOT. DOT reduced plasma Ang-II activity and plasma ET concentration. They couldalso significantly reduce the left ventricular hypertrophy and cardiac mass index. Conclusions Our results suggest that DOT may have an antihypertensive effect on hypertension by inhibit ET-converting enzyme and antioxidant activity, which warrant further exploration. PMID:24447776

  13. Phosphorylated S6K1 (Thr389) is a molecular adipose tissue marker of altered glucose tolerance.

    PubMed

    Moreno-Navarrete, José María; Ortega, Francisco; Sánchez-Garrido, Miguel Ángel; Sabater, Mònica; Ricart, Wifredo; Zorzano, Antonio; Tena-Sempere, Manuel; Fernández-Real, José Manuel

    2013-01-01

    Molecular tissue markers of altered glucose metabolism will be useful as potential targets for antidiabetic drugs. S6K1 is a downstream signal of insulin action. We aimed to evaluate (pThr389)S6K1 and total S6K1 levels in human and rat fat depots as candidate markers of altered glucose metabolism. (pThr389)S6K1 and total S6K1 levels were measured using enzyme linked immune sorbent assay (ELISA) in 49 adipose tissue samples from subjects with morbid obesity and in 18 peri-renal white adipose tissue samples from rats. The effects of high glucose and rosiglitazone have been explored in human preadipocytes. (pThr389)S6K1/(total)S6K1 in subcutaneous adipose tissue was significantly increased subjects with Type 2 diabetes (0.78 ± 0.26 vs. 0.55 ± 0.14, P=.02) and associated with fasting glucose (r=0.46, P=.04) and glycated hemoglobin (r=0.63, P=.02) in SAT. Similar associations with fasting glucose (r=0.43, P=.03) and IRS1 (r=-0.41, P=.04) gene expression were found in visceral adipose tissue. In addition, rat experiments confirmed the higher (pThr389)S6K1/totalS6K1 levels in adipose tissue in association with obesity-associated metabolic disturbances. (pThr389)S6K1/totalS6K1 was validated using western blot in rat adipose tissue. Both ELISA and western blot data significantly correlated (r=0.85, P=.005). In human preadipocytes, high glucose medium led to increased (pThr389)S6K1/total S6K1 levels in comparison with normal glucose medium, which was significantly decreased under rosiglitazone administration. In conclusion, in human and rat adipose tissue, phosphorylated S6K1 is a marker for increased glucose levels.

  14. Virtual endocasts of Eocene Paramys (Paramyinae): oldest endocranial record for Rodentia and early brain evolution in Euarchontoglires

    PubMed Central

    Amador-Mughal, Farrah

    2016-01-01

    Understanding the pattern of brain evolution in early rodents is central to reconstructing the ancestral condition for Glires, and for other members of Euarchontoglires including Primates. We describe the oldest virtual endocasts known for fossil rodents, which pertain to Paramys copei (Early Eocene) and Paramys delicatus (Middle Eocene). Both specimens of Paramys have larger olfactory bulbs and smaller paraflocculi relative to total endocranial volume than later occurring rodents, which may be primitive traits for Rodentia. The encephalization quotients (EQs) of Pa. copei and Pa. delicatus are higher than that of later occurring (Oligocene) Ischyromys typus, which contradicts the hypothesis that EQ increases through time in all mammalian orders. However, both species of Paramys have a lower relative neocortical surface area than later rodents, suggesting neocorticalization occurred through time in this Order, although to a lesser degree than in Primates. Paramys has a higher EQ but a lower neocortical ratio than any stem primate. This result contrasts with the idea that primates were always exceptional in their degree of overall encephalization and shows that relative brain size and neocortical surface area do not necessarily covary through time. As such, these data contradict assumptions made about the pattern of brain evolution in Euarchontoglires. PMID:26817776

  15. Trypanosoma (Megatrypanum) lainsoni n. sp. from Mesomys hispidus (Rodentia: Echimyidae) in Brazil: trypomastigotes described from experimentally infected laboratory mice.

    PubMed

    Naiff, Roberto Daibes; Barrett, Toby Vincent

    2013-01-01

    We report the detection, isolation and description of Trypanosoma (Megatrypanum) lainsoni n. sp. from a caviomorph rodent, Mesomys hispidus (Rodentia: Echimyidae), obtained in the Rio Negro region of the state of Amazonas, in northern Brazil. Laboratory-bred white mice (Mus musculus) and rats (Rattus rattus) were inoculated with large numbers of culture forms by intraperitoneal route, and trypomastigotes appeared in their blood 3-8 days post-inoculation. One single epimastigote was also found in Mus musculus. Similar attempts to infect Rattus norvegicus, hamsters (Mesocricetus auratus), the opossum Didelphis marsupialis, the anteater Tamandua tetradactyla and triatomine bugs were unsuccessful, following six months of observations and microscopic examinations of blood films and blood cultures. As we have found no previous record of a Trypanosoma (Megatrypanum) species naturally infecting a member of the family Echimyidae, or any other caviomorph rodent, we conclude that this is the first time such an infection has been reported. The new species is unusual in the subgenus for its infectivity to laboratory mice.

  16. Virtual endocasts of Eocene Paramys (Paramyinae): oldest endocranial record for Rodentia and early brain evolution in Euarchontoglires.

    PubMed

    Bertrand, Ornella C; Amador-Mughal, Farrah; Silcox, Mary T

    2016-01-27

    Understanding the pattern of brain evolution in early rodents is central to reconstructing the ancestral condition for Glires, and for other members of Euarchontoglires including Primates. We describe the oldest virtual endocasts known for fossil rodents, which pertain to Paramys copei (Early Eocene) and Paramys delicatus (Middle Eocene). Both specimens of Paramys have larger olfactory bulbs and smaller paraflocculi relative to total endocranial volume than later occurring rodents, which may be primitive traits for Rodentia. The encephalization quotients (EQs) of Pa. copei and Pa. delicatus are higher than that of later occurring (Oligocene) Ischyromys typus, which contradicts the hypothesis that EQ increases through time in all mammalian orders. However, both species of Paramys have a lower relative neocortical surface area than later rodents, suggesting neocorticalization occurred through time in this Order, although to a lesser degree than in Primates. Paramys has a higher EQ but a lower neocortical ratio than any stem primate. This result contrasts with the idea that primates were always exceptional in their degree of overall encephalization and shows that relative brain size and neocortical surface area do not necessarily covary through time. As such, these data contradict assumptions made about the pattern of brain evolution in Euarchontoglires.

  17. A phylogeographic study of the endemic rodent Eliurus carletoni (Rodentia: Nesomyinae) in an ecological transition zone of Northern Madagascar.

    PubMed

    Rakotoarisoa, Jean-Eric; Raheriarisena, Martin; Goodman, Steven M

    2013-01-01

    We conducted a mitochondrial phylogeographic study of the endemic dry forest rodent Eliurus carletoni (Rodentia: Nesomyinae) in an ecological transition zone of northern Madagascar (Loky-Manambato) and 2 surrounding regions (Ankarana and Analamerana). The main goal was to assess the evolutionary consequences on this taxon of the complex landscape features and Quaternary ecological vicissitudes. Three haplogroups were identified from the 215 specimens obtained from 15 populations. High levels of genetic diversity and significant genetic differentiation among populations were observed. The different geographical subdivisions of the study area by regions, by river catchment zones, and the physical distance between populations are not correlated with genetic patterns. In contrast, population structure is mostly explained by the geographic distribution of the samples among existing forest blocks. E. carletoni experienced a genetic bottleneck between 18 750 and 7500 years BP, which correlates with periods when moister climates existed on the island. Overall, our data suggest that the complex genetic patterns of E. carletoni can be explained by Quaternary climatic vicissitudes that resulted in habitat fluctuations between dry and humid forests, as well as subsequent human-induced fragmentation of forest habitat.

  18. Habitat use by Oryzomys subflavus (Rodentia) in an open shrubland formation in Restinga de Jurubatiba National Park, RJ, Brazil.

    PubMed

    Bergallo, H G; Luz, J L; Raíces, D S; Hatano, F H; Martins-Hatano, F

    2005-11-01

    The Restinga de Jurubatiba has at least 10 plant formations, including open Clusia shrubland. This formation is composed of dense shrubs of many shapes and sizes, where Clusia hilariana is one of the most important plant species. Shrublands with Clusia (CC) are poorer in plant species and less dense than shrublands without Clusia (SC). Oryzomys subflavus (Rodentia) is the most abundant small mammal species in the open Clusia shrubland. We tested the hypothesis that the abundance of rodents would increase with the size of the patch and would be higher in SC shrublands. Rodents were captured, marked and released in three 780-m-long transects. At each capture site, we evaluated the shape of the shrubland patch, calculated the area and noted the category of the shrubland. Using ANCOVA, we ascertained whether the abundance of Oryzomys subflavus increased with the sampled area and used CC and SC shrublands differently. We also verified if the size of patches used by rodents varies in the same frequency as the size of available shrublands. Rodent abundance was found to increase significantly with the area. There were no differences in the size of the patches used by rodents and the frequency of the size of available patches. This finding indicates that O. subflavus, in the study area, is a generalist species that uses its habitat according to availability.

  19. FimH adhesin of Escherichia coli K1 type 1 fimbriae activates BV-2 microglia

    SciTech Connect

    Lee, Jongseok; Shin, Sooan; Teng, C.-H.; Hong, Suk Jin; Kim, Kwang Sik . E-mail: kwangkim@jhmi.edu

    2005-09-02

    The generation of intense inflammation in the subarachnoid space in response to meningitis-causing bacteria contributes to brain dysfunction and neuronal injury in bacterial meningitis. Microglia, the major immune effector cells in the central nervous system (CNS), become activated by bacterial components to produce proinflammatory immune mediators. In this study, we showed that FimH adhesin, a tip component of type 1 fimbriae of meningitis-causing Escherichia coli K1, activated the murine microglial cell line, BV-2, which resulted in the production of nitric oxide and the release of tumor necrosis factor-{alpha}. Mitogen-activated protein kinases, ERK and p-38, and nuclear factor-{kappa}B were involved in FimH adhesin-mediated microglial activation. These findings suggest that FimH adhesin contributes to the CNS inflammatory response by virtue of activating microglia in E. coli meningitis.

  20. Genomic Comparison of Escherichia coli K1 Strains Isolated from the Cerebrospinal Fluid of Patients with Meningitis †

    PubMed Central

    Yao, Yufeng; Xie, Yi; Kim, Kwang Sik

    2006-01-01

    Escherichia coli is a major cause of enteric/diarrheal diseases, urinary tract infections, and sepsis. E. coli K1 is the leading gram-negative organism causing neonatal meningitis, but the microbial basis of E. coli K1 meningitis is incompletely understood. Here we employed comparative genomic hybridization to investigate 11 strains of E. coli K1 isolated from the cerebrospinal fluid (CSF) of patients with meningitis. These 11 strains cover the majority of common O serotypes in E. coli K1 isolates from CSF. Our data demonstrated that these 11 strains of E. coli K1 can be categorized into two groups based on their profile for putative virulence factors, lipoproteins, proteases, and outer membrane proteins. Of interest, we showed that some open reading frames (ORFs) encoding the type III secretion system apparatus were found in group 2 strains but not in group 1 strains, while ORFs encoding the general secretory pathway are predominant in group 1 strains. These findings suggest that E. coli K1 strains isolated from CSF can be divided into two groups and these two groups of E. coli K1 may utilize different mechanisms to induce meningitis. PMID:16552050

  1. Amino acid availability regulates S6K1 and protein synthesis in avian insulin-insensitive QM7 myoblasts.

    PubMed

    Tesseraud, Sophie; Bigot, Karine; Taouis, Mohammed

    2003-04-10

    The regulation of S6K1 by nutritional status and insulin has been recently reported in vivo in chicken muscle despite the relative insulin resistance of this tissue as estimated by phosphatidylinositol 3-kinase (PI3-kinase) activity. The present work aimed to study the impact of amino acids on S6K1 activity in quail muscle (QM7) myoblasts. Firstly, we characterized S6K1 in QM7 cells and demonstrated the absence of insulin receptors in these cells. Secondly, we showed that amino acids in the absence of insulin induced S6K1 phosphorylation on Thr389 and concomitantly increased its enzymatic activity. Amino acid-induced S6K1 activation was inhibited by LY294002 (PI3-kinase inhibitor) and rapamycin (inhibitor of the mammalian target of rapamycin, mTOR), suggesting the involvement of an avian homolog of mTOR. The availability of individual amino acids (methionine or leucine) regulated S6K1 phosphorylation on Thr389 and QM7 protein synthesis. In conclusion, amino acids regulate S6K1 phosphorylation and activity in QM7 cells through the mTOR/PI3-kinase pathway in an insulin-independent manner.

  2. Treatment of a long-acting anticoagulant rodenticide poisoning cohort with vitamin K1 during the maintenance period.

    PubMed

    Long, Jianhai; Peng, Xiaobo; Luo, Yuan; Sun, Yawei; Lin, Guodong; Wang, Yongan; Qiu, Zewu

    2016-12-01

    Currently, there are few guidelines for the use of vitamin K1 in the maintenance treatment of long-acting anticoagulant rodenticide (LAAR) poisonings. We explored factors in the treatment of LAAR poisoning during the maintenance period in order to suggest feasible treatment models.Data from 24 cases of anticoagulant rodenticide poisoning in our hospital were collected from January 2013 to May 2016. The patients' sex, age, coagulation function, total time from poisoning to treatment with vitamin K1 (prehospital time), vitamin K1 sustained treatment time (VKSTT), anticoagulant rodenticide category, and specific poison dosage were collected. Multivariate analysis was used to evaluate the correlation between vitamin K1 dosage and other factors during the maintenance period.Only VKSTT (partial regression coefficient -1.133, 0.59, P = 0.035) had an obvious influence on the therapeutic dose of vitamin K1 required during the maintenance period.After an initial pulse therapy, the bleeding and coagulation functions were stabilized, and the patients were subsequently treated with vitamin K1 during the maintenance period. Over time, the maintenance dose of vitamin K1 (10-120 mg/d, intravenous drip) was gradually decreased and was not related to toxicant concentration.

  3. Treatment of a long-acting anticoagulant rodenticide poisoning cohort with vitamin K1 during the maintenance period

    PubMed Central

    Long, Jianhai; Peng, Xiaobo; Luo, Yuan; Sun, Yawei; Lin, Guodong; Wang, Yongan; Qiu, Zewu

    2016-01-01

    Abstract Currently, there are few guidelines for the use of vitamin K1 in the maintenance treatment of long-acting anticoagulant rodenticide (LAAR) poisonings. We explored factors in the treatment of LAAR poisoning during the maintenance period in order to suggest feasible treatment models. Data from 24 cases of anticoagulant rodenticide poisoning in our hospital were collected from January 2013 to May 2016. The patients’ sex, age, coagulation function, total time from poisoning to treatment with vitamin K1 (prehospital time), vitamin K1 sustained treatment time (VKSTT), anticoagulant rodenticide category, and specific poison dosage were collected. Multivariate analysis was used to evaluate the correlation between vitamin K1 dosage and other factors during the maintenance period. Only VKSTT (partial regression coefficient −1.133, 0.59, P = 0.035) had an obvious influence on the therapeutic dose of vitamin K1 required during the maintenance period. After an initial pulse therapy, the bleeding and coagulation functions were stabilized, and the patients were subsequently treated with vitamin K1 during the maintenance period. Over time, the maintenance dose of vitamin K1 (10–120 mg/d, intravenous drip) was gradually decreased and was not related to toxicant concentration. PMID:28002326

  4. SOFIA (+FORCAST) Infrared Spectrophotometry of Comet C/2012 K1 (PanStarrs)

    NASA Astrophysics Data System (ADS)

    Woodward, Charles E.; Kelley, Michael S.P.; Wooden, Diane H.; Harker, David E.; De Buizer, James M.; Gicquel, Adeline

    2014-11-01

    Observing and modeling the properties of small, primitive bodies in the solar system whose origins lie beyond the frost line (> 5 AU) provides critical insight into the formation of the first Solar System solids and establishes observation constraints for planetary system formation invoking migration - the ‘Grand Tack’ epoch followed by the ‘Nice Model’ events - that yielded terrestrial planets in the habitable zone. The characteristics of comet dust can provide evidence to validate the new, emerging picture of small body populations - including comet families - resulting from planetary migration in the early Solar System. Here we present preliminary results of infrared 8 to 27 micron spectrophotometric observations of comet C/2012 K1 (PanStarrs), a dynamically new (1/a0 < 50e-6) Oort Cloud comet, conducted with the NASA’s Stratospheric Observatory for Infrared Astronomy (SOFIA) facility during a series of three flights over the period from 2014 June 06-11 UT. During this interval comet C/2012 K1 (PanStarrs) was at a heliocentric distance of ~1.64 AU and a geocentric distance of ~1.74 AU (pre-perihelion). As a "new" comet (first inner solar system passage), the coma grain population may be extremely pristine, unencumbered by a rime and insufficiently irradiated by the Sun to carbonize its surface organics. We will discuss the derived coma grain properties inferred from modeling of the spectral energy distribution derived from the SOFIA (+FORCAST) data and highlight our preliminary conclusions. Continued observations of comets, especially dynamically young Oort Cloud targets, in the 5-37 micron infrared spectral range accessible with SOFIA (+FORCAST) will provide key observational clues to ascertaining the origins of silicates within our protoplanetary disk, and will serve to place our early disk evolution within the context of other circumstellar disks observed today that may contain the seeds of rocky, terrestrial planets.

  5. EPRS is a critical mTORC1-S6K1 effector that influences adiposity in mice.

    PubMed

    Arif, Abul; Terenzi, Fulvia; Potdar, Alka A; Jia, Jie; Sacks, Jessica; China, Arnab; Halawani, Dalia; Vasu, Kommireddy; Li, Xiaoxia; Brown, J Mark; Chen, Jie; Kozma, Sara C; Thomas, George; Fox, Paul L

    2017-02-16

    Metabolic pathways that contribute to adiposity and ageing are activated by the mammalian target of rapamycin complex 1 (mTORC1) and p70 ribosomal protein S6 kinase 1 (S6K1) axis. However, known mTORC1-S6K1 targets do not account for observed loss-of-function phenotypes, suggesting that there are additional downstream effectors of this pathway. Here we identify glutamyl-prolyl-tRNA synthetase (EPRS) as an mTORC1-S6K1 target that contributes to adiposity and ageing. Phosphorylation of EPRS at Ser999 by mTORC1-S6K1 induces its release from the aminoacyl tRNA multisynthetase complex, which is required for execution of noncanonical functions of EPRS beyond protein synthesis. To investigate the physiological function of EPRS phosphorylation, we generated Eprs knock-in mice bearing phospho-deficient Ser999-to-Ala (S999A) and phospho-mimetic (S999D) mutations. Homozygous S999A mice exhibited low body weight, reduced adipose tissue mass, and increased lifespan, similar to S6K1-deficient mice and mice with adipocyte-specific deficiency of raptor, an mTORC1 constituent. Substitution of the Eprs(S999D) allele in S6K1-deficient mice normalized body mass and adiposity, indicating that EPRS phosphorylation mediates S6K1-dependent metabolic responses. In adipocytes, insulin stimulated S6K1-dependent EPRS phosphorylation and release from the multisynthetase complex. Interaction screening revealed that phospho-EPRS binds SLC27A1 (that is, fatty acid transport protein 1, FATP1), inducing its translocation to the plasma membrane and long-chain fatty acid uptake. Thus, EPRS and FATP1 are terminal mTORC1-S6K1 axis effectors that are critical for metabolic phenotypes.

  6. New cross-bridged cyclam derivative CB-TE1K1P, an improved bifunctional chelator for copper radionuclides.

    PubMed

    Zeng, Dexing; Ouyang, Qin; Cai, Zhengxin; Xie, Xiang-Qun; Anderson, Carolyn J

    2014-01-04

    A new cross-bridged cyclam chelator, CB-TE1K1P, was developed for copper-based radiopharmaceuticals, and this chelator can be labelled with (64)Cu under mild conditions in high specific activity. DBCO-PEG4-CB-TE1K1P was synthesized for conjugation to proteins, while Dde-CB-TE1K1P((t)Bu2)-OH was synthesized for solid-phase peptide synthesis. Examples of the conjugation chemistry, radiolabelling and serum stability of each are presented.

  7. 75 FR 42831 - Proposed Collection; Comment Request for Form 1065, Schedule C, Schedule D, Schedule K-1...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-07-22

    ... Items), Schedule L (Balance Sheets per Books), Schedule M-1 (Reconciliation of Income (Loss) per Books.... (Schedule K-1), Balance Sheets per Books (Schedule L), Reconciliation of Income (Loss) per Books With...

  8. Acetaldehyde at clinically relevant concentrations inhibits inward rectifier potassium current I(K1) in rat ventricular myocytes.

    PubMed

    Bébarová, M; Matejovič, P; Šimurdová, M; Šimurda, J

    2015-01-01

    Considering the effects of alcohol on cardiac electrical behavior as well as the important role of the inward rectifier potassium current I(K1) in arrhythmogenesis, this study was aimed at the effect of acetaldehyde, the primary metabolite of ethanol, on I(K1) in rat ventricular myocytes. Acetaldehyde induced a reversible inhibition of I(K1) with IC(50) = 53.7+/-7.7 microM at -110 mV; a significant inhibition was documented even at clinically-relevant concentrations (at 3 microM by 13.1+/-3.0 %). The inhibition was voltage-independent at physiological voltages above -90 mV. The I(K1) changes under acetaldehyde may contribute to alcohol-induced alterations of cardiac electrophysiology, especially in individuals with a genetic defect of aldehyde dehydrogenase where the acetaldehyde level may be elevated.

  9. Mitogen-independent phosphorylation of S6K1 and decreased ribosomal S6 phosphorylation in senescent human fibroblasts.

    PubMed

    Zhang, H; Hoff, H; Marinucci, T; Cristofalo, V J; Sell, C

    2000-08-25

    The p70 ribosomal S6 kinase (S6K1) is rapidly activated following growth factor stimulation of quiescent fibroblasts and inhibition of this enzyme results in a G(1) arrest. Phosphorylation of the ribosomal S6 protein by S6K1 regulates the translation of both ribosomal proteins and initiation factors, leading to an increase in protein synthesis. We have examined the activation of S6K1 in human fibroblasts following mitogen stimulation. In early passage fibroblasts S6K1 is activated following serum stimulation as evidenced by increased kinase activity and site-specific phosphorylation. In contrast, site-specific phosphorylation of S6K1 at Thr421/Ser424 is diminished in senescent fibroblast cultures. A second phosphorylation site within S6K1 (Ser411) is phosphorylated even in the absence of serum stimulation and the enzyme shows increased phosphorylation as judged by decreased electrophoretic mobility. Inhibitor studies indicate that this phosphorylation is dependent upon the mammalian target of rapamycin, PI 3-kinase, and the MAPK pathway. In order to understand the consequences of the altered phosphorylation of the S6K1, we examined the phosphorylation state of the ribosomal S6 protein. In early passage fibroblasts the ribosomal S6 protein is phosphorylated upon serum stimulation while the phosphorylation of the ribosomal S6 protein is drastically reduced in senescent fibroblasts. These results suggest that the intracellular regulators of S6K1 are altered during replicative senescence leading to a deregulation of the enzyme and a loss of ribosomal S6 phosphorylation.

  10. Approximating common fixed points of asymptotically quasi-nonexpansive mappings by a k+1-step iterative scheme with error terms

    NASA Astrophysics Data System (ADS)

    Xiao, Jian-Zhong; Sun, Jing; Huang, Xuan

    2010-02-01

    In this paper a k+1-step iterative scheme with error terms involving k+1 asymptotically quasi-nonexpansive mappings is studied. In usual Banach spaces, some sufficient and necessary conditions are given for the iterative scheme to approximate a common fixed point. In uniformly convex Banach spaces, power equicontinuity for a mapping is introduced and a series of new convergence theorems are established. Several known results in the current literature are extended and refined.

  11. Molecular characterization and expression profile of MAP2K1ip1/MP1 gene from tiger shrimp, Penaeus monodon.

    PubMed

    Yang, Lishi; Liu, Xianjun; Huang, Jianhua; Yang, Qibin; Qiu, Lihua; Liu, Wenjing; Jiang, Shigui

    2012-05-01

    MAPK kinase 1 interacting protein 1 (MAP2K1ip1) is an important scaffold proteins of the mitogen-activated protein kinase (MAPK) pathway that form an active signaling module and enhance the specificity and spatiality of MAPK signaling. In the present study, we identified and characterized a MAP2K1ip1 cDNA from tiger shrimp Penaeus monodon (designated as PmMAP2K1ip1). The open reading frame of PmMAP2K1ip1 is 372 bp encoding 123 amino-acid residues with a MAPK interaction domain. The predicted PmMAP2Kip1 protein is 13.6 KDa with the theoretical isoelectric point of 6.3. PmMAP2K1ip1 shared the highest amino acid with Nasonia vitripennis and Strongylocentrotus purpuratus, at 48% and 47.5%, respectively. Phylogenic analysis shows PmMAP2Kip1 is clustering with SpMAP2Kip1, and close to the group of MAP2Kip1s from insect. Furthermore, semiquantitative RT-PCR revealed PmMAP2Kip1 is widely distributed in most examined tissues except nerve, and high expressed in ovary, hemocyte, intestines and hepatopancreas. Meanwhile, PmMAP2k1ip1 is expressed ubiquitously during larval and sex gland development, and keep a high level at the initial development stage. Quantitative real time RT-PCR revealed PmMAP2K1ip1 were up-regulated by lipopolysaccharide and peptidoglycan (PGN) in haemocyte. These data reveal MAP2K1ip1 is a multifunction protein that involved development and immune response. It is benefit to characterize other MAPK signal genes and elucidate the molecular regulation mechanism of MAPK signaling in tiger shrimp.

  12. The mitochondrial alternative oxidase pathway protects the photosynthetic apparatus against photodamage in Rumex K-1 leaves

    PubMed Central

    2012-01-01

    Background It is known that excess reducing equivalents in the form of NADPH in chloroplasts can be transported via shuttle machineries, such as the malate-oxaloacetate (OAA) shuttle, into the mitochondria, where they are efficiently oxidised by the mitochondrial alternative oxidase (AOX) respiratory pathway. Therefore, it has been speculated that the AOX pathway may protect plants from photoinhibition, but the mechanism by which this protection occurs remains to be elucidated. Results The observation that the malate-OAA shuttle activity and the AOX pathway capacity increased markedly after intense light treatment in Rumex K-1 leaves indicates that excess NADPH was transported from the chloroplasts and oxidised by the AOX pathway. The inhibition of the AOX pathway by salicylhydroxamic acid (SHAM) caused the over-reduction of the photosystem I (PSI) acceptor side, as indicated by the increases in the extent of reduction of P700+. Furthermore, the photosynthetic linear electron flow was restricted, which was indicated by the decreases in the PSII electron transport rate (ETR) and the photosynthetic O2 evolution rate. The restriction of the photosynthetic linear electron flow, which generates the thylakoid ΔpH, inevitably decreased the de-epoxidation of the xanthophyll cycle (ΔPRI). Therefore, the induction of non-photochemical quenching (NPQ) was suppressed when the AOX pathway was inhibited. The effect of the inhibition of the AOX pathway on NPQ induction was less at 20 mM NaHCO3 than at 1 mM NaHCO3. The suppression of NPQ induction by the inhibition of the AOX pathway was also observed during the induction phase of photosynthesis. In addition, the inhibition of the AOX pathway increased the accumulation of hydrogen peroxide (H2O2), suggesting that the AOX pathway functions as an antioxidant mechanism. Conclusions The inhibition of the AOX pathway resulted in the rapid accumulation of NADPH in the chloroplasts, which caused the over-reduction of the PSI acceptor

  13. Lactobacillus rhamnosus GG supernatant enhance neonatal resistance to systemic Escherichia coli K1 infection by accelerating development of intestinal defense

    PubMed Central

    He, Xiaolong; Zeng, Qing; Puthiyakunnon, Santhosh; Zeng, Zhijie; Yang, Weijun; Qiu, Jiawen; Du, Lei; Boddu, Swapna; Wu, Tongwei; Cai, Danxian; Huang, Sheng-He; Cao, Hong

    2017-01-01

    The objective of this study was to determine whether Lactobacillus rhamnosus GG culture supernatant (LCS) has a preventive effect against gut-derived systemic neonatal Escherichia coli (E. coli) K1 infection. The preventive effects were evaluated in human colonic carcinoma cell line Caco-2 and neonatal rat models. Our in vitro results showed that LCS could block adhesion, invasion and translocation of E. coli K1 to Caco-2 monolayer via up-regulating mucin production and maintaining intestinal integrity. In vivo experiments revealed that pre-treatment with LCS significantly decrease susceptibility of neonatal rats to oral E. coli K1 infection as reflected by reduced bacterial intestinal colonization, translocation, dissemination and systemic infections. Further, we found that LCS treated neonatal rats have higher intestinal expressions of Ki67, MUC2, ZO-1, IgA, mucin and lower barrier permeability than those in untreated rats. These results indicated that LCS could enhance neonatal resistance to systemic E. coli K1 infection via promoting maturation of neonatal intestinal defense. In conclusions, our findings suggested that LCS has a prophylactic effect against systemic E. coli K1 infection in neonates. Future studies aimed at identifying the specific active ingredients in LCS will be helpful in developing effective pharmacological strategies for preventing neonatal E. coli K1 infection. PMID:28262688

  14. Introduction of a carbon paste electrode based on nickel carbide for investigation of interaction between warfarin and vitamin K1.

    PubMed

    Torkashvand, Maryam; Gholivand, Mohammad Bagher; Taherpour, Avat Arman; Boochani, Arash; Akhtar, Arsalan

    2017-03-01

    In this paper a novel electrochemical sensor based on nickel carbide (Ni3C) nanoparticles as a new modifier was constructed. Ni3C nanoparticle was synthesized and characterized by scanning electron microscopy, X-ray diffraction and first-principles study. Electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) studies confirmed the electrode modification. Afterwards, the new electrode for the first time was used for interaction study between vitamin K1 and warfarin as an anticoagulant drug by differential pulse voltammetry. The adduct formation between the drug and vitamin K1 was improved by decreasing in anodic peak current of warfarin in the presence of different amounts of vitamin K1. The binding constant between warfarin and vitamin K1 was obtained by voltammetric and UV-vis and fluorescence spectroscopic methods. The molecular modeling method was also performed to explore the structural features and binding mechanism of warfarin to vitamin K1. The different aspects of modeling of vitamin K1 and warfarin and their adduct structures confirmed the adduct formation by hydrogen bonding.

  15. Resolution of structure of PIP5K1A reveals molecular mechanism for its regulation by dimerization and dishevelled

    PubMed Central

    Hu, Jian; Yuan, Qianying; Kang, Xue; Qin, Yuanbo; Li, Lin; Ha, Ya; Wu, Dianqing

    2015-01-01

    Type I phosphatidylinositol phosphate kinase (PIP5K1) phosphorylates the head group of phosphatidylinositol 4-phosphate (PtdIns4P) to generate PtdIns4,5P2, which plays important roles in a wide range of cellular functions including Wnt signalling. However, the lack of its structural information has hindered the understanding of its regulation. Here we report the crystal structure of the catalytic domain of zebrafish PIP5K1A at 3.3 Å resolution. This molecule forms a side-to-side dimer. Mutagenesis study of PIP5K1A reveals two adjacent interfaces for the dimerization and interaction with the DIX domain of the Wnt signalling molecule dishevelled. Although these interfaces are located distally to the catalytic/substrate-binding site, binding to these interfaces either through dimerization or the interaction with DIX stimulates PIP5K1 catalytic activity. DIX binding additionally enhances PIP5K1 substrate binding. Thus, this study elucidates regulatory mechanisms for this lipid kinase and provides a paradigm for the understanding of PIP5K1 regulation by their interacting molecules. PMID:26365782

  16. Structure Function Studies of Vaccinia Virus Host Range Protein K1 Reveal a Novel Functional Surface for Ankyrin Repeat Proteins

    SciTech Connect

    Li, Yongchao; Meng, Xiangzhi; Xiang, Yan; Deng, Junpeng

    2010-06-15

    Poxvirus host tropism at the cellular level is regulated by virus-encoded host range proteins acting downstream of virus entry. The functioning mechanisms of most host range proteins are unclear, but many contain multiple ankyrin (ANK) repeats, a motif that is known for ligand interaction through a concave surface. We report here the crystal structure of one of the ANK repeat-containing host range proteins, the vaccinia virus K1 protein. The structure, at a resolution of 2.3 {angstrom}, showed that K1 consists entirely of ANK repeats, including seven complete ones and two incomplete ones, one each at the N and C terminus. Interestingly, Phe82 and Ser83, which were previously shown to be critical for K1's function, are solvent exposed and located on a convex surface, opposite the consensus ANK interaction surface. The importance of this convex surface was further supported by our additional mutagenesis studies. We found that K1's host range function was negatively affected by substitution of either Asn51 or Cys47 and completely abolished by substitution of both residues. Cys47 and Asn51 are also exposed on the convex surface, spatially adjacent to Phe82 and Ser83. Altogether, our data showed that K1 residues on a continuous convex ANK repeat surface are critical for the host range function, suggesting that K1 functions through ligand interaction and does so with a novel ANK interaction surface.

  17. Scanner performance predictor and optimizer in further low-k1 lithography

    NASA Astrophysics Data System (ADS)

    Aoyama, Hajime; Nakashima, Toshiharu; Ogata, Taro; Kudo, Shintaro; Kita, Naonori; Ikeda, Junji; Matsui, Ryota; Yamamoto, Hajime; Sukegawa, Ayako; Makino, Katsushi; Murayama, Masayuki; Masaki, Kazuo; Matsuyama, Tomoyuki

    2014-03-01

    Due to the importance of errors in lithography scanners, masks, and computational lithography in low-k1 lithography, application software is used to simultaneously reduce them. We have developed "Masters" application software, which is all-inclusive term of critical dimension uniformity (CDU), optical proximity effect (OPE), overlay (OVL), lens control (LNS), tool maintenance (MNT) and source optimization for wide process window (SO), for compensation of the issues on imaging and overlay. In this paper, we describe the more accurate and comprehensive solution of OPE-Master, LNS-Master and SO-Master with functions of analysis, prediction and optimization. Since OPE-Master employed a rigorous simulation, a root cause of error in OPE matching was found out. From the analysis, we had developed an additional knob and evaluated a proof-of- concept for the improvement. Influence of thermal issues on projection optics is evaluated with a heating prediction, and an optimization with scanner knobs on an optimized source taken into account mask 3D effect for obtaining usable process window. Furthermore, we discuss a possibility of correction for reticle expansion by heating comparing calculation and measurement.

  18. Sewage sludge hazardous assessment: chemical evaluation and cytological effects in CHO-k1 cells.

    PubMed

    Bonomo, M M; Morozesk, M; Duarte, I D; Rocha, L D; Fernandes, M N; Matsumoto, S T

    2016-06-01

    Application of sewage sludge in agricultural lands is a growing practice in several countries due to its numerous benefits to soil and crops, where chemical and pathogen levels are determined by corresponding legislation. However, the presence of contaminants in residues must always be controlled before application due to their dangerous effects over the ecosystem and potential risks to human health. The main objective of this study was to integrate biological and chemical analysis in order to help elucidating the residue potential toxic, cytotoxic, and mutagenic effects. We evaluate samples of sewage sludge before and after the sanitizing treatment with lime in cytokinesis-block assay using CHO-k1 culture cells. The sanitizing treatment promoted a decrease in pathogen levels, which is the main purpose of this process. Even with chemical levels below the established by environmental agencies, results showed sewage sludge ability to enhance genotoxic and mutagenic effects, proving that residue should be handled with caution in order to minimize its environmental and human risk.

  19. Characterization of a Novel Thermostable O-Acetylserine Sulfhydrylase from Aeropyrum pernix K1

    PubMed Central

    Mino, Koshiki; Ishikawa, Kazuhiko

    2003-01-01

    An O-acetylserine sulfhydrylase (OASS) from the hyperthermophilic archaeon Aeropyrum pernix K1, which shares the pyridoxal 5′-phosphate binding motif with both OASS and cystathionine β-synthase (CBS), was cloned and expressed by using Escherichia coli Rosetta(DE3). The purified protein was a dimer and contained pyridoxal 5′-phosphate. It was shown to be an enzyme with CBS activity as well as OASS activity in vitro. The enzyme retained 90% of its activity after a 6-h incubation at 100°C. In the O-acetyl-l-serine sulfhydrylation reaction, it had a pH optimum of 6.7, apparent Km values for O-acetyl-l-serine and sulfide of 28 and below 0.2 mM, respectively, and a rate constant of 202 s−1. In the l-cystathionine synthetic reaction, it showed a broad pH optimum in the range of 8.1 to 8.8, apparent Km values for l-serine and l-homocysteine of 8 and 0.51 mM, respectively, and a rate constant of 0.7 s−1. A. pernix OASS has a high activity in the l-cysteine desulfurization reaction, which produces sulfide and S-(2,3-hydroxy-4-thiobutyl)-l-cysteine from l-cysteine and dithiothreitol. PMID:12644499

  20. Role of F225 in O-phosphoserine sulfhydrylase from Aeropyrum pernix K1.

    PubMed

    Takeda, Emi; Kunimoto, Kohei; Kawai, Yoshito; Kataoka, Misumi; Ishikawa, Kazuhiko; Nakamura, Takashi

    2016-09-01

    O-Phosphoserine sulfhydrylase (OPSS) synthesizes cysteine from O-phospho-L-serine (OPS) and sulfide. We have determined the three-dimensional structures of OPSS from hyperthermophilic archaeon Aeropyrum pernix K1 (ApOPSS) in complex with aminoacrylate intermediate (AA) formed from pyridoxal 5'-phosphate with OPS or in complex with cysteine and compared them with that of ApOPSS. We found an orientational change of F225 at the active-site entrance and constructed an F225A mutant to examine its activities and AA stability and clarify the role of F225 in ApOPSS. The OPS and O-acetyl-L-serine (OAS) sulfhydrylase activities of the F225A mutant decreased by 4.2- and 15-fold compared to those of the wild-type (wt) ApOPSS, respectively. The ability of OPS and OAS to form AA also decreased by 12- and 27-fold, respectively. AA was less stable in the F225A mutant than in the wt ApOPSS. Simulated docking showed that leaving groups, such as phosphate and acetate, were oriented to the inside of the active site in the F225A mutant, whereas they were oriented to the entrance in the wt ApOPSS. These results suggest that F225 in ApOPSS plays important roles in maintaining the hydrophobic environment of AA from solvent water and in controlling the orientation of leaving groups.

  1. Catalog of type specimens of recent mammals: Rodentia (Sciuromorpha and Castorimorpha) in the National Museum of Natural History, Smithsonian Institution

    USGS Publications Warehouse

    Fisher, Robert D.; Ludwig, Craig A.

    2012-01-01

    The type collection of Recent mammals in the Division of Mammals, National Museum of Natural History, Smithsonian Institution, contains 843 specimens bearing names of 820 species group taxa of Rodentia (Sciuromorpha and Castorimorpha) as of July 2011. This catalog presents a list of these holdings, which comprise 798 holotypes, 14 lectotypes, seven syntypes (30 specimens), and one neotype. In addition, we include three holotypes and 10 specimens that are part of syntype series that should be in the collection but cannot be found and three syntypes that were originally in this collection but are now known to be in other collections. One specimen that no longer has name-bearing status is included for the record. Forty-one of the names are new since the last type catalog. One new lectotype is designated. Suborders and families are listed as in Wilson and Reeder. Within families, currently recognized genera are arranged alphabetically. Within each currently recognized genus, accounts are arranged alphabetically by original published name. Information in each account includes original name and abbreviated citation thereto, current name if other than original, citation for first use of current name combination for the taxon (or new name combination if used herein for the first time), type designation, U.S. National Museum catalog number(s), preparation, age and sex, type locality, date of collection and name of collector, collector’s original number, and comments or additional information as appropriate. Digital photographs of each specimen serve as a condition report and will be linked to each electronic specimen record.

  2. Description of a new species of Ixodes Latreille, 1795 (Acari: Ixodidae) and redescription of I. lasallei Méndez & Ortiz, 1958, parasites of agoutis and pacas (Rodentia: Dasyproctidae, Cuniculidae) in Central and South America.

    PubMed

    Apanaskevich, Dmitry A; Bermúdez, Sergio E

    2017-05-01

    Ixodes bocatorensis n. sp. (Acari: Ixodidae), is described based on adults ex agoutis (Rodentia: Dasyproctidae), pacas (Rodentia: Cuniculidae) and "tapir and sloth" (Perissodactyla: Tapiridae and Pilosa) from Colombia, Panama and Venezuela. Adults of I. bocatorensis n. sp. are similar to those of I. lasallei Méndez & Ortiz, 1958 but can be distinguished by the scutum dimensions, punctation pattern, gnathosoma and palpi measurements and their ratios, basis capituli anterior angle and shape of the spur of palpal segment I ventrally. For comparative purposes the female of I. lasallei is redescribed and the true male of this species is described for the first time. Studied adults of I. lasallei were found on agoutis, pacas and ocelot (Carnivora: Felidae) in Colombia, Peru and Venezuela.

  3. K6PC-5, a novel sphingosine kinase 1 (SphK1) activator, alleviates dexamethasone-induced damages to osteoblasts through activating SphK1-Akt signaling.

    PubMed

    Ji, Feng; Mao, Li; Liu, Yuanyuan; Cao, Xiaojian; Xie, Yue; Wang, Shouguo; Fei, Haodong

    2015-03-13

    Long-term glucocorticoid usage is a common cause of non-traumatic femoral head osteonecrosis. Glucocorticoids (i.e. dexamethasone (Dex)) could directly induce damages to osteoblasts. In the current study, we investigated the potential activity of K6PC-5 [N-(1,3-dihydroxyisopropyl)-2-hexyl-3-oxo-decanamide], a novel sphingosine kinase 1 (SphK1) activator, against this process. Our data revealed that both osteoblastic-like MC3T3-E1 cells and primary murine osteoblasts were responsible to K6PC-5. K6PC-5 activated SphK1, increased sphingosine-1-phosphate (S1P) production and induced Akt phosphorylation in cultured osteoblasts. Functionally, K6PC-5 protected osteoblasts from Dex-induced apoptosis and necrosis. Such signaling and functional effects by K6PC-5 were prevented by the SphK1 inhibitor N,N-dimethylsphingosine (DMS), and by SphK1-siRNAs. On the other hand, exogenously-added S1P activated Akt and reduced Dex-induced osteoblast damages. LY294002 and MK-2206, two established Akt inhibitors, alleviated K6PC-5- or S1P-mediated osteoblast protection against Dex. Together, our results suggest that K6PC-5 alleviates Dex-induced osteoblast injuries through activating SphK1-Akt signaling. K6PC-5 might be further investigated in animal or clinical studies for its anti-glucocorticoids-associated osteonecrosis potential.

  4. Final report on the EURAMET.PR-K1.a-2009 comparison of spectral irradiance 250 nm—2500 nm

    NASA Astrophysics Data System (ADS)

    Goodman, Teresa; Servantes, William; Woolliams, Emma; Sperfeld, Peter; Simionescu, Mihai; Blattner, Peter; Källberg, Stefan; Khlevnoy, Boris; Dekker, Paul

    2015-01-01

    This report gives the results of the EURAMET.PR-K1.a-2009 comparison of spectral irradiance over the wavelength range 250 nm—2500 nm. Seven laboratories took part, including the pilot. In general the results are consistent, with a few exceptions as explained in the report. The EURAMET.PR-K1.a key comparison detailed in this report was carried out to establish the degrees of equivalence for the participating European laboratories with respect to the Key Comparison Reference Value (KCRV) of the CCPR-K1.a key comparison. The EURAMET.PR-K1.a key comparison was piloted by the National Physical Laboratory (NPL), who also acted as pilot for the CCPR-K1.a key comparison; a further linkage to the KCRV of the CCPR-K1.a key comparison was provided through the participation of the Physikalisch-Technische Bundesanstalt (PTB) in both comparisons. The other participants were: National Institute of Metrology of Romania (INM-RO), Federal Office of Metrology (METAS), VSL Dutch Metrology Institute (VSL), SP Technical Research Institute of Sweden (SP) and All Russian Institute for Optical and Physical Measurements (VNIIOFI). Measurements were made by each laboratory at 44 designated wavelengths, or a subset of these wavelengths. The link laboratories made measurements at all 44 wavelengths. For the purposes of analysis each wavelength has been treated independently, as for the CCPR K1.a comparison. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCPR, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

  5. Adipocyte-specific deletion of Ip6k1 reduces diet-induced obesity by enhancing AMPK-mediated thermogenesis

    PubMed Central

    Zhu, Qingzhang; Ghoshal, Sarbani; Rodrigues, Ana; Gao, Su; Asterian, Alice; Kamenecka, Theodore M.; Barrow, James C.

    2016-01-01

    Enhancing energy expenditure (EE) is an attractive strategy to combat obesity and diabetes. Global deletion of Ip6k1 protects mice from diet-induced obesity (DIO) and insulin resistance, but the tissue-specific mechanism by which IP6K1 regulates body weight is unknown. Here, we have demonstrated that IP6K1 regulates fat accumulation by modulating AMPK-mediated adipocyte energy metabolism. Cold exposure led to downregulation of Ip6k1 in murine inguinal and retroperitoneal white adipose tissue (IWAT and RWAT) depots. Adipocyte-specific deletion of Ip6k1 (AdKO) enhanced thermogenic EE, which protected mice from high-fat diet–induced weight gain at ambient temperature (23°C), but not at thermoneutral temperature (30°C). AdKO-induced increases in thermogenesis also protected mice from cold-induced decreases in body temperature. UCP1, PGC1α, and other markers of browning and thermogenesis were elevated in IWAT and RWAT of AdKO mice. Cold-induced activation of sympathetic signaling was unaltered, whereas AMPK was enhanced, in AdKO IWAT. Moreover, beige adipocytes from AdKO IWAT displayed enhanced browning, which was diminished by AMPK depletion. Furthermore, we determined that IP6 and IP6K1 differentially regulate upstream kinase-mediated AMPK stimulatory phosphorylation in vitro. Finally, treating mildly obese mice with the IP6K inhibitor TNP enhanced thermogenesis and inhibited progression of DIO. Thus, IP6K1 regulates energy metabolism via a mechanism that could potentially be targeted in obesity. PMID:27701146

  6. Colibactin Contributes to the Hypervirulence of pks+ K1 CC23 Klebsiella pneumoniae in Mouse Meningitis Infections

    PubMed Central

    Lu, Min-Chi; Chen, Ying-Tsong; Chiang, Ming-Ko; Wang, Yao-Chen; Hsiao, Pei-Yi; Huang, Yi-Jhen; Lin, Ching-Ting; Cheng, Ching-Chang; Liang, Chih-Lung; Lai, Yi-Chyi

    2017-01-01

    Klebsiella pneumoniae is the most common pathogen of community-acquired meningitis in Taiwan. However, the lack of a physiologically relevant meningitis model for K. pneumoniae has impeded research into its pathogenesis mechanism. Based on the core genome MLST analyses, the hypervirulent K1 K. pneumoniae strains, which are etiologically implicated in adult meningitis, mostly belong to a single clonal complex, CC23. Some K1 CC23 K. pneumoniae strains carry a gene cluster responsible for colibactin production. Colibactin is a small genotoxic molecule biosynthesized by an NRPS-PKS complex, which is encoded by genes located on the pks island. Compared to other hypervirulent K. pneumoniae which primarily infect the liver, the colibactin-producing (pks+) K1 CC23 strains had significant tropism toward the brain of BALB/c mice. We aimed in this study to develop a physiologically relevant meningitis model with the use of pks+ K1 CC23 K. pneumoniae. Acute meningitis was successfully induced in adult BALB/c male mice through orogastric, intranasal, and intravenous inoculation of pks+ K1 CC23 K. pneumoniae. Besides the typical symptoms of bacterial meningitis, severe DNA damages, and caspase 3-independent cell death were elicited by the colibactin-producing K1 CC23 K. pneumoniae strain. The deletion of clbA, which abolished the production of colibactin, substantially hindered K. pneumoniae hypervirulence in the key pathogenic steps toward the development of meningitis. Our findings collectively demonstrated that colibactin was necessary but not sufficient for the meningeal tropism of pks+ K1 CC23 K. pneumoniae, and the mouse model established in this study can be applied to identify other virulence factors participating in the development of this life-threatening disease.

  7. Spatially Resolved Spectroscopy Across HD189733 (K1V) Using Exoplanet Transits

    NASA Astrophysics Data System (ADS)

    Gustavsson, Martin; Dravins, Dainis; Ludwig, Hans-Günter

    2016-06-01

    For testing 3-dimensional models of stellar atmospheres, spectroscopy across spatially resolved stellar surfaces would be desired with a spectral resolution of(R = 100,000) or more. Hydrodynamic models predict variations in line profile shapes, strengths, wavelength positions and asymmetries. These variations vary systematically between disk center and limb and as a function of line strength, excitation potential and wavelength region. However, except for a few supergiants and the Sun, current telescopes are not yet capable of resolving any stellar surfaces. One alternative method to resolve distant stellar surfaces, feasible already now, is differential spectroscopy of transiting exoplanet systems. By subtracting in-transit spectra from the spectrum outside of transit, the spectra from stellar surface portions temporarily hidden behind the planet can be disentangled. Since transiting planets cover only a small portion of the stellar surface, the method requires a very high signal-to-noise ratio, obtainable by averaging numerous similar spectral lines. We apply such differential spectroscopy on the 7.7 mag K1V star HD 189733 ('Alopex'*); its transiting planet covers ˜ 3% of its host star's surface, which is the deepest known transit among the brighter systems. Archival data from the ESO HARPS spectrometerare used to construct averaged profiles of photospheric Fe I lines, with the aim of comparing spatially resolved profiles to analogous synthetic line profiles computed from the 3-dimensional hydrodynamic CO5BOLD model. * We refer to HD 189733 as 'Alopex' (from the Greek 'αλɛπού'), denoting a fox related to the one that gave name to its constellation of Vulpecula.

  8. Development of a High-Throughput Screening Assay to Identify Inhibitors of the Lipid Kinase PIP5K1C.

    PubMed

    Wright, Brittany D; Simpson, Catherine; Stashko, Michael; Kireev, Dmitri; Hull-Ryde, Emily A; Zylka, Mark J; Janzen, William P

    2015-06-01

    Phosphatidylinositol 4-phosphate 5-kinases (PIP5Ks) regulate a variety of cellular processes, including signaling through G protein-coupled receptors (GPCRs), endocytosis, exocytosis, and cell migration. These lipid kinases synthesize phosphatidylinositol 4,5-bisphosphate (PIP2) from phosphatidylinositol 4-phosphate [PI(4)P]. Because small-molecule inhibitors of these lipid kinases did not exist, molecular and genetic approaches were predominantly used to study PIP5K1 regulation of these cellular processes. Moreover, standard radioisotope-based lipid kinase assays cannot be easily adapted for high-throughput screening. Here, we report a novel, high-throughput, microfluidic mobility shift assay to identify inhibitors of PIP5K1C. This assay uses fluorescently labeled phosphatidylinositol 4-phosphate as the substrate and recombinant human PIP5K1C. Our assay exhibited high reproducibility, had a calculated adenosine triphosphate Michaelis constant (Km) of 15 µM, performed with z' values >0.7, and was used to screen a kinase-focused library of ~4700 compounds. From this screen, we identified several potent inhibitors of PIP5K1C, including UNC3230, a compound that we recently found can reduce nociceptive sensitization in animal models of chronic pain. This novel assay will allow continued drug discovery efforts for PIP5K1C and can be adapted easily to screen additional lipid kinases.

  9. Development of a high-throughput screening assay to identify inhibitors of the lipid kinase PIP5K1C

    PubMed Central

    Wright, Brittany D.; Simpson, Catherine; Stashko, Michael; Kireev, Dmitri; Hull-Ryde, Emily A.; Zylka, Mark J.; Janzen, William P.

    2015-01-01

    Phosphatidylinositol 4-phosphate 5-kinases (PIP5Ks) regulate a variety of cellular processes including signaling through G protein-coupled receptors (GPCRs), endocytosis, exocytosis, and cell migration. These lipid kinases synthesize phosphatidylinositol 4,5-bisphosphate (PIP2) from phosphatidylinositol 4-phosphate [PI(4)P]. Since small molecule inhibitors of these lipid kinases did not exist, molecular and genetic approaches were predominantly used to study PIP5K1 regulation of these cellular processes. Moreover, standard radioisotope-based lipid kinase assays cannot be easily adapted for high-throughput screening. Here, we report a novel high-throughput microfluidic mobility shift assay to identify inhibitors of PIP5K1C. This assay utilizes fluorescently labeled phosphatidylinositol 4-phosphate as the substrate and recombinant human PIP5K1C. Our assay exhibited high reproducibility, had a calculated ATP Km of 15 µM, performed with z’ values >0.7, and was used to screen a kinase-focused library of ~4,700 compounds. From this screen, we identified several potent inhibitors of PIP5K1C, including UNC3230, a compound that we recently found can reduce nociceptive sensitization in animal models of chronic pain. This novel assay will allow continued drug discovery efforts for PIP5K1C and can be easily adapted to screen additional lipid kinases. PMID:25534829

  10. Gene-Environment Interactions Target Mitogen-activated Protein 3 Kinase 1 (MAP3K1) Signaling in Eyelid Morphogenesis*

    PubMed Central

    Mongan, Maureen; Meng, Qinghang; Wang, Jingjing; Kao, Winston W.-Y.; Puga, Alvaro; Xia, Ying

    2015-01-01

    Gene-environment interactions determine the biological outcomes through mechanisms that are poorly understood. Mouse embryonic eyelid closure is a well defined model to study the genetic control of developmental programs. Using this model, we investigated how exposure to dioxin-like environmental pollutants modifies the genetic risk of developmental abnormalities. Our studies reveal that mitogen-activated protein 3 kinase 1 (MAP3K1) signaling is a focal point of gene-environment cross-talk. Dioxin exposure, acting through the aryl hydrocarbon receptor (AHR), blocked eyelid closure in genetic mutants in which MAP3K1 signaling was attenuated but did not disturb this developmental program in either wild type or mutant mice with attenuated epidermal growth factor receptor or WNT signaling. Exposure also markedly inhibited c-Jun phosphorylation in Map3k1+/− embryonic eyelid epithelium, suggesting that dioxin-induced AHR pathways can synergize with gene mutations to inhibit MAP3K1 signaling. Our studies uncover a novel mechanism through which the dioxin-AHR axis interacts with the MAP3K1 signaling pathways during fetal development and provide strong empirical evidence that specific gene alterations can increase the risk of developmental abnormalities driven by environmental pollutant exposure. PMID:26109068

  11. Multiplex assessment of serum cytokine and chemokine levels in idiopathic morphea and vitamin K1-induced morphea.

    PubMed

    Cox, Lori Ann; Webster, Guy F; Piera-Velazquez, Sonsoles; Jimenez, Sergio A

    2017-02-20

    The levels of 63 cytokines, chemokines, and growth factors were measured in the serum of four patients with idiopathic morphea and of one patient with vitamin K1-induced morphea employing a multiplex assay to identify the role of inflammatory/immunologic events in their pathogenesis. Full-thickness skin biopsies of affected skin were analyzed by histopathology. Luminex assays for 63 cytokines, chemokines, and growth factors were performed in the sera from four patients with idiopathic morphea and in two different samples of serum obtained in two separate occasions from one patient with vitamin K1-induced morphea. The serum values of numerous inflammatory cytokines and growth factors including IL-2, IL-4, IL-6, and IFNβ were markedly increased in the serum of patients with idiopathic morphea, whereas, these values were normal in the serum of the patient with vitamin K1-induced morphea. In contrast, serum eotaxin levels were greater than threefold higher in the patient with vitamin K1-induced morphea compared to patients with idiopathic morphea. The results demonstrated remarkable increases in the levels of numerous cytokines and chemokines in the serum samples of all patients with idiopathic morphea indicative of a prominent role of inflammatory/immunologic events in its pathogenesis. The results also showed statistically significant differences between idiopathic morphea and vitamin K1-induced morphea suggesting that their development involves different pathogenetic mechanisms.

  12. Heads or Tails: Genotyping of Hepatitis C Virus Concerning the 2k/1b Circulating Recombinant Form

    PubMed Central

    Schuermans, Wim; Orlent, Hans; Desombere, Isabelle; Descheemaeker, Patrick; Van Vlierberghe, Hans; Geerts, Anja; Verhelst, Xavier; Reynders, Marijke; Padalko, Elizaveta

    2016-01-01

    As different hepatitis C virus (HCV) genotypes respond differently to initiated therapy, correct HCV genotyping is essential. A potential risk for misclassification of the intergenotypic HCV circulating recombinant form (CRF) 2k/1b strains exists, depending on the genotyping method used. The aim was to investigate the differences in HCV genotyping methods with regard to CRF 2k/1b and to gain insight in the prevalence of the CRF 2k/1b. Genotyping results by Versant HCV Genotype Assay were compared with nonstructural protein 5B (NS5B) sequencing. In total, from November 2001 until March 2015, 3296 serum samples were analyzed by Versant HCV Genotype Assay. As misclassified CRF is harbored among HCV genotype 2, we further focused our search on 142 (4.3%) samples positive for HCV genotype 2. On 116 (81.7%) retrieved samples, the NS5B sequencing was performed. Twelve out of the 116 retrieved samples (10.3%) were classified as CRF 2k/1b by sequencing of the NS5B region. Ten of these 12 samples were originally misclassified as genotype 2a or 2c, while 2 of them were misclassified as genotype 2. Our results show that the current prevalence of CRF 2k/1b is underestimated. The importance of correct HCV genotyping is emphasized, considering the tailored choice of treatment regimen and overall prognosis. PMID:27563879

  13. KEY COMPARISON: Key comparison of stainless steel 1 kg mass standards: COOMET.M.M-K1

    NASA Astrophysics Data System (ADS)

    Spurný, Robert; Kolozinska, Irina; Snegov, Viktor; Evsievich, Ludmila; Borys, Michael; Milkamanavičiene, Ilona

    2010-01-01

    The COOMET.M.M-K1 key comparison is an international COOMET comparison of 1 kg stainless steel mass standards, using two travelling artefacts from SMU, Slovakia. Thanks to the participation of the PTB in this comparison, it was made possible to link the results to those of the corresponding CCM key comparison, namely CCM.M-K1, thus adding new values of degrees of equivalence for BelGIM (Belarus), NSC IM (Ukraine), PTB (Germany), SMU (Slovakia), VMT/VMC (Lithuania) and VNIIM (Russia), on the CCM.M-K1 graph of equivalence. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCM, according to the provisions of the CIPM Mutual Recognition Arrangement (MRA).

  14. Aerobic growth of Anoxybacillus pushchinoensis K1(T): emended descriptions of A. pushchinoensis and the genus Anoxybacillus

    NASA Technical Reports Server (NTRS)

    Pikuta, Elena; Cleland, David; Tang, Jane

    2003-01-01

    In this work, corrections are made to the descriptions of the species Anoxybacillus pushchinoensis corrig. and the genus ANOXYBACILLUS: Experiments to determine the relationship of A. pushchinoensis K1(T) to oxygen showed that it was capable of aerobic growth, but preferred to grow anaerobically. During aerobic growth, the redox indicator resazurin was reduced as a result of hydrogen gas production. The facultatively anaerobic nature of K1(T) was ascertained by cultivation in aerobic liquid medium, where growth began at the bottom of the tube. The anaerobic nature of K1(T) was also indicated by a negative catalase reaction. This work is submitted to correct the description of the species A. pushchinoensis from obligate anaerobe to aerotolerant anaerobe and to emend the description of the genus Anoxybacillus from obligate anaerobes or facultative anaerobes to aerotolerant anaerobes or facultative anaerobes.

  15. The chromosome complement of Acomys spp. (Rodentia, Muridae) from Oursi, Burkina Faso--the ancestral karyotype of the cahirinus-dimidiatus group?

    PubMed

    Volobouev, V; Gautun, J C; Sicard, B; Tranier, M

    1996-11-01

    We present here data on chromosome banding analysis (R- and C-bands) of Acomys sp. (Rodentia, Muridae) from Oursi, Burkina Faso, characterized by 2n = FN = 68 and comparison of its banding patterns with those of Acomys dimidiatus from Saudi Arabia (2n = 38, FN = 70), studied previously. The study revealed complete homology between acrocentric chromosomes of Acomys sp. and chromosome arms of 16 pairs of metacentric and two pairs of acrocentric chromosomes of A. dimidiatus. In addition to monobrachial homology, one tandem translocation accompanied by a centromeric shift was identified in the karyotype of the latter species. The data obtained show that karyotypes of all the species of the Acomys cahirinus-dimidiatus group studied previously may be derived from that of Acomys sp. from Oursl by means of numerous non-homologous Rb translocations and 1-2 tandem transiocations, and thus its karyotype may be considered as ancestral for the cahirinus-dimidiatus group.

  16. Third lineage of rodent eimerians: morphology, phylogeny and re-description of Eimeria myoxi (Apicomplexa: Eimeriidae) from Eliomys quercinus (Rodentia: Gliridae).

    PubMed

    Kvičerová, Jana; Mikeš, Václav; Hypša, Václav

    2011-09-01

    Coccidian oocysts from feces of 46 individuals of the garden dormouse, Eliomys quercinus (Rodentia: Gliridae), were morphologically and molecularly characterized. Both morphological and sequence data (18S rDNA and ORF 470) showed low variability, indicating that all samples represent a single species. By comparison with published morphological descriptions of coccidia from glirid rodents, we determined that the samples represent Eimeria myoxi. Molecular data suggest that this species does not fall within the 2 known rodent-specific groups but branches as a third independent lineage. However, its exact position in respect to other eimerian clusters could not be established due to the lack of phylogenetic information at this taxonomic level for the 18S rRNA and ORF 470 genes. Based on these results, we provide a re-description of Eimeria myoxi, which contains morphological and molecular characteristics sufficient for its further unequivocal identification.

  17. The role of the subterranean rodent Ctenomys talarum (Rodentia: Octodontidae) in the life cycle of Taenia taeniaeformis (Cestoda: Taeniidae) in urban environments.

    PubMed

    Rossin, Alejandra; Malizia, Ana I; Denegri, Guillermo M

    2004-06-10

    This work is the first report of subterranean rodent Ctenomys talarum (Rodentia: Octodontidae) as intermediate host of Taenia taeniaeformis in urban areas of Mar de Cobo (Buenos Aires Province, Argentina) and to experimentally reproduce in domestic dogs the adult stage of this parasite. Prevalence, mean abundance and mean intensity of infection with T. taeniaeformis larvae in the liver and peritoneal cavity of C. talarum were 64%, 15.3 and 9.8, respectively. Ten adults of T. taeniaeformis were obtained from experimentally infected dogs. Information about the role of subterranean rodents in the life cycle of this parasite is also given. The above mentioned data indicate that T. taeniaeformis is a frequent parasite of this species of rodents, at least within the study area. Also explanations for the high prevalence of larval forms of this parasite in C. talarum populations are given.

  18. The role of genomic islands in Escherichia coli K1 interactions with intestinal and kidney epithelial cells.

    PubMed

    Yousuf, Farzana Abubakar; Rafiq, Sahar; Siddiqui, Ruqaiyyah; Khan, Naveed Ahmed

    2016-04-01

    The completion of Escherichia coli K1 genome has identified several genomic islands that are present in meningitis-causing E. coli RS218 but absent in the non-pathogenic E. coli MG1655. In this study, the role of various genomic islands in E. coli K1 interactions with intestinal epithelial cells (Caco-2) and kidney epithelial cells (MA104) was determined. Using association assays, invasion assays, and intracellular survival assays, the findings revealed that the genomic island deletion mutants of RS218 related to P fimbriae, S fimbriae, F17-like fimbriae, non-fimbrial adhesins, Hek and hemagglutinin, protein secretion system (T1SS for hemolysin; T2SS; T5SS for antigen 43), Iro system and hmu system), invasins (CNF1, IbeA), toxins (α-hemolysin), K1 capsule biosynthesis, metabolism (d-serine catabolism, dihydroxyacetone, glycerol, and glyoxylate metabolism), prophage genes, showed reduced interactions with both cell types. Next, we determined the role of various genomic islands in E. coli K1 resistance to serum. When exposed to the normal human serum, the viability of the genomic island deletion mutants related to adhesins such as S fimbriae, P fimbriae, F17-like fimbriae, non-fimbrial adhesins, Hek and hemagglutinin, antigen 43 and T5SS for antigen 43, T2SS, and T1SS for hemolysin, Iro system and hmu system, prophage genes, metabolism (sugar metabolism and d-serine catabolism), K1 capsule biosynthesis, and invasins such as CNF1 was affected, suggesting their role in bacteremia. The characterization of these genomic islands should reveal mechanisms of E. coli K1 pathogenicity that could be of value as therapeutic targets.

  19. PtK1 cells contain a nondiffusible, dominant factor that makes the Golgi apparatus resistant to brefeldin A

    PubMed Central

    1991-01-01

    Brefeldin A (BFA) was shown in earlier studies of numerous cell types to inhibit secretion, induce enzymes of the Golgi stacks to redistribute into the ER, and to cause the Golgi cisternae to disappear. Here, we demonstrate that the PtK1 line of rat kangaroo kidney cells is resistant to BFA. The drug did not disrupt the morphology of the Golgi complex in PtK1 cells, as judged by immunofluorescence using antibodies to 58- (58K) and 110-kD (beta-COP) Golgi proteins, and by fluorescence microscopy of live cells labeled with C6-NBD-ceramide. In addition, BFA did not inhibit protein secretion, not alter the kinetics or extent of glycosylation of the vesicular stomatitis virus (VSV) glycoprotein (G-protein) in VSV- infected PtK1 cells. To explore the mechanism of resistance to BFA, PtK1 cells were fused with BFA-sensitive CV-1 cells that had been infected with a recombinant SV-40 strain containing the gene for VSV G- protein and, at various times following fusion, the cultures were exposed to BFA. Shortly after cell fusion, heterokaryons contained one Golgi complex associated with each nucleus. Golgi membranes derived from CV-1 cells were sensitive to BFA, whereas those of PtK1 origin were BFA resistant. A few hours after fusion, most heterokaryons contained a single, large Golgi apparatus that was resistant to BFA and contained CV-1 galactosyltransferase. In unfused cells that had been perforated using nitrocellulose filters, retention of beta-COP on the Golgi was optimal in the presence of cytosol, ATP, and GTP. In perforated cell models of the BFA-sensitive MA104 line, BFA caused beta- COP to be released from the Golgi complex in the presence of nucleotides, and either MA104 or PtK1 cytosol. In contrast, when perforated PtK1 cells were incubated with BFA, nucleotides, and cytosol from either cell type, beta-COP remained bound to the Golgi complex. We conclude that PtK1 cells contain a nondiffusible factor, which is located on or very close to the Golgi complex, and

  20. Common Core English and Language Arts K-1 Exemplar Text Set: A Critical Content Analysis of Cultural Representations

    ERIC Educational Resources Information Center

    McCaffrey, Megan

    2014-01-01

    With the implementation of Common Core State Standards (CCSS) in over forty states, teachers are putting into practice the CCSS text exemplars of text complexity. Of particular concern for the purpose of this research are the kindergarten and first grade (K-1) read aloud and independent text exemplar lists. While not intended as core reading…

  1. 78 FR 23981 - Proposed Collection; Comment Request for Form 1041 and Related Schedules D, J, and K-1

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-04-23

    ... Internal Revenue Service Proposed Collection; Comment Request for Form 1041 and Related Schedules D, J, and... Form 1041 and related Schedules D, J, and K-1, U.S. Income Tax Return for Estates and Trusts. DATES... and Trusts (Form 1041), Capital Gains and Losses (Schedule D), Accumulation Distribution for...

  2. 75 FR 10018 - Proposed Collection; Comment Request for Form 1041 and Related Schedules D, J, and K-1

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-03-04

    ... Internal Revenue Service Proposed Collection; Comment Request for Form 1041 and Related Schedules D, J, and... Form 1041 and related Schedules D, J, and K-1, U.S. Income Tax Return for Estates and Trusts. DATES... Trusts (Form 1041), Capital Gains and Losses (Schedule D), Accumulation Distribution for Certain...

  3. SphK1 inhibitor II (SKI-II) inhibits acute myelogenous leukemia cell growth in vitro and in vivo.

    PubMed

    Yang, Li; Weng, Wei; Sun, Zhi-Xin; Fu, Xian-Jie; Ma, Jun; Zhuang, Wen-Fang

    2015-05-15

    Previous studies have identified sphingosine kinase 1 (SphK1) as a potential drug target for treatment of acute myeloid leukemia (AML). In the current study, we investigated the potential anti-leukemic activity of a novel and specific SphK1 inhibitor, SKI-II. We demonstrated that SKI-II inhibited growth and survival of human AML cell lines (HL-60 and U937 cells). SKI-II was more efficient than two known SphK1 inhibitors SK1-I and FTY720 in inhibiting AML cells. Meanwhile, it induced dramatic apoptosis in above AML cells, and the cytotoxicity by SKI-II was almost reversed by the general caspase inhibitor z-VAD-fmk. SKI-II treatment inhibited SphK1 activation, and concomitantly increased level of sphingosine-1-phosphate (S1P) precursor ceramide in AML cells. Conversely, exogenously-added S1P protected against SKI-II-induced cytotoxicity, while cell permeable short-chain ceramide (C6) aggravated SKI-II's lethality against AML cells. Notably, SKI-II induced potent apoptotic death in primary human AML cells, but was generally safe to the human peripheral blood mononuclear cells (PBMCs) isolated from healthy donors. In vivo, SKI-II administration suppressed growth of U937 leukemic xenograft tumors in severe combined immunodeficient (SCID) mice. These results suggest that SKI-II might be further investigated as a promising anti-AML agent.

  4. Inhibition of diabetic-cataract by vitamin K1 involves modulation of hyperglycemia-induced alterations to lens calcium homeostasis.

    PubMed

    Sai Varsha, M K N; Raman, Thiagarajan; Manikandan, Ramar

    2014-11-01

    This study investigated the potential of vitamin K1 against streptozotocin-induced diabetic cataract in Wistar rats. A single, intraperitoneal injection of streptozotocin (STZ) (35 mg/kg) resulted in hyperglycemia, accumulation of sorbitol and formation of advanced glycation end product (AGE) in eye lens. Hyperglycemia in lens also resulted in superoxide anion and hydroxyl radical generation and less reduced glutathione suggesting oxidative stress in lens. Hyperglycemia also resulted in increase in lens Ca2+ and significant inhibition of lens Ca2+ ATPase activity. These changes were associated with cataract formation in diabetic animals. By contrast treatment of diabetic rats with vitamin K1 (5 mg/kg, sc, twice a week) resulted in animals with partially elevated blood glucose and with transparent lenses having normal levels of sorbitol, AGE, Ca2+ ATPase, Ca2+, and oxidative stress. Vitamin K 1 may function to protect against cataract formation in the STZ induced diabetic rat by affecting the homeostasis of blood glucose and minimizing subsequent oxidative and osmotic stress. Thus, these results show that Vitamin K1 inhibits diabetic-cataract by modulating lens Ca2+ homeostasis and its hypoglycemic effect through its direct action on the pancreas.

  5. Glucocorticoids limit acute lung inflammation in concert with inflammatory stimuli by induction of SphK1

    PubMed Central

    Vettorazzi, Sabine; Bode, Constantin; Dejager, Lien; Frappart, Lucien; Shelest, Ekaterina; Klaßen, Carina; Tasdogan, Alpaslan; Reichardt, Holger M.; Libert, Claude; Schneider, Marion; Weih, Falk; Henriette Uhlenhaut, N.; David, Jean-Pierre; Gräler, Markus; Kleiman, Anna; Tuckermann, Jan P.

    2015-01-01

    Acute lung injury (ALI) is a severe inflammatory disease for which no specific treatment exists. As glucocorticoids have potent immunosuppressive effects, their application in ALI is currently being tested in clinical trials. However, the benefits of this type of regimen remain unclear. Here we identify a mechanism of glucocorticoid action that challenges the long-standing dogma of cytokine repression by the glucocorticoid receptor. Contrarily, synergistic gene induction of sphingosine kinase 1 (SphK1) by glucocorticoids and pro-inflammatory stimuli via the glucocorticoid receptor in macrophages increases circulating sphingosine 1-phosphate levels, which proves essential for the inhibition of inflammation. Chemical or genetic inhibition of SphK1 abrogates the therapeutic effects of glucocorticoids. Inflammatory p38 MAPK- and mitogen- and stress-activated protein kinase 1 (MSK1)-dependent pathways cooperate with glucocorticoids to upregulate SphK1 expression. Our findings support a critical role for SphK1 induction in the suppression of lung inflammation by glucocorticoids, and therefore provide rationales for effective anti-inflammatory therapies. PMID:26183376

  6. Can the Brigance K & 1 Screen Detect Cognitive/Academic Giftedness When Used with Preschoolers from Economically Disadvantaged Backgrounds?

    ERIC Educational Resources Information Center

    Mantzicopoulos, Panayota Y.

    2000-01-01

    The accuracy of the Brigance K&1 Screen in the early identification of Head Start children with possible cognitive/academic giftedness was explored with 134 children, 13 of whom were identified as potentially gifted on the K-ABC. These potentially gifted children also performed significantly better on the Brigance than did other children.…

  7. Clothe the Soldier Prototype K1 Load Carriage System Design Assessment using the APLCS Load Carriage Simulator

    DTIC Science & Technology

    2006-03-01

    bending . It also demonstrated superior forward flexion stiffness which is correlated to good combined functional ratings where large movements are...precision potentiometer, pack resistance to forward flexion, lateral bending and torsion were evaluated. The CTS Prototype Pack K1 was compared to...map for iliac (hip) region.................................................................... 20 Figure 3.3.1. Torsional moment as a function of

  8. 76 FR 7847 - Glenn A. Baxter, Application To Renew License for Amateur Radio Service Station K1MAN

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-02-11

    ... interfering with radio communications; (b) To determine whether Glenn A. Baxter willfully and/or repeatedly... From the Federal Register Online via the Government Publishing Office FEDERAL COMMUNICATIONS COMMISSION Glenn A. Baxter, Application To Renew License for Amateur Radio Service Station K1MAN...

  9. Chronic proliferative rhinitis associated with Salmonella enterica subspecies diarizonae serovar 61:k:1, 5, (7) in sheep in Spain.

    PubMed

    Lacasta, D; Ferrer, L M; Ramos, J J; Bueso, J P; Borobia, M; Ruiz de Arcaute, M; Figueras, L; González-Sainz, J M; De Las Heras, M

    2012-11-01

    A chronic proliferative rhinitis in sheep associated with Salmonella enterica subspecies diarizonae serovar 61:k:1, 5, (7) is described. Ten adult sheep from eight traditionally managed Rasa Aragonesa flocks located in Aragon (Spain) were studied. Clinical signs began with bilateral thick mucus nasal discharge, wheezing and snoring and progressed to partial or complete obstruction of the nostrils. Necropsy examination revealed swollen ventral nasal turbinates with a roughened mucosal surface partially covered by small polyps. Histopathology revealed chronic proliferative rhinitis with a predominant population of neutrophils that infiltrated the mucosal epithelium. Plasma cells and macrophages were present in the lamina propria. Organisms expressing Salmonella antigen immunohistochemically were detected within epithelial cells. Salmonella enterica subspecies diarizonae serovar 61:k:1, 5, (7) was isolated as a sole microorganism from nasal swabs taken from five animals. The implication of finding this bacterium in various diseases in sheep and its role as a potential zoonosis are discussed.

  10. An improved high-quality draft genome sequence of Carnobacterium inhibens subsp. inhibens strain K1(T).

    PubMed

    Nicholson, Wayne L; Davis, Christina L; Shapiro, Nicole; Huntemann, Marcel; Clum, Alicia; Reddy, T B K; Pillay, Manoj; Markowitz, Victor; Varghese, Neha; Pati, Amrita; Ivanova, Natalia; Kyrpides, Nikos; Woyke, Tanja

    2016-01-01

    Despite their ubiquity and their involvement in food spoilage, the genus Carnobacterium remains rather sparsely characterized at the genome level. Carnobacterium inhibens K1(T) is a member of the Carnobacteriaceae family within the class Bacilli. This strain is a Gram-positive, rod-shaped bacterium isolated from the intestine of an Atlantic salmon. The present study determined the genome sequence and annotation of Carnobacterium inhibens K1(T). The genome comprised 2,748,608 bp with a G + C content of 34.85 %, which included 2621 protein-coding genes and 116 RNA genes. The strain contained five contigs corresponding to presumptive plasmids of sizes: 19,036; 24,250; 26,581; 65,272; and 65,904 bp.

  11. A tale of tails: Sialidase is key to success in a model of phage therapy against K1-capsulated Escherichia coli

    SciTech Connect

    Bull, J.J.; Vimr, E.R.; Molineux, I.J.

    2010-03-01

    Prior studies treating mice infected with Escherichia coli O18:K1:H7 observed that phages requiring the K1 capsule for infection (K1-dep) were superior to capsule-independent (K1-ind) phages. We show that three K1-ind phages all have low fitness when grown on cells in serum whereas fitnesses of four K1-dep phages were high. The difference is serum-specific, as fitnesses in broth overlapped. Sialidase activity was associated with all K1-dep virions tested but no K1-ind virions, a phenotype supported by sequence analyses. Adding endosialidase to cells infected with K1-ind phage increased fitness in serum by enhancing productive infection after adsorption. We propose that virion sialidase activity is the primary determinant of high fitness on cells grown in serum, and thus in a mammalian host. Although the benefit of sialidase is specific to K1-capsulated bacteria, this study may provide a scientific rationale for selecting phages for therapeutic use in many systemic infections.

  12. Cyclic ichthyosis with epidermolytic hyperkeratosis: A phenotype conferred by mutations in the 2B domain of keratin K1.

    PubMed Central

    Sybert, V P; Francis, J S; Corden, L D; Smith, L T; Weaver, M; Stephens, K; McLean, W H

    1999-01-01

    Bullous congenital ichthyosiform erythroderma (BCIE) is characterized by blistering and erythroderma in infancy and by erythroderma and ichthyosis thereafter. Epidermolytic hyperkeratosis is a hallmark feature of light and electron microscopy. Here we report on four individuals from two families with a unique clinical disorder with histological findings of epidermolytic hyperkeratosis. Manifesting erythema and superficial erosions at birth, which improved during the first few months of life, affected individuals later developed palmoplantar hyperkeratosis with patchy erythema and scale elsewhere on the body. Three affected individuals exhibit dramatic episodic flares of annular, polycyclic erythematous plaques with scale, which coalesce to involve most of the body surface. The flares last weeks to months. In the interim periods the skin may be normal, except for palmoplantar hyperkeratosis. Abnormal keratin-filament aggregates were observed in suprabasal keratinocytes from both probands, suggesting that the causative mutation might reside in keratin K1 or keratin K10. In one proband, sequencing of K1 revealed a heterozygous mutation, 1436T-->C, predicting a change of isoleucine to threonine in the highly conserved helix-termination motif. In the second family, a heterozygous mutation, 1435A-->T, was found in K1, predicting an isoleucine-to-phenylalanine substitution in the same codon. Both mutations were excluded in both a control population and all unaffected family members tested. These findings reveal that a clinical phenotype distinct from classic BCIE but with similar histology can result from K1 mutations and that mutations at this codon give rise to a clinically unique condition. PMID:10053007

  13. Growth and dielectric properties of KTiOPO4 and K1 - x Rb x TiOPO4 crystals

    NASA Astrophysics Data System (ADS)

    Gurtovoĭ, V. G.; Sheleg, A. U.; Guretskiĭ, S. A.; Kalanda, N. A.

    2008-07-01

    Methods of growth of KTiOPO4 and K1 - x Rb x TiOPO4 crystals of high optical quality have been optimized. The dielectric properties (permittivity and conductivity) of the crystals grown have been investigated at frequencies from 102 to 106 Hz in the temperature range from 100 to 350 K, along the [001] crystallographic direction. It is established that partial substitution of K+ ions with Rb+ ions leads to a decrease in the permittivity and conductivity.

  14. Influence of ozone on the susceptibility of Escherichia coli K1 to the bactericidal action of serum.

    PubMed

    Jankowski, S; Cisowska, A; Doroszkiewicz, W

    1996-01-01

    The susceptibility to the bactericidal action of normal bovine serum of twenty two Escherichia coli K1 strains, isolated from the urine of patients with urinary tract infections, was determined. Only four strains were resistant. Ozonization of bacterial suspensions enhanced the sensitivity of the strains to the action of both normal serum and a serum in which the alternative pathway of complement activation was thermally blocked.

  15. High prevalence of MAP2K1 mutations in variant and IGHV4-34-expressing hairy-cell leukemias.

    PubMed

    Waterfall, Joshua J; Arons, Evgeny; Walker, Robert L; Pineda, Marbin; Roth, Laura; Killian, J Keith; Abaan, Ogan D; Davis, Sean R; Kreitman, Robert J; Meltzer, Paul S

    2014-01-01

    To understand the genetic mechanisms driving variant and IGHV4-34-expressing hairy-cell leukemias, we performed whole-exome sequencing of leukemia samples from ten affected individuals, including six with matched normal samples. We identified activating mutations in the MAP2K1 gene (encoding MEK1) in 5 of these 10 samples and in 10 of 21 samples in a validation set (overall frequency of 15/31), suggesting potential new strategies for treating individuals with these diseases.

  16. Quantum cosmology on (k = -1)-Friedmann-Robertson-Walker Universe evolving from stiff matter era to the dust dominated one

    NASA Astrophysics Data System (ADS)

    Dariescu, Marina-Aura; Dariescu, Ciprian

    2017-01-01

    This work is devoted to the spatially open Friedmann-Robertson-Walker (FRW) Universe evolving from the stiff matter era to the dust dominated one. Within the quantum analysis based on the Wheeler-DeWitt equation, we derive the wave function of the (k = -1)-FRW Universe with combined matter sources. On the classical level, one has to deal with the Friedmann equation which leads on a dependence of the scale function on time generally expressed from functional relations involving elliptic integrals.

  17. Arsenite induces cell transformation by reactive oxygen species, AKT, ERK1/2, and p70S6K1

    SciTech Connect

    Carpenter, Richard L.; Jiang, Yue; Jing, Yi; He, Jun; Rojanasakul, Yon; Liu, Ling-Zhi; Jiang, Bing-Hua

    2011-10-28

    Highlights: Black-Right-Pointing-Pointer Chronic exposure to arsenite induces cell proliferation and transformation. Black-Right-Pointing-Pointer Arsenite-induced transformation increases ROS production and downstream signalings. Black-Right-Pointing-Pointer Inhibition of ROS levels via catalase reduces arsenite-induced cell transformation. Black-Right-Pointing-Pointer Interruption of AKT, ERK, or p70S6K1 inhibits arsenite-induced cell transformation. -- Abstract: Arsenic is naturally occurring element that exists in both organic and inorganic formulations. The inorganic form arsenite has a positive association with development of multiple cancer types. There are significant populations throughout the world with high exposure to arsenite via drinking water. Thus, human exposure to arsenic has become a significant public health problem. Recent evidence suggests that reactive oxygen species (ROS) mediate multiple changes to cell behavior after acute arsenic exposure, including activation of proliferative signaling and angiogenesis. However, the role of ROS in mediating cell transformation by chronic arsenic exposure is unknown. We found that cells chronically exposed to sodium arsenite increased proliferation and gained anchorage-independent growth. This cell transformation phenotype required constitutive activation of AKT, ERK1/2, mTOR, and p70S6K1. We also observed these cells constitutively produce ROS, which was required for the constitutive activation of AKT, ERK1/2, mTOR, and p70S6K1. Suppression of ROS levels by forced expression of catalase also reduced cell proliferation and anchorage-independent growth. These results indicate cell transformation induced by chronic arsenic exposure is mediated by increased cellular levels of ROS, which mediates activation of AKT, ERK1/2, and p70S6K1.

  18. Comparative Genomic Analysis Shows That Avian Pathogenic Escherichia coli Isolate IMT5155 (O2:K1:H5; ST Complex 95, ST140) Shares Close Relationship with ST95 APEC O1:K1 and Human ExPEC O18:K1 Strains

    PubMed Central

    Pan, Zihao; Hu, Lin; Wang, Shaohui; Wang, Haojin; Leung, Frederick C.; Dai, Jianjun; Fan, Hongjie

    2014-01-01

    Avian pathogenic E. coli and human extraintestinal pathogenic E. coli serotypes O1, O2 and O18 strains isolated from different hosts are generally located in phylogroup B2 and ST complex 95, and they share similar genetic characteristics and pathogenicity, with no or minimal host specificity. They are popular objects for the study of ExPEC genetic characteristics and pathogenesis in recent years. Here, we investigated the evolution and genetic blueprint of APEC pathotype by performing phylogenetic and comparative genome analysis of avian pathogenic E. coli strain IMT5155 (O2:K1:H5; ST complex 95, ST140) with other E. coli pathotypes. Phylogeny analyses indicated that IMT5155 has closest evolutionary relationship with APEC O1, IHE3034, and UTI89. Comparative genomic analysis showed that IMT5155 and APEC O1 shared significant genetic overlap/similarities with human ExPEC dominant O18:K1 strains (IHE3034 and UTI89). Furthermore, the unique PAI I5155 (GI-12) was identified and found to be conserved in APEC O2 serotype isolates. GI-7 and GI-16 encoding two typical T6SSs in IMT5155 might be useful markers for the identification of ExPEC dominant serotypes (O1, O2, and O18) strains. IMT5155 contained a ColV plasmid p1ColV5155, which defined the APEC pathotype. The distribution analysis of 10 sequenced ExPEC pan-genome virulence factors among 47 sequenced E. coli strains provided meaningful information for B2 APEC/ExPEC-specific virulence factors, including several adhesins, invasins, toxins, iron acquisition systems, and so on. The pathogenicity tests of IMT5155 and other APEC O1:K1 and O2:K1 serotypes strains (isolated in China) through four animal models showed that they were highly virulent for avian colisepticemia and able to cause septicemia and meningitis in neonatal rats, suggesting zoonotic potential of these APEC O1:K1 and O2:K1 isolates. PMID:25397580

  19. Inositol hexakisphosphate kinase 1 (IP6K1) activity is required for cytoplasmic dynein-driven transport

    PubMed Central

    Chanduri, Manasa; Rai, Ashim; Malla, Aushaq Bashir; Wu, Mingxuan; Fiedler, Dorothea; Mallik, Roop; Bhandari, Rashna

    2016-01-01

    Inositol pyrophosphates, such as diphosphoinositol pentakisphosphate (IP7), are conserved eukaryotic signaling molecules that possess pyrophosphate and monophosphate moieties. Generated predominantly by inositol hexakisphosphate kinases (IP6Ks), inositol pyrophosphates can modulate protein function by posttranslational serine pyrophosphorylation. Here, we report inositol pyrophosphates as novel regulators of cytoplasmic dynein-driven vesicle transport. Mammalian cells lacking IP6K1 display defects in dynein-dependent trafficking pathways, including endosomal sorting, vesicle movement, and Golgi maintenance. Expression of catalytically active but not inactive IP6K1 reverses these defects, suggesting a role for inositol pyrophosphates in these processes. Endosomes derived from slime mold lacking inositol pyrophosphates also display reduced dynein-directed microtubule transport. We demonstrate that Ser51 in the dynein intermediate chain (IC) is a target for pyrophosphorylation by IP7, and this modification promotes the interaction of the IC N-terminus with the p150Glued subunit of dynactin. IC–p150Glued interaction is decreased, and IC recruitment to membranes is reduced in cells lacking IP6K1. Our study provides the first evidence for the involvement of IP6Ks in dynein function and proposes that inositol pyrophosphate-mediated pyrophosphorylation may act as a regulatory signal to enhance dynein-driven transport. PMID:27474409

  20. Cytotoxic effects of zearalenone and its metabolites and antioxidant cell defense in CHO-K1 cells.

    PubMed

    Tatay, Elena; Font, Guillermina; Ruiz, Maria-Jose

    2016-10-01

    Zearalenone (ZEA) and its metabolites (α-zearalenol; α-ZOL, β-zearalenol; β-ZOL) are secondary metabolites of Fusarium fungi that produce cell injury. The present study explores mycotoxin-induced cell damage and cellular protection mechanisms in CHO-K1 cells. Cytotoxicity has been determined by reactive oxygen species (ROS) production and DNA damage. ROS production was determined using the fluorescein assay and DNA strand breakage by comet assay. Intracellular protection systems were glutathione (GSH), glutathione peroxidase (GPx), catalase (CAT) and superoxide dismutase (SOD). The results demonstrated that all mycotoxins increased the ROS levels up to 5.3-fold the control levels in CHO-K1 cells. Zearalenone metabolites, but not ZEA, increased DNA damage 43% (α-ZOL) and 28% (β-ZOL) compared to control cells. The GSH levels decreased from 18% to 36%. The GPx and SOD activities respectively increased from 26% to 62% and from 23% to 69% in CHO-K1 cells, whereas CAT activity decreased from 14% to 52%. In addition, intracellular ROS production was induced by ZEA and its metabolites. The endogenous antioxidant system components GSH, GPx and SOD were activated against ZEA and its metabolites. These antioxidant system components thus could contribute to decrease cell injury by ZEA and its metabolites.

  1. Piperazine-2,3-dicarboxylic acid Derivatives as Dual Antagonists of NMDA and GluK1-Containing Kainate Receptors

    PubMed Central

    Irvine, Mark W.; Costa, Blaise M.; Dlaboga, Daniel; Culley, Georgia; Hulse, Richard; Scholefield, Caroline L.; Atlason, Palmi; Fang, Guangyu; Eaves, Richard; Morley, Richard; Mayo-Martin, Maria B.; Amici, Mascia; Bortolotto, Zuner A.; Donaldson, Lucy; Collingridge, Graham L.; Molnár, Elek; Monaghan, Daniel T.; Jane, David E.

    2011-01-01

    Competitive N-methyl-D-aspartate receptor (NMDAR) antagonists bind to the GluN2 subunit, of which there are four types (GluN2A-D). We report that some N1-substituted derivatives of cis-piperazine-2,3-dicarboxylic acid display improved relative affinity for GluN2C and GluN2D versus GluN2A and GluN2B. These derivatives also display subtype-selectivity among the more distantly related kainate receptor family. Compounds 18i and (−)-4 were the most potent kainate receptor antagonists and 18i was selective for GluK1 versus GluK2, GluK3 and AMPA receptors. Modeling studies revealed structural features required for activity at GluK1 subunits and suggested that S674 was vital for antagonist activity. Consistent with this hypothesis, replacing the equivalent residue in GluK3 (alanine) with a serine imparts 18i antagonist activity. Antagonists with dual GluN2D and GluK1 antagonist activity may have beneficial effects in various neurological disorders. Consistent with this idea, antagonist 18i (30 mg/Kg i.p.) showed antinociceptive effects in an animal model of mild nerve injury. PMID:22111545

  2. Vitamin K-containing dietary supplements: comparison of synthetic vitamin K1 and natto-derived menaquinone-7.

    PubMed

    Schurgers, Leon J; Teunissen, Kirsten J F; Hamulyák, Karly; Knapen, Marjo H J; Vik, Hogne; Vermeer, Cees

    2007-04-15

    Vitamin K is a cofactor in the production of blood coagulation factors (in the liver), osteocalcin (in bone), and matrix Gla protein (cartilage and vessel wall). Accumulating evidence suggests that for optimal bone and vascular health, relatively high intakes of vitamin K are required. The synthetic short-chain vitamin K(1) is commonly used in food supplements, but recently the natural long-chain menaquinone-7 (MK-7) has also become available as an over-the-counter (OTC) supplement. The purpose of this paper was to compare in healthy volunteers the absorption and efficacy of K(1) and MK-7. Serum vitamin K species were used as a marker for absorption and osteocalcin carboxylation as a marker for activity. Both K(1) and MK-7 were absorbed well, with peak serum concentrations at 4 hours after intake. A major difference between the 2 vitamin K species is the very long half-life time of MK-7, resulting in much more stable serum levels, and accumulation of MK-7 to higher levels (7- to 8-fold) during prolonged intake. MK-7 induced more complete carboxylation of osteocalcin, and hematologists should be aware that preparations supplying 50 mug/d or more of MK-7 may interfere with oral anticoagulant treatment in a clinically relevant way.

  3. Immunity to killer toxin K1 is connected with the Golgi-to-vacuole protein degradation pathway.

    PubMed

    Valis, K; Masek, T; Novotná, D; Pospísek, M; Janderová, B

    2006-01-01

    Killer strains of Saccharomyces cerevisiae producing killer toxin K1 kill sensitive cells but are resistant to their own toxin. It is assumed that in the producer, an effective interaction between the external toxin and its plasma membrane receptor or the final effector is not possible on the grounds of a conformation change of the receptor or its absence in a membrane. Therefore, it is possible that some mutants with defects in intracellular protein transport and degradation can show a suicidal phenotype during K1 toxin production. We have examined these mutants in a collection of S. cerevisiae strains with deletions in various genes transformed by the pYX213+M1 vector carrying cDNA coding for the K1 toxin under the control of the GAL1 promoter. Determination of the quantity of dead cells in colony population showed that (1) the toxin production from the vector did not support full immunity of producing cells, (2) the suicidal phenotype was not connected with a defect in endocytosis or autophagy, (3) deletants in genes VPS1, VPS23, VPS51 and VAC8 required for the protein degradation pathway between the Golgi body and the vacuole exhibited the highest mortality. These results suggest that interacting molecule(s) on the plasma membrane in the producer might be diverted from the secretion pathway to degradation in the vacuole.

  4. Distinct Roles of ComK1 and ComK2 in Gene Regulation in Bacillus cereus

    PubMed Central

    Mirończuk, Aleksandra M.; Maňu, Amagoia; Kuipers, Oscar P.; Kovács, Ákos T.

    2011-01-01

    The B. subtilis transcriptional factor ComK regulates a set of genes coding for DNA uptake from the environment and for its integration into the genome. In previous work we showed that Bacillus cereus expressing the B. subtilis ComK protein is able to take up DNA and integrate it into its own genome. To extend our knowledge on the effect of B. subtilis ComK overexpression in B. cereus we first determined which genes are significantly altered. Transcriptome analysis showed that only part of the competence gene cluster is significantly upregulated. Two ComK homologues can be identified in B. cereus that differ in their respective homologies to other ComK proteins. ComK1 is most similar, while ComK2 lacks the C-terminal region previously shown to be important for transcription activation by B. subtilis ComK. comK1 and comK2 overexpression and deletion studies using transcriptomics techniques showed that ComK1 enhances and ComK2 decreases expression of the comG operon, when B. subtilis ComK was overexpressed simultaneously. PMID:21747963

  5. Structural insights into conformational changes of a cyclic nucleotide-binding domain in solution from Mesorhizobium loti K1 channel

    PubMed Central

    Schünke, Sven; Stoldt, Matthias; Lecher, Justin; Kaupp, U. Benjamin; Willbold, Dieter

    2011-01-01

    Cyclic nucleotide-sensitive ion channels, known as HCN and CNG channels, are activated by binding of ligands to a domain (CNBD) located on the cytoplasmic side of the channel. The underlying mechanisms are not well understood. To elucidate the gating mechanism, structures of both the ligand-free and -bound CNBD are required. Several crystal structures of the CNBD from HCN2 and a bacterial CNG channel (MloK1) have been solved. However, for HCN2, the cAMP-free and -bound state did not reveal substantial structural rearrangements. For MloK1, structural information for the cAMP-free state has only been gained from mutant CNBDs. Moreover, in the crystal, the CNBD molecules form an interface between dimers, proposed to be important for allosteric channel gating. Here, we have determined the solution structure by NMR spectroscopy of the cAMP-free wild-type CNBD of MloK1. A comparison of the solution structure of cAMP-free and -bound states reveals large conformational rearrangement on ligand binding. The two structures provide insights on a unique set of conformational events that accompany gating within the ligand-binding site. PMID:21430265

  6. Loss of plastoglobule kinases ABC1K1 and ABC1K3 causes conditional degreening, modified prenyl-lipids, and recruitment of the jasmonic acid pathway

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Plastoglobules (PGs) are plastid lipid-protein particles. This study examines the function of PG-localized kinases ABC1K1 and ABC1K3 in Arabidopsis thaliana. Several lines of evidence suggested that ABC1K1 and ABC1K3 form a protein complex. Null mutants for both genes (abc1k1 and abc1k3) and the dou...

  7. Immunoreceptor tyrosine-based activation motif-dependent signaling by Kaposi's sarcoma-associated herpesvirus K1 protein: effects on lytic viral replication.

    PubMed

    Lagunoff, M; Lukac, D M; Ganem, D

    2001-07-01

    The Kaposi's sarcoma-associated herpesvirus (KSHV) K1 gene encodes a polypeptide bearing an immunoreceptor tyrosine-based activation motif (ITAM) that is constitutively active for ITAM-based signal transduction. Although ectopic overexpression of K1 in cultured fibroblasts can lead to growth transformation, in vivo this gene is primarily expressed in lymphoid cells undergoing lytic infection. Here we have examined function of K1 in the setting of lytic replication, through the study of K1 mutants lacking functional ITAMs. Expression of such mutants in BJAB cells cotransfected with wild-type K1 results in dramatic inhibition of K1 signal transduction, as judged by impaired activation of Syk kinase and phospholipase C-gamma2 as well as by diminished expression of a luciferase reporter gene dependent upon K1-induced calcium and Ras signaling. Thus, the mutants behave as dominantly acting inhibitors of K1 function. To assess the role of K1 in lytic replication, we introduced these K1 mutants into BCBL-1 cells, a B-cell lymphoma line latently infected with KSHV, and induced lytic replication by ectopic expression of the KSHV ORF50 transactivator. Expression of lytic cycle genes was diminished up to 80% in the presence of a K1 dominant negative mutant. These inhibitory effects could be overridden by tetradecanoyl phorbol acetate treatment, indicating that inhibition was not due to irreversible cell injury and suggesting that other signaling events could bypass the block. We conclude that ITAM-dependent signaling by K1 is not absolutely required for lytic reactivation but functions to modestly augment lytic replication in B cells, the natural reservoir of KSHV.

  8. Randomized, placebo-controlled trial of K1 acupoint acustimulation to prevent cisplatin- or oxaliplatin-induced nausea

    PubMed Central

    Shen, Yehua; Liu, Luming; Chiang, Joseph S.; Meng, Zhiqiang; Garcia, M. Kay; Chen, Zhen; Peng, Huiting; Bei, Wenying; Zhao, Qi; Spelman, Amy R.; Cohen, Lorenzo

    2014-01-01

    Background More than 70% of cancer patients experience chemotherapy-induced nausea and vomiting (CINV). We examined the effects of electrostimulation of the K1 acupoint located on the sole of the foot, as it is thought to have potential to control CINV. Methods In this trial, 103 patients diagnosed with primary or metastatic liver cancer were recruited before trans-catheter arterial infusion (TAI) of cisplatin (CDDP) or oxaliplatin (OXA) and randomized to group A (N=51; treated with the antiemetic tropisetron and acustimulation at the K1 acupoint for 20 minutes, 1-2 hours before TAI on the first day and then daily for the subsequent 5 days) or group B (N=53; treated with tropisetron and electrostimulation at a placebo point on the heel). The rate, intensity, and duration of nausea and vomiting were collected at baseline and then daily for 5 days after TAI. Quality of life was assessed daily using the MD Anderson Symptom Inventory (MDASI) and the EuroQoL scale. Results No differences were found between groups A and B in the incidence and degree of nausea or vomiting on day 1 or the consecutive 5 days. Patients in group A had better EuroQoL scores than did patients in group B (A: 72.83 versus B: 65.94, P = 0.04) on day 4 but not on the other days. No group differences were noted at any time point for MDASI scores. Conclusions Electrostimulation of K1 combined with antiemetics did not result in initial prevention of CDDP- or OXA-induced nausea or vomiting. PMID:25204437

  9. Vaccinia virus K1L protein mediates host-range function in RK-13 cells via ankyrin repeat and may interact with a cellular GTPase-activating protein.

    PubMed

    Bradley, Ritu R; Terajima, Masanori

    2005-12-01

    The K1L protein of vaccinia virus is required for its growth in certain cell lines (RK-13 and human). The cowpox host-range protein CP77 has been shown to complement K1L function in RK-13 cells, despite a lack of homology between the two proteins except for ankyrin repeats. We investigated the role of ankyrin repeats of K1L protein in RK-13 cells. The growth of a recombinant vaccinia virus, with K1L gene mutated in the most conserved ankyrin repeat, was severely impaired. Infection with the mutant virus caused shutdown of cellular and viral protein synthesis early in infection. We also investigated the interaction of K1L protein with cellular proteins and found that K1L interacts with the rabbit homologue of human ACAP2, a GTPase-activating protein with ankyrin repeats. Our result suggests the importance of ankyrin repeat for host-range function of K1L in RK-13 cells and identifies ACAP2 as a cellular protein, which may be interacting with K1L.

  10. Characterization of PF-4708671, a novel and highly specific inhibitor of p70 ribosomal S6 kinase (S6K1).

    PubMed

    Pearce, Laura R; Alton, Gordon R; Richter, Daniel T; Kath, John C; Lingardo, Laura; Chapman, Justin; Hwang, Catherine; Alessi, Dario R

    2010-10-15

    S6K1 (p70 ribosomal S6 kinase 1) is activated by insulin and growth factors via the PI3K (phosphoinositide 3-kinase) and mTOR (mammalian target of rapamycin) signalling pathways. S6K1 regulates numerous processes, such as protein synthesis, growth, proliferation and longevity, and its inhibition has been proposed as a strategy for the treatment of cancer and insulin resistance. In the present paper we describe a novel cell-permeable inhibitor of S6K1, PF-4708671, which specifically inhibits the S6K1 isoform with a Ki of 20 nM and IC50 of 160 nM. PF-4708671 prevents the S6K1-mediated phosphorylation of S6 protein in response to IGF-1 (insulin-like growth factor 1), while having no effect upon the PMA-induced phosphorylation of substrates of the highly related RSK (p90 ribosomal S6 kinase) and MSK (mitogen- and stress-activated kinase) kinases. PF-4708671 was also found to induce phosphorylation of the T-loop and hydrophobic motif of S6K1, an effect that is dependent upon mTORC1 (mTOR complex 1). PF-4708671 is the first S6K1-specific inhibitor to be reported and will be a useful tool for delineating S6K1-specific roles downstream of mTOR.

  11. Carbamates: A study on genotoxic, cytotoxic, and apoptotic effects induced in Chinese hamster ovary (CHO-K1) cells.

    PubMed

    Soloneski, Sonia; Kujawski, Maciej; Scuto, Anna; Larramendy, Marcelo L

    2015-08-01

    In vitro effects of the carbamates pirimicarb and zineb and their formulations Aficida® (50% pirimicarb) and Azzurro® (70% zineb), respectively, were evaluated in Chinese hamster ovary (CHO-K1) cells. Whereas the cytokinesis-blocked micronucleus cytome assay was employed to test for genotoxicity, MTT, neutral red (NR), and apoptosis evaluation were used as tests for estimating cell viability and succinic dehydrogenase activity, respectively. Concentrations tested were 10-300 μg/ml for pirimicarb and Aficida®, and 1-50 μg/ml for zineb and Azzurro®. All compounds were able to increase the frequency of micronuclei. A marked reduction in the nuclear division index was observed after treatment with 5 μg/ml of zineb and Azzurro® and 10 μg/ml of Azzurro®. Alterations in the cellular morphology not allowing the recognition of binucleated cells exposed to 300 μg/ml pirimicarb and Aficida® as well as 10-50 μg/ml zineb and Azzurro®. All four compounds induced inhibition of both cell viability and succinic dehydrogenase activity and trigger apoptosis in CHO-K1 cells, at least when exposed for 24 h. The data herein demonstrate the genotoxic and cytotoxic effects exerted by these carbamates and reveal the potential risk factor of these pesticides, still extensively used worldwide, for both human health and the environment.

  12. Lipopeptides from the Banyan Endophyte, Bacillus subtilis K1: Mass Spectrometric Characterization of a Library of Fengycins

    NASA Astrophysics Data System (ADS)

    Pathak, Khyati V.; Keharia, Haresh; Gupta, Kallol; Thakur, Suman S.; Balaram, Padmanabhan

    2012-10-01

    Mass spectrometric analysis of a banyan endophyte, Bacillus subtilis K1, extract showing broad spectrum antifungal activity revealed a complex mixture of lipopeptides, iturins, surfactins, and fengycins. Fractionation by reversed-phase high performance liquid chromatography (HPLC) facilitated a detailed analysis of fengycin microheterogeneity. Matrix assisted laser desorption ionization (MALDI) and electrospray ionization (ESI) mass spectrometric studies permitted the identification of several new fengycin variants. Four major sites of heterogeneity are identified: (1) N-terminus β-hydroxy fatty acid moiety, where chain length variation and the presence of unsaturation occur, (2) position 6 (Ala/Val/Ile/Leu), (3) position 10 (Val/Ile) within the macrocyclic ring, and (4) Gln to Glu replacement at position 8, resulting in fengycin variants that differ in mass by 1 Da. Diagnostic fragment ions provide a quick method for localizing the sites of variation in the macrocycle or the linear segment. Subsequent establishment of the sequences is achieved by MS/MS analysis of linear fengycin species produced by hydrolysis of the macrocyclic lactone. Unsaturation in the fatty acid chain and the presence of linear precursors in the B. subtilis K1 extract are also established by mass spectrometry. The anomalous distribution of intensities within isotopic multiplets is a diagnostic for Gln/Glu replacements. High resolution mass spectrometry facilitates the identification of fengycin species differing by 1 Da by localizing the variable position (Gln8/Glu8) in the fengycin variants.

  13. Characterization of a novel human type II epithelial keratin K1b, specifically expressed in eccrine sweat glands.

    PubMed

    Langbein, Lutz; Rogers, Michael A; Praetzel, Silke; Cribier, Bernard; Peltre, Bernard; Gassler, Nikolaus; Schweizer, Jürgen

    2005-09-01

    In this study, we show that a novel human type II epithelial keratin, K1b, is exclusively expressed in luminal duct cells of eccrine sweat glands. Taking this luminal K1b expression as a reference, we have used antibodies against a plethora of epithelial keratins to systematically investigate their expression in the secretory globule and the two-layered sweat duct, which was divided into the intraglandular, intradermal, and intraepidermal (acrosyringium) segments, the latter being further subdivided into the sweat duct ridge and upper intraepidermal duct. We show that (i) each of the eccrine sweat gland tissue compartments expresses their own keratin patterns, (ii) the peripheral and luminal duct layers exhibit a sequential keratin expression, with both representing self-renewing cell layers, (iii) the intradermal duct and the sweat duct ridge display hitherto unknown length variations, and (iv) out of all cell layers, the luminal cell layer is the most robust layer and expresses the highest number of keratins, these being concentrated at the apical side of the cells to form the cuticle. We provide evidence that the cellular and intercellular properties of the peripheral and the luminal layers reflect adaptations to different functions.

  14. Gating of the MlotiK1 potassium channel involves large rearrangements of the cyclic nucleotide-binding domains

    PubMed Central

    Mari, Stefania A.; Pessoa, João; Altieri, Stephen; Hensen, Ulf; Thomas, Lise; Morais-Cabral, João H.; Müller, Daniel J.

    2011-01-01

    Cyclic nucleotide-regulated ion channels are present in bacteria, plants, vertebrates, and humans. In higher organisms, they are closely involved in signaling networks of vision and olfaction. Binding of cAMP or cGMP favors the activation of these ion channels. Despite a wealth of structural and studies, there is a lack of structural data describing the gating process in a full-length cyclic nucleotide-regulated channel. We used high-resolution atomic force microscopy (AFM) to directly observe the conformational change of the membrane embedded bacterial cyclic nucleotide-regulated channel MlotiK1. In the nucleotide-bound conformation, the cytoplasmic cyclic nucleotide-binding (CNB) domains of MlotiK1 are disposed in a fourfold symmetric arrangement forming a pore-like vestibule. Upon nucleotide-unbinding, the four CNB domains undergo a large rearrangement, stand up by ∼1.7 nm, and adopt a structurally variable grouped conformation that closes the cytoplasmic vestibule. This fully reversible conformational change provides insight into how CNB domains rearrange when regulating the potassium channel. PMID:22135457

  15. Effects of Iron-Oxide Nanoparticle Surface Chemistry on Uptake Kinetics and Cytotoxicity in CHO-K1 Cells

    PubMed Central

    Hanot, Camille C.; Choi, Young Suk; Anani, Tareq B.; Soundarrajan, Dharsan; David, Allan E.

    2015-01-01

    Superparamagnetic iron-oxide nanoparticles (SPIONs) show great promise for multiple applications in biomedicine. While a number of studies have examined their safety profile, the toxicity of these particles on reproductive organs remains uncertain. The goal of this study was to evaluate the cytotoxicity of starch-coated, aminated, and PEGylated SPIONs on a cell line derived from Chinese Hamster ovaries (CHO-K1 cells). We evaluated the effect of particle diameter (50 and 100 nm) and polyethylene glycol (PEG) chain length (2k, 5k and 20k Da) on the cytotoxicity of SPIONs by investigating cell viability using the tetrazolium dye 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and sulforhodamine B (SRB) assays. The kinetics and extent of SPION uptake by CHO-K1 cells was also studied, as well as the resulting generation of intracellular reactive oxygen species (ROS). Cell toxicity profiles of SPIONs correlated strongly with their cellular uptake kinetics, which was strongly dependent on surface properties of the particles. PEGylation caused a decrease in both uptake and cytotoxicity compared to aminated SPIONs. Interestingly, 2k Da PEG-modifed SPIONs displayed the lowest cellular uptake and cytotoxicity among all studied particles. These results emphasize the importance of surface coatings when engineering nanoparticles for biomedical applications. PMID:26729108

  16. Cell alterations but no DNA strand breaks induced in vitro by cylindrospermopsin in CHO K1 cells.

    PubMed

    Fessard, Valérie; Bernard, Cécile

    2003-10-01

    Cylindrospermopsin has been involved in some cyanobacterial blooms associated with animal and human intoxications in different countries. Liver is the main target organ even though thymus and kidney are also affected. Its toxic effect has been shown to be induced by protein synthesis inhibition. However, further research about its toxicological potential is required, as revealed by the U.S. Unregulated Contaminant Monitoring Regulation (US UCMR) meeting in 2001. Induction of DNA damage by cylindrospermopsin has been reported by some authors either by a direct effect on DNA or by an indirect effect on associated macromolecules. This study focused on evaluating its in vitro genotoxic potential using the comet assay coupled to various cell alteration measurements. No DNA damage was detected by the alkaline comet assay on Chinese hamster ovary (CHO) K1 cells after 24 h of treatment with cylindrospermopsin concentrations of 0.5 and 1 microg mL(-1). However, inhibition of cell growth was noticed as well as cell blebbing and rounding. These morphological effects were linked to cytoskeletal reorganization (mainly microfilaments) but not to apoptosis. This study concluded that cylindrospermopsin does not obviously react directly with DNA in CHO K1 cells. But the hypothesis of its metabolization into a genotoxic product must be explored further.

  17. BIPM comparison BIPM.RI(II)-K1.Th-228 of activity measurements of the radionuclide 228Th

    NASA Astrophysics Data System (ADS)

    Michotte, C.; Ratel, G.; Courte, S.; Lucas, L.; Kossert, K.; Nähle, O.; Ott, O.

    2016-01-01

    Since 1986, two national metrology institutes (NMI) have submitted two samples of known activity of 228Th to the International Reference System (SIR) for activity comparison at the Bureau International des Poids et Mesures (BIPM), with comparison identifier BIPM.RI(II)-K1.Th-228. The values of the activity submitted were about 300 kBq and 2 MBq. A key comparison reference value (KCRV) has been evaluated for the first time for 228Th. There is only one result remaining in the BIPM.RI(II)-K1.Th-228 comparison, the 1986 NIST result being outdated. The degrees of equivalence between each equivalent activity measured in the SIR and the KCRV have been calculated and the results are given in the form of a table. A graphical presentation is also given. Main text To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCRI, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

  18. The Composition of Comet C/2012 K1 (PanSTARRS) and the Distribution of Primary Volatile Abundances among Comets

    NASA Astrophysics Data System (ADS)

    Roth, Nathan X.; Gibb, Erika L.; Bonev, Boncho P.; DiSanti, Michael A.; Mumma, Michael J.; Villanueva, Geronimo L.; Paganini, Lucas

    2017-04-01

    On 2014 May 22 and 24 we characterized the volatile composition of the dynamically new Oort cloud comet C/2012 K1 (PanSTARRS) using the long-slit, high resolution (λ/Δλ ≈ 25,000) near-infrared echelle spectrograph (NIRSPEC) at the 10 m Keck II telescope on Maunakea, Hawaii. We detected fluorescent emission from six primary volatiles (H2O, HCN, CH4, C2H6, CH3OH, and CO). Upper limits were derived for C2H2, NH3, and H2CO. We report rotational temperatures, production rates, and mixing ratios (relative to water). Compared with median abundance ratios for primary volatiles in other sampled Oort cloud comets, trace gas abundance ratios in C/2012 K1 (PanSTARRS) for CO and HCN are consistent, but CH3OH and C2H6 are enriched while H2CO, CH4, and possibly C2H2 are depleted. When placed in context with comets observed in the near-infrared to date, the data suggest a continuous distribution of abundances of some organic volatiles (HCN, C2H6, CH3OH, CH4) among the comet population. The level of “enrichment” or “depletion” in a given comet does not necessarily correlate across all molecules sampled, suggesting that chemical diversity among comets may be more complex than the simple organics-enriched, organics-normal, and organics-depleted framework.

  19. Body shape and life style of the extinct Balearic dormouse Hypnomys (Rodentia, Gliridae): new evidence from the study of associated skeletons.

    PubMed

    Bover, Pere; Alcover, Josep A; Michaux, Jacques J; Hautier, Lionel; Hutterer, Rainer

    2010-12-31

    Hypnomys is a genus of Gliridae (Rodentia) that occurred in the Balearic Islands until Late Holocene. Recent finding of a complete skeleton of the chronospecies H. morpheus (Late Pleistocene-Early Holocene) and two articulated skeletons of H. cf. onicensis (Late Pliocene) allowed the inference of body size and the calculation of several postcranial indexes. We also performed a Factorial Discriminant Analysis (FDA) in order to evaluate locomotory behaviour and body shape of the taxa. Using allometric models based on skull and tooth measurements, we calculated a body weight between 173 and 284 g for H. morpheus, and direct measurements of articulated skeletons yielded a Head and Body Length (HBL) of 179 mm and a Total Body Length of 295 mm for this species. In addition to the generally higher robustness of postcranial bones already recorded by previous authors, H. morpheus, similar to Canariomys tamarani, another extinct island species, displayed elongated zygopodium bones of the limbs and a wider distal humerus and femur than in an extant related taxon, Eliomys quercinus. Indexes indicated that Hypnomys was more terrestrial and had greater fossorial abilities than E. quercinus. This was also corroborated by a Discriminant Analysis, although no clear additional inference of locomotory abilities could be calculated.

  20. New families of site-specific repetitive DNA sequences that comprise constitutive heterochromatin of the Syrian hamster (Mesocricetus auratus, Cricetinae, Rodentia).

    PubMed

    Yamada, Kazuhiko; Kamimura, Eikichi; Kondo, Mariko; Tsuchiya, Kimiyuki; Nishida-Umehara, Chizuko; Matsuda, Yoichi

    2006-02-01

    We molecularly cloned new families of site-specific repetitive DNA sequences from BglII- and EcoRI-digested genomic DNA of the Syrian hamster (Mesocricetus auratus, Cricetrinae, Rodentia) and characterized them by chromosome in situ hybridization and filter hybridization. They were classified into six different types of repetitive DNA sequence families according to chromosomal distribution and genome organization. The hybridization patterns of the sequences were consistent with the distribution of C-positive bands and/or Hoechst-stained heterochromatin. The centromeric major satellite DNA and sex chromosome-specific and telomeric region-specific repetitive sequences were conserved in the same genus (Mesocricetus) but divergent in different genera. The chromosome-2-specific sequence was conserved in two genera, Mesocricetus and Cricetulus, and a low copy number of repetitive sequences on the heterochromatic chromosome arms were conserved in the subfamily Cricetinae but not in the subfamily Calomyscinae. By contrast, the other type of repetitive sequences on the heterochromatic chromosome arms, which had sequence similarities to a LINE sequence of rodents, was conserved through the three subfamilies, Cricetinae, Calomyscinae and Murinae. The nucleotide divergence of the repetitive sequences of heterochromatin was well correlated with the phylogenetic relationships of the Cricetinae species, and each sequence has been independently amplified and diverged in the same genome.

  1. Body Shape and Life Style of the Extinct Balearic Dormouse Hypnomys (Rodentia, Gliridae): New Evidence from the Study of Associated Skeletons

    PubMed Central

    Bover, Pere; Alcover, Josep A.; Michaux, Jacques J.; Hautier, Lionel; Hutterer, Rainer

    2010-01-01

    Hypnomys is a genus of Gliridae (Rodentia) that occurred in the Balearic Islands until Late Holocene. Recent finding of a complete skeleton of the chronospecies H. morpheus (Late Pleistocene-Early Holocene) and two articulated skeletons of H. cf. onicensis (Late Pliocene) allowed the inference of body size and the calculation of several postcranial indexes. We also performed a Factorial Discriminant Analysis (FDA) in order to evaluate locomotory behaviour and body shape of the taxa. Using allometric models based on skull and tooth measurements, we calculated a body weight between 173 and 284 g for H. morpheus, and direct measurements of articulated skeletons yielded a Head and Body Length (HBL) of 179 mm and a Total Body Length of 295 mm for this species. In addition to the generally higher robustness of postcranial bones already recorded by previous authors, H. morpheus, similar to Canariomys tamarani, another extinct island species, displayed elongated zygopodium bones of the limbs and a wider distal humerus and femur than in an extant related taxon, Eliomys quercinus. Indexes indicated that Hypnomys was more terrestrial and had greater fossorial abilities than E. quercinus. This was also corroborated by a Discriminant Analysis, although no clear additional inference of locomotory abilities could be calculated. PMID:21209820

  2. Catalog of type specimens of recent mammals: orders Didelphimorpha through Chiroptera (Excluding Rodentia) in the National Museum of Natural History, Smithsonian Institution

    USGS Publications Warehouse

    Fisher, Robert D.; Ludwig, Craig A.

    2015-01-01

    The type collection of Recent Mammals in the Division of Mammals, National Museum of Natural History, Smithsonian Institution, contains 820 specimens bearing names of 809 species-group taxa of Didelphimorphia through Chiroptera, excluding Rodentia, as of June 2014. This catalog presents an annotated list of these holdings comprised of 788 holotypes, 26 lectotypes, 11 syntypes (22 specimens), and 4 neotypes. Included are several specimens that should be in the collection but cannot be found or are now known to be in other collections. One hundred and twenty-seven of the names are new since the last type catalog covering these orders, Poole and Schantz (1942). Five specimens reported in Poole and Schantz (1942) were subsequently sent to the Vertebrate Paleontology collection and are not included here. Orders and families are ordered as in Wilson and Reeder (2005); within families, currently recognized genera are arranged alphabetically; within each currently recognized genus, accounts are arranged alphabetically by original published name. Information in each account includes original name and abbreviated citation thereto, current name if other than original, citation for first use of current name combination for the taxon (or new name combination if used herein for the first time), type designation, U.S. National Museum catalog number(s), preparation, age and sex, date of collection and collector, original collector number, type locality, and remarks as appropriate. Digital photographs of each specimen will serve as a condition report and will be attached to each electronic specimen record.

  3. Catalog of type specimens of recent mammals: Rodentia (Myomorpha, Anomaluromorpha, and Hystricomorpha) in the National Museum of Natural History, Smithsonian Institution

    USGS Publications Warehouse

    Fisher, Robert D.; Ludwig, Craig A.

    2014-01-01

    The type collection of Recent mammals in the Division of Mammals, National Museum of Natural History, Smithsonian Institution, contains 945 specimens bearing names of 931 species-group taxa of Rodentia (Myomorpha, Anomaluromorpha, and Hystricomorpha) as of August 2013. This catalog presents an annotated list of these holdings comprised of 905 holotypes, 16 lectotypes, 8 syntypes (48 specimens), and 2 neotypes. In addition, we include 44 specimens that are part of syntype series that should be in the collection but cannot be found or are now known to be in other collections. One hundred and ten of the names are new since the last type catalog covering these suborders A lectotype for Mus peruvianus Peale, 1848, is newly designated herein. Nine specimens previously reported were subsequently sent to the vertebrate paleontology collection and are not included here. Suborders and families are ordered as in Carleton and Musser; within families, currently recognized genera are arranged alphabetically; within each currently recognized genus, accounts are arranged alphabetically by original published name. Information in each account includes original name and abbreviated citation thereto, current name if other than original, citation for first use of current name combination for the taxon (or new name combination if used herein for the first time), type designation, U.S. National Museum catalog number(s), preparation, age and sex, date of collection and collector, original collector number, type locality, and remarks as appropriate. Digital photographs of each specimen will serve as a condition report and will be attached to each electronic specimen record.

  4. Analysis of gamasid mites (Acari: Mesostigmata) associated with the Asian house rat, Rattus tanezumi (Rodentia: Muridae) in Yunnan Province, southwest China.

    PubMed

    Huang, Li-Qin; Guo, Xian-Guo; Speakman, John R; Dong, Wen-Ge

    2013-05-01

    During a survey lasting from 1990 to 2008, we captured 4,113 Asian house rats, Rattus tanezumi Temminck 1844 (Rodentia: Muridae) from 28 counties of Yunnan Province in southwestern China. From these rats, a total of 19,304 gamasid mites (Acari: Mesostigmata) were collected and identified as comprising 50 different species. The species diversity of gamasid mites from this single rat species is higher than that reported previously from multiple hosts within a given geographical region. Of the 50 mite species, 31 species belonged to ectoparasites and 19 species belonged to free-living mites. The species diversity of the mites from rats trapped outdoors was much higher than from rats trapped indoors. The parameter K from the negative binomial distribution was used to measure the spatial distribution patterns of the dominant mite species and revealed that all the mites had an aggregated distribution among the rat hosts. Most mite species showed a predominantly female-biased population structure with many more females than males.

  5. Involvement of NarK1 and NarK2 Proteins in Transport of Nitrate and Nitrite in the Denitrifying Bacterium Pseudomonas aeruginosa PAO1

    PubMed Central

    Sharma, Vandana; Noriega, Chris E.; Rowe, John J.

    2006-01-01

    Two transmembrane proteins were tentatively classified as NarK1 and NarK2 in the Pseudomonas genome project and hypothesized to play an important physiological role in nitrate/nitrite transport in Pseudomonas aeruginosa. The narK1 and narK2 genes are located in a cluster along with the structural genes for the nitrate reductase complex. Our studies indicate that the transcription of all these genes is initiated from a single promoter and that the gene complex narK1K2GHJI constitutes an operon. Utilizing an isogenic narK1 mutant, a narK2 mutant, and a narK1K2 double mutant, we explored their effect on growth under denitrifying conditions. While the ΔnarK1::Gm mutant was only slightly affected in its ability to grow under denitrification conditions, both the ΔnarK2::Gm and ΔnarK1K2::Gm mutants were found to be severely restricted in nitrate-dependent, anaerobic growth. All three strains demonstrated wild-type levels of nitrate reductase activity. Nitrate uptake by whole-cell suspensions demonstrated both the ΔnarK2::Gm and ΔnarK1K2::Gm mutants to have very low yet different nitrate uptake rates, while the ΔnarK1::Gm mutant exhibited wild-type levels of nitrate uptake. Finally, Escherichia coli narK rescued both the ΔnarK2::Gm and ΔnarK1K2::Gm mutants with respect to anaerobic respiratory growth. Our results indicate that only the NarK2 protein is required as a nitrate/nitrite transporter by Pseudomonas aeruginosa under denitrifying conditions. PMID:16391109

  6. Plum Pox Virus 6K1 Protein Is Required for Viral Replication and Targets the Viral Replication Complex at the Early Stage of Infection

    PubMed Central

    Cui, Hongguang

    2016-01-01

    ABSTRACT The potyviral RNA genome encodes two polyproteins that are proteolytically processed by three viral protease domains into 11 mature proteins. Extensive molecular studies have identified functions for the majority of the viral proteins. For example, 6K2, one of the two smallest potyviral proteins, is an integral membrane protein and induces the endoplasmic reticulum (ER)-originated replication vesicles that target the chloroplast for robust viral replication. However, the functional role of 6K1, the other smallest protein, remains uncharacterized. In this study, we developed a series of recombinant full-length viral cDNA clones derived from a Canadian Plum pox virus (PPV) isolate. We found that deletion of any of the short motifs of 6K1 (each of which ranged from 5 to 13 amino acids), most of the 6K1 sequence (but with the conserved sequence of the cleavage sites being retained), or all of the 6K1 sequence in the PPV infectious clone abolished viral replication. The trans expression of 6K1 or the cis expression of a dislocated 6K1 failed to rescue the loss-of-replication phenotype, suggesting the temporal and spatial requirement of 6K1 for viral replication. Disruption of the N- or C-terminal cleavage site of 6K1, which prevented the release of 6K1 from the polyprotein, either partially or completely inhibited viral replication, suggesting the functional importance of the mature 6K1. We further found that green fluorescent protein-tagged 6K1 formed punctate inclusions at the viral early infection stage and colocalized with chloroplast-bound viral replicase elements 6K2 and NIb. Taken together, our results suggest that 6K1 is required for viral replication and is an important viral element of the viral replication complex at the early infection stage. IMPORTANCE Potyviruses account for more than 30% of known plant viruses and consist of many agriculturally important viruses. The genomes of potyviruses encode two polyproteins that are proteolytically

  7. Final report, ongoing key comparison BIPM.QM-K1, ozone at ambient level, comparison with ISCIII (December 2014)

    NASA Astrophysics Data System (ADS)

    Viallon, Joële; Moussay, Philippe; Idrees, Faraz; Wielgosz, Robert; Sanchez, Carmen; Morillo Gomez, Pilar

    2015-01-01

    As part of the ongoing key comparison BIPM.QM-K1, a comparison has been performed between the ozone national standard of the Instituto de Salud Carlos III (ISCIII) and the common reference standard of the key comparison, maintained by the Bureau International des Poids et Mesures (BIPM). The instruments have been compared over a nominal ozone amount-of-substance fraction range of 0 nmol/mol to 500 nmol/mol. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by CCM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

  8. KEY COMPARISON: BIPM comparison BIPM.RI(II)-K1.Rn-222 of activity measurements of the radionuclide 222Rn

    NASA Astrophysics Data System (ADS)

    Ratel, G.; Michotte, C.; Bochud, F. O.

    2004-01-01

    Since 2001, a national metrology institute, the Institut de Radiophysique Appliquée (IRA), Switzerland, has submitted two samples of known activity of 222Rn to the International Reference System (SIR) for activity comparison at the Bureau International des Poids et Mesures (BIPM). The activities ranged from about 13 kBq to 370 kBq. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by Section II of the Consultative Committee for Ionizing Radiation (CCRI(II)), with comparison identifier BIPM.RI(II)-K1.Rn-222, according to the provisions of the Mutual Recognition Arrangement (MRA).

  9. Biological Functions of the Novel Collectins CL-L1, CL-K1, and CL-P1

    PubMed Central

    Ohtani, Katsuki; Suzuki, Yasuhiko; Wakamiya, Nobutaka

    2012-01-01

    Collectins are characterized by a collagen-like sequence and a carbohydrate recognition domain and are members of the vertebrate C-type lectin superfamily. Recently, “novel collectins”, different from “classical collectins” consisting of mannan-binding lectin (MBL) and surfactant proteins A and D (SP-A and SP-D), have been found by reverse genetics. These “novel collectins” consist of collectin liver 1 (CL-L1), collectin kidney 1 (CL-K1), and collectin placenta 1 (CL-P1) and are encoded by three separate genes. Experimental findings on human and animal collectins have shown that both novel collectins and classical collectins play an important role in innate immunity. Based on our recent results and those of others, in this paper, we summarize the new biological functions of these novel collectins in embryonic morphogenesis and development. PMID:22570530

  10. Final report, ongoing key comparison BIPM.QM-K1: Ozone at ambient level, comparison with KRISS (March 2012)

    NASA Astrophysics Data System (ADS)

    Viallon, Joële; Moussay, Philippe; Idrees, Faraz; Wielgosz, Robert; Lee, Sangil; Lee, Jae Y.; Woo, Jin C.; Kim, Byung M.

    2012-01-01

    As part of the ongoing key comparison BIPM.QM-K1, a comparison has been performed between the ozone national standard of Korea maintained by the Korea Research Institute of Standards and Science (KRISS) and the common reference standard of the key comparison, maintained by the Bureau International des Poids et Mesures (BIPM). The instruments have been compared over a nominal ozone amount-of-substance fraction range of 0 nmol/mol to 500 nmol/mol. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

  11. Final report, ongoing key comparison BIPM.QM-K1: Ozone at ambient level, comparison with EMPA (June 2012)

    NASA Astrophysics Data System (ADS)

    Viallon, Joële; Moussay, Philippe; Idrees, Faraz; Wielgosz, Robert; Zellweger, Christoph

    2012-01-01

    As part of the ongoing key comparison BIPM.QM-K1, a comparison has been performed between the ozone standard of the World Meteorological Organisation maintained by the Swiss Federal Laboratories for Materials Testing and Research (EMPA) and the common reference standard of the key comparison, maintained by the Bureau International des Poids et Mesures (BIPM). The instruments have been compared over a nominal ozone amount-of-substance fraction range of 0 nmol/mol to 500 nmol/mol. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

  12. Final report, ongoing key comparison BIPM.QM-K1: Ozone at ambient level, comparison with FMI (February 2012)

    NASA Astrophysics Data System (ADS)

    Viallon, Joële; Moussay, Philippe; Idrees, Faraz; Wielgosz, Robert; Walden, Jari; Kuronen, Pirjo

    2012-01-01

    As part of the ongoing key comparison BIPM.QM-K1, a comparison has been performed between the ozone national standard of Finland maintained by the Finnish Meteorological Institute (FMI) and the common reference standard of the key comparison, maintained by the Bureau International des Poids et Mesures (BIPM). The instruments have been compared over a nominal ozone amount-of-substance fraction range of 0 nmol/mol to 500 nmol/mol. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

  13. Final report on ongoing key comparison BIPM.QM-K1, ozone at ambient level, comparison with VNIIM (June 2014)

    NASA Astrophysics Data System (ADS)

    Viallon, Joële; Moussay, Philippe; Idrees, Faraz; Wielgosz, Robert; Konopelko, Leonid; Kustikov, Yuri

    2015-01-01

    As part of the ongoing key comparison BIPM.QM-K1, a comparison has been performed between the ozone national standard of the D I Mendeleyev Institute for Metrology (VNIIM) and the common reference standard of the key comparison, maintained by the Bureau International des Poids et Mesures (BIPM). The instruments have been compared over a nominal ozone amount-of-substance fraction range of 0 nmol/mol to 500 nmol/mol. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

  14. Final report on ongoing key comparison BIPM.QM-K1, ozone at ambient level, comparison with NIM (July 2014)

    NASA Astrophysics Data System (ADS)

    Viallon, Joële; Moussay, Philippe; Idrees, Faraz; Wielgosz, Robert; Zhou, Zeyi

    2015-01-01

    As part of the ongoing key comparison BIPM.QM-K1, a comparison has been performed between the ozone standard of China maintained by the National Institute of Metrology (NIM) and the common reference standard of the key comparison, maintained by the Bureau International des Poids et Mesures (BIPM). The instruments have been compared over a nominal ozone amount-of-substance fraction range of 0 nmol/mol to 500 nmol/mol. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

  15. Final report, ongoing key comparison BIPM.QM-K1: Ozone at ambient level, comparison with LNE (May 2012)

    NASA Astrophysics Data System (ADS)

    Viallon, Joële; Moussay, Philippe; Idrees, Faraz; Wielgosz, Robert; Macé, Tatiana; Couette, Jérome

    2012-01-01

    As part of the ongoing key comparison BIPM.QM-K1, a comparison has been performed between the ozone national standard of France maintained by the Laboratoire National de métrologie et d'Essais (LNE) and the common reference standard of the key comparison, maintained by the Bureau International des Poids et Mesures (BIPM). The instruments have been compared over a nominal ozone amount-of-substance fraction range of 0 nmol/mol to 500 nmol/mol. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

  16. Final report, ongoing key comparison BIPM.QM-K1: Ozone at ambient level, comparison with UBA (March 2012)

    NASA Astrophysics Data System (ADS)

    Viallon, Joële; Moussay, Philippe; Idrees, Faraz; Wielgosz, Robert; Stummer, Volker; Schinz, Verena

    2012-01-01

    As part of the ongoing key comparison BIPM.QM-K1, a comparison has been performed between the ozone national standard of Germany maintained by the Umweltbundesamt (UBA) and the common reference standard of the key comparison, maintained by the Bureau International des Poids et Mesures (BIPM). The instruments have been compared over a nominal ozone amount-of-substance fraction range of 0 nmol/mol to 500 nmol/mol. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

  17. Identification of IRS-1 Ser-1101 as a target of S6K1 in nutrient- and obesity-induced insulin resistance.

    PubMed

    Tremblay, Frédéric; Brûlé, Sophie; Hee Um, Sung; Li, Yu; Masuda, Kohei; Roden, Michael; Sun, Xiao Jian; Krebs, Michael; Polakiewicz, Roberto D; Thomas, George; Marette, André

    2007-08-28

    S6K1 has emerged as a critical signaling component in the development of insulin resistance through phosphorylation and inhibition of IRS-1 function. This effect can be triggered directly by nutrients such as amino acids or by insulin through a homeostatic negative-feedback loop. However, the role of S6K1 in mediating IRS-1 phosphorylation in a physiological setting of nutrient overload is unresolved. Here we show that S6K1 directly phosphorylates IRS-1 Ser-1101 in vitro in the C-terminal domain of the protein and that mutation of this site largely blocks the ability of amino acids to suppress IRS-1 tyrosine and Akt phosphorylation. Consistent with this finding, phosphorylation of IRS-1 Ser-1101 is increased in the liver of obese db/db and wild-type, but not S6K1(-/-), mice maintained on a high-fat diet and is blocked by siRNA knockdown of S6K1 protein. Finally, infusion of amino acids in humans leads to the concomitant activation of S6K1, phosphorylation of IRS-1 Ser-1101, a reduction in IRS-1 function, and insulin resistance in skeletal muscle. These findings indicate that nutrient- and hormonal-dependent activation of S6K1 causes insulin resistance in mice and humans, in part, by mediating IRS-1 Ser-1101 phosphorylation.

  18. Identification of IRS-1 Ser-1101 as a target of S6K1 in nutrient- and obesity-induced insulin resistance

    PubMed Central

    Tremblay, Frédéric; Brûlé, Sophie; Hee Um, Sung; Li, Yu; Masuda, Kohei; Roden, Michael; Sun, Xiao Jian; Krebs, Michael; Polakiewicz, Roberto D.; Thomas, George; Marette, André

    2007-01-01

    S6K1 has emerged as a critical signaling component in the development of insulin resistance through phosphorylation and inhibition of IRS-1 function. This effect can be triggered directly by nutrients such as amino acids or by insulin through a homeostatic negative-feedback loop. However, the role of S6K1 in mediating IRS-1 phosphorylation in a physiological setting of nutrient overload is unresolved. Here we show that S6K1 directly phosphorylates IRS-1 Ser-1101 in vitro in the C-terminal domain of the protein and that mutation of this site largely blocks the ability of amino acids to suppress IRS-1 tyrosine and Akt phosphorylation. Consistent with this finding, phosphorylation of IRS-1 Ser-1101 is increased in the liver of obese db/db and wild-type, but not S6K1−/−, mice maintained on a high-fat diet and is blocked by siRNA knockdown of S6K1 protein. Finally, infusion of amino acids in humans leads to the concomitant activation of S6K1, phosphorylation of IRS-1 Ser-1101, a reduction in IRS-1 function, and insulin resistance in skeletal muscle. These findings indicate that nutrient- and hormonal-dependent activation of S6K1 causes insulin resistance in mice and humans, in part, by mediating IRS-1 Ser-1101 phosphorylation. PMID:17709744

  19. The role of PI3K/AKT-related PIP5K1α and the discovery of its selective inhibitor for treatment of advanced prostate cancer.

    PubMed

    Semenas, Julius; Hedblom, Andreas; Miftakhova, Regina R; Sarwar, Martuza; Larsson, Rikard; Shcherbina, Liliya; Johansson, Martin E; Härkönen, Pirkko; Sterner, Olov; Persson, Jenny L

    2014-09-02

    Nitrogen-containing heterocyclic compounds are an important class of molecules that are commonly used for the synthesis of candidate drugs. Phosphatidylinositol-4-phosphate 5-kinase-α (PIP5Kα) is a lipid kinase, similar to PI3K. However, the role of PIP5K1α in oncogenic processes and the development of inhibitors that selectively target PIP5K1α have not been reported. In the present study we report that overexpression of PIP5K1α is associated with poor prognosis in prostate cancer and correlates with an elevated level of the androgen receptor. Overexpression of PIP5K1α in PNT1A nonmalignant cells results in an increased AKT activity and an increased survival, as well as invasive malignant phenotype, whereas siRNA-mediated knockdown of PIP5K1α in aggressive PC-3 cells leads to a reduced AKT activity and an inhibition in tumor growth in xenograft mice. We further report a previously unidentified role for PIP5K1α as a druggable target for our newly developed compound ISA-2011B using a high-throughput KINOMEscan platform. ISA-2011B was discovered during our synthetic studies of C-1 indol-3-yl substituted 1,2,3,4-tetrahydroisoquinolines via a Pictet-Spengler approach. ISA-2011B significantly inhibits growth of tumor cells in xenograft mice, and we show that this is mediated by targeting PIP5K1α-associated PI3K/AKT and the downstream survival, proliferation, and invasion pathways. Further, siRNA-mediated knockdown of PIP5K1α exerts similar effects on PC3 cells as ISA-2011B treatment, significantly inhibiting AKT activity, increasing apoptosis and reducing invasion. Thus, PIP5K1α has high potential as a drug target, and compound ISA-2011B is interesting for further development of targeted cancer therapy.

  20. The role of PI3K/AKT-related PIP5K1α and the discovery of its selective inhibitor for treatment of advanced prostate cancer

    PubMed Central

    Semenas, Julius; Hedblom, Andreas; Miftakhova, Regina R.; Sarwar, Martuza; Larsson, Rikard; Shcherbina, Liliya; Johansson, Martin E.; Härkönen, Pirkko; Sterner, Olov; Persson, Jenny L.

    2014-01-01

    Nitrogen-containing heterocyclic compounds are an important class of molecules that are commonly used for the synthesis of candidate drugs. Phosphatidylinositol-4-phosphate 5-kinase-α (PIP5Kα) is a lipid kinase, similar to PI3K. However, the role of PIP5K1α in oncogenic processes and the development of inhibitors that selectively target PIP5K1α have not been reported. In the present study we report that overexpression of PIP5K1α is associated with poor prognosis in prostate cancer and correlates with an elevated level of the androgen receptor. Overexpression of PIP5K1α in PNT1A nonmalignant cells results in an increased AKT activity and an increased survival, as well as invasive malignant phenotype, whereas siRNA-mediated knockdown of PIP5K1α in aggressive PC-3 cells leads to a reduced AKT activity and an inhibition in tumor growth in xenograft mice. We further report a previously unidentified role for PIP5K1α as a druggable target for our newly developed compound ISA-2011B using a high-throughput KINOMEscan platform. ISA-2011B was discovered during our synthetic studies of C-1 indol-3-yl substituted 1,2,3,4-tetrahydroisoquinolines via a Pictet-Spengler approach. ISA-2011B significantly inhibits growth of tumor cells in xenograft mice, and we show that this is mediated by targeting PIP5K1α-associated PI3K/AKT and the downstream survival, proliferation, and invasion pathways. Further, siRNA-mediated knockdown of PIP5K1α exerts similar effects on PC3 cells as ISA-2011B treatment, significantly inhibiting AKT activity, increasing apoptosis and reducing invasion. Thus, PIP5K1α has high potential as a drug target, and compound ISA-2011B is interesting for further development of targeted cancer therapy. PMID:25071204

  1. Multiplex-PCR assay for identification of Klebsiella pneumoniae isolates carrying the cps loci for K1 and K2 capsule biosynthesis.

    PubMed

    Gierczyński, Rafał; Jagielski, Marek; Rastawicki, Waldemar; Kałuzewski, Stanisław

    2007-01-01

    Multiplex-PCR assay for identification of Klebsiella pneumoniae isolates carrying gene clusters for biosynthesis of capsular polysaccharide (CPS) types K1 and K2 was developed. Genes wzc and orf10 of the cps cluster were applied as K1 and K2 specific markers respectively. The assay specificity was confirmed using 147 isolates of Klebsiella spp. including 77 K-antigen reference strains. The multiplex-PCR assay was found simple and cost-effective tool for identification of K. pneumoniae clinical isolates of K1 and K2 geno-serotypes.

  2. Rodentia and lagomorpha

    USGS Publications Warehouse

    Sheffield, S.R.; Sawicka-Kapusta, K.; Cohen, J.B.; Rattner, B.A.; Shore, Richard F.; Rattner, Barnett A.

    2001-01-01

    This comprehensive review examines the extensive literature on wild rodents and lagomorphs as biomonitors of environmental contamination. This chapter covers studies dealing with exposure and effects of environmental contaminants on rodent and lagomorph species, including pesticides (organochlorines, organophosphorus and carbamate compounds, herbicides, plant growth regulators, fungicides, and rodenticides), other organic chemicals, metals, radionuclides, and other miscellaneous contaminants. Many research needs become evident when reviewing ecotoxicological data for rodents and lagomorphs, the most striking being the paucity of information on rodent families other than Muridae (mice and rats). While our ability to qualitatively extrapolate effects observed in laboratory studies to field situations is good for a variety of contaminants, quantitative predictions of dose-response relationships are poor because inter-specific variation and differences in exposure patterns between laboratory and wild species to toxicants are for the most part unknown. More sophisticated comparative toxicity studies need to be undertaken that build on previous work in order to develop a database of information, to account for and model differences in exposure pathways, to document interactions among multiple stressors, to generate data establishing thresholds, critical concentrations, and diagnostic guidelines, and even to develop physiologically-based toxicokinetic models. Such efforts may enhance our ability to predict effects on wild populations, including threatened and endangered species.

  3. Targeted suppression of AR-V7 using PIP5K1α inhibitor overcomes enzalutamide resistance in prostate cancer cells

    PubMed Central

    Sarwar, Martuza; Semenas, Julius; Miftakhova, Regina; Simoulis, Athanasios; Robinson, Brian; Wingren, Anette Gjörloff; Mongan, Nigel P.; Heery, David M.; Johnsson, Heather; Abrahamsson, Per-Anders; Dizeyi, Nishtman; Luo, Jun; Persson, Jenny L.

    2016-01-01

    One mechanism of resistance of prostate cancer (PCa) to enzalutamide (MDV3100) treatment is the increased expression of AR variants lacking the ligand binding-domain, the best characterized of which is AR-V7. We have previously reported that Phosphatidylinositol-4-phosphate 5-kinase alpha (PIP5Kα), is a lipid kinase that links to CDK1 and AR pathways. The discovery of PIP5Kα inhibitor highlight the potential of PIP5K1α as a drug target in PCa. In this study, we show that AR-V7 expression positively correlates with PIP5K1α in tumor specimens from PCa patients. Overexpression of AR-V7 increases PIP5K1α, promotes rapid growth of PCa in xenograft mice, whereas inhibition of PIP5K1α by its inhibitor ISA-2011B suppresses the growth and invasiveness of xenograft tumors overexpressing AR-V7. PIP5K1α is a key co-factor for both AR-V7 and AR, which are present as protein-protein complexes predominantly in the nucleus of PCa cells. In addition, PIP5K1α and CDK1 influence AR-V7 expression also through AKT-associated mechanism dependent on PTEN-status. ISA-2011B disrupts protein stabilization of AR-V7 which is dependent on PIP5K1α, leading to suppression of invasive growth of AR-V7-high tumors in xenograft mice. Our study suggests that combination of enzalutamide and PIP5K1α may have a significant impact on refining therapeutic strategies to circumvent resistance to antiandrogen therapies. PMID:27588408

  4. Binding mode of an α-amino acid-linked quinoxaline-2,3-dione analogue at glutamate receptor subtype GluK1.

    PubMed

    Demmer, Charles S; Møller, Charlotte; Brown, Patricia M G E; Han, Liwei; Pickering, Darryl S; Nielsen, Birgitte; Bowie, Derek; Frydenvang, Karla; Kastrup, Jette S; Bunch, Lennart

    2015-06-17

    Two α-amino acid-functionalized quinoxalines, 1a (CNG-10301) and 1b (CNG-10300), of a quinoxaline moiety coupled to an amino acid moiety were designed, synthesized, and characterized pharmacologically. While 1a displayed low affinity at native AMPA, KA, and NMDA receptors, and at homomeric GluK1,3 receptors, the affinity for GluK2 was in the midmicromolar range (Ki = 136 μM), 1b displayed low to midmicromolar range binding affinity at all the iGluRs (Ki = 9-126 μM). In functional experiments (outside-out patches excised from transfected HEK293T cells), 100 μM 1a partially blocked GluK1 (33% peak response), while GluK2 was unaffected (96% peak response). Furthermore, 1a was shown not to be an agonist at GluK1 and GluK2 at 100 μM. On the other hand, 100 μM 1b fully antagonized GluK1 (8% peak response) but only partially blocked GluK2 (33% peak response). An X-ray structure at 2.3 Å resolution of 1b in the GluK1-LBD (ligand-binding domain) disclosed an unexpected binding mode compared to the predictions made during the design phase; the quinoxaline moiety remains to act as an amino acid bioisostere, but the amino acid moiety is oriented into a new area within the GluK1 receptor. The structure of the GluK1-LBD with 1b showed a large variation in domain openings of the three molecules from 25° to 49°, demonstrating that the GluK1-LBD is capable of undergoing major domain movements.

  5. Activation of SphK1 by K6PC-5 Inhibits Oxygen–Glucose Deprivation/Reoxygenation-Induced Myocardial Cell Death

    PubMed Central

    Shao, Jun-jie; Peng, Yi; Wang, Li-ming; Wang, Jian-kai

    2015-01-01

    In the current study, we evaluated the potential effect of a novel sphingosine kinase 1 (SphK1) activator, K6PC-5, on oxygen–glucose deprivation (OGD)/reoxygenation-induced damages to myocardial cells. We demonstrated that K6PC-5 increased intracellular sphingosine-1-phosphate (S1P) content and remarkably inhibited OGD/reoxygenation-induced death of myocardial cells (H9c2/HL-1 lines and primary murine myocardiocytes). SphK1 inhibitors, B-5354c and SKI-II, or SphK1-siRNA knockdown not only aggregated OGD/reoxygenation-induced cytotoxicity but also nullified the cytoprotection by K6PC-5. On the other hand, overexpression of SphK1 alleviated H9c2 cell death by OGD/reoxygenation, and K6PC-5-mediated cytoprotection was also enhanced in SphK1 overexpressed cells. Molecularly, OGD/reoxygenation activated the mitochondrial death pathway, evidenced by reactive oxygen species (ROS) production, mitochondrial membrane potential reduction, and p53-cyclophilin D (Cyp-D) association, which were all alleviated by K6PC-5 or overexpression of SphK1, but exacerbated by SphK1 knockdown. Furthermore, OGD/reoxygenation induced prodeath ceramide production in myocardial cells, which was largely suppressed by K6PC-5. In the meantime, adding a cell-permeable short-chain ceramide (C6) mimicked OGD/reoxygenation actions and induced ROS production and the mitochondrial death pathway in myocardial cells. Together, we conclude that K6PC-5 inhibits OGD/reoxygenation-induced myocardial cell death probably through activating SphK1. The results of the study indicate a potential benefit of K6PC-5 on ischemic heart disease. PMID:26308910

  6. The Escherichia coli argW-dsdCXA Genetic Island Is Highly Variable, and E. coli K1 Strains Commonly Possess Two Copies of dsdCXA

    PubMed Central

    Moritz, Rebecca L.; Welch, Rodney A.

    2006-01-01

    The genome sequences of Escherichia coli pathotypes reveal extensive genetic variability in the argW-dsdCXA island. Interestingly, the archetype E. coli K1 neonatal meningitis strain, strain RS218, has two copies of the dsdCXA genes for d-serine utilization at the argW and leuX islands. Because the human brain contains d-serine, an epidemiological study emphasizing K1 isolates surveyed the dsdCXA copy number and function. Forty of 41 (97.5%) independent E. coli K1 isolates could utilize d-serine. Southern blot hybridization revealed physical variability within the argW-dsdC region, even among 22 E. coli O18:K1:H7 isolates. In addition, 30 of 41 K1 strains, including 21 of 22 O18:K1:H7 isolates, had two dsdCXA loci. Mutational analysis indicated that each of the dsdA genes is functional in a rifampin-resistant mutant of RS218, mutant E44. The high percentage of K1 strains that can use d-serine is in striking contrast to our previous observation that only 4 of 74 (5%) isolates in the diarrheagenic E. coli (DEC) collection have this activity. The genome sequence of diarrheagenic E. coli isolates indicates that the csrRAKB genes for sucrose utilization are often substituted for dsdC and a portion of dsdX present at the argW-dsdCXA island of extraintestinal isolates. Among DEC isolates there is a reciprocal pattern of sucrose fermentation versus d-serine utilization. The ability to use d-serine is a trait strongly selected for among E. coli K1 strains, which have the ability to infect a wide range of extraintestinal sites. Conversely, diarrheagenic E. coli pathotypes appear to have substituted sucrose for d-serine as a potential nutrient. PMID:17088369

  7. Activation of SphK1 by K6PC-5 Inhibits Oxygen-Glucose Deprivation/Reoxygenation-Induced Myocardial Cell Death.

    PubMed

    Shao, Jun-jie; Peng, Yi; Wang, Li-ming; Wang, Jian-kai; Chen, Xin

    2015-11-01

    In the current study, we evaluated the potential effect of a novel sphingosine kinase 1 (SphK1) activator, K6PC-5, on oxygen-glucose deprivation (OGD)/reoxygenation-induced damages to myocardial cells. We demonstrated that K6PC-5 increased intracellular sphingosine-1-phosphate (S1P) content and remarkably inhibited OGD/reoxygenation-induced death of myocardial cells (H9c2/HL-1 lines and primary murine myocardiocytes). SphK1 inhibitors, B-5354c and SKI-II, or SphK1-siRNA knockdown not only aggregated OGD/reoxygenation-induced cytotoxicity but also nullified the cytoprotection by K6PC-5. On the other hand, overexpression of SphK1 alleviated H9c2 cell death by OGD/reoxygenation, and K6PC-5-mediated cytoprotection was also enhanced in SphK1 overexpressed cells. Molecularly, OGD/reoxygenation activated the mitochondrial death pathway, evidenced by reactive oxygen species (ROS) production, mitochondrial membrane potential reduction, and p53-cyclophilin D (Cyp-D) association, which were all alleviated by K6PC-5 or overexpression of SphK1, but exacerbated by SphK1 knockdown. Furthermore, OGD/reoxygenation induced prodeath ceramide production in myocardial cells, which was largely suppressed by K6PC-5. In the meantime, adding a cell-permeable short-chain ceramide (C6) mimicked OGD/reoxygenation actions and induced ROS production and the mitochondrial death pathway in myocardial cells. Together, we conclude that K6PC-5 inhibits OGD/reoxygenation-induced myocardial cell death probably through activating SphK1. The results of the study indicate a potential benefit of K6PC-5 on ischemic heart disease.

  8. Targeted suppression of AR-V7 using PIP5K1α inhibitor overcomes enzalutamide resistance in prostate cancer cells.

    PubMed

    Sarwar, Martuza; Semenas, Julius; Miftakhova, Regina; Simoulis, Athanasios; Robinson, Brian; Gjörloff Wingren, Anette; Mongan, Nigel P; Heery, David M; Johnsson, Heather; Abrahamsson, Per-Anders; Dizeyi, Nishtman; Luo, Jun; Persson, Jenny L

    2016-09-27

    One mechanism of resistance of prostate cancer (PCa) to enzalutamide (MDV3100) treatment is the increased expression of AR variants lacking the ligand binding-domain, the best characterized of which is AR-V7. We have previously reported that Phosphatidylinositol-4-phosphate 5-kinase alpha (PIP5Kα), is a lipid kinase that links to CDK1 and AR pathways. The discovery of PIP5Kα inhibitor highlight the potential of PIP5K1α as a drug target in PCa. In this study, we show that AR-V7 expression positively correlates with PIP5K1α in tumor specimens from PCa patients. Overexpression of AR-V7 increases PIP5K1α, promotes rapid growth of PCa in xenograft mice, whereas inhibition of PIP5K1α by its inhibitor ISA-2011B suppresses the growth and invasiveness of xenograft tumors overexpressing AR-V7. PIP5K1α is a key co-factor for both AR-V7 and AR, which are present as protein-protein complexes predominantly in the nucleus of PCa cells. In addition, PIP5K1α and CDK1 influence AR-V7 expression also through AKT-associated mechanism dependent on PTEN-status. ISA-2011B disrupts protein stabilization of AR-V7 which is dependent on PIP5K1α, leading to suppression of invasive growth of AR-V7-high tumors in xenograft mice. Our study suggests that combination of enzalutamide and PIP5K1α may have a significant impact on refining therapeutic strategies to circumvent resistance to antiandrogen therapies.

  9. APMP key comparison for the measurement of air kerma for 60Co (APMP.RI(I)-K1.1)

    NASA Astrophysics Data System (ADS)

    Webb, D. V.; Lee, J.-H.; Budiantari, C. T.; Laban, J.; Saito, N.; Srimanoroth, S.; Khaled, N. E.

    2016-01-01

    The results are reported for an APMP.R(I)-K1.1 comparison that extends the regional comparison of standards for air kerma APMP.R(I)-K1 to several laboratories unable to participate earlier. The comparison was conducted with the goal of supporting the relevant calibration and measurement capabilities (CMCs) planned for publication by the participant laboratories. The comparison was conducted by the pilot laboratory, the Australian Radiation Protection and Nuclear Safety (ARPANSA), Australia, supported by the Institute of Nuclear Energy Research (INER), Taiwan, in a modified ring-shaped arrangement from September 2009 to November 2010, in parallel with an APMP.R(I)-K4 comparison being piloted by the INER. The laboratories that took part in the comparison were the ARPANSA, the Centre of Technology of Radiation Safety and Metrology (PTKMR-BATAN), Indonesia, the Division of Radiation and Medical Devices (DMSC), Thailand, the INER, the National Centre for Radiation Science (NCRS), New Zealand, the National Institute for Standards (NIS), Egypt and the National Metrology Institute of Japan (NMIJ/AIST), Japan. The two primary laboratories, ARPANSA and NMIJ, were chosen as the linking laboratories. Three ionization chambers were used as transfer instruments to be calibrated in terms of air kerma in 60Co radiotherapy beams. The comparison result is based on the ratio between the air kerma calibration coefficients (NK) determined by the participants and the mean of the results of the linking laboratories. The mean comparison ratio was found to be within 0.5 % of the key comparison reference value KCRV. The largest deviation between any two comparison ratios for the three chambers in terms of air kerma was 2.0 %. An analysis of the participant uncertainty budgets enabled the calculation of degrees of equivalence (DoE) in terms of the deviations of the results and their associated uncertainties. As a result of this APMP comparison, the BIPM key comparison database (KCDB) should

  10. TOR and S6K1 promote translation reinitiation of uORF-containing mRNAs via phosphorylation of eIF3h.

    PubMed

    Schepetilnikov, Mikhail; Dimitrova, Maria; Mancera-Martínez, Eder; Geldreich, Angèle; Keller, Mario; Ryabova, Lyubov A

    2013-04-17

    Mammalian target-of-rapamycin (mTOR) triggers S6 kinase (S6K) activation to phosphorylate targets linked to translation in response to energy, nutrients, and hormones. Pathways of TOR activation in plants remain unknown. Here, we uncover the role of the phytohormone auxin in TOR signalling activation and reinitiation after upstream open reading frame (uORF) translation, which in plants is dependent on translation initiation factor eIF3h. We show that auxin triggers TOR activation followed by S6K1 phosphorylation at T449 and efficient loading of uORF-mRNAs onto polysomes in a manner sensitive to the TOR inhibitor Torin-1. Torin-1 mediates recruitment of inactive S6K1 to polysomes, while auxin triggers S6K1 dissociation and recruitment of activated TOR instead. A putative target of TOR/S6K1-eIF3h-is phosphorylated and detected in polysomes in response to auxin. In TOR-deficient plants, polysomes were prebound by inactive S6K1, and loading of uORF-mRNAs and eIF3h was impaired. Transient expression of eIF3h-S178D in plant protoplasts specifically upregulates uORF-mRNA translation. We propose that TOR functions in polysomes to maintain the active S6K1 (and thus eIF3h) phosphorylation status that is critical for translation reinitiation.

  11. Functional differentiation of trailing and leading forelimbs during locomotion on the ground and on a horizontal branch in the European red squirrel (Sciurus vulgaris, Rodentia).

    PubMed

    Schmidt, André

    2011-06-01

    Mammalian locomotion is characterized by the frequent use of in-phase gaits in which the footfalls of the left and right fore- or hindlimbs are unevenly spaced in time. Although previous studies have identified a functional differentiation between the first limb (trailing limb) and the second limb (leading limb) to touch the ground during terrestrial locomotion, the influence of a horizontal branch on limb function has never been explored. To determine the functional differences between trailing and leading forelimbs during locomotion on the ground and on a horizontal branch, X-ray motion analysis and force measurements were carried out in two European red squirrels (Sciurus vulgaris, Rodentia). The differences observed between trailing and leading forelimbs were minimal during terrestrial locomotion, where both limbs fulfill two functions and go through a shock-absorbing phase followed by a generating phase. During locomotion on a horizontal branch, European red squirrels reduce speed and all substrate reaction forces transmitted may be due to the reduction of vertical oscillation of the center of mass. Further adjustments during locomotion on a horizontal branch differ significantly between trailing and leading forelimbs and include limb flexion, lead intervals, limb protraction and vertical displacement of the scapular pivot. Consequently, trailing and leading forelimbs perform different functions. Trailing forelimbs function primarily as shock-absorbing elements, whereas leading forelimbs are characterized by a high level of stiffness. This functional differentiation indicates that European red squirrels 'test' the substrate for stability with the trailing forelimb, while the leading forelimb responds to or counteracts swinging or snapping branches.

  12. The mammalian 2'-5' oligoadenylate synthetase gene family: evidence for concerted evolution of paralogous Oas1 genes in Rodentia and Artiodactyla.

    PubMed

    Perelygin, Andrey A; Zharkikh, Andrey A; Scherbik, Svetlana V; Brinton, Margo A

    2006-10-01

    Multiple 2'-5' oligoadenylate (2-5A) synthetases are important components of innate immunity in mammals. Gene families encoding these proteins have previously been studied mainly in humans and mice. To reconstruct the evolution of this gene family in mammals, a search for additional 2-5A synthetase genes was performed in rat, cattle, pig, and dog. Twelve 2'-5' oligoadenylate synthetase (Oas) genes were identified in the rat genome, including eight Oas1 genes, two Oas1 pseudogenes, single copies of Oas2 and Oas3, and two Oas-like genes, Oasl1 and Oasl2. Four OAS genes were detected in the pig genome and five OAS genes were found in both the cattle and dog genomes. An OAS3 gene was not found in either the cattle or the pig genome. While two tandemly duplicated OAS-like (OASL) genes were identified in the dog genome, only a single OASL orthologue was found in both the cattle and the pig genomes. The bovine and porcine OASL genes contain premature stop codons and encode truncated proteins, which lack the typical C-terminal double ubiquitin domains. The cDNA sequences of the rat, cattle, pig, and dog OAS genes were amplified, sequenced and compared with each other and with those in the human, mouse, horse, and chicken genomes. Evidence of concerted evolution of paralogous 2'-5' oligoadenylate synthetase 1 genes was obtained in rodents (Rodentia) and even-toed ungulates (Artiodactyla). Calculations using the nonparametric Kolmogorov-Smirnov test suggested that the homogenization of paralogous OAS1 sequences was due to gene conversion rather than stabilizing selection.

  13. A Golgi study of the sixth layer of the cerebral cortex. I. The lissencephalic brain of Rodentia, Lagomorpha, Insectivora and Chiroptera.

    PubMed Central

    Ferrer, I; Fabregues, I; Condom, E

    1986-01-01

    A study of the morphological characteristics of the neurons in layer VI of the cerebral cortex was carried out using the rapid Golgi method in several lissencephalic species including Rodentia (rat, mouse, vole (Microtus agrestis) and hamster), Lagomorpha (rabbit), Insectivora (hedgehog) and in the Chiroptera the dwarf bat (Pipistrellus pipistrellus). There was a basic uniformity in the structure of the sixth layer. Main neuronal types in lamina VIa were large pyramidal neurons, triangular or atypical pyramidal cells, multiapical pyramidal neurons, inverted pyramids, fusiform neurons, Martinotti cells and bi-tufted cells. Main neuronal types in lamina VIb were medium sized, flattened pyramids, large and small horizontal neurons, horizontal pyramidal cells, fan shaped neurons and multipolar spinous neurons with long descending axons. Sparsely spinous and spine-free multipolar neurons with short axons were present in the two laminae of layer VI, but sparsely spinous neurons with axons similar to those found in basket cells of other layers of the cortex were observed mainly in lamina VIa. Neuronal subsystems were tentatively classified on the basis of the course of the axons. Pyramidal neurons, fusiform neurons, multiapical pyramidal cells, inverted pyramidal cells, fan shaped neurons and multipolar neurons with large descending axons were interpreted as being the main source of long projection and association connections. Large horizontal neurons were interpreted as possible ipsilateral association neurons because the horizontal course of the axons over long distances followed the boundary of the deeper region of the sixth layer. Three intracortical (association) subsystems were included. Axons of Martinotti cells and collateral ascending axons of pyramidal neurons (including multiapical pyramidal neurons) formed the ascending interlaminar fibrillary subsystem. Axons of small horizontal cells and horizontal collaterals of pyramidal neurons formed the horizontal

  14. RNA polymerase activity in PtK1 micronuclei containing individual chromosomes: an in vitro and in situ study

    SciTech Connect

    Labidi, B.; Gregoire, M.; Frackowiak, S.; Hernandez-Verdun, D.; Bouteille, M.

    1987-03-01

    Micronuclei have been induced by colchicine in rat kangaroo (Potorous tridactylis) PtK1 cells. The synthesis of RNA was investigated both in isolated micronuclei by quantifying RNA polymerase activities at different ionic strengths with or without inhibitors, and in micronucleated cells by radioautography after (/sup 3/H)uridine pulse labeling. In vitro transcription shows that isolated micronuclei are able to take up (/sup 3/H)UTP. The rate curves of incorporation are close to those of isolated diploid nuclei, though the level of incorporation was relatively lower (65-70%) than control nuclei. This indicates that micronuclei react to the ionic environment and to inhibitors in the same manner as described for many species of isolated diploid nuclei. The labelling distributions plotted from radioautographs show that micronuclei were able to efficiently incorporate the hot precursor. Furthermore, for short pulses there is no homogeneity in the labelling density among the different micronuclei and there is no correlation between the labelling intensity and the size of micronuclei. After 60-min pulse time, there is an enhanced uptake of (/sup 3/H)uridine and all the micronuclei exhibit considerable labelling, although less than control cells. Thus, the micronuclei exhibit some characteristic RNA transcriptional activity in situ as well as after isolation. This material should be a particular interesting model with which to study the physiological activity and the role of each individual interphasic chromosome.

  15. Interference harmonics and rigorous EM spectrum analysis method for low-k1 CD Bossung tilt correction

    NASA Astrophysics Data System (ADS)

    Chou, Shuo-Yen; Ng, Hoi-Tou; Chen, Yi-Yin; Lee, Chien-Fu; Liu, Ru-Gun; Gau, Tsai-Sheng

    2013-04-01

    This paper discusses the CD Bossung tilt phenomena in low-k1 lithography using interference harmonics and rigorous EM spectrum analysis. Interference harmonics analysis is introduced to explain the interaction of diffraction orders in the focal region leading to this abnormal CD behavior. This method decomposes the vector image formula into a superposition of cosine components to describe the interference of diffraction orders. The symmetry properties of components of an optical projection system were investigated to find out three potential sources for the asymmetric Bossung behavior, namely mask 3D (M3D) effect, lens aberration, and wafer reflectivity. Under good lens aberration and substrate reflectivity controls, the M3D effect accounts for most of the CD Bossung tilt. A rigorous EM mask spectral analysis was performed to reveal the impact of mask topography on the near-field intensity of mask transmission and the far-field image formation. From the analysis, the asymmetric phase distribution in the mask spectrum is the root cause for CD Bossung tilt. Using both the interference harmonics and the rigorous EM spectrum analysis, the effect of various resolution enhancement techniques (RET) to the Bossung tilt is also studied to find the best RET combination for M3D immunity. In addition, a pupil optimization algorithm based on these two analyses is proposed to generate the phase compensation map for M3D effect counteraction.

  16. Ligand-induced structural changes in the cyclic nucleotide-modulated potassium channel MloK1

    NASA Astrophysics Data System (ADS)

    Kowal, Julia; Chami, Mohamed; Baumgartner, Paul; Arheit, Marcel; Chiu, Po-Lin; Rangl, Martina; Scheuring, Simon; Schröder, Gunnar F.; Nimigean, Crina M.; Stahlberg, Henning

    2014-01-01

    Cyclic nucleotide-modulated ion channels are important for signal transduction and pacemaking in eukaryotes. The molecular determinants of ligand gating in these channels are still unknown, mainly because of a lack of direct structural information. Here we report ligand-induced conformational changes in full-length MloK1, a cyclic nucleotide-modulated potassium channel from the bacterium Mesorhizobium loti, analysed by electron crystallography and atomic force microscopy. Upon cAMP binding, the cyclic nucleotide-binding domains move vertically towards the membrane, and directly contact the S1-S4 voltage sensor domains. This is accompanied by a significant shift and tilt of the voltage sensor domain helices. In both states, the inner pore-lining helices are in an ‘open’ conformation. We propose a mechanism in which ligand binding can favour pore opening via a direct interaction between the cyclic nucleotide-binding domains and voltage sensors. This offers a simple mechanistic hypothesis for the coupling between ligand gating and voltage sensing in eukaryotic HCN channels.

  17. An enzyme-linked immunosorbent assay (ELISA) for quantification of human collectin 11 (CL-11, CL-K1).

    PubMed

    Selman, L; Henriksen, M L; Brandt, J; Palarasah, Y; Waters, A; Beales, P L; Holmskov, U; Jørgensen, T J D; Nielsen, C; Skjodt, K; Hansen, S

    2012-01-31

    Collectin 11 (CL-11), also referred to as collectin kidney 1 (CL-K1), is a pattern recognition molecule that belongs to the collectin group of proteins involved in innate immunity. It interacts with glycoconjugates on pathogen surfaces and has been found in complex with mannose-binding lectin-associated serine protease 1 (MASP-1) and/or MASP-3 in circulation. Mutation in the CL-11 gene was recently associated with the developmental syndrome 3MC. In the present study, we established and thoroughly validated a sandwich enzyme-linked immunosorbent assay (ELISA) based on two different monoclonal antibodies. The assay is highly sensitive, specific and shows excellent quantitative characteristics such as reproducibility, dilution linearity and recovery (97.7-104%). The working range is 0.15-34 ng/ml. The CL-11 concentration in two CL-11-deficient individuals affected by the 3MC syndrome was determined to be below 2.1 ng/ml. We measured the mean serum CL-11 concentration to 284 ng/ml in 100 Danish blood donors, with a 95% confidence interval of 269-299 ng/ml. There was no significant difference in the CL-11 concentration measured in matched serum and plasma samples. Storage of samples and repeated freezing and thawing to a certain extent did not influence the ELISA. This ELISA offers a convenient and reliable method for studying CL-11 levels in relation to a variety of human diseases and syndromes.

  18. Ligand-induced structural changes in the cyclic nucleotide-modulated potassium channel MloK1

    PubMed Central

    Kowal, Julia; Chami, Mohamed; Baumgartner, Paul; Arheit, Marcel; Chiu, Po-Lin; Rangl, Martina; Scheuring, Simon; Schröder, Gunnar F.; Nimigean, Crina M.; Stahlberg, Henning

    2014-01-01

    Cyclic nucleotide-modulated ion channels are important for signal transduction and pacemaking in eukaryotes. The molecular determinants of ligand gating in these channels are still unknown, mainly because of a lack of direct structural information. Here we report ligand-induced conformational changes in full-length MloK1, a cyclic nucleotide-modulated potassium channel from the bacterium Mesorhizobium loti, analysed by electron crystallography and atomic force microscopy. Upon cAMP binding, the cyclic nucleotide-binding domains move vertically towards the membrane, and directly contact the S1–S4 voltage sensor domains. This is accompanied by a significant shift and tilt of the voltage sensor domain helices. In both states, the inner pore-lining helices are in an ‘open’ conformation. We propose a mechanism in which ligand binding can favour pore opening via a direct interaction between the cyclic nucleotide-binding domains and voltage sensors. This offers a simple mechanistic hypothesis for the coupling between ligand gating and voltage sensing in eukaryotic HCN channels. PMID:24469021

  19. Structure and function of collectin liver 1 (CL-L1) and collectin 11 (CL-11, CL-K1).

    PubMed

    Selman, Lana; Hansen, Soren

    2012-09-01

    The collectins are a group of innate immune proteins structurally characterized by their content of a carbohydrate recognition domain and a collagen-like region. Collectin liver 1 (CL-L1) and collectin 11 (CL-11, alias collectin kidney 1, CL-K1) are the more recently described members of this group. Their genomic organization and protein structure reveal many similarities. However, CL-11 is a serum protein, whereas CL-L1 appears to be restricted to the cytosol of cells such as hepatocytes. Specificity analyses of the CRDs reveal some differences in their preferences for saccharides: CL-11 binds most avidly to l-fucose and d-mannose, whereas CL-L1 shows preference for d-mannose, d-fucose, N-acetylglucosamine, and surprisingly also d-galactose. CL-11 binds to various microorganisms including Escherichia coli, Candida albicans and Influenza A virus. Polymorphisms in the CL-11 gene (COLEC11) leading to deficiencies have recently been identified as causative for 3MC syndrome. The 3MC syndrome is associated with a wide spectrum of developmental features including facial dysmorphism, cognitive impairment, hearing loss and vesicorenal anomalies. Similar polymorphic associations were reported for the mannan-binding lectin-associated serine protease 3 (MASP-3), and falls into line with the observation that CL-11 is found in circulating complexes with MASP-1/3. These findings suggest dual or overlapping functions of CL-11 in innate immunity and in fetal development.

  20. Surface modification by argon plasma treatment improves antioxidant defense ability of CHO-k1 cells on titanium surfaces.

    PubMed

    de Queiroz, Jana Dara Freires; Leal, Angélica Maria de Sousa; Terada, Maysa; Agnez-Lima, Lucymara Fassarela; Costa, Isolda; Pinto, Nadja Cristhina de Souza; de Medeiros, Silvia Regina Batistuzzo

    2014-04-01

    Titanium is one of the most used materials in implants and changes in its surface can modify the cellular functional response to better implant fixation. An argon plasma treatment generates a surface with improved mechanical proprieties without modifying its chemical composition. Oxidative stress induced by biomaterials is considered one of the major causes of implant failure and studies in this field are fundamental to evaluate the biocompatibility of a new material. Therefore, in this work, induction of oxidative stress by titanium surfaces subjected to plasma treatment (PTTS) was evaluated. The viability of CHO-k1 cells was higher on PTTS discs. Cells grown on titanium surfaces are subjected to intracellular oxidative stress. Titanium discs subjected to the plasma treatment induced less oxidative stress than the untreated ones, which resulted in improved cellular survival. These were associated with improved cellular antioxidant response in Plasma Treated Titanium Surface (PTTS). Furthermore, a decrease in protein and DNA oxidative damage was observed on cells grown on the roughed surface when compared to the smooth one. In conclusion, our data suggest that the treatment of titanium with argon plasma may improve its biocompatible, thus improving its performance as implants or as a scaffold in tissue engineering.

  1. Thermostability and reactivity in organic solvent of O-phospho-L-serine sulfhydrylase from hyperthermophilic archaeon Aeropyrum pernix K1.

    PubMed

    Nakamura, Takashi; Asai, Shinji; Nakata, Kaori; Kunimoto, Kohei; Oguri, Masateru; Ishikawa, Kazuhiko

    2015-01-01

    O-phospho-l-serine sulfhydrylase (OPSS) from archaeon Aeropyrum pernix K1 is able to synthesize l-cysteine even at 80 °C. In this article, we compared thermal stability and reactivity in organic solvent of OPSS with those of O-acetyl-l-serine sulfhydrylase B (OASS-B) from Escherichia coli. As a result, the thermostability of OPSS was much higher than that of OASS-B. Moreover, the activity of OPSS increased in the reaction mixture containing the organic solvent, such as N, N'-dimethyl formamide and 1,4-dioxane, whereas that of OASS-B gradually decreased as the content of organic solvent increased. From the crystal structural analysis, the intramolecular electrostatic interactions of N-terminal domain in OPSS seemed to be correlated with the tolerance of OPSS to high temperature and organic solvent. These results indicate that OPSS is more superior to OASS-B for the industrial production of l-cysteine and unnatural amino acids that are useful pharmaceuticals in the presence of organic solvent.

  2. Identification of Arg-12 in the active site of Escherichia coli K1 CMP-sialic acid synthetase.

    PubMed Central

    Stoughton, D M; Zapata, G; Picone, R; Vann, W F

    1999-01-01

    Escherichia coli K1 CMP-sialic acid synthetase catalyses the synthesis of CMP-sialic acid from CTP and sialic acid. The active site of the 418 amino acid E. coli enzyme was localized to its N-terminal half. The bacterial CMP-sialic acid synthetase enzymes have a conserved motif, IAIIPARXXSKGLXXKN, at their N-termini. Several basic residues have been identified at or near the active site of the E. coli enzyme by chemical modification and site-directed mutagenesis. Only one of the lysines in the N-terminal motif, Lys-21, appears to be essential for activity. Mutation of Lys-21 in the N-terminal motif results in an inactive enzyme. Furthermore, Arg-12 of the N-terminal motif appears to be an active-site residue, based on the following evidence. Substituting Arg-12 with glycine or alanine resulted in inactive enzymes, indicating that this residue is required for enzymic activity. The Arg-12-->Lys mutant was partially active, demonstrating that a positive charge is required at this site. Steady-state kinetic analysis reveals changes in k(cat), K(m) and K(s) for CTP, which implicates Arg-12 in catalysis and substrate binding. PMID:10510306

  3. Systemic signalling in photosynthetic induction of Rumex K-1 (Rumex patientia × Rumex tianschaious) leaves.

    PubMed

    Hou, Fei; Jin, Li-Qiao; Zhang, Zi-Shan; Gao, Hui-Yuan

    2015-04-01

    The rapid induction of photosynthesis is critical for plants under light-fleck environment. Most previous studies about photosynthetic induction focused upon single leaf, but they did not consider the systemic integrity of plant. Here, we verified whether systemic signalling is involved in photosynthetic induction. Rumex K-1 (Rumex patientia × Rumex tianschaious) plants were grown under light-fleck condition. After whole night dark adaptation, different numbers of leaves (system leaf or SL) were pre-illuminated with light, and then the photosynthetic induction of other leaves (target leaf or TL) was investigated. This study showed that the pre-illumination of SL promoted photosynthetic induction in TL. This promotion was independent of the number of SL, the light intensity on SL and the distance between SL and TL, indicating that this systemic signalling is non-dose-dependent. More interestingly, the photosynthetic induction was promoted by only the pre-illumination of morphological upper leaf rather than the pre-illumination of morphological lower leaf, indicating that the transfer of this signal is directional. The results showed that the transfer of this systemic signalling depends upon the phloem. This systemic signalling helps plants to use light energy more efficiently under light flecks.

  4. Cell growth stimulating effect of Ganoderma lucidum spores and their potential application for Chinese hamster ovary K1 cell cultivation.

    PubMed

    Li, Ding; Zhong, Qi; Liu, Tingting; Wang, Jufang

    2016-06-01

    In this work, water-soluble extracts of Ganoderma lucidum spores (Gls), a Chinese medicinal herb that possesses cell growth stimulating function, were found to be an effective growth factor for Chinese hamster ovary (CHO) cell cultivation. The Gls extract was prepared and supplemented to CHO K1 cell culture media with various serum levels. Our results obtained from both the static culture and the spinner-flask suspension culture showed that use of small-amount Gls extract effectively promoted cell growth and suppressed cell apoptosis induced by serum deprivation with normal cell cycle maintained in a low-serum medium. The low-serum medium containing 1 % (v/v) fetal bovine serum (FBS) and 0.01 % (w/v) Gls extract showed a comparable performance on both cell growth and fusion protein productivity with the conventional CHO culture medium containing 10 % (v/v) FBS and a commercial serum-free medium. This is the first study of the potential of Gls extracts for use as an alternative cell growth factor and nutrient for CHO cells. The findings have presented a new approach to economic cultivation of CHO cells for therapeutic protein production.

  5. Occurrence of chromosome- or plasmid-mediated aerobactin iron transport systems and hemolysin production among clonal groups of human invasive strains of Escherichia coli K1.

    PubMed

    Valvano, M A; Silver, R P; Crosa, J H

    1986-04-01

    The incidence of the aerobactin system and the genetic location of aerobactin genes were investigated in Escherichia coli K1 neonatal isolates belonging to different clonal groups. A functional aerobactin system was found in all members of the O7 MP3, O1 MP5, O1 MP9, and O18 MP9 clonal groups examined and also in K1 strains having O6, O16, and O75 lipopolysaccharide types, which are less frequently associated with neonatal infections. In contrast, the aerobactin system was not detected in strains from the O18 MP6 clone. The combined results of plasmid and colony hybridization experiments showed that the aerobactin genes were located on the chromosome in the majority (75%) of the aerobactin-producing K1 isolates, the genetic location of the aerobactin genes was closely correlated with the outer membrane protein profile rather than the O lipopolysaccharide type, the K1 strains harboring a chromosome-mediated aerobactin system did not possess colicin V genes, and five of six K1 isolates possessing a plasmid-borne aerobactin system contained colicin V genes which were located on the same plasmids carrying the aerobactin genes. The comparison of hemolysin production with possession of the aerobactin system in virulent clones of E. coli K1 strains showed that all of the aerobactin-producing strains from the O18 MP9 and O7 MP3 clonal groups did not synthesize hemolysin, whereas 11 of 12 aerobactin-nonproducing O18 MP6 isolates were hemolytic. Of the K1 strains examined, 92.5% possessed either the aerobactin system or the ability to produce hemolysin or both.

  6. Sulfhydryl compounds inhibit the cyto- and geno-toxicity of o-phenylphenol metabolites in CHO-K1 cells.

    PubMed

    Tayama, S; Nakagawa, Y

    1991-01-01

    The effects of cysteine and reduced glutathione (GSH) on the genotoxicity of o-phenylphenol (OPP) and its metabolites, phenylhydroquinone (PHQ) and phenylbenzoquinone (PBQ), were examined using the frequency of sister-chromatid exchanges (SCEs) and chromosome aberrations in CHO-K1 cells as parameters. Cytotoxic (cell-progression delay) and cytogenetic effects induced by a 3-h treatment with OPP, PHQ (100 micrograms/ml) or PBQ (50 micrograms/ml) with S9 mix after a 27-h expression time were inhibited by cysteine or GSH (3-10 mM). Materials corresponding to the cysteine or GSH adducts were found by HPLC in each incubation mixture. In the culture without S9 mix, PHQ and PBQ showed severe cytotoxicity since no metaphases could be obtained at doses over 25 and 5 micrograms/ml, respectively, and the sulfhydryl compounds inhibited the toxicity by the formation of adducts with PBQ and by inhibiting the formation of PBQ in the case of PHQ. With PHQ, the sulfhydryl compounds appeared to inhibit autooxidation. However, the sulfhydryl compounds did not inhibit the cytotoxic and cytogenetic effects caused by OPP in the cell mixture without S9 mix, but on the contrary intensified them. No adduct formation was detected in the incubation solution. On the basis of these results, it is considered that electrophilic quinone (PBQ) and/or semiquinone (phenylsemiquinone, PSQ) radicals, capable of binding to nucleophilic small molecules (such as cysteine and GSH) or (biological) macromolecules, are produced from metabolite PHQ in metabolic oxidation of OPP, and induce cyto- and geno-toxic effects in the cells. The cyto- and geno-toxic effects of OPP itself to the cells are clearly independent of any electrophilic radical reaction.

  7. The K1 Capsular Polysaccharide of Acinetobacter baumannii Strain 307-0294 Is a Major Virulence Factor ▿

    PubMed Central

    Russo, Thomas A.; Luke, Nicole R.; Beanan, Janet M.; Olson, Ruth; Sauberan, Shauna L.; MacDonald, Ulrike; Schultz, L. Wayne; Umland, Timothy C.; Campagnari, Anthony A.

    2010-01-01

    Acinetobacter baumannii is a pathogen of increasing medical importance with a propensity to be multidrug resistant, thereby making treatment challenging. Little is known of virulence traits in A. baumannii. To identify virulence factors and potential drug targets, random transposon (Tn) mutants derived from the A. baumannii strain AB307-0294 were screened to identify genes essential for growth in human ascites fluid in vitro, an inflammatory exudative fluid. These studies led to the identification of two genes that were predicted to be required for capsule polymerization and assembly. The first, ptk, encodes a putative protein tyrosine kinase (PTK), and the second, epsA, encodes a putative polysaccharide export outer membrane protein (EpsA). Monoclonal antibodies used in flow cytometric and Western analyses confirmed that these genes are required for a capsule-positive phenotype. A capsule-positive phenotype significantly optimized growth in human ascites fluid, survival in human serum, and survival in a rat soft tissue infection model. Importantly, the clearance of the capsule-minus mutants AB307.30 (ptk mutant, capsule minus) and AB307.45 (epsA mutant, capsule minus) was complete and durable. These data demonstrated that the K1 capsule from AB307-0294 was an important protectin. Further, these data suggested that conserved proteins, which contribute to the capsule-positive phenotype, are potential antivirulence drug targets. Therefore, the results from this study have important biologic and translational implications and, to the best of our knowledge, are the first to address the role of capsule in the pathogenesis of A. baumannii infection. PMID:20643860

  8. Effects of lunar and mars dust simulants on HaCaT keratinocytes and CHO-K1 fibroblasts

    NASA Astrophysics Data System (ADS)

    Rehders, Maren; Grosshäuser, Bianka B.; Smarandache, Anita; Sadhukhan, Annapurna; Mirastschijski, Ursula; Kempf, Jürgen; Dünne, Matthias; Slenzka, Klaus; Brix, Klaudia

    2011-04-01

    Exposure to lunar dust during Apollo missions resulted in occasional reports of ocular, respiratory and dermal irritations which showed that lunar dust has a risk potential for human health. This is caused by its high reactivity as well as its small size, leading to a wide distribution also inside habitats. Hence, detailed information regarding effects of extraterrestrial lunar dusts on human health is required to best support future missions to moon, mars or other destinations. In this study, we used several methods to assess the specific effects of extraterrestrial dusts onto mammalian skin by exposing HaCaT keratinocytes and CHO-K1 fibroblasts to dusts simulating lunar or mars soils. These particular cell types were chosen because the skin protects the human body from potentially harmful substances and because a well orchestrated program ensures proper wound healing. Keratinocytes and fibroblasts were exposed to the dusts for different durations of time and their effects on morphology and viability of the cells were determined. Cytotoxicity was measured using the MTT assay and by monitoring culture impedance, while phalloidin staining of the actin cytoskeleton was performed to address structural integrity of the cells which was also investigated by propidium iodide intake. It was found that the effects of the two types of dust simulants on the different features of both cell lines varied to a considerable extent. Moreover, proliferation of HaCaT keratinocytes, as analyzed by Ki67 labeling, was suppressed in sub-confluent cultures exposed to lunar dust simulant. Furthermore, experimental evidence is provided for a delay in regeneration of keratinocyte monolayers from scratch-wounding when exposed to lunar dust simulant. The obtained results will facilitate further investigations of dust exposure during wound healing and will ease risk assessment studies e.g., for lunar lander approaches. The investigations will help to determine safety measures to be taken during

  9. Effects of lunar and mars dust on HaCaT keratinocytes and CHO-K1 fibroblasts

    NASA Astrophysics Data System (ADS)

    Brix, Klaudia; Slenzka, Klaus; Rehders, Maren; Sadhukhan, Annapurna; Mistry, Rima; Duenne, Matthias; Kempf, Juergen

    Exposure to lunar dust during Apollo missions resulted in occasional reports of ocular, respira-tory and dermal irritations which showed that lunar dust has a risk potential for human health. This is caused by its high reactivity as well as its small size, leading to a wide distribution also inside habitats. Hence, detailed information regarding effects of lunar dust on human health is required to best support future missions to moon. In this study, we used different methods to assess the specific effects of lunar dust onto mammalian skin by exposing HaCaT keratinocytes and CHO-K1 fibroblasts to dusts simulating lunar or mars soils. These particular cell types were chosen because the skin protects the human body from potentially harmful substances and since a well orchestrated program ensures proper repair in cases of wounding. Keratinocytes and fibroblasts were exposed to the dusts for different durations of time and their effects on morphology, metabolic state, survival and proliferation of the cells were determined. Cytotoxi-city and proliferation were measured using the MTT assay, metabolic activity was analyzed by vital staining of mitochondria, and phalloidin staining of the actin cytoskeleton was performed to address structural integrity of the cells. It was found that the effects of the two types of soils on the different features of both cell lines varied to considerable extent, and that lunar and mars dust were specific in their effects. The obtained results will facilitate detailed inves-tigations of dust exposure during wound healing and will ease risk assessment studies for e.g. lunar lander approaches. The investigations will help to assess the risks and to determine safety measures to be taken during extraterrestrial expeditions in order to minimize risks to human health associated with exposure of human skin to dust contaminants.

  10. KEY COMPARISON: Final report on CIPM key comparison CCM.D-K1: density measurements of a silicon sphere

    NASA Astrophysics Data System (ADS)

    Fujii, Kenichi; Bettin, Horst; Peuto, Anna; Chang, Kyung-Ho; Richard, Philippe; Jacques, Claude; Matilla Vicente, Carmen; Becerra, Luis Omar

    2006-01-01

    This report describes the results on a CIPM key comparison of solid density measurements, which was carried out through July 2001 to May 2003. This CIPM key comparison, designated as CCM.D-K1, was coordinated by the National Metrology Institute of Japan (NMIJ, Japan), Swiss Federal Office of Metrology and Accreditation (METAS, Switzerland) and National Research Council Canada (NRC, Canada). These three national metrology institutes (NMIs) formed a pilot group to determine the technical protocol for this key comparison. A total of eight NMIs, namely NMIJ, Physikalisch-Technische Bundesanstalt (PTB, Germany), Istituto di Metrologia 'G Colonnetti' (IMGC, Italy), Korea Research Institute of Standards and Science (KRISS, Korea), METAS, NRC, Centro Español de Metrologia (CEM, Spain) and National Center of Metrology (CENAM, Mexico), participated in this key comparison. A 1 kg single-crystal silicon sphere, prepared by NMIJ, was circulated to each of the NMIs as a travelling standard. Each NMI determined the mass, volume and density of the travelling standard with respect to the mass standard and solid density standard of each NMI by mass measurement and hydrostatic weighing. The reference value of the density was determined with a relative expanded uncertainty of 2.9 × 10-7. When the degrees of equivalence were evaluated by differences from the reference value, the differences for the mass, volume and density were almost equal to or less than expanded uncertainties of the differences, showing a good equivalence of the capabilities for the solid density measurement at the participating NMIs. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCM, according to the provisions of the CIPM Mutual Recognition Arrangement (MRA).

  11. An enzyme-linked immunosorbent assay (ELISA) for quantification of human collectin 11 (CL-11, CL-K1)

    PubMed Central

    Selman, L.; Henriksen, M.L.; Brandt, J.; Palarasah, Y.; Waters, A.; Beales, P.L.; Holmskov, U.; Jørgensen, T.J.D.; Nielsen, C.; Skjodt, K.; Hansen, S.

    2012-01-01

    Collectin 11 (CL-11), also referred to as collectin kidney 1 (CL-K1), is a pattern recognition molecule that belongs to the collectin group of proteins involved in innate immunity. It interacts with glycoconjugates on pathogen surfaces and has been found in complex with mannose-binding lectin-associated serine protease 1 (MASP-1) and/or MASP-3 in circulation. Mutation in the CL-11 gene was recently associated with the developmental syndrome 3MC. In the present study, we established and thoroughly validated a sandwich enzyme-linked immunosorbent assay (ELISA) based on two different monoclonal antibodies. The assay is highly sensitive, specific and shows excellent quantitative characteristics such as reproducibility, dilution linearity and recovery (97.7–104%). The working range is 0.15–34 ng/ml. The CL-11 concentration in two CL-11-deficient individuals affected by the 3MC syndrome was determined to be below 2.1 ng/ml. We measured the mean serum CL-11 concentration to 284 ng/ml in 100 Danish blood donors, with a 95% confidence interval of 269–299 ng/ml. There was no significant difference in the CL-11 concentration measured in matched serum and plasma samples. Storage of samples and repeated freezing and thawing to a certain extent did not influence the ELISA. This ELISA offers a convenient and reliable method for studying CL-11 levels in relation to a variety of human diseases and syndromes. PMID:22301270

  12. Intraperitoneal prophylaxis with CpG oligodeoxynucleotides protects neutropenic mice against intracerebral Escherichia coli K1 infection

    PubMed Central

    2014-01-01

    Background Prophylaxis with unmethylated cytosine phosphate guanidine (CpG) oligodeoxynucleotides (ODN) protects against several systemic experimental infections. Escherichia coli is a major cause of Gram-negative neonatal bacterial meningitis and also causes meningitis and meningoencephalitis in older and immunocompromised patients. Methods Wild-type (wt) and Toll-like receptor 9 (TLR9)-deficient mice were rendered neutropenic by intraperitoneal administration of the anti-Ly-6G monoclonal antibody. Immunocompetent and neutropenic mice received intraperitoneal CpG ODN or vehicle 72 h prior to induction of E. coli K1 meningoencephalitis. Results Pre-treatment with CpG ODN significantly increased survival of neutropenic wt mice from 33% to 75% (P = 0.0003) but did not protect neutropenic TLR9-/- mice. The protective effect of CpG ODN was associated with an enhanced production of interleukin (IL)-12/IL-23p40 with sustained increased levels in serum and spleen at least for 17 days after conditioning compared to buffer-treated animals. CpG-treated neutropenic wt mice showed reduced bacterial concentrations and increased recruitment of Ly6ChighCCR2+ monocytes in brain and spleen 42 h after infection. The levels of macrophage inflammatory protein 1α (MIP-1α) and interferon gamma (IFN-γ) in spleen were higher 42 h after infection in CpG-treated compared to buffer-treated neutropenic animals. In immunocompetent mice, prophylaxis with CpG ODN did not significantly increase survival compared to the buffer group (60% vs. 45%, P = 0.2). Conclusions These findings suggest that systemic administration of CpG ODN may help to prevent bacterial CNS infections in immunocompromised individuals. PMID:24456653

  13. Fast intracellular dissolution and persistent cellular uptake of silver nanoparticles in CHO-K1 cells: implication for cytotoxicity.

    PubMed

    Jiang, Xiumei; Miclăuş, Teodora; Wang, Liming; Foldbjerg, Rasmus; Sutherland, Duncan S; Autrup, Herman; Chen, Chunying; Beer, Christiane

    2015-03-01

    Toxicity of silver nanoparticles (Ag NPs) has been reported both in vitro and in vivo. However, the intracellular stability and chemical state of Ag NPs are still not very well studied. In this work, we systematically investigated the cellular uptake pathways, intracellular dissolution and chemical species, and cytotoxicity of Ag NPs (15.9 ± 7.6 nm) in Chinese hamster ovary cell subclone K1 cells, a cell line recommended by the OECD for genotoxicity studies. Quantification of intracellular nanoparticle uptake and ion release was performed through inductively coupled plasma mass spectrometry. X-ray absorption near-edge structure (XANES) was employed to assess the chemical state of intracellular silver. The toxic potential of Ag NPs and Ag(+) was evaluated by cell viability, reactive oxygen species (ROS) production and live-dead cell staining. The results suggest that cellular uptake of Ag NPs involves lipid-raft-mediated endocytosis and energy-independent diffusion. The degradation study shows that Ag NPs taken up into cells dissolved quickly and XANES results directly indicated that the internalized Ag was oxidized to Ag-O- species and then stabilized in silver-sulfur (Ag-S-) bonds within the cells. Subsequent cytotoxicity studies show that Ag NPs decrease cell viability and increase ROS production. Pre-incubation with N-acetyl-L-cysteine, an efficient antioxidant and Ag(+) chelator, diminished the cytotoxicity caused by Ag NPs or Ag(+) exposure. Our study suggests that the cytotoxicity mechanism of Ag NPs is related to the intracellular release of silver ions, followed by their binding to SH-groups, presumably coming from amino acids or proteins, and affecting protein functions and the antioxidant defense system of cells.

  14. Investigation of superparamagnetic (Fe3O4) nanoparticles and magnetic field exposures on CHO-K1 cell line

    NASA Astrophysics Data System (ADS)

    Coker, Zachary; Estlack, Larry; Hussain, Saber; Choi, Tae-Youl; Ibey, Bennett L.

    2016-03-01

    Rapid development in nanomaterial synthesis and functionalization has led to advanced studies in actuation and manipulation of cellular functions for biomedical applications. Often these actuation techniques employ externally applied magnetic fields to manipulate magnetic nanomaterials inside cell bodies in order to drive or trigger desired effects. While cellular interactions with low-frequency magnetic fields and nanoparticles have been extensively studied, the fundamental mechanisms behind these interactions remain poorly understood. Additionally, modern investigations on these concurrent exposure conditions have been limited in scope, and difficult to reproduce. This study presents an easily reproducible method of investigating the biological impact of concurrent magnetic field and nanoparticle exposure conditions using an in-vitro CHO-K1 cell line model, with the purpose of establishing grounds for in-depth fundamental studies of the mechanisms driving cellular-level interactions. Cells were cultured under various nanoparticle and magnetic field exposure conditions from 0 to 500 μg/ml nanoparticle concentrations, and DC, 50 Hz, or 100 Hz magnetic fields with 2.0 mT flux density. Cells were then observed by confocal fluorescence microscopy, and subject to biological assays to determine the effects of concurrent extreme-low frequency magnetic field and nanoparticle exposures on cellnanoparticle interactions, such as particle uptake and cell viability by MTT assay. Current results indicate little to no variation in effect on cell cultures based on magnetic field parameters alone; however, it is clear that deleterious synergistic effects of concurrent exposure conditions exist based on a significant decrease in cell viability when exposed to high concentrations of nanoparticles and concurrent magnetic field.

  15. A novel O-phospho-L-serine sulfhydrylation reaction catalyzed by O-acetylserine sulfhydrylase from Aeropyrum pernix K1.

    PubMed

    Mino, Koshiki; Ishikawa, Kazuhiko

    2003-09-11

    O-Acetylserine sulfhydrylase (OASS), a pyridoxal 5'-phosphate (PLP)-dependent enzyme, catalyzes the synthesis of L-cysteine from O-acetyl-L-serine and sulfide. O-Acetyl-L-serine is labile at high temperatures at which hyperthermophilic archaea live. Herein, a study of the substrate specificity of OASS from Aeropyrum pernix K1 with respect to O-acetyl-L-serine in L-cysteine synthesis is described. L-Azaserine, 3-chloro-L-alanine, and O-phospho-L-serine reacted with A. pernix OASS in a PLP-dependent manner. Sulfhydrylation reactions using these substrates reached a maximum in the pH range between 7.3 and 8.1. L-Azaserine and O-phospho-L-serine were found to be heat-stable substrates. The presence of FeCl3 or NiCl2 strongly inhibited the O-acetyl-L-serine sulfhydrylation reaction, whereas the O-phospho-L-serine sulfhydrylation reaction was only slightly inhibited. Kinetic analyses revealed that the O-phospho-L-serine sulfhydrylation reaction as well as the O-acetyl-L-serine sulfhydrylation reaction for A. pernix OASS followed a ping-pong bi-bi mechanism. In the case of the O-phospho-L-serine sulfhydrylation reaction at 85 degrees C, the K(m) values for O-phospho-L-serine and sulfide, and the rate constant were 250 mM, 12.5 mM, and 14000 s(-1), respectively. The reactivity of O-phospho-L-serine in the L-cysteine synthetic reaction provides a key for understanding the biosynthesis of L-cysteine by hyperthermophilic archaea. This is the first report of an enzyme that catalyzes the O-phospho-L-serine sulfhydrylation reaction.

  16. Structural analysis of the substrate recognition mechanism in O-phosphoserine sulfhydrylase from the hyperthermophilic archaeon Aeropyrum pernix K1.

    PubMed

    Nakamura, Takashi; Kawai, Yoshito; Kunimoto, Kohei; Iwasaki, Yuka; Nishii, Kaoru; Kataoka, Misumi; Ishikawa, Kazuhiko

    2012-09-07

    L-Cysteine is synthesized from O-acetyl-L-serine (OAS) and sulfide by O-acetylserine sulfhydrylase (OASS; EC 2.5.1.47) in plants and bacteria. O-phosphoserine sulfhydrylase (OPSS; EC 2.5.1.65) is a novel enzyme from the hyperthermophilic aerobic archaeon Aeropyrum pernix K1 (2003). OPSS can use OAS or O-phospho-L-serine (OPS) to synthesize L-cysteine. To elucidate the mechanism of the substrate specificity of OPSS, we analyzed three-dimensional structures of the active site of the enzyme. The active-site lysine (K127) of OPSS forms an internal Schiff base with pyridoxal 5'-phosphate. Therefore, crystals of the complexes formed by the K127A mutant with the external Schiff base of pyridoxal 5'-phosphate with either OPS or OAS were prepared and examined by X-ray diffraction analysis. In contrast to that observed for OASS, no significant difference was seen in the overall structure between the free and complexed forms of OPSS. The side chains of T152, S153, and Q224 interacted with the carboxylate of the substrates, as a previous study has suggested. The side chain of R297 has been proposed to recognize the phosphate group of OPS. Surprisingly, however, the position of R297 was significantly unchanged in the complex of the OPSS K127A mutant with the external Schiff base, allowing enough space for an interaction with OPS. The positively charged environment around the entrance of the active site including S153 and R297 is important for accepting negatively charged substrates such as OPS.

  17. Inhibition of topoisomerase II{alpha} activity in CHO K1 cells by 2-[(aminopropyl)amino]ethanethiol (WR-1065)

    SciTech Connect

    Grdina, D.J. |; Constantinou, A.; Shigematsu, N.; Murley, J.S.

    1993-06-01

    The aminothiol 2-[(aminopropyl)amino]ethanethiol (WR-1065) is the active thiol of the clinically studied radioprotective agent S-2-(3-aminopropylamino)ethylphosphorothioic acid (WR-2721). WR-1065 is an effective radiation protector and antimutagenic agent when it is administered 30 min prior to radiation exposure to Chinese hamster ovary Kl cells at a concentration of 4 mM. Under these exposure conditions, topoisomerase (topo) I and II activities and associated protein contents were measured in the K1 cell line using the DNA relaxation assay, the P4 unknotting assay, and immunoblotting, respectively. WR-1065 was ineffective in modifying topo I activity, but it did reduce topo IIa activity by an average of 50 percent. The magnitude of topo IIa protein content, however, was not affected by these exposure conditions. Cell cycle effects were monitored by the method of flow cytometry. Exposure of cells to 4 mM WR-1065 for a period of up to 6 h resulted in a buildup of cells in the G2 compartment. However, in contrast to topo II inhibitors used in chemotherapy, WR-1065 is an effective radioprotector agent capable of protecting against both radiation-induced cell lethality and mutagenesis. One of several mechanisms of radiation protection attributed to aminothiol compounds such as WR-1065 has been their ability to affect endogenous enzymatic reactions involved in DNA synthesis, repair, and cell cycle progression. These results are consistent with such a proposed mechanism and demonstrate in particular a modifying effect by 2-[(aminopropyl)amino]ethanethiol on type II topoisomerase, which is involved in DNA synthesis.

  18. C-terminal region of GADD34 regulates eIF2α dephosphorylation and cell proliferation in CHO-K1 cells.

    PubMed

    Otsuka, Ryo; Harada, Nagakatsu; Aoki, Shouhei; Shirai, Kanna; Nishitsuji, Kazuchika; Nozaki, Ayane; Hatakeyama, Adzumi; Shono, Masayuki; Mizusawa, Noriko; Yoshimoto, Katsuhiko; Nakaya, Yutaka; Kitahata, Hiroshi; Sakaue, Hiroshi

    2016-01-01

    GADD34 is a member of a growth arrest and DNA damage (GADD)-inducible gene family. Here, we established a novel Chinese hamster ovary (CHO)-K1-derived cell line, CHO-K1-G34M, which carries a nonsense mutation (termed the Q525X mutation) in the GADD34 gene. The Q525X mutant protein lacks the C-terminal 66 amino acids required for GADD34 to bind to and activate protein phosphatase 1 (PP1). We investigated the effects of GADD34 with or without the Q525X mutation on the phosphorylation status of PP1 target proteins, including the α subunit of eukaryotic initiation factor 2 (eIF2α) and glycogen synthase kinase 3β (GSK3β). CHO-K1-G34M cells had higher levels of eIF2α phosphorylation compared to the control CHO-K1-normal cells both in the presence and absence of endoplasmic reticulum stress. Overexpression of the wild-type GADD34 protein in CHO-K1-normal cells largely reduced eIF2α phosphorylation, while overexpression of the Q525X mutant did not produce similar reductions. Meanwhile, neither wild type nor Q525X mutation of GADD34 affected the GSK3β phosphorylation status. GADD34 also did not affect the canonical Wnt signaling pathway downstream of GSK3β. Cell proliferation rates were higher, while expression levels of the cyclin-dependent kinase inhibitor p21 were lower in CHO-K1-G34M cells compared to the CHO-K1-normal cells. The GADD34 Q525X mutant had a reduced ability to inhibit cell proliferation and enhance p21 expression of the CHO-K1-normal cells compared to the wild-type GADD34 protein. These results suggest that the GADD34 protein C-terminal plays important roles in regulating not only eIF2α dephosphorylation but also cell proliferation in CHO-K1 cells.

  19. [Occurrence of selected loci of the cps region for capsule K1 or K2 synthesis in epidemic strains of Klebsiella pneumoniae isolated from infants in Poland].

    PubMed

    Gierczyński, Rafał; Kałuzewski, Stanisław; Zasada, Aleksandra A; Rastawicki, Waldemar; Jagielski, Marek

    2005-01-01

    Number of 60 epidemic strains of K. pneumoniae and 15 isolated occasionally from stool samples were tested for presence of 4 and 11 selected loci of the cps cluster of K1 and K2. Following open reading frames (ORFs): 1, 2, 3, 4, 7, 9, 10, 14, 15 (gnd) of cps K2 strain Chedid and genes magA, gmd, wzc, wca of cps K1 strain DTS were searched by PCR in tested and reference strains O1:K1 A5054 and O1:K2 B5055. The ORFs 1 to 3 and ORF15 were detected in both the reference and epidemic strains as well as in the greatest majority of the occasional isolates. Thus, such ORFs were found K. pneumoniae cps common domains, while tested ORFs 4 to 14 were observed in strain B5055 and 11 epidemic isolates from patients of the same hospital ward. Only exception was a single strain O3 occasionally isolated from faeces. The tested genes of cps K1 were detected only in strain A5054 and in two O3 occasional isolates from faeces. Interestingly, these genes as well ORFs 4 to 14 were detected together in the appropriate reference and tested strains with only two exceptions. Therefore, the cps sector occupied by ORFs 4 to 14 was found as group-specific domain. The occurrence ratio of cps K2 group-specific loci among epidemic strains from infants was 18%, while the K1 group-specific loci were absent.

  20. Myocoptid mites (Acariformes: Myocoptidae) of the fauna of the former USSR.

    PubMed

    Bochkov, Andre V

    2016-11-17

    Mites of the family Myocoptidae (Acariformes: Sarcoptoidea) of the former USSR are revised based on the collection of the Zoological Institute RAS (St. Petersburg, Russia). Seventeen described species are recorded. Four species are described as new for science: Trichoecius dubininae sp. nov. from Lasiopodomys gregalis (Pallas) (Rodentia: Cricetidae) from Russia (Chelyabinsk Prov., type locality), Kazakhstan and Kirghizia, T. lemmus sp. nov. from Lemmus sibiricus (Kerr) (Rodentia: Cricetidae) from Russia (Taymyr peninsula), T. meriones sp. nov. from Meriones meridianus (Pallas) (Rodentia: Muridae) from Turkmenia, and Myocoptes meriones sp. nov. from Meriones tamariscinus (type host) from Kirghizia (type locality) and Meriones libycus (Lichtenstein) from Tajikistan. In addition, the presence of 7 more species in the fauna of the former USSR is considered highly probable. Myocoptids of the former USSR represent all 6 genera currently recognized in the family. Keys to species, their hosts and localities are provided and all data are summarized in tabular format. A new name Apocalyptoides nom. nov. is proposed for the preoccupied generic name Apocalypsis Bochkov, 2010 not Butler (1876).

  1. Heterologous expression of metK1-sp and afsR-sp in Streptomyces venezuelae for the production of pikromycin.

    PubMed

    Maharjan, Sushila; Oh, Tae-Jin; Lee, Hei Chan; Sohng, Jae Kyung

    2008-09-01

    Two regulator genes, metK1-sp and afsR-sp, from Streptomyces peucetius ATCC 27952 were heterologously expressed in S. venezuelae ATCC 15439, to produce 14-membered pikromycin antibiotics. The production of pikromycin was increased by 1.6-fold and 2.6-fold by the expression of metK1-sp and afsR-sp, respectively. The overexpression of metK1-sp and afsR-sp in S. venezuelae stimulated the expression of the pathway-specific regulatory genes, pikD and ketosynthase, as demonstrated by RT-PCR. The elevated transcripts of the pikD and ketosynthase genes were consistent with the enhanced production of pikromycin.

  2. Dimerization of Nitrophorin 4 at Low pH and Comparison to the K1A Mutant of Nitrophorin 1

    PubMed Central

    2015-01-01

    Nitrophorin 4, one of the four NO-carrying heme proteins from the salivary glands of Rhodnius prolixus, forms a homodimer at pH 5.0 with a Kd of ∼8 μM. This dimer begins to dissociate at pH 5.5 and is completely dissociated to monomer at pH 7.3, even at 3.7 mM. The dimer is significantly stabilized by binding NO to the heme and at pH 7.3 would require dilution to well below 0.2 mM to completely dissociate the NP4-NO homodimer. The primary techniques used for investigating the homodimer and the monomer–dimer equilibrium were size-exclusion fast protein liquid chromatography at pH 5.0 and 1H{15N} heteronuclear single-quantum coherence spectroscopy as a function of pH and concentration. Preparation of site-directed mutants of NP4 (A1K, D30A, D30N, V36A/D129A/L130A, K38A, R39A, K125A, K125E, D132A, L133V, and K38Q/R39Q/K125Q) showed that the N-terminus, D30, D129, D132, at least one heme propionate, and, by association, likely also E32 and D35 are involved in the dimerization. The “closed loop” form of the A–B and G–H flexible loops of monomeric NP4, which predominates in crystal structures of the monomeric protein reported at pH 5.6 but not at pH 7.5 and which involves all of the residues listed above except D132, is required for dimer formation. Wild-type NP1 does not form a homodimer, but NP1(K1A) and native N-terminal NP1 form dimers in the presence of NO. The homodimer of NP1, however, is considerably less stable than that of NP4 in the absence of NO. This suggests that additional aspartate or glutamate residues present in the C-terminal region of NP4, but not NP1, are also involved in stabilizing the dimer. PMID:25489673

  3. DNA-DSB in CHO-K1 cells induced by heavy-ions: Break rejoining and residual damage (GSI)

    NASA Technical Reports Server (NTRS)

    Taucher-Scholz, G.; Heilmann, J.; Becher, G.; Kraft, G.

    1994-01-01

    DNA double strand breaks (DSB's) are the critical lesions involved in cellular effects of ionizing radiation. Therefore, the evaluation of DSB induction in mammalian cells after heavy ion irradiation is an essential task for the assessment of high-LET radiation risk in space. Of particular interest has been the question of how the biological efficiency for the cellular inactivation endpoint relates to the initial lesions (DSBs) at varying LETs. For cell killing, an increased Relative Biological Efficiency (RBE) has been determined for highLET radiation around 100-200 keV/mu m. At higher LET, the RBE's decrease again to values below one for the very heavy particles. At GSI, DSB-induction was measured in CHO-K1 cells following irradiation with accelerated particles covering a wide LET range. The electrophoretic elution of fragmented DNA out of agarose plugs in a constant electrical field was applied for the detection of DSB's. The fraction of DNA retained was determined considering the relative intensities of ethidium bromide fluorescence in the well and in the gel lane. Dose-effect curves were established, from which the RBE for DSB induction was calculated at a fraction of 0.7 of DNA retained In summary, these rejoining studies are in line with an enhanced severity of the DNA DSB's at higher LET's, resulting in a decreased repairability of the induced lesions. However, no information concerning the fidelity of strand breaks rejoining is provided in these studies. To assess correct rejoining of DNA fragments an experimental system involving individual DNA hybridization bands has been set up. In preliminary experiments Sal I generated DNA fragments of 0.9 Mbp were irradiated with xrays and incubated for repair However, restitution of the original signals was not observed, probably due to the high radiation dose necessary for breakage of a fragment of this size. A banding pattern with NotI hybridization signals in a higher MW range (3Mbp) has been obtained by varying

  4. Deletion of C7L and K1L Genes Leads to Significantly Decreased Virulence of Recombinant Vaccinia Virus TianTan

    PubMed Central

    Liu, Zheng; Wang, Shuhui; Zhang, Qicheng; Tian, Meijuan; Hou, Jue; Wang, Rongmin; Liu, Chang; Ji, Xu; Liu, Ying; Shao, Yiming

    2013-01-01

    The vaccinia virus TianTan (VTT) has been modified as an HIV vaccine vector in China and has shown excellent performance in immunogenicity and safety. However, its adverse effects in immunosuppressed individuals warrant the search for a safer vector in the following clinic trails. In this study, we deleted the C7L and K1L genes of VTT and constructed six recombinant vaccinia strains VTT△C7L, VTT△K1L, VTT△C7LK1L, VTKgpe△C7L, VTKgpe△K1L and VTT△C7LK1L-gag. The pathogenicity and immunogenicity of these recombinants were evaluated in mouse and rabbit models. Comparing to parental VTT, VTT△C7L and VTT△K1L showed significantly decreased replication capability in CEF, Vero, BHK-21 and HeLa cell lines. In particular, replication of VTT△C7LK1L decreased more than 10-fold in all four cell lines. The virulence of all these mutants were decreased in BALB/c mouse and rabbit models; VTT△C7LK1L once again showed the greatest attenuation, having resulted in no evident damage in mice and erythema of only 0.4 cm diameter in rabbits, compared to 1.48 cm for VTT. VTKgpe△C7L, VTKgpe△K1L and VTT△C7LK1L-gag elicited as strong cellular and humoral responses against HIV genes as did VTKgpe, while humoral immune response against the vaccinia itself was reduced by 4-8-fold. These data show that deletion of C7L and K1L genes leads to significantly decreased virulence without compromising animal host immunogenicity, and may thus be key to creating a more safe and effective HIV vaccine vector. PMID:23840887

  5. SphK1 inhibitor II (SKI-II) inhibits acute myelogenous leukemia cell growth in vitro and in vivo

    SciTech Connect

    Yang, Li; Weng, Wei; Sun, Zhi-Xin; Fu, Xian-Jie; Ma, Jun Zhuang, Wen-Fang

    2015-05-15

    Previous studies have identified sphingosine kinase 1 (SphK1) as a potential drug target for treatment of acute myeloid leukemia (AML). In the current study, we investigated the potential anti-leukemic activity of a novel and specific SphK1 inhibitor, SKI-II. We demonstrated that SKI-II inhibited growth and survival of human AML cell lines (HL-60 and U937 cells). SKI-II was more efficient than two known SphK1 inhibitors SK1-I and FTY720 in inhibiting AML cells. Meanwhile, it induced dramatic apoptosis in above AML cells, and the cytotoxicity by SKI-II was almost reversed by the general caspase inhibitor z-VAD-fmk. SKI-II treatment inhibited SphK1 activation, and concomitantly increased level of sphingosine-1-phosphate (S1P) precursor ceramide in AML cells. Conversely, exogenously-added S1P protected against SKI-II-induced cytotoxicity, while cell permeable short-chain ceramide (C6) aggravated SKI-II's lethality against AML cells. Notably, SKI-II induced potent apoptotic death in primary human AML cells, but was generally safe to the human peripheral blood mononuclear cells (PBMCs) isolated from healthy donors. In vivo, SKI-II administration suppressed growth of U937 leukemic xenograft tumors in severe combined immunodeficient (SCID) mice. These results suggest that SKI-II might be further investigated as a promising anti-AML agent. - Highlights: • SKI-II inhibits proliferation and survival of primary and transformed AML cells. • SKI-II induces apoptotic death of AML cells, but is safe to normal PBMCs. • SKI-II is more efficient than two known SphK1 inhibitors in inhibiting AML cells. • SKI-II inhibits SphK1 activity, while increasing ceramide production in AML cells. • SKI-II dose-dependently inhibits U937 xenograft growth in SCID mice.

  6. Deletion of C7L and K1L genes leads to significantly decreased virulence of recombinant vaccinia virus TianTan.

    PubMed

    Liu, Zheng; Wang, Shuhui; Zhang, Qicheng; Tian, Meijuan; Hou, Jue; Wang, Rongmin; Liu, Chang; Ji, Xu; Liu, Ying; Shao, Yiming

    2013-01-01

    The vaccinia virus TianTan (VTT) has been modified as an HIV vaccine vector in China and has shown excellent performance in immunogenicity and safety. However, its adverse effects in immunosuppressed individuals warrant the search for a safer vector in the following clinic trails. In this study, we deleted the C7L and K1L genes of VTT and constructed six recombinant vaccinia strains VTT△C7L, VTT△K1L, VTT△C7LK1L, VTKgpe△C7L, VTKgpe△K1L and VTT△C7LK1L-gag. The pathogenicity and immunogenicity of these recombinants were evaluated in mouse and rabbit models. Comparing to parental VTT, VTT△C7L and VTT△K1L showed significantly decreased replication capability in CEF, Vero, BHK-21 and HeLa cell lines. In particular, replication of VTT△C7LK1L decreased more than 10-fold in all four cell lines. The virulence of all these mutants were decreased in BALB/c mouse and rabbit models; VTT△C7LK1L once again showed the greatest attenuation, having resulted in no evident damage in mice and erythema of only 0.4 cm diameter in rabbits, compared to 1.48 cm for VTT. VTKgpe△C7L, VTKgpe△K1L and VTT△C7LK1L-gag elicited as strong cellular and humoral responses against HIV genes as did VTKgpe, while humoral immune response against the vaccinia itself was reduced by 4-8-fold. These data show that deletion of C7L and K1L genes leads to significantly decreased virulence without compromising animal host immunogenicity, and may thus be key to creating a more safe and effective HIV vaccine vector.

  7. A liver abscess deprived a healthy adult of eyesight: endogenous endophthalmitis associated with a pyogenic liver abscess caused by serotype K1 Klebsiella pneumonia.

    PubMed

    Maruno, Takahisa; Ooiwa, Yoko; Takahashi, Ken; Kodama, Yuzo; Takakura, Shunji; Ichiyama, Satoshi; Chiba, Tsutomu

    2013-01-01

    Klebsiella pneumonia usually causes urinary tract infections, pneumonia, and other infectious diseases in hospitalized and immunocompromised patients. Among the types of Klebsiella pneumonia, serotype K1 is known to be a highly virulent pathogen. We herein report the case of a healthy 63-year-old man with a pyogenic liver abscess and bilateral endogenous endophthalmitis caused by serotype K1 Klebsiella pneumonia. Although the patient received percutaneous abscess drainage and antibiotic therapy, he lost his eyesight. To improve the poor prognoses of ocular complications, providing both an earlier diagnosis and treatment is critical.

  8. PPIP5K1 modulates ligand competition between diphosphoinositol polyphosphates and PtdIns(3,4,5)P3 for polyphosphoinositide-binding domains.

    PubMed

    Gokhale, Nikhil A; Zaremba, Angelika; Janoshazi, Agnes K; Weaver, Jeremy D; Shears, Stephen B

    2013-08-01

    We describe new signalling consequences for PPIP5K1 (diphosphoinositol pentakisphosphate kinase type 1)-mediated phosphorylation of InsP6 and 5-InsP7 to 1-InsP7 and InsP8. In NIH 3T3 cells, either hyperosmotic stress or receptor activation by PDGF (platelet-derived growth factor) promoted translocation of PPIP5K1 from the cytoplasm to the plasma membrane. The PBD1 (polyphosphoinositide-binding domain) in PPIP5K1 recapitulated that translocation. Mutagenesis of PBD1 to reduce affinity for PtdIns(3,4,5)P3 prevented translocation. Using surface plasmon resonance, we found that PBD1 association with vesicular PtdIns(3,4,5)P3 was inhibited by InsP6 and diphosphoinositol polyphosphates. However, the inhibition by PPIP5K1 substrates (IC50: 5-InsP7=5 μM and InsP6=7 μM) was substantially more potent than that of the PPIP5K1 products (IC50: InsP8=32 μM and 1-InsP7=43 μM). This rank order of ligand competition with PtdIns(3,4,5)P3 was also exhibited by the PH (pleckstrin homology) domains of Akt (also known as protein kinase B), GRP1 (general receptor for phosphoinositides 1) and SIN1 (stress-activated protein kinase-interaction protein 1). We propose that, in vivo, PH domain binding of InsP6 and 5-InsP7 suppresses inappropriate signalling ('noise') from stochastic increases in PtdIns(3,4,5)P3. That restraint may be relieved by localized depletion of InsP6 and 5-InsP7 at the plasma membrane following PPIP5K1 recruitment. We tested this hypothesis in insulin-stimulated L6 myoblasts, using mTOR (mechanistic/mammalian target of rapamycin)-mediated phosphorylation of Akt on Ser473 as a readout for SIN1-mediated translocation of mTORC (mTOR complex) 2 to the plasma membrane [Zoncu, Efeyan and Sabatini (2011) Nat. Rev. Mol. Cell Biol. 12, 21-35]. Knockdown of PPIP5K1 expression was associated with a 40% reduction in Ser473 phosphorylation. A common feature of PtdIns(3,4,5)P3-based signalling cascades may be their regulation by PPIP5K1.

  9. Capture of syncytin-Mar1, a Fusogenic Endogenous Retroviral Envelope Gene Involved in Placentation in the Rodentia Squirrel-Related Clade

    PubMed Central

    Redelsperger, François; Cornelis, Guillaume; Vernochet, Cécile; Tennant, Bud C.; Catzeflis, François; Mulot, Baptiste; Heidmann, Odile; Dupressoir, Anne

    2014-01-01

    ABSTRACT Syncytin genes are fusogenic envelope protein (env) genes of retroviral origin that have been captured for a function in placentation. Within rodents, two such genes have previously been identified in the mouse-related clade, allowing a demonstration of their essential role via knockout mice. Here, we searched for similar genes in a second major clade of the Rodentia order, the squirrel-related clade, taking advantage of the complete sequencing of the ground squirrel Ictidomys tridecemlineatus genome. In silico search for env genes with full coding capacity identified several candidate genes with one displaying placenta-specific expression, as revealed by quantitative reverse transcription-PCR analysis of a large panel of tissues. This gene belongs to a degenerate endogenous retroviral element, with recognizable hallmarks of an integrated provirus. Cloning of the gene in an expression vector for ex vivo cell-cell fusion and pseudotype assays demonstrated fusogenicity on a large panel of mammalian cells. In situ hybridization on placenta sections showed specific expression in domains where trophoblast cells fuse into a syncytiotrophoblast at the fetomaternal interface, consistent with a role in syncytium formation. Finally, we show that the gene is conserved among the tribe Marmotini, thus dating its capture back to about at least 25 million years ago, with evidence for purifying selection and conservation of fusogenic activity. This gene that we named syncytin-Mar1 is distinct from all seven Syncytin genes identified to date in eutherian mammals and is likely to be a major effector of placentation in its related clade. IMPORTANCE Syncytin genes are fusogenic envelope genes of retroviral origin, ancestrally captured for a function in placentation. Within rodents, two such genes had been previously identified in the mouse-related clade. Here, in the squirrel-related rodent clade, we identified the envelope gene of an endogenous retrovirus with all the

  10. Capture of syncytin-Mar1, a fusogenic endogenous retroviral envelope gene involved in placentation in the Rodentia squirrel-related clade.

    PubMed

    Redelsperger, François; Cornelis, Guillaume; Vernochet, Cécile; Tennant, Bud C; Catzeflis, François; Mulot, Baptiste; Heidmann, Odile; Heidmann, Thierry; Dupressoir, Anne

    2014-07-01

    Syncytin genes are fusogenic envelope protein (env) genes of retroviral origin that have been captured for a function in placentation. Within rodents, two such genes have previously been identified in the mouse-related clade, allowing a demonstration of their essential role via knockout mice. Here, we searched for similar genes in a second major clade of the Rodentia order, the squirrel-related clade, taking advantage of the complete sequencing of the ground squirrel Ictidomys tridecemlineatus genome. In silico search for env genes with full coding capacity identified several candidate genes with one displaying placenta-specific expression, as revealed by quantitative reverse transcription-PCR analysis of a large panel of tissues. This gene belongs to a degenerate endogenous retroviral element, with recognizable hallmarks of an integrated provirus. Cloning of the gene in an expression vector for ex vivo cell-cell fusion and pseudotype assays demonstrated fusogenicity on a large panel of mammalian cells. In situ hybridization on placenta sections showed specific expression in domains where trophoblast cells fuse into a syncytiotrophoblast at the fetomaternal interface, consistent with a role in syncytium formation. Finally, we show that the gene is conserved among the tribe Marmotini, thus dating its capture back to about at least 25 million years ago, with evidence for purifying selection and conservation of fusogenic activity. This gene that we named syncytin-Mar1 is distinct from all seven Syncytin genes identified to date in eutherian mammals and is likely to be a major effector of placentation in its related clade. Importance: Syncytin genes are fusogenic envelope genes of retroviral origin, ancestrally captured for a function in placentation. Within rodents, two such genes had been previously identified in the mouse-related clade. Here, in the squirrel-related rodent clade, we identified the envelope gene of an endogenous retrovirus with all the features of a

  11. KEY COMPARISON: Final report on CCPR K1-a: Spectral irradiance from 250 nm to 2500 nm

    NASA Astrophysics Data System (ADS)

    Woolliams, Emma R.; Fox, Nigel P.; Cox, Maurice G.; Harris, Peter M.; Harrison, Neil J.

    2006-01-01

    The CCPR K1-a key comparison of spectral irradiance (from 250 nm to 2500 nm) was carried out to meet the requirements of the Mutual Recognition Arrangement by 13 participating national metrology institutes (NMIs). Because of the fragile nature of the tungsten halogen lamps used as comparison artefacts, the comparison was arranged as a star comparison with three lamps per participant. NPL (United Kingdom) piloted the comparison and, by measuring all lamps, provided a link between participants' measurements. The other participants were BNM-INM (France), CENAM (Mexico), CSIRO (Australia), HUT (Finland), IFA-CSIC (Spain), MSL-IRL (New Zealand), NIM (China), NIST (United States of America), NMIJ (Japan), NRC (Canada), PTB (Germany) and VNIIOFI (Russian Federation). Before the analysis was completed and the results known, the pilot discussed with each participant which lamp measurements should be included as representative of their comparison. As a consequence of this check, at least one measurement was excluded from one third of the lamps because of changes due to transportations. The comparison thus highlighted the difficulty regarding the availability of suitable transfer standards for the dissemination of spectral irradiance. The use of multiple lamps and multiple measurements ensured sufficient redundancy that all participants were adequately represented. In addition, during this pre-draft A phase all participants had the opportunity to review the uncertainty budgets and methods of all other participants. This new process helped to ensure that all submitted results and their associated uncertainties were evaluated in a consistent manner. The comparison was analysed using a model-based method which regarded each lamp as having a stable spectral irradiance and the measurements made by an NMI as systematically influenced by a factor that applies to all that NMI's measurements. The aim of the analysis was to estimate the systematic factor for each NMI. Across the

  12. Mach's principle: Exact frame-dragging via gravitomagnetism in perturbed Friedmann-Robertson-Walker universes with K=({+-}1,0)

    SciTech Connect

    Schmid, Christoph

    2009-03-15

    We show that there is exact dragging of the axis directions of local inertial frames by a weighted average of the cosmological energy currents via gravitomagnetism for all linear perturbations of all Friedmann-Robertson-Walker (FRW) universes and of Einstein's static closed universe, and for all energy-momentum-stress tensors and in the presence of a cosmological constant. This includes FRW universes arbitrarily close to the Milne Universe and the de Sitter universe. Hence the postulate formulated by Ernst Mach about the physical cause for the time-evolution of inertial axes is shown to hold in general relativity for linear perturbations of FRW universes. - The time-evolution of local inertial axes (relative to given local fiducial axes) is given experimentally by the precession angular velocity {omega}-vector{sub gyro} of local gyroscopes, which in turn gives the operational definition of the gravitomagnetic field: B-vector{sub g}{identical_to}-2{omega}-vector{sub gyro}. The gravitomagnetic field is caused by energy currents J-vector{sub {epsilon}} via the momentum constraint, Einstein's G{sup 0-}circumflex{sub i-circumflex} equation, (-{delta}+{mu}{sup 2})A-vector{sub g}=-16{pi}G{sub N}J-vector{sub {epsilon}} with B-vector{sub g}=curl A-vector{sub g}. This equation is analogous to Ampere's law, but it holds for all time-dependent situations. {delta} is the de Rham-Hodge Laplacian, and {delta}=-curl curl for the vorticity sector in Riemannian 3-space. - In the solution for an open universe the 1/r{sup 2}-force of Ampere is replaced by a Yukawa force Y{sub {mu}}(r)=(-d/dr)[(1/R)exp(-{mu}r)], form-identical for FRW backgrounds with K=(-1,0). Here r is the measured geodesic distance from the gyroscope to the cosmological source, and 2{pi}R is the measured circumference of the sphere centered at the gyroscope and going through the source point. The scale of the exponential cutoff is the H-dot radius, where H is the Hubble rate, dot is the derivative with respect to

  13. Selective synthesis and labeling of the polysialic acid capsule in Escherichia coli K1 strains with mutations in nanA and neuB.

    PubMed Central

    Vimr, E R

    1992-01-01

    The enzymes required for polysialic acid capsule synthesis in Escherichia coli K1 are encoded by region 2 neu genes of the multigenic kps cluster. To facilitate analysis of capsule synthesis and translocation, an E. coli K1 strain with mutations in nanA and neuB, affecting sialic acid degradation and synthesis, respectively, was constructed by transduction. The acapsular phenotype of the mutant was corrected in vivo by exogenous addition of sialic acid. By blocking sialic acid degradation, the nanA mutation allows intracellular metabolite accumulation, while the neuB mutation prevents dilution by the endogenous sialic acid pool and allows capsule synthesis to be controlled experimentally by the exogenous addition of sialic acid to the growth medium. Complementation was detected by bacteriophage K1F adsorption or infectivity assays. Polysialic acid translocation was observed within 2 min after addition of sialic acid to the growth medium, demonstrating the rapidity in vivo of sialic acid transport, activation, and polymerization and translocation of polysaccharide to the cell surface. Phage adsorption was not inhibited by chloramphenicol, demonstrating that de novo protein synthesis was not required for polysialic acid synthesis or translocation at 37 degrees C. Exogenous radiolabeled sialic acid was incorporated exclusively into capsular polysaccharide. The polymeric nature of the labeled capsular material was confirmed by gel permeation chromatography and susceptibility of sialyl polymers to K1F endo-N-acylneuraminidase. The ability to experimentally manipulate capsule expression provides new approaches for investigating polysialic acid synthesis and membrane translocation mechanisms. PMID:1400168

  14. Rapidly fatal bacteremic pneumonia caused by Klebsiella pneumoniae with K1 hypermucoviscosity phenotype in a previously healthy young man receiving levofloxacin treatment.

    PubMed

    Chuang, Tzu-Yi; Lin, Chou-Jui; Chi, Chun-Lin; Liu, An-Yu; Lee, Shih-Wei; Lin, T L; Wang, Jin-Town; Hsueh, Po-Ren

    2009-10-01

    Fatal bacteremic Klebsiella pneumoniae pneumonia is commonly encountered in alcoholic and diabetic patients. This report describes a previously healthy young man with rapidly fatal bacteremic pneumonia caused by K. pneumoniae serotype K1, complicated by septic shock and multiple organ dysfunction.

  15. Rapamycin-enhanced mitomycin C-induced apoptotic death is mediated through the S6K1-Bad-Bak pathway in peritoneal carcinomatosis.

    PubMed

    Song, X; Dilly, A-K; Kim, S-Y; Choudry, H A; Lee, Y J

    2014-06-05

    Peritoneal carcinomatosis (PC) is the most common secondary cancerous disease, and more effective novel regimens are needed. In this study, we identified a novel combination treatment for PC, chemotherapeutic agent mitomycin C in combination with mTOR (mammalian target of rapamycin) inhibitor rapamycin. We observed that the combination of mitomycin C and rapamycin induced synergistic cytotoxicity and apoptosis, which was mediated through an increase in caspase activation. The combination of mitomycin C and rapamycin inactivated p70 S6 ribosomal kinase (S6K1) and dephosphorylated Bad, leading to dissociation of Bcl-xL from Bak, which resulted in Bak oligomerization, mitochondria dysfunction and cytochrome c release. PF-4708671, a S6K1-specific inhibitor, enhanced the combination treatment-induced apoptosis, whereas S6K1 E389 DeltaCT-HA (S6K1 active form) dramatically decreased the induction of apoptosis. In addition, the combination treatment significantly inhibited LS174T intraperitoneal tumor growth in vivo. This study provides a preclinical rationale for apoptosis induction linked with the mTOR pathway through a combination of chemotherapeutic agents and mTOR inhibitor, and will support this combinatorial strategy to PC patients.

  16. Whole-Genome Sequences of the Archetypal K1 Escherichia coli Neonatal Isolate RS218 and Contemporary Neonatal Bacteremia Clinical Isolates SCB11, SCB12, and SCB15.

    PubMed

    Day, Michael W; Jackson, Lydgia A; Akins, Darrin R; Dyer, David W; Chavez-Bueno, Susana

    2015-02-26

    Neonatal bacteremia Escherichia coli strains commonly belong to the K1 capsular type. Their ability to cause invasive neonatal disease appears to be determined by other virulence factors that have yet to be identified. We report here the genome sequences of four E. coli neonatal bacteremia isolates, including that of the archetypal strain RS218.

  17. Co-purification of microsomal epoxide hydrolase with the warfarin-sensitive vitamin K1 oxide reductase of the vitamin K cycle.

    PubMed

    Guenthner, T M; Cai, D; Wallin, R

    1998-01-15

    Vitamin K1 oxide reductase activity has been partially purified from rat liver microsomes. A three-step procedure produced a preparation in which warfarin-sensitive vitamin K1 oxide reductase activity was 118-fold enriched over the activity in intact rat liver microsomes. A major component of the multi-protein mixture was identified as a 50 kDa protein that strongly cross-reacts with antiserum prepared against homogeneous rat liver microsomal epoxide hydrolase. The reductase preparation also had a high level or epoxide hydrolase activity against two xenobiotic epoxide substrates. The K(m) values for hydrolysis by the reductase preparation were similar to those for homogeneous microsomal epoxide hydrolase itself, and the specific hydrolase activities of the reductase preparation were 25-35% of the specific activities measured for the homogeneous hydrolase preparation. Antibodies prepared against homogeneous microsomal epoxide hydrolase inhibited up to 80% of reductase activity of the reductase preparation. Homogeneous microsomal epoxide hydrolase had no vitamin K1 oxide reductase activity. This evidence suggests that microsomal epoxide hydrolase, or a protein that is very similar to it, is a major functional component of a multi-protein complex that is responsible for vitamin K1 oxide reduction in rat liver microsomes.

  18. Risk Assessment of Head Start Children with the Brigance K&1 Screen: Differential Performance by Sex, Age, and Predictive Accuracy for Early School Achievement and Special Education Placement.

    ERIC Educational Resources Information Center

    Mantzicopoulos, Panayota

    1999-01-01

    Examined differences in performance as well as reliability and validity indices for 256 Head Start children screened with Brigance K&1 screen. Found high overall test consistency, but considerable variability across subscales. Classification analyses established that the Brigance was not completely accurate in predicting early school…

  19. Role of mTORC1-S6K1 signaling pathway in regulation of hematopoietic stem cell and acute myeloid leukemia.

    PubMed

    Ghosh, Joydeep; Kapur, Reuben

    2017-03-22

    Dysregulation of the mechanistic target of rapamycin complex 1 (mTORC1)-p70 ribosomal protein kinase 1 (S6K1) signaling pathway occurs frequently in acute myeloid leukemia (AML) patients. This pathway also plays a critical role in maintaining normal cellular processes. Given the importance of leukemia stem cells (LSC) in the development of minimal residual disease (MRD), it is critical to use therapeutic interventions that target LSC population to prevent disease relapse. mTORC1-S6K1 pathway has been identified as an important regulator of hematopoietic stem cell (HSC) and LSC functions. Both HSC and LSC functions require regulation of key cellular processes including proliferation, metabolism and autophagy, which are regulated by mTORC1 pathway. Despite mTORC1-S6K1 pathway being a critical regulator of AML initiation and progression, inhibitors of this pathway alone have yielded mixed results in clinical studies. Recent studies have identified strategies to develop new mTORC1-S6K1 inhibitors like RapaLink-1, which could circumvent the drug resistance observed in AML cells as well as in LSC. In this article, we review recent advances made in identifying the role of different components of this pathway in the regulation of HSC and LSC along with possible therapeutic approaches.

  20. α-Melanocyte stimulating hormone attenuates dexamethasone-induced osteoblast damages through activating melanocortin receptor 4-SphK1 signaling.

    PubMed

    Guo, Shiguang; Xie, Yue; Fan, Jian-bo; Ji, Feng; Wang, Shouguo; Fei, Haodong

    2016-01-08

    Long-term glucocorticoid (GC) usage may cause non-traumatic femoral head osteonecrosis. Dexamethasone (Dex) is shown to exert potent cytotoxic effect to osteoblasts. Here, we investigated the potential activity of α-melanocyte stimulating hormone (α-MSH) against the process. Our data revealed that pretreatment of α-MSH significantly inhibited Dex-induced apoptosis and necrosis in both osteoblastic-like MC3T3-E1 cells and primary murine osteoblasts. Melanocortin receptor 4 (MC4R) acts as the receptor of α-MSH in mediating its actions in osteoblasts. The MC4R antagonist SHU9119, or shRNA-mediated knockdown of MC4R, almost abolished α-MSH-induced activation of downstream signalings (Akt and Erk1/2) and its pro-survival effect in osteoblasts. Further studies showed that α-MSH activated MC4R downstream sphingosine kinase 1 (SphK1) and increased cellular sphingosine-1-phosphate (S1P) content in MC3T3-E1 cells and primary murine osteoblasts, which were blocked by SHU9119 or MC4R shRNAs. SphK1 inhibition by the its inhibitor N,N-dimethylsphingosine (DMS), or SphK1 knockdown by targeted-shRNAs, largely attenuated α-MSH-mediated osteoblast protection against Dex. Together, these results suggest that α-MSH alleviates Dex-induced damages to cultured osteoblasts through activating MC4R-SphK1 signaling.

  1. In silico predicted structural and functional insights of all missense mutations on 2B domain of K1/K10 causing genodermatoses

    PubMed Central

    Banerjee, Santasree; Wu, Qian; Ying, Yuyi; Li, Yanni; Shirota, Matsuyuki; Neculai, Dante; Li, Chen

    2016-01-01

    The K1 and K10 associated genodermatoses are characterized by clinical symptoms of mild to severe redness, blistering and hypertrophy of the skin. In this paper, we set out to computationally investigate the structural and functional effects of missense mutations on the 2B domain of K1/K10 heterodimer and its consequences in disease phenotype. We modeled the structure of the K1/K10 heterodimer based on crystal structures for the human homolog K5/K14 heterodimer, and identified that the missense mutations exert their effects on stability and assembly competence of the heterodimer by altering physico-chemical properties, interatomic interactions, and inter-residue atomic contacts. Comparative structural analysis between all the missense mutations and SNPs showed that the location and physico-chemical properties of the substituted amino acid are significantly correlated with phenotypic variations. In particular, we find evidence that a particular SNP (K10, p.E443K) is a pathogenic nsSNP which disrupts formation of the hydrophobic core and destabilizes the heterodimer through the loss of interatomic interactions. Our study is the first comprehensive report analyzing the mutations located on 2B domain of K1/K10 heterodimeric coiled-coil complex. PMID:27421141

  2. Emergence of KPC-producing Klebsiella pneumoniae hypervirulent clone of capsular serotype K1 that belongs to sequence type 11 in Mainland China.

    PubMed

    Wei, Dan-Dan; Wan, La-Gen; Deng, Qiong; Liu, Yang

    2016-06-01

    KPC-2 has been rarely reported in hypervirulent Klebsiella pneumoniae strains. Here, we describe a KPC-2-producing K. pneumoniae hypervirulent clone of capsular serotype K1 belonging to sequence type 11. The presence of KPC carbapenemase in hypervirulent clone could mark an evolutionary step toward its establishment as major nosocomial pathogen.

  3. Evodiamine Induces Apoptosis and Enhances TRAIL-Induced Apoptosis in Human Bladder Cancer Cells through mTOR/S6K1-Mediated Downregulation of Mcl-1

    PubMed Central

    Zhang, Tao; Qu, Shanna; Shi, Qi; He, Dalin; Jin, Xunbo

    2014-01-01

    The tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), either alone or in combination with other anti-cancer agents, has been considered as a new strategy for anti-cancer therapy. In this study, we demonstrated that evodiamine, a quinolone alkaloid isolated from the fruit of Evodia fructus, induced apoptosis and enhanced TRAIL-induced apoptosis in human bladder cancer cells. To elucidate the underlying mechanism, we found that evodiamine significantly reduced the protein levels of Mcl-1 in 253J and T24 bladder cancer cells, and overexpression of this molecule attenuated the apoptosis induced by evodiamine alone, or in combination with TRAIL. Further experiments revealed that evodiamine did not affect the mRNA level, proteasomal degradation and protein stability of Mcl-1. On the other hand, evodiamine inhibited the mTOR/S6K1 pathway, which usually regulates protein translation; moreover, knockdown of S6K1 with small interfering RNA (siRNA) effectively reduced Mcl-1 levels, indicating evodiamine downregulates c-FLIP through inhibition of mTOR/S6K1 pathway. Taken together, our results indicate that evodiamine induces apoptosis and enhances TRAIL-induced apoptosis possibly through mTOR/S6K1-mediated downregulation of Mcl-1; furthermore, these findings provide a rationale for the combined application of evodiamine with TRAIL in the treatment of bladder cancer. PMID:24566141

  4. Rapamycin-enhanced mitomycin C-induced apoptotic death is mediated through the S6K1–Bad–Bak pathway in peritoneal carcinomatosis

    PubMed Central

    Song, X; Dilly, A-K; Kim, S-Y; Choudry, H A; Lee, Y J

    2014-01-01

    Peritoneal carcinomatosis (PC) is the most common secondary cancerous disease, and more effective novel regimens are needed. In this study, we identified a novel combination treatment for PC, chemotherapeutic agent mitomycin C in combination with mTOR (mammalian target of rapamycin) inhibitor rapamycin. We observed that the combination of mitomycin C and rapamycin induced synergistic cytotoxicity and apoptosis, which was mediated through an increase in caspase activation. The combination of mitomycin C and rapamycin inactivated p70 S6 ribosomal kinase (S6K1) and dephosphorylated Bad, leading to dissociation of Bcl-xL from Bak, which resulted in Bak oligomerization, mitochondria dysfunction and cytochrome c release. PF-4708671, a S6K1-specific inhibitor, enhanced the combination treatment-induced apoptosis, whereas S6K1 E389 DeltaCT-HA (S6K1 active form) dramatically decreased the induction of apoptosis. In addition, the combination treatment significantly inhibited LS174T intraperitoneal tumor growth in vivo. This study provides a preclinical rationale for apoptosis induction linked with the mTOR pathway through a combination of chemotherapeutic agents and mTOR inhibitor, and will support this combinatorial strategy to PC patients. PMID:24901052

  5. Measurement of Vitamin K1 in Commercial Canola Cultivars from Growing Locations in North and South America Using High-Performance Liquid Chromatography-Tandem Mass Spectrometry.

    PubMed

    Claussen, Fred A; Taylor, Mary L; Breeze, Matthew L; Liu, Kang

    2015-01-27

    Plant oils, including canola oil, are considered to be major sources of vitamin K as the second most substantial contributor of vitamin K to the human diet. Green leafy vegetables are the largest source of vitamin K. However, the effects of environment and germplasm on vitamin K levels in harvested canola seed have not been extensively investigated. To better understand these relationships, harvested canola seed from a range of diverse cultivars grown in different geographical locations in North and South America was assessed for levels of vitamin K. The analytical method developed to perform this measurement was based on C30 reversed-phase HPLC that could distinguish the biologically active trans-vitamin K1 from the inactive cis-isomer. Results demonstrated that for the majority of the canola cultivars evaluated, those cultivated in the North American sites had higher average vitamin K1 levels than those cultivated in the South American sites. Not all of the cultivars exhibited differences in response to the environment, suggesting that individual cultivar genetics also played a role in the variability of vitamin K1 levels observed in canola seed. Results from this study suggest that cultivar and environmental effects influence vitamin K1 levels in canola seed and provide a context to assess compositional variability of new cultivars.

  6. A Novel Saccharomyces cerevisiae Killer Strain Secreting the X Factor Related to Killer Activity and Inhibition of S. cerevisiae K1, K2 and K28 Killer Toxins.

    PubMed

    Melvydas, Vytautas; Bružauskaitė, Ieva; Gedminienė, Genovaitė; Šiekštelė, Rimantas

    2016-09-01

    It was determined that Kx strains secrete an X factor which can inhibit all known Saccharomyces cerevisiae killer toxins (K1, K2, K28) and some toxins of other yeast species-the phenomenon not yet described in the scientific literature. It was shown that Kx type yeast strains posess a killer phenotype producing small but clear lysis zones not only on the sensitive strain α'1 but also on the lawn of S. cerevisiae K1, K2 and K28 type killer strains at temperatures between 20 and 30 °C. The pH at which killer/antikiller effect of Kx strain reaches its maximum is about 5.0-5.2. The Kx yeast were identified as to belong to S. cerevisiae species. Another newly identified S. cerevisiae killer strain N1 has killer activity but shows no antikilller properties against standard K1, K2 and K28 killer toxins. The genetic basis for Kx killer/antikiller phenotype was associated with the presence of M-dsRNA which is bigger than M-dsRNA of standard S. cerevisiae K1, K2, K28 type killer strains. Killer and antikiller features should be encoded by dsRNA. The phenomenon of antikiller (inhibition) properties was observed against some killer toxins of other yeast species. The molecular weight of newly identified killer toxins which produces Kx type strains might be about 45 kDa.

  7. Identification of a Dual Inhibitor of Janus Kinase 2 (JAK2) and p70 Ribosomal S6 Kinase1 (S6K1) Pathways*

    PubMed Central

    Byun, Sanguine; Lim, Semi; Mun, Ji Young; Kim, Ki Hyun; Ramadhar, Timothy R.; Farrand, Lee; Shin, Seung Ho; Thimmegowda, N. R.; Lee, Hyong Joo; Frank, David A.; Clardy, Jon; Lee, Sam W.; Lee, Ki Won

    2015-01-01

    Bioactive phytochemicals can suppress the growth of malignant cells, and investigation of the mechanisms responsible can assist in the identification of novel therapeutic strategies for cancer therapy. Ginger has been reported to exhibit potent anti-cancer effects, although previous reports have often focused on a narrow range of specific compounds. Through a direct comparison of various ginger compounds, we determined that gingerenone A selectively kills cancer cells while exhibiting minimal toxicity toward normal cells. Kinase array screening revealed JAK2 and S6K1 as the molecular targets primarily responsible for gingerenone A-induced cancer cell death. The effect of gingerenone A was strongly associated with relative phosphorylation levels of JAK2 and S6K1, and administration of gingerenone A significantly suppressed tumor growth in vivo. More importantly, the combined inhibition of JAK2 and S6K1 by commercial inhibitors selectively induced apoptosis in cancer cells, whereas treatment with either agent alone did not. These findings provide rationale for dual targeting of JAK2 and S6K1 in cancer for a combinatorial therapeutic approach. PMID:26242912

  8. A chronic increase in physical activity inhibits fed-state mTOR/S6K1 signaling and reduces IRS-1 serine phosphorylation in rat skeletal muscle.

    PubMed

    Glynn, Erin L; Lujan, Heidi L; Kramer, Victoria J; Drummond, Micah J; DiCarlo, Stephen E; Rasmussen, Blake B

    2008-02-01

    A chronic increase in physical activity and (or) endurance training can improve insulin sensitivity in insulin-resistant skeletal muscle. Cellular mechanisms responsible for the development of insulin resistance are unclear, though one proposed mechanism is that nutrient overload chronically increases available energy, over-activating the mammalian target of rapamycin (mTOR) and ribosomal S6 kinase 1 (S6K1) signaling pathway leading to increased phosphorylation of serine residues on insulin receptor substrate-1 (IRS-1). The objective of this study was to determine if increased physical activity would inhibit mTOR/S6K1 signaling and reduce IRS-1 serine phosphorylation in rat skeletal muscle. Soleus muscle was collected from fed male Sprague-Dawley sedentary rats (Inactive) and rats with free access to running wheels for 9 weeks (Active). Immunoblotting methods were used to measure phosphorylation status of mTOR, S6K1, IRS-1, and PKB/Akt (protein kinase B/AKT), and total abundance of proteins associated with the mTOR pathway. Muscle citrate synthase activity and plasma insulin and glucose concentrations were measured. Phosphorylation of mTOR (Ser2448), S6K1 (Thr389), and IRS-1 (Ser636-639) was reduced in Active rats (p<0.05). Total protein abundance of mTOR, S6K1, IRS-1, 4E-BP1, eEF2, PKB/Akt and AMPKalpha, and phosphorylation of PKB/Akt were unaffected (p>0.05). Total SKAR protein, a downstream target of S6K1, and citrate synthase activity increased in Active rats (p<0.05), though plasma insulin and glucose levels were unchanged (p>0.05). Reduced mTOR/S6K1 signaling during chronic increases in physical activity may play an important regulatory role in the serine phosphorylation of IRS-1, which should be examined as a potential mechanism for attenuation of insulin resistance associated with increased IRS-1 serine phosphorylation.

  9. Essential role for SphK1/S1P signaling to regulate hypoxia-inducible factor 2α expression and activity in cancer.

    PubMed

    Bouquerel, P; Gstalder, C; Müller, D; Laurent, J; Brizuela, L; Sabbadini, R A; Malavaud, B; Pyronnet, S; Martineau, Y; Ader, I; Cuvillier, O

    2016-03-14

    The sphingosine kinase-1/sphingosine 1-phosphate (SphK1/S1P) signaling pathway has been reported to modulate the expression of the canonical transcription factor hypoxia-inducible HIF-1α in multiple cell lineages. HIF-2α is also frequently overexpressed in solid tumors but its role has been mostly studied in clear cell renal cell carcinoma (ccRCC), the most common form of kidney cancer, where HIF-2α has been established as a driver of a more aggressive disease. In this study, the role of SphK1/S1P signaling with regard to HIF-2α was investigated in various cancer cell models including ccRCC cells. Under hypoxic conditions or in ccRCC lacking a functional von Hippel-Lindau (VHL) gene and expressing high levels of HIF-2α, SphK1 activity controls HIF-2α expression and transcriptional activity through a phospholipase D (PLD)-driven mechanism. SphK1 silencing promotes a VHL-independent HIF-2α loss of expression and activity and reduces cell proliferation in ccRCC. Importantly, downregulation of SphK1 is associated with impaired Akt and mTOR signaling in ccRCC. Taking advantage of a monoclonal antibody neutralizing extracellular S1P, we show that inhibition of S1P extracellular signaling blocks HIF-2α accumulation in ccRCC cell lines, an effect mimicked when the S1P transporter Spns2 or the S1P receptor 1 (S1P1) is silenced. Here, we report the first evidence that the SphK1/S1P signaling pathway regulates the transcription factor hypoxia-inducible HIF-2α in diverse cancer cell lineages notably ccRCC, where HIF-2α has been established as a driver of a more aggressive disease. These findings demonstrate that SphK1/S1P signaling may act as a canonical regulator of HIF-2α expression in ccRCC, giving support to its inhibition as a therapeutic strategy that could contribute to reduce HIF-2 activity in ccRCC.

  10. VPS34 regulates TSC1/TSC2 heterodimer to mediate RheB and mTORC1/S6K1 activation and cellular transformation

    PubMed Central

    Mohan, Nishant; Shen, Yi; Dokmanovic, Milos; Endo, Yukinori; Hirsch, Dianne S.; Wu, Wen Jin

    2016-01-01

    VPS34 is reported to activate S6K1 and is implicated in regulating cell growth, the mechanisms of which remain elusive. Here, we describe novel mechanisms by which VPS34 upregulates mTOR/S6K1 activity via downregulating TSC2 protein and activating RheB activity. Specifically, upregulation of VPS34 lipid kinase increases local production of ptdins(3)p in the plasma membrane, which recruits PIKFYVE, a FYVE domain containing protein, to ptdins(3)p enriched regions of the plasma membrane, where VPS34 forms a protein complex with PIKFYVE and TSC1. This in turn disengages TSC2 from the TSC1/TSC2 heterodimer, leading to TSC2 ubiquitination and degradation. Downregulation of TSC2 promotes the activation of RheB and mTOR/S6K1. When VPS34 lipid kinase activity is increased by introduction of an H868R mutation, ptdins(3)p production at the plasma membrane is dramatically increased, which recruits more PIKFYVE and TSC1 molecules to the plasma membrane. This results in the enhanced TSC2 ubiquitination and degradation, and subsequent activation of RheB and mTORC1/S6K1, leading to oncogenic transformation. The role played by VPS34 in regulating mTOR/S6K1 activity and cellular transformation is underscored by the fact that the VPS34 kinase dead mutant blocks VPS34-induced recruitment of PIKFYVE and TSC1 to the plasma membrane. This study provides mechanistic insight into the cellular function of VPS34 in regulating oncogenic transformation and important indications for identifying VPS34 specific mutations in human cancers. PMID:27409169

  11. Bioluminescent imaging reveals novel patterns of colonization and invasion in systemic Escherichia coli K1 experimental infection in the neonatal rat.

    PubMed

    Witcomb, Luci A; Collins, James W; McCarthy, Alex J; Frankel, Gadi; Taylor, Peter W

    2015-12-01

    Key features of Escherichia coli K1-mediated neonatal sepsis and meningitis, such as a strong age dependency and development along the gut-mesentery-blood-brain course of infection, can be replicated in the newborn rat. We examined temporal and spatial aspects of E. coli K1 infection following initiation of gastrointestinal colonization in 2-day-old (P2) rats after oral administration of E. coli K1 strain A192PP and a virulent bioluminescent derivative, E. coli A192PP-lux2. A combination of bacterial enumeration in the major organs, two-dimensional bioluminescence imaging, and three-dimensional diffuse light imaging tomography with integrated micro-computed tomography indicated multiple sites of colonization within the alimentary canal; these included the tongue, esophagus, and stomach in addition to the small intestine and colon. After invasion of the blood compartment, the bacteria entered the central nervous system, with restricted colonization of the brain, and also invaded the major organs, in line with increases in the severity of symptoms of infection. Both keratinized and nonkeratinized surfaces of esophagi were colonized to a considerably greater extent in susceptible P2 neonates than in corresponding tissues from infection-resistant 9-day-old rat pups; the bacteria appeared to damage and penetrate the nonkeratinized esophageal epithelium of infection-susceptible P2 animals, suggesting the esophagus represents a portal of entry for E. coli K1 into the systemic circulation. Thus, multimodality imaging of experimental systemic infections in real time indicates complex dynamic patterns of colonization and dissemination that provide new insights into the E. coli K1 infection of the neonatal rat.

  12. The collectins CL-L1, CL-K1 and CL-P1, and their roles in complement and innate immunity.

    PubMed

    Hansen, Soren W K; Ohtani, Katsuki; Roy, Nitai; Wakamiya, Nobutaka

    2016-10-01

    Both the complement system and collectins play important roles in our innate immune system. The collectins, which are characterized by their inclusion of a collagen-like region and a calcium-dependent carbohydrate recognition domain, are pattern recognition molecules and include the well characterized proteins mannan-binding lectin (MBL) and the surfactant proteins SP-A/-D. Collectin liver 1 (CL-L1), collectin kidney 1 (CL-K1) and collectin placenta 1 (CL-P1) are the most recently discovered collectins. Although their function is still under investigation, accumulating information suggests that CL-L1, CL-K1 and CL-P1 play important roles in host defense by recognizing a variety of microorganisms and interacting with effector proteins, including complement components. The recent establishment of the existence of CL-K1 in the circulation in form of heteromeric complexes with CL-L1 (known as CL-LK) and its activation of the lectin pathway via MASPs, drew new attention in the complement biology, which was further strengthened by the observed interactions between CL-P1 and CRP-C1q-factor H or properdin. Deficiency of either CL-K1 or MASP-3 has been demonstrated in 3MC syndrome patients with developmental abnormalities, showing that lectin pathway components, regulation and/or activation are essential during the embryonic development; another feature that they most likely share CL-P1. Herein, we discuss the recent characteristics and roles of the collectins CL-L1, CL-K1 and CL-P1 in the complement system, in innate immunity and their possible association with disease development and pathogenesis.

  13. Circulating levels of vitamin K1, menaquinone-4, and menaquinone-7 in healthy elderly Japanese women and patients with vertebral fractures and patients with hip fractures.

    PubMed

    Kawana, K; Takahashi, M; Hoshino, H; Kushida, K

    2001-08-01

    Recently, vitamin K has become increasingly of interest in the bone metabolism field because of its role as a cofactor in the carboxylation of osteocalcin. Although the role of osteocalcin is not clear, noncarboxylated osteocalcin is one risk factor in hip fractures. It has been reported that the circulating levels of vitamin K1 in osteoporotic patients were significantly lower than those of age-matched control subjects. In this study, we measured circulating levels of vitamin K1, menaquinone-4 (MK-4) and menaquinone-7 (MK-7) in 23 normal healthy women aged 52-93 years (mean +/- SD: 80.1 +/- 3.5), 13 female patients with vertebral fractures aged 66-93 years (80.3 +/- 7.8) and 38 female patients with hip fractures aged 76-87 years (79.8 +/- 9.2), (all Japanese), in order to make sure whether these vitamin K levels were different in these three groups. Serum circulating levels of MK-4 was undetectable in most subjects (only one out of 74). Appreciable numbers from these three groups had undetectable levels of MK-7 (52% of the control group, 23% of the vertebral fracture group and 24% of the hip fracture group). Eight subjects from the normal control group (35%) and five patients from the vertebral group (38%) had undetectable levels of vitamin K1. We did not find a significant difference in the measurable levels of vitamin K1, MK-4 and MK-7 in patients with vertebral fractures or patients with hip fractures compared to age-matched normal controls. Undetectable levels of measured vitamin K1, MK-4 and MK-7 in most of subjects may significantly affect the results.

  14. Regulation of miR-29b-1/a transcription and identification of target mRNAs in CHO-K1 cells.

    PubMed

    Muluhngwi, Penn; Richardson, Kirsten; Napier, Joshua; Rouchka, Eric C; Mott, Justin L; Klinge, Carolyn M

    2017-03-15

    miR-29b and miR-29a transcript levels were reported to increase in exponentially growing CHO-K1 cells. Here, we examine the regulation of miR-29b-1/a in CHO-K1 cells. We observed that 4-hydroxytamoxifen (4-OHT) increased pri-miR-29b-1 and pri-miR-29a transcription in CHO-K1 cells by activating endogenous estrogen receptor α (ERα). DICER, an established, bona fide target of miR-29b-1/a, was shown to be regulated by 4-OHT in CHO-K1 cells. We showed that miR-29b-1 and miR-29a serve a repressive role in cell proliferation, migration, invasion, and colony formation in CHO-K1 cells. To identify other targets of miR-29b-1 and miR-29a, RNA sequencing was performed by transfecting cells with anti-miR-29a, which inhibits both miR-29a and miR-29b-1, pre-miR-29b-1, and/or pre-miR-29a. In silico network analysis in MetaCore™ identified common and unique putative gene targets of miR-29b-1 and miR-29a. Pathway analysis of identified putative miR-29 targets were related to cell adhesion, cytoskeletal remodeling, and development. Further inquiry revealed regulation of pathways mediating responses to growth factor stimulus and cell cycle regulation.

  15. Cross-talk between sirtuin and mammalian target of rapamycin complex 1 (mTORC1) signaling in the regulation of S6 kinase 1 (S6K1) phosphorylation.

    PubMed

    Hong, Sungki; Zhao, Bin; Lombard, David B; Fingar, Diane C; Inoki, Ken

    2014-05-09

    p70 ribosomal S6 kinase (S6K1), a major substrate of the mammalian target of rapamycin (mTOR) kinase, regulates diverse cellular processes including protein synthesis, cell growth, and survival. Although it is well known that the activity of S6K1 is tightly coupled to its phosphorylation status, the regulation of S6K1 activity by other post-translational modifications such as acetylation has not been well understood. Here we show that the acetylation of the C-terminal region (CTR) of S6K1 blocks mTORC1-dependent Thr-389 phosphorylation, an essential phosphorylation site for S6K1 activity. The acetylation of the CTR of S6K1 is inhibited by the class III histone deacetylases, SIRT1 and SIRT2. An S6K1 mutant lacking acetylation sites in its CTR shows enhanced Thr-389 phosphorylation and kinase activity, whereas the acetylation-mimetic S6K1 mutant exhibits decreased Thr-389 phosphorylation and kinase activity. Interestingly, relative to the acetylation-mimetic S6K1 mutant, the acetylation-defective mutant displays higher affinity toward Raptor, an essential scaffolding component of mTORC1 that recruits mTORC1 substrates. These observations indicate that sirtuin-mediated regulation of S6K1 acetylation is an additional important regulatory modification that impinges on the mechanisms underlying mTORC1-dependent S6K1 activation.

  16. Inhibition of the SphK1/S1P signaling pathway by melatonin in mice with liver fibrosis and human hepatic stellate cells.

    PubMed

    González-Fernández, Bárbara; Sánchez, Diana I; Crespo, Irene; San-Miguel, Beatriz; Álvarez, Marcelino; Tuñón, María J; González-Gallego, Javier

    2016-11-01

    The sphingosine kinase 1/sphingosine 1-phosphate (SphK1/S1P) system is involved in different pathological processes, including fibrogenesis. Melatonin abrogates activation of hepatic stellate cells (HSCs) and attenuates different profibrogenic pathways in animal models of fibrosis, but it is unknown if protection associates with its inhibitory effect on the SphK1/S1P axis. Mice in treatment groups received carbon tetrachloride (CCl4 ) 5 μL g(-1) body wt i.p. twice a week for 4 or 6 weeks. Melatonin was given at 5 or 10 mg kg(-1)  day(-1) i.p, beginning 2 weeks after the start of CCl4 administration. At both 4 and 6 weeks following CCl4 treatment, liver mRNA levels, protein concentration and immunohistochemical labelling for SphK1 increased significantly. S1P production, and expression of S1P receptor (S1PR)1, S1PR3 and acid sphingomyelinase (ASMase) were significantly elevated. However, there was a decreased expression of S1PR2 and S1P lyase (S1PL). Melatonin attenuated liver fibrosis, as shown by a significant inhibition of the expression of α-smooth muscle actin (α-SMA), transforming growth factor (TGF)-β and collagen (Col) Ι. Furthermore, melatonin inhibited S1P production, lowered expression of SphK1, S1PR1, SP1R3, and ASMase, and increased expression of S1PL. Melatonin induced a reversal of activated human HSCs cell line LX2, as evidenced by a reduction in α-SMA, TGF-β, and Col I expression. Melatonin-treated cells also exhibited an inhibition of the SphK1/S1P axis. Antifibrogenic effect of SphK1 inhibition was confirmed by treatment of LX2 cells with PF543. Abrogation of the lipid signaling pathway by the indole reveals novel molecular pathways that may account for the protective effect of melatonin in liver fibrogenesis. © 2016 BioFactors, 2016.

  17. Comprehensive Assessment of the Regulons Controlled by the FixLJ-FixK2-FixK1 Cascade in Bradyrhizobium japonicum▿ †

    PubMed Central

    Mesa, Socorro; Hauser, Felix; Friberg, Markus; Malaguti, Emmanuelle; Fischer, Hans-Martin; Hennecke, Hauke

    2008-01-01

    Symbiotic N2 fixation in Bradyrhizobium japonicum is controlled by a complex transcription factor network. Part of it is a hierarchically arranged cascade in which the two-component regulatory system FixLJ, in response to a moderate decrease in oxygen concentration, activates the fixK2 gene. The FixK2 protein then activates not only a number of genes essential for microoxic respiration in symbiosis (fixNOQP and fixGHIS) but also further regulatory genes (rpoN1, nnrR, and fixK1). The results of transcriptome analyses described here have led to a comprehensive and expanded definition of the FixJ, FixK2, and FixK1 regulons, which, respectively, consist of 26, 204, and 29 genes specifically regulated in microoxically grown cells. Most of these genes are subject to positive control. Particular attention was addressed to the FixK2-dependent genes, which included a bioinformatics search for putative FixK2 binding sites on DNA (FixK2 boxes). Using an in vitro transcription assay with RNA polymerase holoenzyme and purified FixK2 as the activator, we validated as direct targets eight new genes. Interestingly, the adjacent but divergently oriented fixK1 and cycS genes shared the same FixK2 box for the activation of transcription in both directions. This recognition site may also be a direct target for the FixK1 protein, because activation of the cycS promoter required an intact fixK1 gene and either microoxic or anoxic, denitrifying conditions. We present evidence that cycS codes for a c-type cytochrome which is important, but not essential, for nitrate respiration. Two other, unexpected results emerged from this study: (i) specifically FixK1 seemed to exert a negative control on genes that are normally activated by the N2 fixation-specific transcription factor NifA, and (ii) a larger number of genes are expressed in a FixK2-dependent manner in endosymbiotic bacteroids than in culture-grown cells, pointing to a possible symbiosis-specific control. PMID:18689489

  18. C-Terminal residues in small potassium channel blockers OdK1 and OSK3 from scorpion venom fine-tune the selectivity.

    PubMed

    Kuzmenkov, Alexey I; Peigneur, Steve; Chugunov, Anton O; Tabakmakher, Valentin M; Efremov, Roman G; Tytgat, Jan; Grishin, Eugene V; Vassilevski, Alexander A

    2017-02-04

    We report isolation, sequencing, and electrophysiological characterization of OSK3 (α-KTx 8.8 in Kalium and Uniprot databases), a potassium channel blocker from the scorpion Orthochirus scrobiculosus venom. Using the voltage clamp technique, OSK3 was tested on a wide panel of 11 voltage-gated potassium channels expressed in Xenopus oocytes, and was found to potently inhibit Kv1.2 and Kv1.3 with IC50 values of ~331nM and ~503nM, respectively. OdK1 produced by the scorpion Odontobuthus doriae differs by just two C-terminal residues from OSK3, but shows marked preference to Kv1.2. Based on the charybdotoxin-potassium channel complex crystal structure, a model was built to explain the role of the variable residues in OdK1 and OSK3 selectivity.

  19. Combining Electron Crystallography and X-ray Crystallography to Study the MlotiK1 Cyclic Nucleotide-Regulated Potassium Channel

    SciTech Connect

    Clayton, G.; Aller, S; Wang, J; Unger, V; Morais-Cabral, J

    2009-01-01

    We have recently reported the X-ray structure of the cyclic nucleotide-regulated potassium channel, MlotiK1. Here we describe the application of both electron and X-ray crystallography to obtain high quality crystals. We suggest that the combined application of these techniques provides a useful strategy for membrane protein structure determination. We also present negative stain projection and cryo-data projection maps. These maps provide new insights about the properties of the MlotiK1 channel. In particular, a comparison of a 9 {angstrom} cryo-data projection with calculated model maps strongly suggests that there is a very weak interaction between the pore and the S1-S4 domains of this 6 TM tetrameric cation channel and that the S1-S4 domains can adopt multiple orientations relative to the pore.

  20. Inhibition of p70S6K1 Activation by Pdcd4 Overcomes the Resistance to an IGF-1R/IR Inhibitor in Colon Carcinoma Cells.

    PubMed

    Zhang, Yan; Wang, Qing; Chen, Li; Yang, Hsin-Sheng

    2015-03-01

    Agents targeting insulin-like growth factor 1 receptor (IGF-1R) are being actively examined in clinical trials. Although there has been some initial success of single-agent targeting IGF-1R, attempts in later studies failed because of resistance. This study aimed to understand the effects of programmed cell death 4 (Pdcd4) on the chemosensitivity of the IGF-1R inhibitor OSI-906 in colorectal cancer cells and the mechanism underlying this impact. Using OSI-906-resistant and -sensitive colorectal cancer cells, we found that the Pdcd4 level directly correlates with cell chemosensitivity to OSI-906. In addition, tumors derived from Pdcd4 knockdown cells resist the growth inhibitory effect of OSI-906 in a colorectal cancer xenograft mouse model. Moreover, Pdcd4 enhances the antiproliferative effect of OSI-906 in resistant cells through suppression of p70S6K1 activation. Knockdown of p70S6K1, but not p70S6K2, significantly increases the chemosensitivity of OSI-906 in cultured colorectal cancer cells. Furthermore, the combination of OSI-906 and PF-4708671, a p70S6K1 inhibitor, efficiently suppresses the growth of OSI-906-resistant colon tumor cells in vitro and in vivo. Taken together, activation of p70S6K1 that is inhibited by Pdcd4 is essential for resistance to the IGF-1R inhibitor in colon tumor cells, and the combinational treatment of OSI-906 and PF-4708671 results in enhanced antiproliferation effects in colorectal cancer cells in vitro and in vivo, providing a novel venue to overcome the resistance to the IGF-1R inhibitor in treating colorectal cancer.

  1. Insulin activation of vacuolar protein sorting 34 mediates localized phosphatidylinositol 3-phosphate production at lamellipodia and activation of mTOR/S6K1.

    PubMed

    Hirsch, Dianne S; Shen, Yi; Dokmanovic, Milos; Yu, Joyce; Mohan, Nishant; Elzarrad, Mohammed Khair; Wu, Wen Jin

    2014-06-01

    The class III phosphatidylinositol 3-kinase, VPS34, phosphorylates the D3 hydroxyl of inositol generating phosphatidylinositol 3-phosphate (ptdins(3)p). Initial studies suggested that ptdins(3)p solely functioned as a component of vesicular and endosomal membranes and that VPS34 did not function in signal transduction. However, VPS34 has recently been shown to be required for insulin-mediated activation of S6 kinase 1 (S6K1). Whether VPS34 activity is directly regulated by insulin is unclear. It is also not known whether VPS34 activity can be spatially restricted in response to extracellular stimuli. Data presented here demonstrate that in response to insulin, VPS34 is activated and translocated to lamellipodia where it produces ptdins(3)p. The localized production of ptdins(3)p is dependent on Src phosphorylation of VPS34. In cells expressing VPS34 with mutations at Y231 or Y310, which are Src-phosphorylation sites, insulin-stimulated VPS34 translocation to the plasma membrane and lamellipodia formation are blocked. mTOR also colocalizes with VPS34 and ptdins(3)p at lamellipodia following insulin-stimulation. In cells expressing the VPS34-Y231F mutant, which blocks lamellipodia formation, mTOR localization at the plasma membrane and insulin-mediated S6K1 activation are reduced. This suggests that mTOR localization at lamellipodia is important for full activation of S6K1 induced by insulin. These data demonstrate that insulin can spatially regulate VPS34 activity through Src-mediated tyrosine phosphorylation and that this membrane localized activity contributes to lamellipodia formation and activation of mTOR/S6K1signaling.

  2. Computational investigation of sphingosine kinase 1 (SphK1) and calcium dependent ERK1/2 activation downstream of VEGFR2 in endothelial cells

    PubMed Central

    Bazzazi, Hojjat; Popel, Aleksander S.

    2017-01-01

    Vascular endothelial growth factor (VEGF) is a powerful regulator of neovascularization. VEGF binding to its cognate receptor, VEGFR2, activates a number of signaling pathways including ERK1/2. Activation of ERK1/2 is experimentally shown to involve sphingosine kinase 1 (SphK1) activation and its calcium-dependent translocation downstream of ERK1/2. Here we construct a rule-based computational model of signaling downstream of VEGFR2, by including SphK1 and calcium positive feedback mechanisms, and investigate their consequences on ERK1/2 activation. The model predicts the existence of VEGF threshold in ERK1/2 activation that can be continuously tuned by cellular concentrations of SphK1 and sphingosine 1 phosphate (S1P). The computer model also predicts powerful effects of perturbations in plasma and ER calcium pump rates and the current through the CRAC channels on ERK1/2 activation dynamics, highlighting the critical role of intracellular calcium in shaping the pERK1/2 signal. The model is then utilized to simulate anti-angiogenic therapeutic interventions targeting VEGFR2-ERK1/2 axis. Simulations indicate that monotherapies that exclusively target VEGFR2 phosphorylation, VEGF, or VEGFR2 are ineffective in shutting down signaling to ERK1/2. By simulating therapeutic strategies that target multiple nodes of the pathway such as Raf and SphK1, we conclude that combination therapy should be much more effective in blocking VEGF signaling to EKR1/2. The model has important implications for interventions that target signaling pathways in angiogenesis relevant to cancer, vascular diseases, and wound healing. PMID:28178265

  3. Regulation of Toll-like receptor 2 interaction with Ecgp96 controls Escherichia coli K1 invasion of brain endothelial cells

    PubMed Central

    Krishnan, Subramanian; Chen, Shuang; Turcatel, Gianluca; Arditi, Moshe; Prasadarao, Nemani V.

    2012-01-01

    SUMMARY The interaction of outer membrane protein A (OmpA) with its receptor, Ecgp96 (a homologue of Hsp90β) is critical for the pathogenesis of E. coli K1 meningitis. Since Hsp90 chaperones Toll-like receptors (TLRs), we examined the role of TLRs in E. coli K1 infection. Herein, we show that newborn TLR2−/− mice are resistant to E. coli K1 meningitis, while TLR4−/− mice succumb to infection sooner. In vitro, OmpA+ E. coli infection selectively upregulates Ecgp96 and TLR2 in human brain microvascular endothelial cells (HBMEC), whereas OmpA− E. coli upregulates TLR4 in these cells. Furthermore, infection with OmpA+ E. coli causes Ecgp96 and TLR2 translocate to the plasma membrane of HBMEC as a complex. Immunoprecipitation studies of the plasma membrane fractions from infected HBMEC reveal that the C-termini of Ecgp96 and TLR2 are critical for OmpA+ E. coli invasion. Knockdown of TLR2 using siRNA results in inefficient membrane translocation of Ecgp96 and significantly reduces invasion. In addition, the interaction of Ecgp96 and TLR2 induces a bipartite signal, one from Ecgp96 through PKC-α while the other from TLR2 through MyD88, ERK1/2 and NF-κB. This bipartite signal ultimately culminates in the efficient production of NO, which in turn promotes E. coli K1 invasion of HBMEC. PMID:22963587

  4. S6K1 and E2FB are in mutually antagonistic regulatory links controlling cell growth and proliferation in Arabidopsis.

    PubMed

    Henriques, Rossana; Magyar, Zoltán; Bögre, László

    2013-06-01

    Plant development is dependent on the coordination between growth and cell proliferation. The nutrient sensing TOR kinase and its downstream target, the 40S ribosomal S6 Kinase, are central controllers of cell growth that were also shown to determine cell size by inhibiting the onset of mitosis in yeast and animal cells. We have shown that the Arabidopsis S6 Kinase1 inhibits cell proliferation through the RBR-E2FB complex. S6K1 interacts with RBR via its N-terminal RBR binding motif, promotes its nuclear localization and consequent RBR-dependent repression of cell cycle genes through E2FB. Here we show that S6K1 and E2FB are in a mutually antagonistic relationship both in their protein abundance and in their activity. We propose that this double inhibitory regulatory connection between S6K1 and E2FB forms a regulatory switch that might be important to determine whether cells divide or grow.

  5. SKAR links pre-mRNA splicing to mTOR/S6K1-mediated enhanced translation efficiency of spliced mRNAs.

    PubMed

    Ma, Xiaoju Max; Yoon, Sang-Oh; Richardson, Celeste J; Jülich, Kristina; Blenis, John

    2008-04-18

    Different protein complexes form on newly spliced mRNA to ensure the accuracy and efficiency of eukaryotic gene expression. For example, the exon junction complex (EJC) plays an important role in mRNA surveillance. The EJC also influences the first, or pioneer round of protein synthesis through a mechanism that is poorly understood. We show that the nutrient-, stress-, and energy-sensing checkpoint kinase, mTOR, contributes to the observed enhanced translation efficiency of spliced over nonspliced mRNAs. We demonstrate that, when activated, S6K1 is recruited to the newly synthesized mRNA by SKAR, which is deposited at the EJC during splicing, and that SKAR and S6K1 increase the translation efficiency of spliced mRNA. Thus, SKAR-mediated recruitment of activated S6K1 to newly processed mRNPs serves as a conduit between mTOR checkpoint signaling and the pioneer round of translation when cells exist in conditions supportive of protein synthesis.

  6. Adenoviral delivery of truncated MMP-8 fused with the hepatocyte growth factor mutant 1K1 ameliorates liver cirrhosis and promotes hepatocyte proliferation

    PubMed Central

    Liu, Jinghua; Li, Jianbo; Fu, Weiwei; Tang, Jiacheng; Feng, Xu; Chen, Jiang; Liang, Yuelong; Jin, Ren’an; Xie, Anyong; Cai, Xiujun

    2015-01-01

    Liver cirrhosis is a chronic liver disease caused by chronic liver injury, which activates hepatic stellate cells (HSCs) and the secretion of extracellular matrix (ECM). Cirrhosis accounts for an extensive level of morbidity and mortality worldwide, largely due to lack of effective treatment options. In this study, we have constructed a fusion protein containing matrix metal-loproteinase 8 (MMP-8) and the human growth factor mutant 1K1 (designated cMMP8-1K1) and delivered it into hepatocytes and in vivo and in cell culture via intravenous injection of fusion protein-harboring adenovirus. In doing so, we found that the cMMP8-1K1 fusion protein promotes the proliferation of hepatocytes, likely resulting from the combined inhibition of type I collagen secretion and the degradation of the ECM in the HSCs. This fusion protein was also observed to ameliorate liver cirrhosis in our mouse model. These changes appear to be linked to changes in downstream gene expression. Taken together, these results suggest a possible strategy for the treatment of liver cirrhosis and additional work is warranted. PMID:26527860

  7. Application of extracellular lipopeptide biosurfactant produced by endophytic Bacillus subtilis K1 isolated from aerial roots of banyan (Ficus benghalensis) in microbially enhanced oil recovery (MEOR).

    PubMed

    Pathak, Khyati V; Keharia, Hareshkumar

    2014-02-01

    Bacillus subtilis K1 isolated from aerial roots of banyan tree secreted mixture of surfactins, iturins and fengycins with high degree of heterogeneity. The extracellular extract consisting of mixture of these cyclic lipopeptides exhibited very good emulsification activity as well as excellent emulsion stability. The culture accumulated maximum surfactant up to 48 h of growth during batch fermentation in Luria broth. The emulsion of hexane, heptane and octane prepared using 48-h-old culture supernatant of B. subtilis K1 remained stable up to 2 days while emulsion of four stroke engine oil remained stable for more than a year. The critical micelle concentration of crude lipopeptide biosurfactant extracted by acid precipitation from 48-h-old fermentation broth of B. subtilis K1 was found to be 20.5 μg/mL. The biosurfactant activity was found to be stable at 100 °C for 2 h, over a pH range of 6-12 h and over an NaCl concentration up to 10 % (w/v). The application of biosurfactant on laboratory scale sand pack column saturated with four stroke engine oil resulted in ~43 % enhanced oil recovery, suggesting its suitability in microbially enhanced oil recovery.

  8. SphK1 mediates hepatic inflammation in a mouse model of NASH induced by high saturated fat feeding and initiates proinflammatory signaling in hepatocytes.

    PubMed

    Geng, Tuoyu; Sutter, Alton; Harland, Michael D; Law, Brittany A; Ross, Jessica S; Lewin, David; Palanisamy, Arun; Russo, Sarah B; Chavin, Kenneth D; Cowart, L Ashley

    2015-12-01

    Steatohepatitis occurs in up to 20% of patients with fatty liver disease and leads to its primary disease outcomes, including fibrosis, cirrhosis, and increased risk of hepatocellular carcinoma. Mechanisms that mediate this inflammation are of major interest. We previously showed that overload of saturated fatty acids, such as that which occurs with metabolic syndrome, induced sphingosine kinase 1 (SphK1), an enzyme that generates sphingosine-1-phosphate (S1P). While data suggest beneficial roles for S1P in some contexts, we hypothesized that it may promote hepatic inflammation in the context of obesity. Consistent with this, we observed 2-fold elevation of this enzyme in livers from humans with nonalcoholic fatty liver disease and also in mice with high saturated fat feeding, which recapitulated the human disease. Mice exhibited activation of NFκB, elevated cytokine production, and immune cell infiltration. Importantly, SphK1-null mice were protected from these outcomes. Studies in cultured cells demonstrated saturated fatty acid induction of SphK1 message, protein, and activity, and also a requirement of the enzyme for NFκB signaling and increased mRNA encoding TNFα and MCP1. Moreover, saturated fat-induced NFκB signaling and elevation of TNFα and MCP1 mRNA in HepG2 cells was blocked by targeted knockdown of S1P receptor 1, supporting a role for this lipid signaling pathway in inflammation in nonalcoholic fatty liver disease.

  9. Adenoviral delivery of truncated MMP-8 fused with the hepatocyte growth factor mutant 1K1 ameliorates liver cirrhosis and promotes hepatocyte proliferation.

    PubMed

    Liu, Jinghua; Li, Jianbo; Fu, Weiwei; Tang, Jiacheng; Feng, Xu; Chen, Jiang; Liang, Yuelong; Jin, Ren'an; Xie, Anyong; Cai, Xiujun

    2015-01-01

    Liver cirrhosis is a chronic liver disease caused by chronic liver injury, which activates hepatic stellate cells (HSCs) and the secretion of extracellular matrix (ECM). Cirrhosis accounts for an extensive level of morbidity and mortality worldwide, largely due to lack of effective treatment options. In this study, we have constructed a fusion protein containing matrix metal-loproteinase 8 (MMP-8) and the human growth factor mutant 1K1 (designated cMMP8-1K1) and delivered it into hepatocytes and in vivo and in cell culture via intravenous injection of fusion protein-harboring adenovirus. In doing so, we found that the cMMP8-1K1 fusion protein promotes the proliferation of hepatocytes, likely resulting from the combined inhibition of type I collagen secretion and the degradation of the ECM in the HSCs. This fusion protein was also observed to ameliorate liver cirrhosis in our mouse model. These changes appear to be linked to changes in downstream gene expression. Taken together, these results suggest a possible strategy for the treatment of liver cirrhosis and additional work is warranted.

  10. Role of hepatocyte S6K1 in palmitic acid-induced endoplasmic reticulum stress, lipotoxicity, insulin resistance and in oleic acid-induced protection.

    PubMed

    Pardo, Virginia; González-Rodríguez, Águeda; Muntané, Jordi; Kozma, Sara C; Valverde, Ángela M

    2015-06-01

    The excess of saturated free fatty acids, such as palmitic acid, that induces lipotoxicity in hepatocytes, has been implicated in the development of non-alcoholic fatty liver disease also associated with insulin resistance. By contrast, oleic acid, a monounsaturated fatty acid, attenuates the effects of palmitic acid. We evaluated whether palmitic acid is directly associated with both insulin resistance and lipoapoptosis in mouse and human hepatocytes and the impact of oleic acid in the molecular mechanisms that mediate both processes. In human and mouse hepatocytes palmitic acid at a lipotoxic concentration triggered early activation of endoplasmic reticulum (ER) stress-related kinases, induced the apoptotic transcription factor CHOP, activated caspase 3 and increased the percentage of apoptotic cells. These effects concurred with decreased IR/IRS1/Akt insulin pathway. Oleic acid suppressed the toxic effects of palmitic acid on ER stress activation, lipoapoptosis and insulin resistance. Besides, oleic acid suppressed palmitic acid-induced activation of S6K1. This protection was mimicked by pharmacological or genetic inhibition of S6K1 in hepatocytes. In conclusion, this is the first study highlighting the activation of S6K1 by palmitic acid as a common and novel mechanism by which its inhibition by oleic acid prevents ER stress, lipoapoptosis and insulin resistance in hepatocytes.

  11. MiR-145 is downregulated in human ovarian cancer and modulates cell growth and invasion by targeting p70S6K1 and MUC1

    SciTech Connect

    Wu, Huijuan; Xiao, ZhengHua; Wang, Ke; Liu, Wenxin; Hao, Quan

    2013-11-29

    Highlights: •MiR-145 is downregulated in human ovarian cancer. •MiR-145 targets p70S6K1 and MUC1. •p70S6K1 and MUC1 are involved in miR-145 mediated tumor cell growth and cell invasion, respectively. -- Abstract: MicroRNAs (miRNAs) are a family of small non-coding RNA molecules that regulate gene expression at post-transcriptional levels. Previous studies have shown that miR-145 is downregulated in human ovarian cancer; however, the roles of miR-145 in ovarian cancer growth and invasion have not been fully demonstrated. In the present study, Northern blot and qRT-PCR analysis indicate that miR-145 is downregulated in ovarian cancer tissues and cell lines, as well as in serum samples of ovarian cancer, compared to healthy ovarian tissues, cell lines and serum samples. Functional studies suggest that miR-145 overexpression leads to the inhibition of colony formation, cell proliferation, cell growth viability and invasion, and the induction of cell apoptosis. In accordance with the effect of miR-145 on cell growth, miR-145 suppresses tumor growth in vivo. MiR-145 is found to negatively regulate P70S6K1 and MUC1 protein levels by directly targeting their 3′UTRs. Importantly, the overexpression of p70S6K1 and MUC1 can restore the cell colony formation and invasion abilities that are reduced by miR-145, respectively. MiR-145 expression is increased after 5-aza-CdR treatment, and 5-aza-CdR treatment results in the same phenotype as the effect of miR-145 overexpression. Our study suggests that miR-145 modulates ovarian cancer growth and invasion by suppressing p70S6K1 and MUC1, functioning as a tumor suppressor. Moreover, our data imply that miR-145 has potential as a miRNA-based therapeutic target for ovarian cancer.

  12. Analysis of the SDS-PAGE patterns of outer membrane proteins from Escherichia coli strains that have lost the ability to form K1 antigen and varied in the susceptibility to normal human serum.

    PubMed

    Cisowska, Agnieszka; Bugla-Płoskońska, Gabriela

    2014-01-01

    We used SDS-polyacrylamide gel electrophoresis to investigate the outer membrane proteins (OMPs) band composition of 19 Escherichia coli K1 strains that have spontaneously lost the ability to form K1 polysaccharide capsule (E. coli K1-) and demonstrated different degrees of susceptibility to the bactericidal action of normal human serum. Presented results showed that there were differences between E. coli K1- strains in OMPs expressing capacity. The analysis performed on OMPs has not revealed a direct association between the different OMPs band composition and the susceptibility of these strains to the serum.

  13. Macrophage receptor with collagenous structure (MARCO) is a dynamic adhesive molecule that enhances uptake of carbon nanotubes by CHO-K1 Cells

    SciTech Connect

    Hirano, Seishiro; Fujitani, Yuji; Furuyama, Akiko; Kanno, Sanae

    2012-02-15

    The toxicity of carbon nanotubes (CNTs), a highly promising nanomaterial, is similar to that of asbestos because both types of particles have a fibrous shape and are biopersistent. Here, we investigated the characteristics of macrophage receptor with collagenous structure (MARCO), a membrane receptor expressed on macrophages that recognizes environmental or unopsonized particles, and we assessed whether and how MARCO was involved in cellular uptake of multi-walled CNTs (MWCNTs). MARCO-transfected Chinese hamster ovary (CHO-K1) cells took up polystyrene beads irrespective of the particle size (20 nm–1 μm). In the culture of MARCO-transfected CHO-K1 cells dendritic structures were observed on the bottom of culture dishes, and the edges of these dendritic structures were continually renewed as the cell body migrated along the dendritic structures. MWCNTs were first tethered to the dendritic structures and then taken up by the cell body. MWCNTs appeared to be taken up via membrane ruffling like macropinocytosis, rather than phagocytosis. The cytotoxic EC{sub 50} value of MWCNTs in MARCO-transfected CHO-K1 cells was calculated to be 6.1 μg/mL and transmission electron microscopic observation indicated that the toxicity of MWCNTs may be due to the incomplete inclusion of MWCNTs by the membrane structure. -- Highlights: ►Carbon nanotubes (CNTs) were tethered to MARCO in vitro. ►CNTs were taken up rapidly into the cell body via MARCO by membrane ruffling. ►The incomplete inclusion of CNTs by membranes caused cytotoxicity.

  14. Hcp family proteins secreted via the type VI secretion system coordinately regulate Escherichia coli K1 interaction with human brain microvascular endothelial cells.

    PubMed

    Zhou, Yan; Tao, Jing; Yu, Hao; Ni, Jinjing; Zeng, Lingbing; Teng, Qihui; Kim, Kwang Sik; Zhao, Guo-Ping; Guo, Xiaokui; Yao, Yufeng

    2012-03-01

    Type VI secretion systems (T6SSs) are involved in the pathogenicity of several gram-negative bacteria. Based on sequence analysis, we found that a cluster of Escherichia coli virulence factors (EVF) encoding a putative T6SS exists in the genome of the meningitis-causing E. coli K1 strain RS218. The T6SS-associated deletion mutants exhibited significant defects in binding to and invasion of human brain microvascular endothelial cells (HBMEC) compared with the parent strain. Hcp family proteins (the hallmark of T6SS), including Hcp1 and Hcp2, were localized in the bacterial outer membrane, but the involvements of Hcp1 and Hcp2 have been shown to differ in E. coli-HBMEC interaction. The deletion mutant of hcp2 showed defects in the bacterial binding to and invasion of HBMEC, while Hcp1 was secreted in a T6SS-dependent manner and induced actin cytoskeleton rearrangement, apoptosis, and the release of interleukin-6 (IL-6) and IL-8 in HBMEC. These findings demonstrate that the T6SS is functional in E. coli K1, and two Hcp family proteins participate in different steps of E. coli interaction with HBMEC in a coordinate manner, e.g., binding to and invasion of HBMEC, the cytokine and chemokine release followed by cytoskeleton rearrangement, and apoptosis in HBMEC. This is the first demonstration of the role of T6SS in meningitis-causing E. coli K1, and T6SS-associated Hcp family proteins are likely to contribute to the pathogenesis of E. coli meningitis.

  15. Fine-scale mapping of the 5q11.2 breast cancer locus reveals at least three independent risk variants regulating MAP3K1.

    PubMed

    Glubb, Dylan M; Maranian, Mel J; Michailidou, Kyriaki; Pooley, Karen A; Meyer, Kerstin B; Kar, Siddhartha; Carlebur, Saskia; O'Reilly, Martin; Betts, Joshua A; Hillman, Kristine M; Kaufmann, Susanne; Beesley, Jonathan; Canisius, Sander; Hopper, John L; Southey, Melissa C; Tsimiklis, Helen; Apicella, Carmel; Schmidt, Marjanka K; Broeks, Annegien; Hogervorst, Frans B; van der Schoot, C Ellen; Muir, Kenneth; Lophatananon, Artitaya; Stewart-Brown, Sarah; Siriwanarangsan, Pornthep; Fasching, Peter A; Ruebner, Matthias; Ekici, Arif B; Beckmann, Matthias W; Peto, Julian; dos-Santos-Silva, Isabel; Fletcher, Olivia; Johnson, Nichola; Pharoah, Paul D P; Bolla, Manjeet K; Wang, Qin; Dennis, Joe; Sawyer, Elinor J; Tomlinson, Ian; Kerin, Michael J; Miller, Nicola; Burwinkel, Barbara; Marme, Frederik; Yang, Rongxi; Surowy, Harald; Guénel, Pascal; Truong, Thérèse; Menegaux, Florence; Sanchez, Marie; Bojesen, Stig E; Nordestgaard, Børge G; Nielsen, Sune F; Flyger, Henrik; González-Neira, Anna; Benitez, Javier; Zamora, M Pilar; Arias Perez, Jose Ignacio; Anton-Culver, Hoda; Neuhausen, Susan L; Brenner, Hermann; Dieffenbach, Aida Karina; Arndt, Volker; Stegmaier, Christa; Meindl, Alfons; Schmutzler, Rita K; Brauch, Hiltrud; Ko, Yon-Dschun; Brüning, Thomas; Nevanlinna, Heli; Muranen, Taru A; Aittomäki, Kristiina; Blomqvist, Carl; Matsuo, Keitaro; Ito, Hidemi; Iwata, Hiroji; Tanaka, Hideo; Dörk, Thilo; Bogdanova, Natalia V; Helbig, Sonja; Lindblom, Annika; Margolin, Sara; Mannermaa, Arto; Kataja, Vesa; Kosma, Veli-Matti; Hartikainen, Jaana M; Wu, Anna H; Tseng, Chiu-chen; Van Den Berg, David; Stram, Daniel O; Lambrechts, Diether; Zhao, Hui; Weltens, Caroline; van Limbergen, Erik; Chang-Claude, Jenny; Flesch-Janys, Dieter; Rudolph, Anja; Seibold, Petra; Radice, Paolo; Peterlongo, Paolo; Barile, Monica; Capra, Fabio; Couch, Fergus J; Olson, Janet E; Hallberg, Emily; Vachon, Celine; Giles, Graham G; Milne, Roger L; McLean, Catriona; Haiman, Christopher A; Henderson, Brian E; Schumacher, Fredrick; Le Marchand, Loic; Simard, Jacques; Goldberg, Mark S; Labrèche, France; Dumont, Martine; Teo, Soo Hwang; Yip, Cheng Har; See, Mee-Hoong; Cornes, Belinda; Cheng, Ching-Yu; Ikram, M Kamran; Kristensen, Vessela; Zheng, Wei; Halverson, Sandra L; Shrubsole, Martha; Long, Jirong; Winqvist, Robert; Pylkäs, Katri; Jukkola-Vuorinen, Arja; Kauppila, Saila; Andrulis, Irene L; Knight, Julia A; Glendon, Gord; Tchatchou, Sandrine; Devilee, Peter; Tollenaar, Robert A E M; Seynaeve, Caroline; Van Asperen, Christi J; García-Closas, Montserrat; Figueroa, Jonine; Chanock, Stephen J; Lissowska, Jolanta; Czene, Kamila; Klevebring, Daniel; Darabi, Hatef; Eriksson, Mikael; Hooning, Maartje J; Hollestelle, Antoinette; Martens, John W M; Collée, J Margriet; Hall, Per; Li, Jingmei; Humphreys, Keith; Shu, Xiao-Ou; Lu, Wei; Gao, Yu-Tang; Cai, Hui; Cox, Angela; Cross, Simon S; Reed, Malcolm W R; Blot, William; Signorello, Lisa B; Cai, Qiuyin; Shah, Mitul; Ghoussaini, Maya; Kang, Daehee; Choi, Ji-Yeob; Park, Sue K; Noh, Dong-Young; Hartman, Mikael; Miao, Hui; Lim, Wei Yen; Tang, Anthony; Hamann, Ute; Torres, Diana; Jakubowska, Anna; Lubinski, Jan; Jaworska, Katarzyna; Durda, Katarzyna; Sangrajrang, Suleeporn; Gaborieau, Valerie; Brennan, Paul; McKay, James; Olswold, Curtis; Slager, Susan; Toland, Amanda E; Yannoukakos, Drakoulis; Shen, Chen-Yang; Wu, Pei-Ei; Yu, Jyh-Cherng; Hou, Ming-Feng; Swerdlow, Anthony; Ashworth, Alan; Orr, Nick; Jones, Michael; Pita, Guillermo; Alonso, M Rosario; Álvarez, Nuria; Herrero, Daniel; Tessier, Daniel C; Vincent, Daniel; Bacot, Francois; Luccarini, Craig; Baynes, Caroline; Ahmed, Shahana; Healey, Catherine S; Brown, Melissa A; Ponder, Bruce A J; Chenevix-Trench, Georgia; Thompson, Deborah J; Edwards, Stacey L; Easton, Douglas F; Dunning, Alison M; French, Juliet D

    2015-01-08

    Genome-wide association studies (GWASs) have revealed SNP rs889312 on 5q11.2 to be associated with breast cancer risk in women of European ancestry. In an attempt to identify the biologically relevant variants, we analyzed 909 genetic variants across 5q11.2 in 103,991 breast cancer individuals and control individuals from 52 studies in the Breast Cancer Association Consortium. Multiple logistic regression analyses identified three independent risk signals: the strongest associations were with 15 correlated variants (iCHAV1), where the minor allele of the best candidate, rs62355902, associated with significantly increased risks of both estrogen-receptor-positive (ER(+): odds ratio [OR] = 1.24, 95% confidence interval [CI] = 1.21-1.27, ptrend = 5.7 × 10(-44)) and estrogen-receptor-negative (ER(-): OR = 1.10, 95% CI = 1.05-1.15, ptrend = 3.0 × 10(-4)) tumors. After adjustment for rs62355902, we found evidence of association of a further 173 variants (iCHAV2) containing three subsets with a range of effects (the strongest was rs113317823 [pcond = 1.61 × 10(-5)]) and five variants composing iCHAV3 (lead rs11949391; ER(+): OR = 0.90, 95% CI = 0.87-0.93, pcond = 1.4 × 10(-4)). Twenty-six percent of the prioritized candidate variants coincided with four putative regulatory elements that interact with the MAP3K1 promoter through chromatin looping and affect MAP3K1 promoter activity. Functional analysis indicated that the cancer risk alleles of four candidates (rs74345699 and rs62355900 [iCHAV1], rs16886397 [iCHAV2a], and rs17432750 [iCHAV3]) increased MAP3K1 transcriptional activity. Chromatin immunoprecipitation analysis revealed diminished GATA3 binding to the minor (cancer-protective) allele of rs17432750, indicating a mechanism for its action. We propose that the cancer risk alleles act to increase MAP3K1 expression in vivo and might promote breast cancer cell survival.

  16. Fine-Scale Mapping of the 5q11.2 Breast Cancer Locus Reveals at Least Three Independent Risk Variants Regulating MAP3K1

    PubMed Central

    Glubb, Dylan M.; Maranian, Mel J.; Michailidou, Kyriaki; Pooley, Karen A.; Meyer, Kerstin B.; Kar, Siddhartha; Carlebur, Saskia; O’Reilly, Martin; Betts, Joshua A.; Hillman, Kristine M.; Kaufmann, Susanne; Beesley, Jonathan; Canisius, Sander; Hopper, John L.; Southey, Melissa C.; Tsimiklis, Helen; Apicella, Carmel; Schmidt, Marjanka K.; Broeks, Annegien; Hogervorst, Frans B.; van der Schoot, C. Ellen; Muir, Kenneth; Lophatananon, Artitaya; Stewart-Brown, Sarah; Siriwanarangsan, Pornthep; Fasching, Peter A.; Ruebner, Matthias; Ekici, Arif B.; Beckmann, Matthias W.; Peto, Julian; dos-Santos-Silva, Isabel; Fletcher, Olivia; Johnson, Nichola; Pharoah, Paul D.P.; Bolla, Manjeet K.; Wang, Qin; Dennis, Joe; Sawyer, Elinor J.; Tomlinson, Ian; Kerin, Michael J.; Miller, Nicola; Burwinkel, Barbara; Marme, Frederik; Yang, Rongxi; Surowy, Harald; Guénel, Pascal; Truong, Thérèse; Menegaux, Florence; Sanchez, Marie; Bojesen, Stig E.; Nordestgaard, Børge G.; Nielsen, Sune F.; Flyger, Henrik; González-Neira, Anna; Benitez, Javier; Zamora, M. Pilar; Arias Perez, Jose Ignacio; Anton-Culver, Hoda; Neuhausen, Susan L.; Brenner, Hermann; Dieffenbach, Aida Karina; Arndt, Volker; Stegmaier, Christa; Meindl, Alfons; Schmutzler, Rita K.; Brauch, Hiltrud; Ko, Yon-Dschun; Brüning, Thomas; Nevanlinna, Heli; Muranen, Taru A.; Aittomäki, Kristiina; Blomqvist, Carl; Matsuo, Keitaro; Ito, Hidemi; Iwata, Hiroji; Tanaka, Hideo; Dörk, Thilo; Bogdanova, Natalia V.; Helbig, Sonja; Lindblom, Annika; Margolin, Sara; Mannermaa, Arto; Kataja, Vesa; Kosma, Veli-Matti; Hartikainen, Jaana M.; Wu, Anna H.; Tseng, Chiu-chen; Van Den Berg, David; Stram, Daniel O.; Lambrechts, Diether; Zhao, Hui; Weltens, Caroline; van Limbergen, Erik; Chang-Claude, Jenny; Flesch-Janys, Dieter; Rudolph, Anja; Seibold, Petra; Radice, Paolo; Peterlongo, Paolo; Barile, Monica; Capra, Fabio; Couch, Fergus J.; Olson, Janet E.; Hallberg, Emily; Vachon, Celine; Giles, Graham G.; Milne, Roger L.; McLean, Catriona; Haiman, Christopher A.; Henderson, Brian E.; Schumacher, Fredrick; Le Marchand, Loic; Simard, Jacques; Goldberg, Mark S.; Labrèche, France; Dumont, Martine; Teo, Soo Hwang; Yip, Cheng Har; See, Mee-Hoong; Cornes, Belinda; Cheng, Ching-Yu; Ikram, M. Kamran; Kristensen, Vessela; Zheng, Wei; Halverson, Sandra L.; Shrubsole, Martha; Long, Jirong; Winqvist, Robert; Pylkäs, Katri; Jukkola-Vuorinen, Arja; Kauppila, Saila; Andrulis, Irene L.; Knight, Julia A.; Glendon, Gord; Tchatchou, Sandrine; Devilee, Peter; Tollenaar, Robert A.E.M.; Seynaeve, Caroline; Van Asperen, Christi J.; García-Closas, Montserrat; Figueroa, Jonine; Chanock, Stephen J.; Lissowska, Jolanta; Czene, Kamila; Klevebring, Daniel; Darabi, Hatef; Eriksson, Mikael; Hooning, Maartje J.; Hollestelle, Antoinette; Martens, John W.M.; Collée, J. Margriet; Hall, Per; Li, Jingmei; Humphreys, Keith; Shu, Xiao-Ou; Lu, Wei; Gao, Yu-Tang; Cai, Hui; Cox, Angela; Cross, Simon S.; Reed, Malcolm W.R.; Blot, William; Signorello, Lisa B.; Cai, Qiuyin; Shah, Mitul; Ghoussaini, Maya; Kang, Daehee; Choi, Ji-Yeob; Park, Sue K.; Noh, Dong-Young; Hartman, Mikael; Miao, Hui; Lim, Wei Yen; Tang, Anthony; Hamann, Ute; Torres, Diana; Jakubowska, Anna; Lubinski, Jan; Jaworska, Katarzyna; Durda, Katarzyna; Sangrajrang, Suleeporn; Gaborieau, Valerie; Brennan, Paul; McKay, James; Olswold, Curtis; Slager, Susan; Toland, Amanda E.; Yannoukakos, Drakoulis; Shen, Chen-Yang; Wu, Pei-Ei; Yu, Jyh-Cherng; Hou, Ming-Feng; Swerdlow, Anthony; Ashworth, Alan; Orr, Nick; Jones, Michael; Pita, Guillermo; Alonso, M. Rosario; Álvarez, Nuria; Herrero, Daniel; Tessier, Daniel C.; Vincent, Daniel; Bacot, Francois; Luccarini, Craig; Baynes, Caroline; Ahmed, Shahana; Healey, Catherine S.; Brown, Melissa A.; Ponder, Bruce A.J.; Chenevix-Trench, Georgia; Thompson, Deborah J.; Edwards, Stacey L.; Easton, Douglas F.; Dunning, Alison M.; French, Juliet D.

    2015-01-01

    Genome-wide association studies (GWASs) have revealed SNP rs889312 on 5q11.2 to be associated with breast cancer risk in women of European ancestry. In an attempt to identify the biologically relevant variants, we analyzed 909 genetic variants across 5q11.2 in 103,991 breast cancer individuals and control individuals from 52 studies in the Breast Cancer Association Consortium. Multiple logistic regression analyses identified three independent risk signals: the strongest associations were with 15 correlated variants (iCHAV1), where the minor allele of the best candidate, rs62355902, associated with significantly increased risks of both estrogen-receptor-positive (ER+: odds ratio [OR] = 1.24, 95% confidence interval [CI] = 1.21–1.27, ptrend = 5.7 × 10−44) and estrogen-receptor-negative (ER−: OR = 1.10, 95% CI = 1.05–1.15, ptrend = 3.0 × 10−4) tumors. After adjustment for rs62355902, we found evidence of association of a further 173 variants (iCHAV2) containing three subsets with a range of effects (the strongest was rs113317823 [pcond = 1.61 × 10−5]) and five variants composing iCHAV3 (lead rs11949391; ER+: OR = 0.90, 95% CI = 0.87–0.93, pcond = 1.4 × 10−4). Twenty-six percent of the prioritized candidate variants coincided with four putative regulatory elements that interact with the MAP3K1 promoter through chromatin looping and affect MAP3K1 promoter activity. Functional analysis indicated that the cancer risk alleles of four candidates (rs74345699 and rs62355900 [iCHAV1], rs16886397 [iCHAV2a], and rs17432750 [iCHAV3]) increased MAP3K1 transcriptional activity. Chromatin immunoprecipitation analysis revealed diminished GATA3 binding to the minor (cancer-protective) allele of rs17432750, indicating a mechanism for its action. We propose that the cancer risk alleles act to increase MAP3K1 expression in vivo and might promote breast cancer cell survival. PMID:25529635

  17. Comparison of five different in vitro assays for assessment of sodium metavanadate cytotoxicity in Chinese hamster ovary cells (CHO-K1 line).

    PubMed

    Zwolak, Iwona

    2015-08-01

    This investigation was undertaken to compare five different in vitro cytotoxicity assays for their power in revealing vanadium-mediated toxicity in Chinese hamster ovary (CHO)-K1 cells. The cells were exposed to sodium metavanadate (NaVO(3)) in the range of 10-1000 µM for 24 h and thereafter the cytotoxic effects of NaVO(3) were measured by colorimetric in vitro assays: the neutral red (NR) test, the 2,3-bis[2-methoxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxyanilide inner salt (XTT) assay, the resazurin assay, the sulforhodamine B (SR-B) assay, and by microscopic assessment of cell viability using the trypan blue (TB) staining method. Among the assays used, the NR test was the most sensitive, since it revealed metavanadate cytotoxicity at the lowest NaVO(3) dose (=50 µM). Also, NaVO(3) cytotoxicity expressed as inhibitory concentration (IC) showed the lowest values for the NR test. Three other tests XTT, resazurin, and SR-B assays showed intermediate sensitivity revealing the cytotoxicity of NaVO(3) at 100 µM. The corresponding IC10 and IC50 values calculated for the XTT, resazurin, and SR-B tests were similar. The TB staining method was the least sensitive, since it recorded metavanadate cytotoxicity at the highest NaVO(3) concentration tested (=600 µM). Based on the cytotoxicity end points measured with the above assays, it can be concluded that lysosomal/Golgi apparatus damage (measured by NR assay) may be the primary effect of NaVO(3) on CHO-K1 cells. The disintegration of mitochondria (assessed with the XTT and resazurin assays) probably follows lysosomal impairment. Plasma membrane permeability (staining with TB) occurs at a late stage of NaVO(3)-induced cytotoxicity on CHO-K1 cells. The results obtained in this research work show that the NR test can be recommended as a very sensitive assay for the assessment of NaVO(3) cytotoxicity in the CHO-K1 cell culture model. Considering the convenience of assay performance along with adequate sensitivity

  18. Extending aggressive low-k1 design rule requirements for 90-nm and 65-nm nodes via simultaneous optimization of NA, illumination, and OPC

    NASA Astrophysics Data System (ADS)

    Roy, Sabita; Van Den Broeke, Douglas J.; Chen, J. F.; Liebchen, Armin; Chen, Ting; Hsu, Stephen D.; Shi, Xuelong; Socha, Robert J.

    2004-05-01

    Under low-k1 patterning constraints, it has been a challenge for the lithography process to meet the aggressive IC design rule requirements for the 90nm and the upcoming 65nm nodes. From the imaging perspective, we see the geometric design rules are largely governed by numerical aperture (NA), illumination settings, and OPC for any resolution enhancement technique (RET) applied mask. We report a case study of exploring a set of process feasible design rule criteria based on a state-of-the-art μProcessor chip that contains three different styles of circuit design - standard library cell (SLC), random logic (RML), and SRAM. To keep the packing density higher for SRAM, the critical criteria for design rules involve achievable minimum pitch, sufficient area of contact-landing pad, minimum line end shortening (LES) to ensure poly endcap, and preferably to have optimum pitch for the placement of Scattering Bars (SB). For RML, the goal is to achieve the printing of ever smaller critical dimension (CD) with a greater CD uniformity control. The SLC should be designed to be comparable with both RML and SRAM devices. Hence, the design rule constraints for CD, space, line end, minimum pitch, and SB placement for SLC cell is critically confined. Unlike the traditional method of assuming a linear scaling for the design rule set, we explore achievable design rule criteria for very low k1 imaging by simultaneously optimizing NA, illumination settings, and OPC (for the optimum placement of SB) for a calibrated process. This is done by analyzing the CD control and the maximum overlapped process window for critical lines, spaces, line ends, and with the respective k1 factor for the three types of circuits. For 90nm node with k1 as low as 0.36, a feasible set of design rules for the μProcessor chip can be obtained using 6% attPSM with 6% exposure latitude at 400nm of overlapped depth of focus. Using the similar approach for the scaled down 65nm 6% attPSM, it resulted inadequate

  19. Final report on the torque key komparison CCM.T-K1.2 measurand torque: 0 N.m, 500 N.m, 1000 N.m

    NASA Astrophysics Data System (ADS)

    Röske, Dirk

    2015-01-01

    The purpose of the CIPM subsequent bilateral comparison CCM.T-K1.2 was to link another participant, namely the National Institute of Metrology (Thailand), in short NIMT, to the CCM.T-K1 torque key comparison. The measuring capabilities up to 1000 N.m of dead-weight torque standard machines with supported lever were investigated. The pilot laboratory was the same in both comparisons—it was the Physikalisch-Technische Bundesanstalt (PTB, Braunschweig, Germany). The same two very stable torque transducers with well-known properties were used as travelling standards. The measurements at the participating laboratory were carried out between November 2007 and February 2008. According to the technical protocol, torque steps of 500 N.m and 1000 N.m had to be measured both in clockwise and anticlockwise directions. Corrections had to be applied to the results reported by the participants taking into account the use of different amplifiers, the creep (due to different loading times of the machines) and the environmental conditions in the laboratories (temperature and relative humidity of the ambient air). The results of the pilot laboratory in this bilateral comparison are in very good agreement with the same results obtained in the CCM.T-K1 comparison. For each of the transducers, the two torque steps and both senses of direction of the torque vector, the key comparison reference value of the CCM.T-K1 was taken, and the results of participant NIMT were calculated with respect to these values. The agreement between the results is very good. The smallest expanded (k = 2) relative uncertainty of the machine stated by the participant is 1 × 10-4. The results of the comparison support this uncertainty statement. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by CCM, according

  20. Analysis of possible genotoxicity of the herbicide flurochloridone and its commercial formulations: Endo III and Fpg alkaline comet assays in Chinese hamster ovary (CHO-K1) cells.

    PubMed

    Soloneski, Sonia; Nikoloff, Noelia; Larramendy, Marcelo L

    2016-02-01

    Cytotoxic and genotoxic effects of flurochloridone (FLC) and its formulations Twin Pack Gold(®) and Rainbow(®) were evaluated in CHO-K1 cells. Using the alkaline single-cell gel electrophoresis (SCGE) assay, we observed that FLC (15 μg/ml), Twin Pack Gold(®) or Rainbow(®) induced primary DNA damage, increasing the frequency of damaged nucleoids. Vitamin E pretreatment did not modify the effect. Decreased cell viability was observed only in Twin Pack Gold(®)-treated cultures and was significantly ameliorated by vitamin E. Post-treatment of herbicide-damaged CHO-K1 cells with the enzymes Endo III or Fpg did not increase FLC-, Twin Pack Gold(®)-, or Rainbow(®)-induced DNA damage. These results demonstrate that neither FLC nor FLC-based formulations induce DNA damage through hydroxyl radical or lipid alkoxyl radical production, and that the induced DNA lesions were not related to oxidative damage at the purine/pyrimidine level. Our observations strongly suggest that the cytotoxic effects observed after Twin Pack Gold(®) exposure are due to the excipients contained within the technical formulation rather than FLC itself.

  1. Phenotypic Heterogeneity in Expression of the K1 Polysaccharide Capsule of Uropathogenic Escherichia coli and Downregulation of the Capsule Genes during Growth in Urine

    PubMed Central

    King, Jane E.; Aal Owaif, Hasan A.; Jia, Jia

    2015-01-01

    Uropathogenic Escherichia coli (UPEC) is the major causative agent of uncomplicated urinary tract infections (UTI). The K1 capsule on the surface of UPEC strains is a key virulence factor, and its expression may be important in the onset and progression of UTI. In order to understand capsule expression in more detail, we analyzed its expression in the UPEC strain UTI89 during growth in rich medium (LB medium) and urine and during infection of a bladder epithelial cell line. Comparison of capsule gene transcription using a chromosomal gfp reporter fusion showed a significant reduction in transcription during growth in urine compared to that during growth in LB medium. When examined at the single-cell level, following growth in both media, capsule gene expression appears to be heterogeneous, with two distinct green fluorescent protein (GFP)-expressing populations. Using anti-K1 antibody, we showed that this heterogeneity in gene expression results in two populations of encapsulated and unencapsulated cells. We demonstrated that the capsule hinders attachment to and invasion of epithelial cells and that the unencapsulated cells within the population preferentially adhere to and invade bladder epithelial cells. We found that once internalized, UTI89 starts to produce capsule to aid in its intracellular survival and spread. We propose that this observed phenotypic diversity in capsule expression is a fitness strategy used by the bacterium to deal with the constantly changing environment of the urinary tract. PMID:25870229

  2. Genetic mutations in the K1 and K10 genes of patients with epidermolytic hyperkeratosis. Correlation between location and disease severity.

    PubMed Central

    Syder, A J; Yu, Q C; Paller, A S; Giudice, G; Pearson, R; Fuchs, E

    1994-01-01

    Epidermolytic hyperkeratosis (EH) is a skin disease caused by mutations in the genes encoding K1 and K10, the differentiation-specific keratins of epidermis. To explore the heterogeneity of mutations and to assess whether a correlation exists between disease severity and the extent to which a mutation interferes with keratin network formation, we determined the genetic bases of four severe incidences of EH and one unusually mild case. Two severe cases have the same mutation, K10-R156:C, at a conserved arginine that we previously showed was mutated to a histidine in two unrelated EH families. An additional severe case has a mutation six residues away, still within the amino end of the alpha-helical rod domain of K10. The other severe case has a mutation in the conserved carboxy end of the K1 rod. In contrast, affected members of the atypically mild family have a mutation just proximal to the conserved carboxy end of the K10 rod. By genetic engineering and gene transfection, we demonstrate that each mutation is functionally responsible for the keratin filament aberrations that are typical of keratinocytes cultured from these patients. Moreover, we show that the mild EH mutation less severely affects filament network formation. Taken together, our studies strengthen the link between filament perturbations, cell fragility, and degeneration. Images PMID:7512983

  3. Determination of processing effects and of storage stability on vitamin K1 (Phylloquinone) in Sea Buckthorn Berries (Hippophaë rhamnoides L. ssp. rhamnoides) and related products.

    PubMed

    Gutzeit, D; Baleanu, G; Winterhalter, P; Jerz, G

    2007-11-01

    Phylloquinone (vitamin K(1)) is the primary dietary source of vitamin K. Processing effects and stability of phylloquinone were investigated during juice and concentrate production from sea buckthorn (Hippophaë rhamnoides) using berries from 2 different growing areas. During industrial juice production the technological processing of the berries caused a loss of about 36% to 54% phylloquinone in the generated juice. The following processing steps leading to the concentrated juice resulted in a complete depletion of phylloquinone. Sea buckthorn berries and juice were stored at 6, 25, and 40 degrees C for up to 7 d to determine the temperature effects on phylloquinone during storage. Content of vitamin K(1) in sea buckthorn berries was affected by storage time and storage temperature. Storage of freshly harvested berries resulted in a significant increase (P < 0.01) of phylloquinone ranging from 21% up to 186% (wet weight). The juices showed almost identical significant degradation (P < 0.01) of phylloquinone of about 18% to 32% at 6, 25, and 40 degrees C indicating that intensity of decomposition is independent of temperature (6 to 40 degrees C) and storage time in the range of consumer storage conditions.

  4. Induction of chromosomal damage in CHO-K1 cells and their repair-deficient mutant XRS5 by x-ray and particle irradiation

    NASA Astrophysics Data System (ADS)

    Nasonova, E.; Ritter, S.; Fomenkova, T.; Kraft, G.

    The cytogenetic effects of X-rays and Au ions were investigated in repair-proficient CHO-K1 cells and their radiosensitive mutant strain xrs5, which shows a defect in the rejoining of DNA double-strand breaks. Both cell lines were synchronized by mitotic shake off, irradiated in G_1-phase with either 250 kV X-rays or 780 MeV/u Au ions (LET: 1150 keV/mum) and chromosome aberrations were analyzed in first post-irradiation metaphases. Isoeffective doses of X-rays for the induction of aberrant cells and aberrations per cell were about 14 times lower for xrs5 than for CHO-K1 cells. After high LET radiation the difference in the cytogenetic response of both cell lines was drastically diminished. Furthermore, the analysis of the aberration types induced by sparsely and densely ionizing radiation showed for both cell lines specific changes in the spectrum of aberration types as LET increases. The experimental results are discussed with respect to the different types of lesions induced by sparsely and densely ionizing radiation.

  5. S6K1 promotes invasiveness of breast cancer cells in a model of metastasis of triple-negative breast cancer

    PubMed Central

    Khotskaya, Yekaterina B; Goverdhan, Aarthi; Shen, Jia; Ponz-Sarvise, Mariano; Chang, Shih-Shin; Hsu, Ming-Chuan; Wei, Yongkun; Xia, Weiya; Yu, Dihua; Hung, Mien-Chie

    2014-01-01

    Breast cancer is the second-leading cause of oncology-related death in US women. Of all invasive breast cancers, patients with tumors lacking expression of the estrogen and progesterone hormone receptors and overexpression of human epidermal growth factor receptor 2 have the poorest clinical prognosis. These referred to as triple-negative breast cancer (TNBC) represent an aggressive form of disease that is marked by early-onset metastasis, high tumor recurrence rate, and low overall survival during the first three years post-diagnosis. In this report, we discuss a novel model of early-onset TNBC metastasis to bone and lungs, derived from MDA-MB-231 cells. Breast cancer cells injected intravenously produced rapid, osteolytic metastases in long bones and spines of athymic nude mice, with concurrent metastasis to lungs, liver, and soft tissues. From the bone metastases, we developed a highly metastatic luciferase-tagged cell line variant named MDA-231-LUC Met. In this report, we demonstrate that the Akt/mTOR/S6K1 axis is hyperactivated in these cells, leading to a dramatic increase in phosphorylation of S6 ribosomal protein at Ser235/236. Lastly, we provide evidence that inhibition of the furthest downstream kinase in the mTOR pathway, S6K1, with a highly specific inhibitor PF-4708671 inhibits cell migration, and thus may provide a potent anti-metastatic adjuvant therapy approach. PMID:25075253

  6. Participation of the NPL in 2008 in the BIPM.RI(II)-K1.Mn-56 comparison of activity measurements of the radionuclide 56Mn

    NASA Astrophysics Data System (ADS)

    Michotte, C.; Ratel, G.; Courte, S.; Keightley, J.

    2015-01-01

    In 2008, the National Physical Laboratory (NPL) submitted one sample of known activity of 56Mn to the International Reference System (SIR) for activity comparison at the Bureau International des Poids et Mesures (BIPM), with comparison identifier BIPM.RI(II)-K1.Mn-56. The value of the activity submitted was about 290 kBq. There is now one result in the BIPM.RI(II)-K1.Mn-56 comparison. The key comparison reference value (KCRV) has been evaluated for the first time using the NPL result and a BIPM result dating November 1978. The degree of equivalence between the NPL equivalent activity measured in the SIR and the KCRV has been calculated and is reported. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCRI, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

  7. Anti-listerial activity of a polymeric film coated with hybrid coatings doped with Enterocin 416K1 for use as bioactive food packaging.

    PubMed

    Iseppi, Ramona; Pilati, Francesco; Marini, Michele; Toselli, Maurizio; de Niederhäusern, Simona; Guerrieri, Elisa; Messi, Patrizia; Sabia, Carla; Manicardi, Giuliano; Anacarso, Immacolata; Bondi, Moreno

    2008-04-30

    In this study, Enterocin 416K1, a bacteriocin produced by Enterococcus casseliflavus IM 416K1, was entrapped in an organic-inorganic hybrid coating applied to a LDPE (low-density polyethylene) film for its potential use in the active food packaging field. The antibacterial activity of the coated film was evaluated against Listeria monocytogenes NCTC 10888 by qualitative modified agar diffusion assay, quantitative determination in listeria saline solution suspension and direct contact with artificially contaminated food samples (frankfurters and fresh cheeses) stored at room and refrigeration temperatures. All investigations demonstrated that enterocin-activated coatings have a good anti-listeria activity. Qualitative tests showed a clear zone of inhibition in the indicator lawn in contact with and around the coated film. During the quantitative antibacterial evaluation the L. monocytogenes viable counts decreased to 1.5 log units compared to the control. The inhibitory capability was confirmed also in food-contact assays. In all food samples packed with coated films we observed a significant decrease in L. monocytogenes viable counts in the first 24 h compared to the control. This difference was generally maintained up to the seventh day and then decreased, with the exception of the cheese samples stored at refrigeration temperature.

  8. NOTCH1, TP53, and MAP2K1 Mutations in Splenic Diffuse Red Pulp Small B-cell Lymphoma Are Associated With Progressive Disease.

    PubMed

    Martinez, Daniel; Navarro, Alba; Martinez-Trillos, Alejandra; Molina-Urra, Ricardo; Gonzalez-Farre, Blanca; Salaverria, Itziar; Nadeu, Ferran; Enjuanes, Anna; Clot, Guillem; Costa, Dolors; Carrio, Ana; Villamor, Neus; Colomer, Dolors; Martinez, Antonio; Bens, Susanne; Siebert, Reiner; Wotherspoon, Andrew; Beà, Sílvia; Matutes, Estella; Campo, Elias

    2016-02-01

    Splenic diffuse red pulp small B-cell lymphoma (SDRPL) is considered an indolent neoplasm and its pathogenesis is not well known. We investigated the molecular characteristics of 19 SDRPL patients, 5 of them with progressive disease. IGHV genes were mutated in 9/13 (69%). Cytogenetic and molecular studies identified complex karyotypes in 2 cases, and IGH rearrangements in 3, with PAX5 and potentially TCL1 as partners in each one of them. Copy number arrays showed aberrations in 69% of the tumors, including recurrent losses of 10q23, 14q31-q32, and 17p13 in 3, and 9p21 in 2 cases. Deletion of 7q31.3-q32.3 was present in only 1 case and no trisomies 3 or 18 were detected. NOTCH1 and MAP2K1 were mutated in 2 cases each, whereas BRAF, TP53, and SF3B1 were mutated each in single cases. No mutations were found in NOTCH2 or MYD88. Four of the 5 patients with aggressive disease had mutations in NOTCH1 (2 cases), TP53 (1 case), and MAP2K1 (1 case). The progression-free survival of patients with mutated genes was significantly shorter than in the unmutated (P=0.011). These findings show that SDRPL share some mutated genes but not chromosomal alterations, with other splenic lymphomas, that may confer a more aggressive behavior.

  9. Numerical modeling of humic colloid borne americium (III) migration in column experiments using the transport/speciation code K1D and the KICAM model.

    PubMed

    Schüssler, W; Artinger, R; Kim, J I; Bryan, N D; Griffin, D

    2001-02-01

    The humic colloid borne Am(III) transport was investigated in column experiments for Gorleben groundwater/sand systems. It was found that the interaction of Am with humic colloids is kinetically controlled, which strongly influences the migration behavior of Am(III). These kinetic effects have to be taken into account for transport/speciation modeling. The kinetically controlled availability model (KICAM) was developed to describe actinide sorption and transport in laboratory batch and column experiments. Application of the KICAM requires a chemical transport/speciation code, which simultaneously models both kinetically controlled processes and equilibrium reactions. Therefore, the code K1D was developed as a flexible research code that allows the inclusion of kinetic data in addition to transport features and chemical equilibrium. This paper presents the verification of K1D and its application to model column experiments investigating unimpeded humic colloid borne Am migration. Parmeters for reactive transport simulations were determined for a Gorleben groundwater system of high humic colloid concentration (GoHy 2227). A single set of parameters was used to model a series of column experiments. Model results correspond well to experimental data for the unretarded humic borne Am breakthrough.

  10. Numerical modeling of humic colloid borne Americium (III) migration in column experiments using the transport/speciation code K1D and the KICAM model

    NASA Astrophysics Data System (ADS)

    Schüßler, W.; Artinger, R.; Kim, J. I.; Bryan, N. D.; Griffin, D.

    2001-02-01

    The humic colloid borne Am(III) transport was investigated in column experiments for Gorleben groundwater/sand systems. It was found that the interaction of Am with humic colloids is kinetically controlled, which strongly influences the migration behavior of Am(III). These kinetic effects have to be taken into account for transport/speciation modeling. The kinetically controlled availability model (KICAM) was developed to describe actinide sorption and transport in laboratory batch and column experiments. Application of the KICAM requires a chemical transport/speciation code, which simultaneously models both kinetically controlled processes and equilibrium reactions. Therefore, the code K1D was developed as a flexible research code that allows the inclusion of kinetic data in addition to transport features and chemical equilibrium. This paper presents the verification of K1D and its application to model column experiments investigating unimpeded humic colloid borne Am migration. Parameters for reactive transport simulations were determined for a Gorleben groundwater system of high humic colloid concentration (GoHy 2227). A single set of parameters was used to model a series of column experiments. Model results correspond well to experimental data for the unretarded humic borne Am breakthrough.

  11. Effect of Evolutionary Adaption on Xylosidase Activity in Thermotolerant Yeast Isolates Kluyveromyces marxianus NIRE-K1 and NIRE-K3.

    PubMed

    Behera, Shuvashish; Sharma, Nilesh K; Arora, Richa; Kumar, Sachin

    2016-08-01

    Efficient use of xylose along with glucose is necessary for the economic production of lignocellulosic based biofuels. Xylose transporters play an important role in the microorganisms for efficient utilization of xylose. In the present study, a novel method has been developed for a rapid assay of xylose transport activity in the xylose-utilizing isolates and other known yeasts. An assay was conducted to compare the activity of β-xylosidase using p-nitrophenyl-β-D-xylopyranoside (pNPX) in the intact, intracellular, and extracellular yeasts cells showing xylose transporter. Saccharomyces cerevisiae (MTCC 170) showed no xylosidase activity, while little growth was observed in the xylose-containing medium. Although other yeasts, i.e., Kluyveromyces marxianus NIRE-K1 (MTCC 5933), K. marxianus NIRE-K3 (MTCC 5934), and Candida tropicalis (MTCC 230), showed xylosidase activity in intact, intracellular, and extracellular culture. The xylosidase activity in intact cell was higher than that of extracellular and intracellular activity in all the yeast cells. The enzyme activity was higher in case of K. marxianus NIRE-K1 and K. marxianus NIRE-K3 rather than the C. tropicalis. Further, better xylosidase activity was observed in adapted K. marxianus cells which were 2.79-28.46 % higher than that of native (non-adapted) strains, which indicates the significant improvement in xylose transportation.

  12. SERENDIPITOUS DETECTION OF X-RAY EMISSION FROM THE HOT BORN-AGAIN CENTRAL STAR OF THE PLANETARY NEBULA K 1-16

    SciTech Connect

    Montez, Rodolfo Jr.; Kastner, Joel H. E-mail: jhk@cis.rit.edu

    2013-03-20

    We report the serendipitous detection of point-like X-ray emission from the hot, PG1159-type central star of the planetary nebula (CSPN) K 1-16 by the XMM-Newton and Chandra X-Ray Observatories. The CSPN lies superimposed on a galaxy cluster that includes an X-ray-bright quasar, but we have successfully isolated the CSPN X-ray emission from the strong diffuse background contributed by the quasar and intracluster gas. We have modeled the XMM-Newton and Chandra X-ray data, taking advantage of the contrasting detection efficiencies of the two observatories to better constrain the low-energy spectral response of Chandra's Advanced CCD Imaging Spectrometer. We find that the CSPN X-ray spectrum is well characterized by the combination of a non-local thermodynamic equilibrium model atmosphere with T{sub *} {approx} 135 kK and a carbon-rich, optically thin thermal plasma with T{sub X} {approx} 1 MK. These results for X-ray emission from the K 1-16 CSPN, combined with those obtained for other PG1159-type objects, lend support to the 'born-again' scenario for Wolf-Rayet and PG1159 CSPNe, wherein a late helium shell flash dredges up carbon-rich intershell material and ejects this material into the circumstellar environment.

  13. Analysis of vitamin K1 in fruits and vegetables using accelerated solvent extraction and liquid chromatography tandem mass spectrometry with atmospheric pressure chemical ionization.

    PubMed

    Jäpelt, Rie Bak; Jakobsen, Jette

    2016-02-01

    The objective of this study was to develop a rapid, sensitive, and specific analytical method to study vitamin K1 in fruits and vegetables. Accelerated solvent extraction and solid phase extraction was used for sample preparation. Quantification was done by liquid chromatography tandem mass spectrometry with atmospheric pressure chemical ionization in selected reaction monitoring mode with deuterium-labeled vitamin K1 as an internal standard. The precision was estimated as the pooled estimate of three replicates performed on three different days for spinach, peas, apples, banana, and beetroot. The repeatability was 5.2% and the internal reproducibility was 6.2%. Recovery was in the range 90-120%. No significant difference was observed between the results obtained by the present method and by a method using the same principle as the CEN-standard i.e. liquid-liquid extraction and post-column zinc reduction with fluorescence detection. Limit of quantification was estimated to 0.05 μg/100g fresh weight.

  14. Synthesis, biological evaluation, and structure-activity relationships of N-benzoyl-2-hydroxybenzamides as agents active against P. falciparum (K1 strain), Trypanosomes, and Leishmania.

    PubMed

    Stec, Jozef; Huang, Qingqing; Pieroni, Marco; Kaiser, Marcel; Fomovska, Alina; Mui, Ernest; Witola, William H; Bettis, Samuel; McLeod, Rima; Brun, Reto; Kozikowski, Alan P

    2012-04-12

    In our efforts to identify novel chemical scaffolds for the development of new antiprotozoal drugs, a compound library was screened against Toxoplasma gondii tachyzoites with activity discovered for N-(4-ethylbenzoyl)-2-hydroxybenzamide 1a against T. gondii as described elsewhere. Synthesis of a compound set was guided by T. gondii SAR with 1r found to be superior for T. gondii , also active against Thai and Sierra Leone strains of Plasmodium falciparum , and with superior ADMET properties as described elsewhere. Herein, synthesis methods and details of the chemical analysis of the compounds in this series are described. Further, this series of N-benzoyl-2-hydroxybenzamides was repurposed for testing against four other protozoan parasites: Trypanosoma brucei rhodesiense , Trypanosoma cruzi , Leishmania donovani , and P. falciparum (K1 isolate). Structure-activity analyses led to the identification of compounds in this set with excellent antileishmanial activity (compound 1d). Overall, compound 1r was the best and had activity 21-fold superior to that of the standard antimalarial drug chloroquine against the K1 P. falciparum isolate.

  15. A novel regulatory element (E77) isolated from CHO‐K1 genomic DNA enhances stable gene expression in Chinese hamster ovary cells

    PubMed Central

    Kang, Shin‐Young; Kim, Yeon‐Gu; Kang, Seunghee; Lee, Hong Weon

    2016-01-01

    Abstract Vectors flanked by regulatory DNA elements have been used to generate stable cell lines with high productivity and transgene stability; however, regulatory elements in Chinese hamster ovary (CHO) cells, which are the most widely used mammalian cells in biopharmaceutical production, are still poorly understood. We isolated a novel gene regulatory element from CHO‐K1 cells, designated E77, which was found to enhance the stable expression of a transgene. A genomic library was constructed by combining CHO‐K1 genomic DNA fragments with a CMV promoter‐driven GFP expression vector, and the E77 element was isolated by screening. The incorporation of the E77 regulatory element resulted in the generation of an increased number of clones with high expression, thereby enhancing the expression level of the transgene in the stable transfectant cell pool. Interestingly, the E77 element was found to consist of two distinct fragments derived from different locations in the CHO genome shotgun sequence. High and stable transgene expression was obtained in transfected CHO cells by combining these fragments. Additionally, the function of E77 was found to be dependent on its site of insertion and specific orientation in the vector construct. Our findings demonstrate that stable gene expression mediated by the CMV promoter in CHO cells may be improved by the isolated novel gene regulatory element E77 identified in the present study. PMID:26762773

  16. Amino acid-dependent signaling via S6K1 and MYC is essential for regulation of rDNA transcription

    PubMed Central

    Kang, Jian; Kusnadi, Eric P.; Ogden, Allison J.; Hicks, Rodney J.; Bammert, Lukas; Kutay, Ulrike; Hung, Sandy; Sanij, Elaine; Hannan, Ross D.; Hannan, Katherine M.; Pearson, Richard B.

    2016-01-01

    Dysregulation of RNA polymerase I (Pol I)-dependent ribosomal DNA (rDNA) transcription is a consistent feature of malignant transformation that can be targeted to treat cancer. Understanding how rDNA transcription is coupled to the availability of growth factors and nutrients will provide insight into how ribosome biogenesis is maintained in a tumour environment characterised by limiting nutrients. We demonstrate that modulation of rDNA transcription initiation, elongation and rRNA processing is an immediate, co-regulated response to altered amino acid abundance, dependent on both mTORC1 activation of S6K1 and MYC activity. Growth factors regulate rDNA transcription initiation while amino acids modulate growth factor-dependent rDNA transcription by primarily regulating S6K1-dependent rDNA transcription elongation and processing. Thus, we show for the first time amino acids regulate rRNA synthesis by a distinct, post-initiation mechanism, providing a novel model for integrated control of ribosome biogenesis that has implications for understanding how this process is dysregulated in cancer. PMID:27385002

  17. Serendipitous Detection of X-Ray Emission from the Hot Born-again Central Star of the Planetary Nebula K 1-16

    NASA Astrophysics Data System (ADS)

    Montez, Rodolfo, Jr.; Kastner, Joel H.

    2013-03-01

    We report the serendipitous detection of point-like X-ray emission from the hot, PG1159-type central star of the planetary nebula (CSPN) K 1-16 by the XMM-Newton and Chandra X-Ray Observatories. The CSPN lies superimposed on a galaxy cluster that includes an X-ray-bright quasar, but we have successfully isolated the CSPN X-ray emission from the strong diffuse background contributed by the quasar and intracluster gas. We have modeled the XMM-Newton and Chandra X-ray data, taking advantage of the contrasting detection efficiencies of the two observatories to better constrain the low-energy spectral response of Chandra's Advanced CCD Imaging Spectrometer. We find that the CSPN X-ray spectrum is well characterized by the combination of a non-local thermodynamic equilibrium model atmosphere with T sstarf ~ 135 kK and a carbon-rich, optically thin thermal plasma with T X ~ 1 MK. These results for X-ray emission from the K 1-16 CSPN, combined with those obtained for other PG1159-type objects, lend support to the "born-again" scenario for Wolf-Rayet and PG1159 CSPNe, wherein a late helium shell flash dredges up carbon-rich intershell material and ejects this material into the circumstellar environment.

  18. A novel regulatory element (E77) isolated from CHO-K1 genomic DNA enhances stable gene expression in Chinese hamster ovary cells.

    PubMed

    Kang, Shin-Young; Kim, Yeon-Gu; Kang, Seunghee; Lee, Hong Weon; Lee, Eun Gyo

    2016-05-01

    Vectors flanked by regulatory DNA elements have been used to generate stable cell lines with high productivity and transgene stability; however, regulatory elements in Chinese hamster ovary (CHO) cells, which are the most widely used mammalian cells in biopharmaceutical production, are still poorly understood. We isolated a novel gene regulatory element from CHO-K1 cells, designated E77, which was found to enhance the stable expression of a transgene. A genomic library was constructed by combining CHO-K1 genomic DNA fragments with a CMV promoter-driven GFP expression vector, and the E77 element was isolated by screening. The incorporation of the E77 regulatory element resulted in the generation of an increased number of clones with high expression, thereby enhancing the expression level of the transgene in the stable transfectant cell pool. Interestingly, the E77 element was found to consist of two distinct fragments derived from different locations in the CHO genome shotgun sequence. High and stable transgene expression was obtained in transfected CHO cells by combining these fragments. Additionally, the function of E77 was found to be dependent on its site of insertion and specific orientation in the vector construct. Our findings demonstrate that stable gene expression mediated by the CMV promoter in CHO cells may be improved by the isolated novel gene regulatory element E77 identified in the present study.

  19. Loss of Plastoglobule Kinases ABC1K1 and ABC1K3 Causes Conditional Degreening, Modified Prenyl-Lipids, and Recruitment of the Jasmonic Acid Pathway[W

    PubMed Central

    Lundquist, Peter K.; Poliakov, Anton; Giacomelli, Lisa; Friso, Giulia; Appel, Mason; McQuinn, Ryan P.; Krasnoff, Stuart B.; Rowland, Elden; Ponnala, Lalit; Sun, Qi; van Wijk, Klaas J.

    2013-01-01

    Plastoglobules (PGs) are plastid lipid-protein particles. This study examines the function of PG-localized kinases ABC1K1 and ABC1K3 in Arabidopsis thaliana. Several lines of evidence suggested that ABC1K1 and ABC1K3 form a protein complex. Null mutants for both genes (abc1k1 and abc1k3) and the double mutant (k1 k3) displayed rapid chlorosis upon high light stress. Also, k1 k3 showed a slower, but irreversible, senescence-like phenotype during moderate light stress that was phenocopied by drought and nitrogen limitation, but not cold stress. This senescence-like phenotype involved degradation of the photosystem II core and upregulation of chlorophyll degradation. The senescence-like phenotype was independent of the EXECUTER pathway that mediates genetically controlled cell death from the chloroplast and correlated with increased levels of the singlet oxygen–derived carotenoid β-cyclocitral, a retrograde plastid signal. Total PG volume increased during light stress in wild type and k1 k3 plants, but with different size distributions. Isolated PGs from k1 k3 showed a modified prenyl-lipid composition, suggesting reduced activity of PG-localized tocopherol cyclase (VTE1), and was consistent with loss of carotenoid cleavage dioxygenase 4. Plastid jasmonate biosynthesis enzymes were recruited to the k1 k3 PGs but not wild-type PGs, while pheophytinase, which is involved in chlorophyll degradation, was induced in k1 k3 and not wild-type plants and was localized to PGs. Thus, the ABC1K1/3 complex contributes to PG function in prenyl-lipid metabolism, stress response, and thylakoid remodeling. PMID:23673981

  20. Vimentin, a Novel NF-κB Regulator, Is Required for Meningitic Escherichia coli K1-Induced Pathogen Invasion and PMN Transmigration across the Blood-Brain Barrier

    PubMed Central

    Zhang, Bao; Liu, Li-Qun; Wu, Xuedong; Mor-Vaknin, Nirit; Markovitz, David M.; Cao, Hong; Zhou, Yan-Hong

    2016-01-01

    Background NF-κB activation, pathogen invasion, polymorphonuclear leukocytes (PMN) transmigration (PMNT) across the blood-brain barrier (BBB) are the pathogenic triad hallmark features of bacterial meningitis, but the mechanisms underlying these events remain largely unknown. Vimentin, which is a novel NF-κB regulator, is the primary receptor for the major Escherichia coli K1 virulence factor IbeA that contributes to the pathogenesis of neonatal bacterial sepsis and meningitis (NSM). We have previously shown that IbeA-induced NF-κB signaling through its primary receptor vimentin as well as its co-receptor PTB-associated splicing factor (PSF) is required for pathogen penetration and leukocyte transmigration across the BBB. This is the first in vivo study to demonstrate how vimentin and related factors contributed to the pathogenic triad of bacterial meningitis. Methodology/Principal Findings The role of vimentin in IbeA+ E. coli K1-induced NF-κB activation, pathogen invasion, leukocyte transmigration across the BBB has now been demonstrated by using vimentin knockout (KO) mice. In the in vivo studies presented here, IbeA-induced NF-κB activation, E. coli K1 invasion and polymorphonuclear neutrophil (PMN) transmigration across the BBB were significantly reduced in Vim-/- mice. Decreased neuronal injury in the hippocampal dentate gyrus was observed in Vim-/- mice with meningitis. The major inflammatory regulator α7 nAChR and several signaling molecules contributing to NF-κB activation (p65 and p-CamKII) were significantly reduced in the brain tissues of the Vim-/- mice with E. coli meningitis. Furthermore, Vim KO resulted in significant reduction in neuronal injury and in α7 nAChR-mediated calcium signaling. Conclusion/Significance Vimentin, a novel NF-κB regulator, plays a detrimental role in the host defense against meningitic infection by modulating the NF-κB signaling pathway to increase pathogen invasion, PMN recruitment, BBB permeability and neuronal

  1. Final report on the ongoing key comparison BIPM.QM-K1: Ozone at ambient level, comparison with ISCIII (December 2012)

    NASA Astrophysics Data System (ADS)

    Viallon, Joële; Moussay, Philippe; Idrees, Faraz; Wielgosz, Robert; Morillo Gomez, Pilar; Sánchez, Carmen

    2013-01-01

    As part of the ongoing key comparison BIPM.QM-K1, a comparison has been performed between the ozone national standard of the Instituto de Salud Carlos III (ISCIII) and the common reference standard of the key comparison, maintained by the Bureau International des Poids et Mesures (BIPM). The instruments have been compared over a nominal ozone amount-of-substance fraction range of 0 nmol/mol to 500 nmol/mol. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

  2. Final report on the ongoing key comparison BIPM.QM-K1: Ozone at ambient level, comparison with ISCIII (December 2010)

    NASA Astrophysics Data System (ADS)

    Viallon, Joële; Moussay, Philippe; Idrees, Faraz; Wielgosz, Robert; Morillo Gomez, Pilar; Sánchez, Carmen

    2011-01-01

    As part of the ongoing key comparison BIPM.QM-K1, a comparison has been performed between the ozone national standard of the Instituto de Salud Carlos III (ISCIII) and the common reference standard of the key comparison, maintained by the Bureau International des Poids et Mesures (BIPM). The instruments have been compared over a nominal ozone amount-of-substance fraction range of 0 nmol/mol to 500 nmol/mol. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (MRA).

  3. KEY COMPARISON: Final report, on-going key comparison BIPM.QM-K1: Ozone at ambient level, comparison with ISCIII, 2007

    NASA Astrophysics Data System (ADS)

    Viallon, Joële; Moussay, Philippe; Wielgosz, Robert; Morillo Gomez, Pilar; Sánchez Blaya, Carmen

    2009-01-01

    As part of the on-going key comparison BIPM.QM-K1, a comparison has been performed between the ozone national standard of the Instituto de Salud Carlos III (ISCIII) and the common reference standard of the key comparison, maintained by the Bureau International des Poids et Mesures (BIPM). The instruments have been compared over a nominal ozone mole fraction range of 0 nmol/mol to 500 nmol/mol. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (MRA).

  4. Investigation into the evolution of the structure of K1-x LixTa1-y NbyO3 single crystals under variations in temperature

    NASA Astrophysics Data System (ADS)

    Borisov, S. A.; Vakhrushev, S. B.; Koroleva, E. Yu.; Naberezhnov, A. A.; Syrnikov, P. P.; Simkin, V. G.; Kutnjak, Z.; Egami, T.; Dmowski, W.; Piekarz, P.

    2007-05-01

    The evolution of the structure of K1-x LixTa1-y NbyO3 single crystals with x = 0.001, y = 0.026, and 1900 ppm Cu (KLTN277) and with x = 0.0014 and y = 0.024 (KLTN123), which exhibit an extremely high permittivity (up to 4 × 105 in the quasi-static regime for the KLTN277 crystal), is investigated in the range from room temperature to 20 K. It is demonstrated that, upon cooling to the lowest temperatures, both crystals retain their cubic structure, but the lattice parameters pass through a minimum at the temperature of the observed anomalies of the dielectric response (˜50 K). In the neutron diffraction pattern of the KLTN123 sample, satellites appear in the vicinity of the (hhh) reflections at temperatures below ˜50 K. These satellites can be associated with the nucleation of the rhombohedral phase.

  5. KEY COMPARISON: Activity measurements of the radionuclide 153Sm for the ANSTO, Australia in the ongoing comparison BIPM.RI(II)-K1.Sm-153

    NASA Astrophysics Data System (ADS)

    Ratel, G.; Michotte, C.; Reinhard, M.; Alexiev, D.; Mo, L.

    2005-01-01

    In 2004, the Australian Nuclear Science and Technology Organisation (ANSTO) submitted two samples of known activity of 153Sm to the International Reference System (SIR). The value of the activity submitted was about 920 MBq. This key comparison result has been added to the matrix of degrees of equivalence in the key comparison database that now contains five results, identifier BIPM.RI(II)-K1.Sm-153. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by Section II of the Consultative Committee for Ionizing Radiation (CCRI(II)), according to the provisions of the Mutual Recognition Arrangement (MRA).

  6. Corrosion cracking of 03N18K1M3TYu and 02N12Kh5M3 maraging steels in chloride solutions

    SciTech Connect

    Pavlov, V.N.; Chumalo, G.V.; Vereshchagin, A.N.; Melekhov, R.K.

    1987-07-01

    The authors investigate the electrochemical behavior in 0.5% NaCl solution and 42% MgCl/sub 2/ solution and the tendency toward corrosion cracking was determined in boiling 0.5% chloride solution of the cobalt-containing maraging steels in the title. Weld specimens and specimens of the base metal of 03N18K1M3TYu steel were tested in 3% NaCl solution for resistance to corrosion cracking. Additional investigations were made of specimens of that steel with previously created fatigue cracks of the base metal and the weld specimens in 3% NaCl solutions, since that steel is a promising material for structures operating in sea water and low concentration chloride solutions.

  7. Fe-vacancy ordering in superconducting K1–xFe2–ySe2: First-principles calculations and Monte Carlo simulations

    DOE PAGES

    Fang, Yong; Tai, Yuan -Yen; Deng, Junkai; ...

    2015-07-20

    Fe vacancies in the 33 K superconductor K1–xFe2–ySe2 show ordering schemes that may be correlated with its superconducting properties. First-principles calculations and kinetic Monte Carlo simulations lead to a very simple model for vacancy ordering. Repulsive dipolar interactions between Fe vacancies show three ground states: amore » $$\\sqrt{8}\\times \\sqrt{10}$$ rhombus-ordered structure for 12.5% vacancies, a $$\\sqrt{5}\\times \\sqrt{5}$$ squared lattice for 20% vacancies, and a $$\\sqrt{5}\\times \\sqrt{5}$$ rhombus-ordered structure for 25% vacancies. Other structural states are derived from these three ground states and may contain additional disordered spatial regions. As a result, the repulsive interaction between Fe vacancies arises from enhanced Fe–Se covalent bonds, which differs from the well-known attractive interaction of Fe vacancies in body-centered cubic Fe.« less

  8. Final report of the ongoing key comparison BIPM.QM-K1: Ozone at ambient level, comparison with JRC (October 2013)

    NASA Astrophysics Data System (ADS)

    Viallon, Joële; Moussay, Philippe; Idrees, Faraz; Wielgosz, Robert; Lagler, Friedrich

    2014-01-01

    As part of the ongoing key comparison BIPM.QM-K1, a comparison has been performed between the ozone standard of the European Commission maintained by the Joint Research Centre (JRC) and the common reference standard of the key comparison, maintained by the Bureau International des Poids et Mesures (BIPM). The instruments have been compared over a nominal ozone amount-of-substance fraction range of 0 nmol/mol to 500 nmol/mol. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

  9. Accumulation of polycyclic aromatic hydrocarbon-induced single strand breaks is attributed to slower rejoining processes by DNA polymerase inhibitor, cytosine arabinoside in CHO-K1 cells

    SciTech Connect

    Park, Jongkun ); Lee, Jungsup; Lee, Hyungho; Choi, Insoon; Park, Sangdai )

    1991-01-01

    The authors demonstrate a successful induction of DNA single breaks in CHO-K1 cells by cocultivation with mouse embryonic fibroblasts (MEF) during exposure to benzo(a)pyrene (BP) or 3-methylcholanthrene (MC). When compared to those induced by methyl methanesulfonate (MMS), the DNA single strand breaks induced by BP and MC were markedly accumulated by post-incubation with cytosine arabinoside (araC) and were much more delayed in their rejoining. These results suggest that the active metabolites of BP or MC produced by cocultivation with MEF or microsomal fraction (S-15) result in the formation of large DNA adducts which require an active participation of DNA polymerase {alpha}({delta}) in the polymerization step of excision repair for their removal.

  10. Routine clinical determination of carotene, vitamin E, vitamin A, 25-hydroxy vitamin D3 and trans-vitamin K1 in human serum by straight phase HPLC.

    PubMed

    Van Haard, P M; Engel, R; Postma, T

    1987-01-01

    A universal extraction procedure is described for fat-soluble vitamins in human serum. Methods are presented for routine quantitative analysis by isocratic straight phase HPLC with UV-detection of (alpha + beta)-carotene, vitamin E (alpha-tocopherol) and vitamin A (all-trans-retinol) in one single run, and of vitamin K1 (trans-phylloquinone) and 25-hydroxy vitamin D3 after sample clean-up using disposable reversed-phase cartridges. The limits of detection, precisions and selectivities of the developed assays are shown to be satisfactory after more than three years' experience. The routine clinical determination of fat-soluble vitamins can be performed in less than 5 mL of serum. Analyses of external quality control and randomly taken outpatient samples are shown to be of great value in assessing laboratory performance.

  11. Regulation of amino acid transport system L by amino acid availability in CHO-K1 cells. A special role for leucine.

    PubMed

    Moreno, A; Lobatón, C D; Oxender, D L

    1985-10-10

    Starvation of CHO-K1 cells for leucine leads to a 3-4-fold increase in transport system L activity, without modification of transport through systems A and ASC. The concentration of leucine must be below 10 microM before the enhancement of transport can be clearly seen. To achieve low concentrations of leucine such as 10 microM, extensive dialysis of fetal calf serum was required. The enhancement of transport was completed after 12-24 h of starvation and was fully reversed within 1 h of re-feeding with leucine. Starvation for isoleucine, valine or phenylalanine also produced an increase in system L transport activity, but the effect was only one half of that seen following leucine starvation.

  12. Final report on the ongoing key comparison BIPM.QM-K1: Ozone at ambient level, comparison with EAA (September 2011)

    NASA Astrophysics Data System (ADS)

    Viallon, Joële; Moussay, Philippe; Idrees, Faraz; Wielgosz, Robert; Froehlich, Marina; Wolf, Andreas

    2011-01-01

    As part of the ongoing key comparison BIPM.QM-K1, a comparison has been performed between the ozone national standard of Austria maintained by the Umweltbundesamt GmbH (EAA) and the common reference standard of the key comparison, maintained by the Bureau International des Poids et Mesures (BIPM). The instruments have been compared over a nominal ozone amount-of-substance fraction range of 0 nmol/mol to 500 nmol/mol. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (MRA).

  13. Final report of the ongoing key comparison BIPM.QM-K1: Ozone at ambient level, comparison with CHMI (September 2013)

    NASA Astrophysics Data System (ADS)

    Viallon, Joële; Moussay, Philippe; Idrees, Faraz; Wielgosz, Robert; Novák, Jiří; Vokoun, Miroslav

    2013-01-01

    As part of the ongoing key comparison BIPM.QM-K1, a comparison has been performed between the ozone standard of the Czech Republic maintained by the Czech Hydrometeorological Institute (CHMI) and the common reference standard of the key comparison, maintained by the Bureau International des Poids et Mesures (BIPM). The instruments have been compared over a nominal ozone amount-of-substance fraction range of 0 nmol/mol to 500 nmol/mol. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

  14. Final report of the ongoing key comparison BIPM.QM-K1: Ozone at ambient level, comparison with METAS (July 2013)

    NASA Astrophysics Data System (ADS)

    Viallon, Joële; Moussay, Philippe; Idrees, Faraz; Wielgosz, Robert; Niederhauser, Bernhard

    2013-01-01

    As part of the ongoing key comparison BIPM.QM-K1, a comparison has been performed between the ozone standard of Switzerland maintained by the Swiss Federal Office of Metrology (METAS) and the common reference standard of the key comparison, maintained by the Bureau International des Poids et Mesures (BIPM). The instruments have been compared over a nominal ozone amount-of-substance fraction range of 0 nmol/mol to 500 nmol/mol. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

  15. Final report of the ongoing key comparison BIPM.QM-K1: Ozone at ambient level, comparison with NIST (February 2013)

    NASA Astrophysics Data System (ADS)

    Viallon, Joële; Moussay, Philippe; Idrees, Faraz; Wielgosz, Robert; Norris, James E.; Guenther, Franck

    2014-01-01

    As part of the ongoing key comparison BIPM.QM-K1, a comparison has been performed between the ozone national standard of the National Institute of Standards and Technology (NIST) and the common reference standard of the key comparison, maintained by the Bureau International des Poids et Mesures (BIPM). The instruments have been compared over a nominal ozone amount-of-substance fraction range of 0 nmol/mol to 500 nmol/mol. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

  16. Final report of the ongoing key comparison BIPM.QM-K1: Ozone at ambient level, comparison with NPL (May 2013)

    NASA Astrophysics Data System (ADS)

    Viallon, Joële; Moussay, Philippe; Idrees, Faraz; Wielgosz, Robert; Sweeney, Bryan; Quincey, Paul

    2014-01-01

    As part of the ongoing key comparison BIPM.QM-K1, a comparison has been performed between the ozone standard of the United Kingdom maintained by the National Physical Laboratory (NPL) and the common reference standard of the key comparison, maintained by the Bureau International des Poids et Mesures (BIPM). The instruments have been compared over a nominal ozone amount-of-substance fraction range of 0 nmol/mol to 500 nmol/mol. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

  17. Final report of the ongoing key comparison BIPM.QM-K1: Ozone at ambient level, comparison with NMISA (July 2012)

    NASA Astrophysics Data System (ADS)

    Viallon, Joële; Moussay, Philippe; Idrees, Faraz; Wielgosz, Robert; Botha, Angelique

    2013-01-01

    As part of the ongoing key comparison BIPM.QM-K1, a comparison has been performed between the ozone standard of the World Meteorological Organization maintained by the National Metrology Institute of South Africa (NMISA) and the common reference standard of the key comparison, maintained by the Bureau International des Poids et Mesures (BIPM). The instruments have been compared over a nominal ozone amount-of-substance fraction range of 0 nmol/mol to 500 nmol/mol. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

  18. Final report on the ongoing key comparison BIPM.QM-K1: Ozone at ambient level, comparison with NMC, A*STAR (December 2010)

    NASA Astrophysics Data System (ADS)

    Viallon, Joële; Moussay, Philippe; Idrees, Faraz; Wielgosz, Robert; Rakowska, Agata; Chye, Teo Chin

    2011-01-01

    As part of the ongoing key comparison BIPM.QM-K1, a comparison has been performed between the ozone national standard of Singapore maintained by the National Metrology Centre, A*STAR (NMC, A*STAR) and the common reference standard of the key comparison, maintained by the Bureau International des Poids et Mesures (BIPM). The instruments have been compared over a nominal ozone amount-of-substance fraction range of 0 nmol/mol to 500 nmol/mol. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (MRA).

  19. Final report, on-going key comparison BIPM.QM-K1: ozone at ambient level, comparison with DMDM, July 2015

    NASA Astrophysics Data System (ADS)

    Viallon, J.; Moussay, P.; Wielgosz, R.; Bebic, J.; Norris, J. E.; Guenther, F.

    2016-01-01

    As part of the on-going key comparison BIPM.QM-K1, a comparison has been performed between the ozone national standard of the Directorate of Measures and Precious Metals (DMDM) and the common reference standard of the key comparison, maintained by the Bureau International des Poids et Mesures (BIPM), via a transfer standard maintained by the National Institute of Standards and Technology (NIST). The instruments have been compared over a nominal ozone amount-of-substance fraction range of 0 nmol/mol to 500 nmol/mol Main text To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

  20. Final report of the ongoing key comparison BIPM.QM-K1: Ozone at ambient level, comparison with NMC, A*STAR, May 2013

    NASA Astrophysics Data System (ADS)

    Viallon, Joële; Moussay, Philippe; Wielgosz, Robert; Li, Hou; Hui, Wendy Liu; Norris, James E.; Guenther, Frank

    2014-01-01

    As part of the on-going key comparison BIPM.QM-K1, a comparison has been performed between the ozone national standard of Singapore maintained by the National Metrology Centre, A*STAR (NMC, A*STAR) and the common reference standard of the key comparison, maintained by the Bureau International des Poids et Mesures (BIPM), via a transfer standard maintained by the National Institute of Standards and Technology (NIST). The instruments have been compared over a nominal ozone amount-of-substance fraction range of 0 nmol/mol to 500 nmol/mol Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

  1. Final report, on-going key comparison BIPM.QM-K1, ozone at ambient level, comparison with NIST, July 2015

    NASA Astrophysics Data System (ADS)

    Viallon, Joële; Moussay, Philippe; Wielgosz, Robert; Hodges, Joe; Norris, James E.

    2017-01-01

    As part of the ongoing key comparison BIPM.QM-K1, a comparison has been performed between the ozone national standard of the National Institute of Standards and Technology (NIST) and the common reference standard of the key comparison, maintained by the Bureau International des Poids et Mesures (BIPM). The instruments have been compared over a nominal ozone amount-of-substance fraction range of 0 nmol/mol to 500 nmol/mol. Main text To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

  2. Final report on the ongoing key comparison BIPM.QM-K1: Ozone at ambient level, comparison with CENICA (October 2010)

    NASA Astrophysics Data System (ADS)

    Viallon, Joële; Moussay, Philippe; Idrees, Faraz; Wielgosz, Robert; Fentanes, Oscar

    2011-01-01

    As part of the ongoing key comparison BIPM.QM-K1, a comparison has been performed between the ozone national standard of the Centro Nacional de Investigación y Capacitación Ambiental (CENICA) and the common reference standard of the key comparison, maintained by the Bureau International des Poids et Mesures (BIPM). The instruments have been compared over a nominal ozone amount-of-substance fraction range of 0 nmol/mol to 500 nmol/mol. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCQM, according to the provisions of the CIPM Mutual Recognition Arrangement (MRA).

  3. Delta I = 1 staggering effect for negative parity rotational bands with K = 1/2 in W/Os/Pt odd-mass nuclei

    NASA Astrophysics Data System (ADS)

    Taha, M. M.

    2015-11-01

    The anomalous negative-parity bands of odd-mass nuclei W/Os/Pt for N = 103 isotones are studied within the framework of particle rotor model (PRM). The phenomenon of Δ I = 1 staggering or signature splitting in energies occurs as one plots the gamma transitional energy over spin (EGOS) versus spin for the 1/2-[521] band originating from N = 5 single particle orbital. The rotational band with K = 1/2 separates into two signature partners. The levels with I = 1/2, 5/2, 9/2,… are displaced relatively to the levels with I = 3/2,7/2,11/2,…. The deviations of the level energies from the rigid rotor values is described by Coriolis coupling.

  4. Induction of 8-hydroxy-2'-deoxyguanosine in CHO-K1 cells exposed to phenyl-hydroquinone, a metabolite of ortho-phenylphenol.

    PubMed

    Nakagawa, Y; Tayama, S

    1996-03-29

    The induction of 8-hydroxy-2'-deoxyguanosine (8-OHdG), an index of oxidative DNA modification, was investigated in CHO-K1 cells exposed to phenyl-hydroquinone (PHQ), a major metabolite of ortho-phenylphenol (OPP), an antimicrobial. Addition of PHQ at a concentration of 50 microM to CHO cell suspensions (10(6) cells/ml) induced slight elevation of intracellular 8-OHdG levels. Pretreatment of CHO cells with 3-amino-1,2,4-triazole (AT, 20 mM) enhanced PHQ-induced 8-OHdG formation which was accompanied by cell death. Pretreatment of CHO-K1 cells with AT (20 mM) and deferoxamine (DeFe, 20 mM) inhibited the formation of 8-OHdG as well as cell death caused by PHQ. Neither AT nor DeFe affected cell viability or the formation of 8-OHdG in untreated CHO cells during the incubation period. The loss of cellular glutathione induced by the addition of PHQ alone was enhanced by the pretreatment of CHO cells with AT or AT plus DeFe. When PHQ was added to AT-pretreated cell suspensions, the concentration of PHQ decreased with time. This decrease was accompanied by the formation of phenyl-benzoquinone (PBQ). These results suggest that the reactive oxygen species derived from autoxidation of PHQ which converts to PBQ via phenyl-semiquinone elicit DNA damage in CHO cells, especially when the activity of cellular catalase is inhibited.

  5. Origin of giant piezoelectric effect in lead-free K1−xNaxTa1−yNbyO3 single crystals

    PubMed Central

    Tian, Hao; Meng, Xiangda; Hu, Chengpeng; Tan, Peng; Cao, Xilong; Shi, Guang; Zhou, Zhongxiang; Zhang, Rui

    2016-01-01

    A series of high-quality, large-sized (maximum size of 16 × 16 × 32 mm3) K1−xNaxTa1−yNbyO3 (x = 0.61, 0.64, and 0.70 and corresponding y = 0.58, 0.60, and 0.63) single crystals were grown using the top-seed solution growth method. The segregation of the crystals, which allowed for precise control of the individual components of the crystals during growth, was investigated. The obtained crystals exhibited excellent properties without being annealed, including a low dielectric loss (0.006), a saturated hysteresis loop, a giant piezoelectric coefficient d33 (d33 = 416 pC/N, determined by the resonance method and d33* = 480 pC/N, measured using a piezo-d33 meter), and a large electromechanical coupling factor, k33 (k33 = 83.6%), which was comparable to that of lead zirconate titanate. The reason the piezoelectric coefficient d33 of K0.39Na0.61Ta0.42Nb0.58O3 was larger than those of the other two crystals grown was elucidated through first-principles calculations. The obtained results indicated that K1−xNaxTa1−yNbyO3 crystals can be used as a high-quality, lead-free piezoelectric material. PMID:27160075

  6. Polarisation observations of H2O JK-1 K1 = 532 - 441 620.701 GHz maser emission with Herschel/HIFI in Orion KL

    NASA Astrophysics Data System (ADS)

    Jones, S. C.; Houde, M.; Harwit, M.; Kidger, M.; Kraus, A.; MCoey, C.; Marston, A.; Melnick, G.; Menten, K. M.; Morris, P.; Teyssier, D.; Tolls, V.

    2014-07-01

    Context. The high intensities and narrow bandwidths exhibited by some astronomical masers make them ideal tools for studying star-forming giant molecular clouds. The water maser transition JK-1K1 = 532-441 at 620.701 GHz can only be observed from above Earth's strongly absorbing atmosphere; its emission has recently been detected from space. Aims: We sought to further characterize the star-forming environment of Orion KL by investigating the linear polarisation of a source emitting a narrow 620.701 GHz maser feature with the heterodyne spectrometer HIFI on board the Herschel Space Observatory. Methods: High-resolution spectral datasets were collected over a thirteen month period beginning in 2011 March, to establish not only the linear polarisation but also the temporal variability of the source. Results: Within a 3σ uncertainty, no polarisation was detected to an upper limit of approximately 2%. These results are compared with coeval linear polarisation measurements of the 22.235 GHz JK-1K1 = 616 - 523 maser line from the Effelsberg 100-m radio telescope, typically a much stronger maser transition. Although strongly polarised emission is observed for one component of the 22.235 GHz maser at 7.2 km s-1, a weaker component at the same velocity as the 620.701 GHz maser at 11.7 km s-1 is much less polarised. Herschel is an ESA space observatory with science instruments provided by European-led Principal Investigator consortia and with significant participation from NASA.Appendices are available in electronic form at http://www.aanda.org

  7. Origin of giant piezoelectric effect in lead-free K1-xNaxTa1-yNbyO3 single crystals.

    PubMed

    Tian, Hao; Meng, Xiangda; Hu, Chengpeng; Tan, Peng; Cao, Xilong; Shi, Guang; Zhou, Zhongxiang; Zhang, Rui

    2016-05-10

    A series of high-quality, large-sized (maximum size of 16 × 16 × 32 mm(3)) K1-xNaxTa1-yNbyO3 (x = 0.61, 0.64, and 0.70 and corresponding y = 0.58, 0.60, and 0.63) single crystals were grown using the top-seed solution growth method. The segregation of the crystals, which allowed for precise control of the individual components of the crystals during growth, was investigated. The obtained crystals exhibited excellent properties without being annealed, including a low dielectric loss (0.006), a saturated hysteresis loop, a giant piezoelectric coefficient d33 (d33 = 416 pC/N, determined by the resonance method and d33(*) = 480 pC/N, measured using a piezo-d33 meter), and a large electromechanical coupling factor, k33 (k33 = 83.6%), which was comparable to that of lead zirconate titanate. The reason the piezoelectric coefficient d33 of K0.39Na0.61Ta0.42Nb0.58O3 was larger than those of the other two crystals grown was elucidated through first-principles calculations. The obtained results indicated that K1-xNaxTa1-yNbyO3 crystals can be used as a high-quality, lead-free piezoelectric material.

  8. Origin of giant piezoelectric effect in lead-free K1‑xNaxTa1‑yNbyO3 single crystals

    NASA Astrophysics Data System (ADS)

    Tian, Hao; Meng, Xiangda; Hu, Chengpeng; Tan, Peng; Cao, Xilong; Shi, Guang; Zhou, Zhongxiang; Zhang, Rui

    2016-05-01

    A series of high-quality, large-sized (maximum size of 16 × 16 × 32 mm3) K1‑xNaxTa1‑yNbyO3 (x = 0.61, 0.64, and 0.70 and corresponding y = 0.58, 0.60, and 0.63) single crystals were grown using the top-seed solution growth method. The segregation of the crystals, which allowed for precise control of the individual components of the crystals during growth, was investigated. The obtained crystals exhibited excellent properties without being annealed, including a low dielectric loss (0.006), a saturated hysteresis loop, a giant piezoelectric coefficient d33 (d33 = 416 pC/N, determined by the resonance method and d33* = 480 pC/N, measured using a piezo-d33 meter), and a large electromechanical coupling factor, k33 (k33 = 83.6%), which was comparable to that of lead zirconate titanate. The reason the piezoelectric coefficient d33 of K0.39Na0.61Ta0.42Nb0.58O3 was larger than those of the other two crystals grown was elucidated through first-principles calculations. The obtained results indicated that K1‑xNaxTa1‑yNbyO3 crystals can be used as a high-quality, lead-free piezoelectric material.

  9. Diminished satellite cell fusion and S6K1 expression in myotubes derived from skeletal muscle of low birth weight neonatal pigs.

    PubMed

    Chen, Ying; Zhu, Haibo; McCauley, Sydney R; Zhao, Lidan; Johnson, Sally E; Rhoads, Robert P; El-Kadi, Samer W

    2017-02-01

    Low birth weight (LBWT) is consistently associated with impaired postnatal muscle growth in mammals. Satellite cell (SC)-mediated myonuclear incorporation precedes protein accumulation in the early stages of postnatal muscle development and growth. The objective of this study was to investigate proliferation and differentiation of SCs and the regulation of protein synthesis signaling in response to insulin-like growth factor (IGF)-I stimulation in SC-derived myotubes of LBWT neonatal pigs. SCs isolated from Longissimus dorsi muscle of LBWT and NBWT pigs (3-d-old, n = 8) were cultured and induced to proliferate and differentiate to myotubes in vitro. On day 3 of differentiation, myotubes were fasted in serum-free media for 3 h and treated with human recombinant R(3)-insulin-like growth factor-I (rh IGF-I) at 0, 25, and 50 ng × mL(-1) for 30 min. There was no difference in proliferation rates of SCs from LBWT and NBWT pigs. However, LBWT SC fusion was 15% lower (P ≤ 0.05) without a difference in MyoD or myogenin mRNA expression in comparison with NBWT pigs, suggesting SCs are not intrinsically different between the two groups. IGF-Ι stimulation at physiological concentrations activated downstream effectors of mTOR similarly in myotubes from LBWT and NBWT pigs. However, abundance of ribosomal protein S6 kinase 1(S6K1) was lower in myotubes of LBWT compared to their NBWT siblings (P ≤ 0.05). These results indicate that the modest reduction in SC fusion and S6K1 expression are not the major contributors to the impaired postnatal muscle growth of LBWT pigs.

  10. Hop rho iso-alpha acids, berberine, vitamin D3 and vitamin K1 favorably impact biomarkers of bone turnover in postmenopausal women in a 14-week trial.

    PubMed

    Holick, Michael F; Lamb, Joseph J; Lerman, Robert H; Konda, Veera R; Darland, Gary; Minich, Deanna M; Desai, Anuradha; Chen, Tai C; Austin, Melissa; Kornberg, Jacob; Chang, Jyh-Lurn; Hsi, Alex; Bland, Jeffrey S; Tripp, Matthew L

    2010-05-01

    Osteoporosis is a major health issue facing postmenopausal women. Increased production of pro-inflammatory cytokines resulting from declining estrogen leads to increased bone resorption. Nutrition can have a positive impact on osteoporosis prevention and amelioration. The objective of this study was to investigate the impact of targeted phytochemicals and nutrients essential for bone health on bone turnover markers in healthy postmenopausal women. In this 14-week, single-blinded, 2-arm placebo-controlled pilot study, all women were instructed to consume a modified Mediterranean-style low-glycemic-load diet and to engage in limited aerobic exercise; 17 randomized to the placebo and 16 to the treatment arm (receiving 200 mg hop rho iso-alpha acids, 100 mg berberine sulfate trihydrate, 500 IU vitamin D(3) and 500 microg vitamin K(1), twice daily). Thirty-two women completed the study. Baseline nutrient intake did not differ between arms. At 14 weeks, the treatment arm exhibited an estimated 31% mean reduction (P = 0.02) in serum osteocalcin (a marker of bone turnover), whereas the placebo arm exhibited a 19% increase (P = 0.03) compared to baseline. Serum 25-hydroxyvitamin D (25(OH)D) increased by 13% (P = 0.24) in the treatment arm and decreased by 25% (P < 0.01) in the placebo arm. The between-arm differences for OC and 25(OH)D were statistically significant. Serum IGF-I was increased in both arms, but the increase was more significant in the treatment arm at 14 weeks (P < 0.01). Treatment with hop rho iso-alpha acids, berberine sulfate trihydrate, vitamin D(3) and vitamin K(1) produced a more favorable bone biomarker profile that supports a healthy bone metabolism.

  11. Meningitic Escherichia coli K1 Penetration and Neutrophil Transmigration Across the Blood–Brain Barrier are Modulated by Alpha7 Nicotinic Receptor

    PubMed Central

    Zheng, Xueye; Wu, Chun-Hua; Jong, Ambrose; Sheard, Michael A.; Shi, Wei; Huang, Sheng-He

    2011-01-01

    Alpha7 nicotinic acetylcholine receptor (nAChR), an essential regulator of inflammation, is abundantly expressed in hippocampal neurons, which are vulnerable to bacterial meningitis. However, it is unknown whether α7 nAChR contributes to the regulation of these events. In this report, an aggravating role of α7 nAChR in host defense against meningitic E. coli infection was demonstrated by using α7-deficient (α7-/-) mouse brain microvascular endothelial cells (BMEC) and animal model systems. As shown in our in vitro and in vivo studies, E. coli K1 invasion and polymorphonuclear neutrophil (PMN) transmigration across the blood-brain barrier (BBB) were significantly reduced in α7-/- BMEC and α7-/- mice. Stimulation by nicotine was abolished in the α7-/- cells and animals. The same blocking effect was achieved by methyllycaconitine (α7 antagonist). The tight junction molecules occludin and ZO-1 were significantly reduced in the brain cortex of wildtype mice infected with E. coli and treated with nicotine, compared to α7-/- cells and animals. Decreased neuronal injury in the hippocampal dentate gyrus was observed in α7-/- mice with meningitis. Proinflammatory cytokines (IL-1β, IL-6, TNFα, MCP-1, MIP-1alpha, and RANTES) and adhesion molecules (CD44 and ICAM-1) were significantly reduced in the cerebrospinal fluids of the α7-/- mice with E. coli meningitis. Furthermore, α7 nAChR is the major calcium channel for nicotine- and E. coli K1-increased intracellular calcium concentrations of mouse BMEC. Taken together, our data suggest that α7 nAChR plays a detrimental role in the host defense against meningitic infection by modulation of pathogen invasion, PMN recruitment, calcium signaling and neuronal inflammation. PMID:21966399

  12. New species and records of mites of the superfamily Sarcoptoidea (Acariformes: Psoroptidia) from mammals in Brazil.

    PubMed

    Bochkov, Andre V; Valim, Michel P

    2016-01-01

    Sixteen species of the superfamily Sarcoptoidea (Acariformes: Psoroptidia) belonging to 10 genera of the families Atopomelidae, Listrophoridae, Chirodiscidae, and Listropsoralgidae are recorded in Brazil. Among them, three species, Prolistrophorus hylaeamys sp. nov. from Hylaeamys laticeps (Lund, 1840) (Cricetidae: Sigmodontinae) from Minas Gerais, Lynxacarus serrafreirei sp. nov. from Galictis cuja (Molina, 1782) (Carnivora: Mustelidae) from Rio de Janeiro (Listrophoridae), and Didelphoecius micoureus sp. nov. (Atopomelidae) from Micoureus paraguayanus (Tate, 1931) (Didelphimorphia: Didelphidae) from Minas Gerais are described as new for science. Three species of the family Listrophoridae, Prolistrophorus bidentatus Fain et Lukoschus, 1984 from Akodon cursor (Winge, 1887) (Rodentia: Cricetidae) (new host), Prolistrophorus ctenomys Fain, 1970 from Ctenomys torquatus Lichtenstein, 1830 (Rodentia: Ctenomyidae) (new host), and Leporacarus sylvilagi Fain, Whitaker et Lukoschus, 1981 from Sylvilagus brasiliensis (Linnaeus, 1758) (Lagomorpha: Leporidae) (new host) -from Minas Gerais and Rio Grande do Sul, and one species of the family Chirodiscidae, Parakosa tadarida McDaniel and Lawrence, 1962 from Molossus molossus (Pallas, 1766) (Chiroptera: Molossidae) are recorded for the first time in Brazil. The previously unknown female of Didelphoecius validus Fain, Zanatta-Coutinho et Fonseca, 1996 (Atopomelidae) from Metachirus nudicaudatus (Geoffroy, 1803) (Didelphimorphia: Didelphidae) from Minas Gerais is described. All data on host-parasite associations of sarcoptoids in Brazil are summarized. Totally, 61 sarcoptoid species of 8 families are recorded in Brazil.

  13. Integral Representation of the Pictorial Proof of Sum of [superscript n][subscript k=1]k[superscript 2] = 1/6n(n+1)(2n+1)

    ERIC Educational Resources Information Center

    Kobayashi, Yukio

    2011-01-01

    The pictorial proof of the sum of [superscript n][subscript k=1] k[superscript 2] = 1/6n(n+1)(2n+1) is represented in the form of an integral. The integral representations are also applicable to the sum of [superscript n][subscript k-1] k[superscript m] (m greater than or equal to 3). These representations reveal that the sum of [superscript…

  14. Effect of scavengers of active oxygen species on cell damage caused in CHO-K1 cells by phenylhydroquinone, an o-phenylphenol metabolite.

    PubMed

    Tayama, S; Nakagawa, Y

    1994-07-01

    Phenylhydroquinone (PHQ), a metabolite of o-phenylphenol (OPP), is easily autoxidized to phenylbenzoquinone (PBQ) via the semiquinone (phenylsemiquinone, PSQ) with concomitant production of superoxide anion radicals (O2-.). We have used scavengers of active oxygen species to examine whether or not O2-. produced during oxidation of PHQ is related to cell damage in CHO-K1 cells. PHQ at 10 micrograms/ml (3-h treatment) induced sister-chromatid exchange (SCE), endoreduplication (ERD) and cell-cycle delay in CHO-K1 cells. These effects were inhibited by catalase (280 U/ml), a scavenger of hydrogen peroxide (H2O2), as well as by the reductants, ascorbate (3 mM) and GSH (1 mM). Mannitol (50 mM), a scavenger of hydroxyl radical (OH.), was ineffective and superoxide dismutase (SOD, 150 U/ml), a scavenger of O2-., or SOD plus catalase rather intensified the toxicity as did aminotriazole (20 mM), an inhibitor of catalase. Analyses of incubation solutions by HPLC showed that the extent of cell damage is correlated with PHQ loss; catalase suppressed PHQ loss, whereas SOD promoted it. The correlation was more clearly seen in the time courses of cell death and PHQ loss during incubation of PHQ with each of the scavengers of active oxygen species. These results show that neither O2-. nor OH. participates in the cell damage, but rather H2O2 generated via dismutation of O2-. may participate, probably by accelerating the autoxidation of PHQ and thus causing an increase in the production of toxic intermediates. In fact, conversion of PHQ to PBQ, a reactive product, was demonstrated during incubation with PHQ in phosphate-buffered saline by following the changes in UV-visible spectra of PHQ. Inclusion of H2O2 (0.2 or 1 mM) in the incubation mixture accelerated the PHQ loss. The present results can be explained in terms of the autoxidation mechanism of hydroquinone proposed by O'Brien (1991). Different from the results in the absence of S9 mix, the cell damage induced by 50 micrograms

  15. Role of p70S6K1-mediated phosphorylation of eIF4B and PDCD4 proteins in the regulation of protein synthesis.

    PubMed

    Dennis, Michael D; Jefferson, Leonard S; Kimball, Scot R

    2012-12-14

    Modulation of mRNA binding to the 40 S ribosomal subunit during translation initiation controls not only global rates of protein synthesis but also regulates the pattern of protein expression by allowing for selective inclusion, or exclusion, of mRNAs encoding particular proteins from polysomes. The mRNA binding step is modulated by signaling through a protein kinase known as the mechanistic target of rapamycin complex 1 (mTORC1). mTORC1 directly phosphorylates the translational repressors eIF4E binding proteins (4E-BP) 1 and 2, releasing them from the mRNA cap binding protein eIF4E, thereby promoting assembly of the eIF4E·eIF4G complex. mTORC1 also phosphorylates the 70-kDa ribosomal protein S6 kinase 1 (p70S6K1), which subsequently phosphorylates eIF4B, and programmed cell death 4 (PDCD4), which sequesters eIF4A from the eIF4E·eIF4G complex, resulting in repressed translation of mRNAs with highly structured 5'-untranslated regions. In the present study, we compared the role of the 4E-BPs in the regulation of global rates of protein synthesis to that of eIF4B and PDCD4. We found that maintenance of eIF4E interaction with eIF4G was not by itself sufficient to sustain global rates of protein synthesis in the absence of mTORC1 signaling to p70S6K1; phosphorylation of both eIF4B and PDCD4 was additionally required. We also found that the interaction of eIF4E with eIF4G was maintained in the liver of fasted rats as well as in serum-deprived mouse embryo fibroblasts lacking both 4E-BP1 and 4E-BP2, suggesting that the interaction of eIF4G with eIF4E is controlled primarily through the 4E-BPs.

  16. A rat model of nerve agent exposure applicable to the pediatric population: The anticonvulsant efficacies of atropine and GluK1 antagonists

    SciTech Connect

    Miller, Steven L.; Aroniadou-Anderjaska, Vassiliki; Figueiredo, Taiza H.; Prager, Eric M.; Almeida-Suhett, Camila P.; Apland, James P.; and others

    2015-04-15

    Inhibition of acetylcholinesterase (AChE) after nerve agent exposure induces status epilepticus (SE), which causes brain damage or death. The development of countermeasures appropriate for the pediatric population requires testing of anticonvulsant treatments in immature animals. In the present study, exposure of 21-day-old (P21) rats to different doses of soman, followed by probit analysis, produced an LD{sub 50} of 62 μg/kg. The onset of behaviorally-observed SE was accompanied by a dramatic decrease in brain AChE activity; rats who did not develop SE had significantly less reduction of AChE activity in the basolateral amygdala than rats who developed SE. Atropine sulfate (ATS) at 2 mg/kg, administered 20 min after soman exposure (1.2 × LD{sub 50}), terminated seizures. ATS at 0.5 mg/kg, given along with an oxime within 1 min after exposure, allowed testing of anticonvulsants at delayed time-points. The AMPA/GluK1 receptor antagonist LY293558, or the specific GluK1 antagonist UBP302, administered 1 h post-exposure, terminated SE. There were no degenerating neurons in soman-exposed P21 rats, but both the amygdala and the hippocampus were smaller than in control rats at 30 and 90 days post-exposure; this pathology was not present in rats treated with LY293558. Behavioral deficits present at 30 days post-exposure, were also prevented by LY293558 treatment. Thus, in immature animals, a single injection of atropine is sufficient to halt nerve agent-induced seizures, if administered timely. Testing anticonvulsants at delayed time-points requires early administration of ATS at a low dose, sufficient to counteract only peripheral toxicity. LY293558 administered 1 h post-exposure, prevents brain pathology and behavioral deficits. - Highlights: • The LD{sub 50} of soman was determined in postnatal-day-21 rats. • Rats with no seizures after 1.2XLD{sub 50} soman had less reduction of AChE in the amygdala. • Atropine sulfate (ATS) at 2 mg/kg, given at 20 min after

  17. Effect of Brazilian propolis (AF-08) on genotoxicity, cytotoxicity and clonogenic death of Chinese hamster ovary (CHO-K1) cells irradiated with (60)Co gamma-radiation.

    PubMed

    Santos, Geyza Spigoti; Tsutsumi, Shigetoshi; Vieira, Daniel Perez; Bartolini, Paolo; Okazaki, Kayo

    2014-03-01

    The present study was conducted in order to evaluate the effect of Brazilian propolis (AF-08; 5, 10, 15, 30, 50, 100, and 200μg/mL) in protecting CHO-K1 cells against genotoxic and cytotoxic damage and clonogenic death induced by (60)Co gamma-radiation (1.0, 2.0, 4.0, and 6.0Gy). For this purpose, three interlinked endpoints were analyzed: induction of DNA damage by use of the micronucleus (MN) test (genotoxic damage), cell viability by means of the MTS assay, and differential staining (cytotoxic damage) and clonogenic death via the colony-formation test (cytotoxic damage). The MN test revealed that propolis alone (5-100μg/mL) was not genotoxic up to 100μg/mL and that 30μg/mL of propolis reduced the radiation-induced DNA damage (∼56% reduction, p<0.05), exhibiting a radio-protective effect on irradiated CHO-K1 cells. On the other hand, analysis of cytotoxicity showed that a concentration of 50μg/mL presented a significant proliferative effect (p<0.001) when associated with radiation, decreasing the percentage of necrotic cells (p<0.01). No mediated cytotoxic effect was found, but the concentration of 200μg/mL was toxic when analyzed at 24 and 48h via the differential staining technique, but not at 72h after irradiation, analyzed with the MTS assay. Differential staining also showed that necrosis was the main death modality in irradiated cells and that apoptosis was induced only at the toxic concentration of propolis (200μg/mL). Concerning the clonogenic capacity, a concentration of 50μg/mL also exhibited a significant stimulating effect on cell proliferation (p<0.001), in agreement with the data from differential staining. Taken together, these data suggest that the use of propolis AF-08 for the prevention of the adverse effects of ionizing radiation is promising. Nevertheless, additional investigations are necessary for a better understanding of potential applications of propolis to improve human health.

  18. KEY COMPARISON: Update of the BIPM comparison BIPM.RI(II)-K1.F-18 of activity measurements of the radionuclide 18F to include the NPL

    NASA Astrophysics Data System (ADS)

    Ratel, G.; Michotte, C.; Woods, M. J.

    2003-01-01

    Since 2001, four national metrology institutes (NMIs) have submitted four samples of known activity of 18F to the International Reference System (SIR) for activity comparison at the Bureau International des Poids et Mesures, the most recent being that of the NPL (UK). The activities ranged from about 1 MBq to 8 MBq. The key comparison reference value (KCRV) has been recalculated to include the latest value and the degrees of equivalence between each equivalent activity measured in the SIR and the key comparison reference value (KCRV) have been calculated and the results are given in the form of a matrix. A graphical presentation is also given. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by Section II of the Consultative Committee for Ionizing Radiation (CCRI(II)), comparison identifier BIPM.RI(II)-K1.F-18, according to the provisions of the Mutual Recognition Arrangement (MRA).

  19. KEY COMPARISON: Update of the BIPM comparison BIPM.RI(II)-K1.F-18 of activity measurements of the radionuclide 18F to include the CIEMAT

    NASA Astrophysics Data System (ADS)

    Ratel, G.; Michotte, C.; García-Toraño, E.; Los Arcos, J.-M.

    2004-01-01

    Since 2001, five national metrology institutes (NMIs) have submitted five samples of known activity of 18F to the International Reference System (SIR) for activity comparison at the Bureau International des Poids et Mesures (BIPM), the most recent being that of the CIEMAT (Spain). The activities ranged from about 1 MBq to 18 MBq. The key comparison reference value (KCRV) has been recalculated to include the latest value and the degrees of equivalence between each equivalent activity measured in the SIR have been calculated and the results are given in the form of a matrix. A graphical presentation is also given for this key comparison with identifier BIPM.RI(II)-K1.F-18. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by Section II of the Consultative Committee for Ionizing Radiation (CCRI(II)), according to the provisions of the Mutual Recognition Arrangement (MRA).

  20. KEY COMPARISON: Update of the BIPM comparison BIPM.RI(II)-K1.F-18 of activity measurements of the radionuclide 18F to include the PTB

    NASA Astrophysics Data System (ADS)

    Ratel, G.; Michotte, C.; Kossert, K.; Janßen, H.

    2006-01-01

    Since 2001, six national metrology institutes (NMIs) have submitted six samples of known activity of 18F to the International Reference System (SIR) for activity comparison at the Bureau International des Poids et Mesures (BIPM), the most recent being that of the PTB (Germany). The activities ranged from about 1 MBq to 18 MBq. The key comparison reference value (KCRV) has been recalculated to include the latest value, with the agreement of the CCRI(II). The degrees of equivalence between each equivalent activity measured in the SIR have been recalculated and the results are given in the form of a matrix. A graphical presentation is also given for this key comparison with identifier BIPM.RI(II)-K1.F-18. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by Section II of the Consultative Committee for Ionizing Radiation (CCRI(II)), according to the provisions of the Mutual Recognition Arrangement (MRA).

  1. Glucose repression of lactose/galactose metabolism in Kluyveromyces lactis is determined by the concentration of the transcriptional activator LAC9 (K1GAL4) [corrected

    PubMed Central

    Zachariae, W; Kuger, P; Breunig, K D

    1993-01-01

    In the budding yeast Kluyveromyces lactis glucose repression of genes involved in lactose and galactose metabolism is primarily mediated by LAC9 (or K1GAL4) the homologue of the well-known Saccharomyces cerevisiae transcriptional activator GAL4. Phenotypic difference in glucose repression existing between natural strains are due to differences in the LAC9 gene (Breunig, 1989, Mol.Gen.Genet. 261, 422-427). Comparison between the LAC9 alleles of repressible and non-repressible strains revealed that the phenotype is a result of differences in LAC9 gene expression. A two-basepair alteration in the LAC9 promoter region produces a promoter-down effect resulting in slightly reduced LAC9 protein levels under all growth conditions tested. In glucose/galactose medium any change in LAC9 expression drastically affects expression of LAC9 controlled genes e.g. those encoding beta-galactosidase or galactokinase revealing a strong dependence of the kinetics of induction on the LAC9 concentration. We propose that in tightly repressible strains the activator concentration drops below a critical threshold that is required for induction to occur. A model is presented to explain how small differences in activator levels are amplified to produce big changes in expression levels of metabolic genes. Images PMID:8441621

  2. Frozen-in correlations in K1-x(NH4)xI mixed crystals: A Raman-scattering study

    NASA Astrophysics Data System (ADS)

    Berret, Jean-François; Sauvajol, Jean-Louis

    1994-06-01

    We report on low-frequency Raman-scattering results obtained on K1-x(NH4)xI mixed crystals and on its deuterated isomorph (x=0.46 and 0.62) as a function of temperature. With decreasing temperature, the Raman susceptibility is dominated by the growth and the narrowing of a strong component around 30 cm-1 and a much weaker one at 44 cm-1. A comparison of the present results with recent inelastic neutron scattering data on the same mixed crystals enabled us to assign the components at 30 and 44 cm-1 to the density of states of transverse and longitudinal acoustic phonons at the Brillouin-zone boundary. For the most prominent band, it is also shown that the drastic increase of the intensity coincides with the onset of freezing of antiferroelectric correlations evidenced by diffuse scattering measurements (TF=45 and 58 K for x=46% and 62%, respectively). The strong activity of the zone-boundary acoustic branches below TF is interpreted in terms of breaking of the local symmetry of the mixed crystal. Finally, the temperature dependences of the main Raman active components identified below 250 cm-1 are also discussed, with particular attention to aspects relevant to the freezing transition and to the orientational disorder.

  3. Superconducting specific-heat jump ΔCel∝Tcβ(β ≈2) for K1-xNaxFe2As2

    NASA Astrophysics Data System (ADS)

    Grinenko, V.; Efremov, D. V.; Drechsler, S.-L.; Aswartham, S.; Gruner, D.; Roslova, M.; Morozov, I.; Nenkov, K.; Wurmehl, S.; Wolter, A. U. B.; Holzapfel, B.; Büchner, B.

    2014-02-01

    We present a systematic study of the electronic specific-heat jump (ΔCel) at the superconducting transition temperature Tc of K1-xNaxFe2As2. Both Tc and ΔCel monotonously decrease with increasing x. The specific heat jump scales approximately with a power law, ΔCel∝Tcβ, with β ≈2 determined by the impurity scattering rate, in contrast to most iron-pnictide superconductors, where the remarkable Bud'ko-Ni-Canfield (BNC) scaling ΔCel∝T3 has been found. Both the T dependence of Cel(T ) in the superconducting state and the nearly quadratic scaling of ΔCel at Tc are well described by the Eliashberg theory for a two-band d-wave superconductor with weak pair breaking due to nonmagnetic impurities. The disorder induced by the Na substitution significantly suppresses the small gaps, leading to gapless states in the slightly disordered superconductor, which results in a large observed residual Sommerfeld coefficient in the superconducting state for x >0.

  4. Three predictions: Comet 67P/Churyumov-Gerasimenko, comet C/2012 K1 PANSTARRS, and comet C/2013 V5 Oukaimeden

    NASA Astrophysics Data System (ADS)

    Ferrín, Ignacio

    2014-09-01

    We make the following predictions: (1) The secular light curve (SLC) of comet 67P/Churyumov-Gerasimenko exhibits a photometric anomaly in magnitude that is present in 1982, 1996, 2002 and 2009. Thus it must be real. We interpret this anomaly as a topographic feature on the surface of the nucleus that may be a field of debris, a region made only of dust or an area of solid stones but in any case it is depleted in volatiles. We predict that images taken by spacecraft Rosetta will show a region morphologically different to the rest of the nucleus, at the pole pointing to the Sun near perihelion. (2) Comet C/2012 K1 PANSTARRS exhibits the same Slope Discontinuity Event (SDE)+magnitude dip after the event than other comets listed in Table 1 most of which disintegrated. This group includes comet C/2012 S1 ISON. Thus, it is reasonable to expect that this comet may disintegrate too. The probability of disintegration of this comet is 27%. (3) Comet C/2013 V5 Oukaimeden exhibits the same SDE+standstill signature exhibited by other comets in Table 1. We predict that there is a 93% probability that this comet will disintegrate. (4) Another purpose of this work is to present evidence to conclude that the SLCs have predictive power.

  5. Investigation of diffuse phase transition in ferroelectric Pb2- x K1+ x Li x Nb5O15 (0 ≤ x ≤ 1.5) ceramics

    NASA Astrophysics Data System (ADS)

    Choukri, E.; Neqali, A.; Abkhar, Z.; Alimoussa, A.; Hajji, L.; Mezzane, D.; Belboukhari, A.; Amjoud, M.; Gagou, Y.; El Marssi, M.; Luk'yanchuk, I.

    2016-06-01

    Substitution of Pb with Li and K in the Pb2KNb5O15 phases leads to a new composition with chemical composition Pb2- x K1+ x Li x Nb5O15 which crystallizes with tetragonal tungsten bronze-type structure. Ferroelectric ceramics with different compositions were synthesized using solid-state reaction and complex dielectric permittivity measurements in these compounds were performed in a frequency and temperature range of 20 Hz-1 MHz and from 25 to 550 °C, respectively. Special attention was paid to the diffuse phase transition (DPT) that occurs close to the Curie temperature. The empirical equation proposed by Santos-Eiras for a phenomenological description of the temperature dependence of the dielectric permittivity (\\varepsilon_{{r}}^' }}) peak is used to calculate some characteristic parameters of DPT. From the results, it must be assumed that these compounds show a diffuse phase transition with non-relaxor behavior. A basic phase diagram showing the evolution of T m function of composition x is deduced from this study.

  6. Virulence patterns and long-range genetic mapping of extraintestinal Escherichia coli K1, K5, and K100 isolates: use of pulsed-field gel electrophoresis.

    PubMed Central

    Ott, M; Bender, L; Blum, G; Schmittroth, M; Achtman, M; Tschäpe, H; Hacker, J

    1991-01-01

    A total of 127 extraintestinal Escherichia coli strains of the capsule serotypes K1, K5, and K100 from human and animal sources were analyzed for DNA sequences specific for the genes for various adhesins (P fimbriae [pap] and P-related sequences [prs], S fimbriae [sfa]/F1C fimbriae [foc], and type I fimbriae [fim]), aerobactin (aer), and hemolysin (hly). The expression of corresponding virulence factors was also tested. Twenty-four selected strains were analyzed by long-range DNA mapping to evaluate their genetic relationships. DNA sequences for the adhesins were often found in strains not expressing them, while strains with hemolysin and aerobactin genes usually did express them. Different isolates of the same serotype often expressed different virulence patterns. The use of virulence-associated gene probes for Southern hybridization with genomic DNA fragments separated by pulsed-field gel electrophoresis revealed that a highly heterogeneous restriction fragment length and hybridization pattern existed even within strains of the same serotype. Long-range DNA mapping is therefore useful for the evaluation of genetic relatedness among individual isolates and facilitates the performance of precise molecular epidemiology. Images PMID:1677349

  7. Comparative study of the cytotoxicity and genotoxicity of kaurenoic acid and its semi-synthetic derivatives methoxy kaurenoic acid and kaurenol in CHO-K1 cells.

    PubMed

    Cano, Bruno Limonti; Moreira, Monique Rodrigues; Goulart, Mirian Oliveira; Dos Santos Gonçalves, Natália; Veneziani, Rodrigo Cassio Sola; Bastos, Jairo Kenupp; Ambrósio, Sérgio Ricardo; Dos Santos, Raquel Alves

    2017-04-01

    The diterpene kaurenoic acid (KA) has vasorelaxant, antimicrobial, anti-tumoural and anti-leishmanial effects. Semi-synthetic derivatives were obtained to achieve more satisfactory responses. The assessment of genotoxicity is part of the toxicological evaluation of therapeutic compound candidates. The present study investigated the cytotoxicity and genotoxicity of KA and its semi-synthetic derivatives methoxy kaurenoic acid (MKA) and kaurenol (KRN) using the CHO-K1 cell line. The cytotoxicity evaluation demonstrated that treatments with 200 and 400 μM KA reduced cellular proliferation to 36.5 and 4.43%, respectively, and that 100 and 200 μM KA reduced the survival fraction (SF) to 48.1 and 5.5%, respectively. MKA and KRN at concentrations of 400 μM reduced proliferation to 81 and 86.8%, respectively, while 100 and 200 μM KRN reduced the SF to 50%, and 200 μM MKA reduced the SF to 74%. No genotoxicity was observed for KA or MKA. However, 100 μM KRN increased the DNA damage index, as detected by comet assay, although a micronucleus assay did not confirm these data. The results demonstrated that KA and its semi-synthetic derivative MKA were not genotoxic when tested at noncytotoxic concentrations, but KRN was genotoxic at the highest concentration that was tested, as demonstrated by the comet assay.

  8. Cephalosporin-induced alteration in hepatic glutathione redox state. A potential mechanism for inhibition of hepatic reduction of vitamin K1,2,3-epoxide in the rat.

    PubMed Central

    Mitchell, M C; Mallat, A; Lipsky, J J

    1990-01-01

    Hypoprothrombinemia is a serious adverse effect of antimicrobial therapy that occurs after administration of some second- and third-generation cephalosporins which contain the methyltetrazole-thiol (MTT) group. Previous studies have shown that in vitro MTT directly inhibits microsomal gamma-carboxylation of a synthetic pentapeptide. Since MTT is a thiocarbamide, a type of compound that can increase oxidation of glutathione, the present studies were carried out to determine whether alterations in hepatic glutathione redox state might interfere with vitamin K metabolism. Dose-related increases in biliary efflux and hepatic concentration of oxidized glutathione (GSSG) occurred after intravenous administration of MTT or MTT-containing antibiotics to rats. This finding suggested that these compounds could alter the hepatic glutathione redox state in vivo. Microsomal reduction of vitamin K epoxide occurred in the presence of 100 microM dithiothreitol (DTT), but was inhibited by preincubation with GSSG at concentrations as low as 10 microM. At higher concentrations of DTT (1.0 mM) inhibition by GSSG persisted, but higher concentrations were required, suggesting that the thiol/disulfide ratio, rather than the absolute concentration of GSSG was important. By contrast, GSSG did not effect microsomal gamma-carboxylation of a pentapeptide, using either vitamin K1 or its hydroquinone as a cofactor. These findings suggest a novel mechanism for the hypoprothrombinemia occurring after administration of MTT-containing antibiotics. PMID:1978724

  9. Mutually exclusive recurrent somatic mutations in MAP2K1 and BRAF support a central role for ERK activation in LCH pathogenesis

    PubMed Central

    Chakraborty, Rikhia; Hampton, Oliver A.; Shen, Xiaoyun; Simko, Stephen J.; Shih, Albert; Abhyankar, Harshal; Lim, Karen Phaik Har; Covington, Kyle R.; Trevino, Lisa; Dewal, Ninad; Muzny, Donna M.; Doddapaneni, Harshavardhan; Hu, Jianhong; Wang, Linghua; Lupo, Philip J.; Hicks, M. John; Bonilla, Diana L.; Dwyer, Karen C.; Berres, Marie-Luise; Poulikakos, Poulikos I.; Merad, Miriam; McClain, Kenneth L.; Wheeler, David A.

    2014-01-01

    Langerhans cell histiocytosis (LCH) is a myeloproliferative disorder characterized by lesions composed of pathological CD207+ dendritic cells with an inflammatory infiltrate. BRAFV600E remains the only recurrent mutation reported in LCH. In order to evaluate the spectrum of somatic mutations in LCH, whole exome sequencing was performed on matched LCH and normal tissue samples obtained from 41 patients. Lesions from other histiocytic disorders, juvenile xanthogranuloma, Erdheim-Chester disease, and Rosai-Dorfman disease were also evaluated. All of the lesions from histiocytic disorders were characterized by an extremely low overall rate of somatic mutations. Notably, 33% (7/21) of LCH cases with wild-type BRAF and none (0/20) with BRAFV600E harbored somatic mutations in MAP2K1 (6 in-frame deletions and 1 missense mutation) that induced extracellular signal-regulated kinase (ERK) phosphorylation in vitro. Single cases of somatic mutations of the mitogen-activated protein kinase (MAPK) pathway genes ARAF and ERBB3 were also detected. The ability of MAPK pathway inhibitors to suppress MAPK kinase and ERK phosphorylation in cell culture and primary tumor models was dependent on the specific LCH mutation. The findings of this study support a model in which ERK activation is a universal end point in LCH arising from pathological activation of upstream signaling proteins. PMID:25202140

  10. Solid solution formation in the new cuprate system (Sr, K) 1- x(Ca, Na) xCuO 2± z (0≤ x≤0.5)

    NASA Astrophysics Data System (ADS)

    Chavira, E.; Hernández, E.; Navarro, O.; Baños, L.; Guzmán, J.

    1999-10-01

    Synthesis and some experimental results of the new cuprate system (Sr,K) 1- x(Ca,Na) xCuO 2± z(0.0≤ x≤1.0) are reported for the first time. We prepared a series of polycrystalline samples of this system with x-steps of 0.1, by solid-state reaction method in air at ambient pressure below melt temperature. It is observed the formation of a solid solution in the range 0≤ x≤0.5 at 750°C. The characterization of the samples by X-ray powder diffraction indicates that the single phase is isostructural to the one for Cu 2SrO 3 compound, reported in the JCPDS-International Center for Diffraction Data (ICDD) file No. 39-0250, which has an orthorhombic unit cell. The compositions in the range 0.6≤ x≤1.0 have an unknown phase and CuO. Further characterization studies by scanning electron microscopy, thermogravimetric and differential thermal analysis are shown.

  11. Simultaneous estimation of vitamin K1 and heparin with low limit of detection using cascaded channels fiber optic surface plasmon resonance.

    PubMed

    Tabassum, Rana; Gupta, Banshi D

    2016-12-15

    We report an approach for the simultaneous estimation of vitamin K1 (VK1) and heparin via cascaded channel multianalyte sensing probe employing fiber optic surface plasmon resonance technique. Cladding from two well separated portions of the fiber is removed and are respectively coated with thin films of silver (channel-1) and copper (channel-2). The nanohybrid of multiwalled carbon nanotube in chitosan is fabricated over silver layer for the sensing of VK1 whereas core shell nanostructure of polybrene@ZnO is coated over copper layer for the sensing of heparin. Spectral interrogation method is used for the characterization of the sensor. Analyte selectivity of both the channels is performed by carrying out experiments using independent solutions of VK1 and heparin. Experiments performed on the solution of the mixture of VK1 and heparin show red shifts in both the channels on changing the concentration of both the analytes in the mixture. The operating range of both VK1 and heparin is from 0 to 10(-3)g/l. The limit of detection of the sensor is 2.66×10(-4)µg/l and 2.88×10(-4)µg/l for VK1 and heparin respectively which are lower than the reported ones. The additional advantages of the present sensor are low cost, possibility of online monitoring and remote sensing.

  12. The interaction of N-glycans in Fcγ receptor I α-chain with Escherichia coli K1 outer membrane protein A for entry into macrophages: experimental and computational analysis.

    PubMed

    Krishnan, Subramanian; Liu, Fan; Abrol, Ravinder; Hodges, Jacqueline; Goddard, William A; Prasadarao, Nemani V

    2014-11-07

    Neonatal meningitis, caused by Escherichia coli K1, is a serious central nervous system disease. We have established that macrophages serve as permissive niches for E. coli K1 to multiply in the host and for attaining a threshold level of bacterial load, which is a prerequisite for the onset of the disease. Here, we demonstrate experimentally that three N-glycans in FcγRIa interact with OmpA of E. coli K1 for binding to and entering the macrophages. Adoptive transfer of FcγRIa(-/-) bone marrow-derived macrophages transfected with FcγRIa into FcγRIa(-/-) newborn mice renders them susceptible to E. coli K1-induced meningitis. In contrast, mice that received bone marrow-derived macrophages transfected with FcγRIa in which N-glycosylation sites 1, 4, and 5 are mutated to alanines exhibit resistance to E. coli K1 infection. Our molecular dynamics and simulation studies predict that N-glycan 5 exhibits strong binding at the barrel site of OmpA formed by loops 3 and 4, whereas N-glycans 1 and 4 interact with loops 1, 3, and 4 of OmpA at tip regions. Molecular modeling data also suggest no role for the IgG binding site in the invasion process. In agreement, experimental mutations in IgG binding site had no effect on the E. coli K1 entry into macrophages in vitro or on the onset of meningitis in newborn mice. Together, this integration of experimental and computational studies reveals how the N-glycans in FcγRIa interact with the OmpA of E. coli K1 for inducing the disease pathogenesis.

  13. Determination of trans vitamin K1 in infant and medical nutritional products using AOAC Method 999.15 with modified preparation and extraction procedures and C30 bonded phase chromatography: single-laboratory validation.

    PubMed

    Schimpf, Karen J; Thompson, Linda B; Schmitz, Daniel J

    2010-01-01

    Modifications were made to AOAC Official Method 999.15 to extend its applicability to specialty infant formulas containing hydrolyzed proteins and free amino acids, and to medical and adult nutritional products. Minor changes to the sample preparation procedure and chromatographic separation improved vitamin K1 recoveries and reduced chromatographic interferences in these types of matrixes. Currently AOAC Method 999.15 is applicable only to the determination of total vitamin K1 (phylloquione) in infant formula and milk (fluid, ready-to-feed, and powdered) containing > 1 microg vitamin K1/100 g solids. AOAC Method 999.15 recoveries of vitamin K1 were improved by altering sample sizes, extraction solvents and amounts, and the reagent addition order and amount of water or aqueous solutions added. The chromatographic separation of vitamin K1 in medical nutritional products containing canola and marine oils was improved, and trans vitamin K1 was separated from the biologically inactive cis isomer in all products with a C30 3 microm column and a 100% methanol mobile phase. With these modifications to the extraction procedure and chromatographic separation, AOAC Method 999.15 demonstrated acceptable precision and accuracy for the quantitation of trans vitamin K1 in specialty infant formulas containing hydrolyzed proteins and free amino acids, and medical and adult nutritional products. A single-laboratory validation of these minor modifications was completed. Fourteen different product matrixes were analyzed during validation. The intermediate precision averaged 4.15% RSD (range 2.52-5.81% RSD), and recovery data averaged 100.1% (range 92.2-109%).

  14. Identification of Restriction Factors by Human Genome-Wide RNA Interference Screening of Viral Host Range Mutants Exemplified by Discovery of SAMD9 and WDR6 as Inhibitors of the Vaccinia Virus K1L−C7L− Mutant

    PubMed Central

    Sivan, Gilad; Ormanoglu, Pinar; Buehler, Eugen C.; Martin, Scott E.

    2015-01-01

    ABSTRACT RNA interference (RNAi) screens intended to identify host factors that restrict virus replication may fail if the virus already counteracts host defense mechanisms. To overcome this limitation, we are investigating the use of viral host range mutants that exhibit impaired replication in nonpermissive cells. A vaccinia virus (VACV) mutant with a deletion of both the C7L and K1L genes, K1L−C7L−, which abrogates replication in human cells at a step prior to late gene expression, was chosen for this strategy. We carried out a human genome-wide small interfering RNA (siRNA) screen in HeLa cells infected with a VACV K1L−C7L− mutant that expresses the green fluorescent protein regulated by a late promoter. This positive-selection screen had remarkably low background levels and resulted in the identification of a few cellular genes, notably SAMD9 and WDR6, from approximately 20,000 tested that dramatically enhanced green fluorescent protein expression. Replication of the mutant virus was enabled by multiple siRNAs to SAMD9 or WDR6. Moreover, SAMD9 and WDR6 clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 knockout HeLa cell lines were permissive for replication of the K1L−C7L− mutant, in agreement with the siRNA data. Expression of exogenous SAMD9 or interferon regulatory factor 1 restricted replication of the K1L−C7L− mutant in the SAMD9−/− cells. Independent interactions of SAMD9 with the K1 and C7 proteins were suggested by immunoprecipitation. Knockout of WDR6 did not reduce the levels of SAMD9 and interactions of WDR6 with SAMD9, C7, and K1 proteins were not detected, suggesting that these restriction factors act independently but possibly in the same innate defense pathway. PMID:26242627

  15. Vertical Extraction Process Implemented at the 118-K-1 Burial Ground for Removal of Irradiated Reactor Debris from Silo Structures - 12431

    SciTech Connect

    Teachout, Douglas B.; Adamson, Clinton J.; Zacharias, Ames

    2012-07-01

    The primary objective of a remediation project is the safe extraction and disposition of diverse waste forms and materials. Remediation of a solid waste burial ground containing reactor hardware and irradiated debris involves handling waste with the potential to expose workers to significantly elevated dose rates. Therefore, a major challenge confronted by any remediation project is developing work processes that facilitate compliant waste management practices while at the same time implementing controls to protect personnel. Traditional burial ground remediation is accomplished using standard excavators to remove materials from trenches and other excavation configurations often times with minimal knowledge of waste that will be encountered at a specific location. In the case of the 118-K-1 burial ground the isotopic activity postulated in historic documents to be contained in vertical cylindrical silos was sufficient to create the potential for a significant radiation hazard to project personnel. Additionally, certain reported waste forms posed an unacceptably high potential to contaminate the surrounding environment and/or workers. Based on process knowledge, waste management requirements, historic document review, and a lack of characterization data it was determined that traditional excavation techniques applied to remediation of vertical silos would expose workers to unacceptable risk. The challenging task for the 118-K-1 burial ground remediation project team then became defining an acceptable replacement technology or modification of an existing technology to complete the silo remediation. Early characterization data provided a good tool for evaluating the location of potential high exposure rate items in the silos. Quantitative characterization was a different case and proved difficult because of the large diameter of the silos and the potential for variable density of attenuating soils and waste forms in the silo. Consequently, the most relevant

  16. UV/Visible Observations of C/2012 K1 PanSTARRS and C/2013 A1 Siding Spring from a Stratospheric Telescope

    NASA Astrophysics Data System (ADS)

    Young, E. F.; Diller, J.; Dinkel, K.; Dischner, Z.; Cheng, A. F.; Hibbitts, C.; Osterman, S. N.

    2014-12-01

    The UV-VIS (Ultra-Violet/Visible) instrument on the BOPPS mission (Balloon Observation Platform for Planetary Science) has two main goals: to demonstrate pointing stability at the 0.1" level, commensurate with the near-space PSF (Point Spread Function) expected from the 80 cm aperture telescope, and to observe targets in wavelengths where the telluric transmission is low. Two potential targets of a BOPPS flight in September 2014 are comets C/2012 K1 PanSTARRS and C/2013 A1 Siding Spring. The UV-VIS science camera has been outfitted with four filters from the NASA's Hale-Bopp filter set: an OH filter at 308 nm, a CN filter at 385 nm and continuum filters at 345 and 445 nm. UV-VIS can potentially measure OH emission from the cometary targets if the BOPPS flight extends past sunset; the sky background due to Rayleigh scattering at 310 nm is too high for daytime observations, even from float altitudes near 35 km. The BOPPS telescope is stabilized at the few arcsecond level by inertial-guidance gyroscopes that control coarse elevation and azimuth reaction wheels. The UV-VIS bench includes a fast guide camera and an FSM (fine steering mirror) - both COTS items - to bridge the pointing gap from a few arcseconds to a tenth of an arcsecond. In two separate hang tests, the guide camera was able to provide a 50 Hz pointing error signal on a star, and the resulting FSM-corrected images were reduced in areal size by 50% (e.g., linear PSF widths reduced from 2.4" to 1.6"). The FSM performance is expected to achieve the 0.1" stability goal in the stratosphere, where the image degradation is caused by motion of the optical system instead of atmospheric turbulence.

  17. N- and 6-O-Sulfated Heparan Sulfates Mediate Internalization of Coxsackievirus B3 Variant PD into CHO-K1 Cells

    PubMed Central

    Zautner, Andreas E.; Jahn, Birgit; Hammerschmidt, Elke; Wutzler, Peter; Schmidtke, Michaela

    2006-01-01

    Recently, it was demonstrated that the coxsackievirus B3 variant PD (CVB3 PD) is able to infect coxsackievirus-adenovirus receptor (CAR)-lacking cells by using heparan sulfates (HS) as additional receptors (A. E. Zautner, U. Korner, A. Henke, C. Badorff, and M. Schmidtke, J. Virol. 77:10071-10077, 2003). For this study, competition experiments with growth factors binding to known HS sequences as well as with specifically desulfated heparins were performed with Chinese hamster ovary cells (CHO-K1) to determine the structural requirements of HS for interaction with CVB3. Hepatocyte growth factor interacting with HS sequences containing [IdUA-GlcNSO3(6OSO3)]n, but not basic fibroblast growth factor binding to [HexUA-GlcNSO3-HexUA-GlcNSO3-IdUA(2OSO3)]n, was shown to compete effectively with CVB3 PD for cell surface HS. Whereas unmodified heparin and 2-O-desulfated heparin strongly inhibited the CVB3 PD-induced cytopathic effect, the antiviral activity was markedly reduced after N-, O- and 6-O-desulfation of heparin. Taken together, these results indicate that 6-O- and N-sulfation of GlcNAc of HS is crucial for HS interaction with CVB3 PD and that the disaccharide [IdUA-GlcNSO3(6OSO3)]n is involved in viral binding. Results from experiments with various inhibitors of endocytic pathways suggest that HS-mediated virus internalization is pH dependent. Despite the fact that CVB3 PD initiates infection about four times slower by making use of HS as a receptor than by using CAR, the time required for a complete viral life cycle in Chinese hamster ovary cells was independent of the utilized receptor. PMID:16775350

  18. Crystal structure and phase transition behavior in (K1-xNax)NbO3-based lead-free piezoelectric ceramic over a wide range of temperatures

    NASA Astrophysics Data System (ADS)

    Yamada, H.; Matsuoka, T.; Kozuka, H.; Yamazaki, M.; Ohbayashi, K.; Ida, T.

    2016-12-01

    The phase transition temperature in a (K, Na)NbO3 (KNN) phase of a KNN-based composite lead-free piezoelectric ceramic with a KTiNbO5 system ((K1-xNax)0.86Ca0.04Li0.02Nb0.85O3-δ-K0.85Ti0.85Nb1.15O5-BaZrO3-Co3O4-Fe2O3-ZnO) is lower than that in an undoped KNN ceramic by approximately 200 °C. We have studied the structural changes around the phase transition by using synchrotron powder X-ray diffraction and transmission electron microscopy. The crystal system of the main KNN phase is assigned to tetragonal as a stable structure at room temperature and does not change to orthorhombic on lowering the temperature all at once. The crystal structure changes from tetragonal to orthorhombic through the successive transition state. The curve of the phase transition temperature from x = 0.33 to 0.75 has a V shape and reaches its lowest value of approximately 0 °C in the vicinity of x = 0.56. From selected-area electron diffraction patterns of the KNN phase, weak superlattice spots owing to the tilt-ordered NbO6 octahedra are observed for x ≥ 0.56. This tilt-ordered NbO6 octahedral phase is formed at the nanometer-scale (nanodomains) in the tetragonal and orthorhombic KNN matrices, regardless of the phase transition. The minimum x to generate the nanodomains is substantially equal to the Na fraction at which the starting temperature of the successive phase transition shifts to the lowest.

  19. [Construction of mammalian cell expression vector for pAcGFP-bFADD fusion protein and its expression in CHO-K1 cell].

    PubMed

    Yang, Runjun; Xu, Shangzhong; Zhang, Lupei; Li, Junya; Gao, Xue

    2008-11-01

    Fas-associated death domain (FADD) is a signal connection protein in Fas/FasL apoptotic path which might play a key role on apoptosis by transferring apoptotic signal. To reveal the intracellular signal transduction molecules involved in the procedure of follicular development in bovine ovary, we cloned FADD gene in bovine ovary tissue with RT-PCR, deleted the termination codon in its cDNA and directionally cloned the amplified FADD gene into eukaryotic expression vector pAcGFP-N1 including AcGFP, successfully constructed the fusion protein recombinant plasmid. After identifying by restrictive enzyme Bgl II/EcoR I and sequencing, transfected pAcGFP-bFADD into CHO-K1 cell mediated by Lipofectamine 2000, observed the expression of AcGFP and detected the transcription and expression of FADD by RT-PCR and Western blotting. The results showed that the cattle FADD was successfully cloned, the pAcGFP-bFADD fusion protein recombinant plasmid was successfully constructed by introducing Bgl II, EcoR I cloning site at two ends of FADD open reading frame and inserting a Kozak sequence before start codon. AcGFP expression was detected as early as 24 h after transfection. The percentage of AcGFP positive cells reached about 65% after 24 h. A 654 bp transcription was amplified by RT-PCR, and 51.4 kD target protein was detected by Western blotting. Construction of pAcGFP-bFADD recombinant plasmid should be helpful for further understanding the mechanism of regulation of FADD on bovine oocytes formation and development.

  20. Multiplex polymerase chain reaction-based deletion analysis of spontaneous, gamma ray- and alpha-induced hprt mutants of CHO-K1 cells.

    PubMed

    Schwartz, J L; Rotmensch, J; Sun, J; An, J; Xu, Z; Yu, Y; Hsie, A W

    1994-11-01

    Independent Chinese hamster ovary (CHO)-K1 cell mutants at the hypoxanthine-guanine phosphoribosyltransferase (hprt) locus were isolated from untreated, 60Co gamma ray- and 212Bi alpha-exposed cells and the genetic changes underlying the mutation determined by multiplex polymerase chain reaction (PCR)-based exon deletion analysis. In the 71 spontaneous mutants analyzed, 77.5% of the clones showed no change in exon number or size, 15.5% showed a loss of a single exon, 4.2% showed a loss of 2-8 exons, and 2.8% showed loss of all nine hprt exons (total gene deletion). Exposure to 6 Gy of gamma rays, which reduced survival levels to 10%, produced a significantly different deletion spectrum that was shifted toward deletions with 45% of the 20 mutants analyzed showing a loss of a single exon and 30% showing a loss of all nine exons. Exposure to 2 Gy alpha radiation from 212Bi, a 220Rn daughter, a dose which also reduced survival levels to about 10%, resulted in a deletion spectrum similar to the gamma-ray spectrum in that more than 75% of the 49 mutants analyzed were deletions. The alpha spectrum, however, was significantly different from both the spontaneous and gamma spectra with 55.1% of the alpha mutants showing a loss of all nine exons, 10.2% showing loss of a single exon, and 14.3% showing loss of 2-8 exons. Thus, alpha-radiation appears to produce larger intragenic deletions than gamma radiation. The results suggest that intragenic deletion size should be considered when low- and high linear energy transfer (LET) mutation spectra are compared.

  1. Thermal Physics of the Inner Coma: ALMA Studies of the Methanol Distribution and Excitation in Comet C/2012 K1 (PanSTARRS)

    NASA Astrophysics Data System (ADS)

    Cordiner, M. A.; Biver, N.; Crovisier, J.; Bockelée-Morvan, D.; Mumma, M. J.; Charnley, S. B.; Villanueva, G.; Paganini, L.; Lis, D. C.; Milam, S. N.; Remijan, A. J.; Coulson, I. M.; Kuan, Y.-J.; Boissier, J.

    2017-03-01

    We present spatially and spectrally resolved observations of CH3OH emission from comet C/2012 K1 (PanSTARRS), using the Atacama Large Millimeter/submillimeter Array on 2014 June 28–29. Two-dimensional maps of the line-of-sight average rotational temperature (T rot) were derived, covering spatial scales 0.″3–1.″8 (corresponding to sky-projected distances ρ ∼ 500–2500 km). The CH3OH column density distributions are consistent with isotropic, uniform outflow from the nucleus, with no evidence for extended sources of CH3OH in the coma. The {T}{rot}(ρ ) radial profiles show a significant drop within a few thousand kilometers of the nucleus, falling from about 60 to 20 K between ρ =0 and 2500 km on June 28, whereas on June 29, T rot fell from about 120 to 40 K between ρ = 0 km and 1000 km. The observed T rot behavior is interpreted primarily as a result of variations in the coma kinetic temperature due to adiabatic cooling of the outflowing gas, as well as radiative cooling of the CH3OH rotational levels. Our excitation model shows that radiative cooling is more important for the J=7-6 transitions (at 338 GHz) than for the K=3-2 transitions (at 252 GHz), resulting in a strongly sub-thermal distribution of levels in the J=7-6 band at ρ ≳ 1000 {km}. For both bands, the observed temperature drop with distance is less steep than predicted by standard coma theoretical models, which suggests the presence of a significant source of heating in addition to the photolytic heat sources usually considered.

  2. Polymerase chain reaction-deletion analysis of spontaneous, gamma ray-, and alpha-induced hprt mutants of CHO-K1 cells.

    SciTech Connect

    Schwartz, J. L.; Rotmensch, J.; Sun, J.; An, J.; Xu, Z.; Yu, Y.; Hsie, A. W.; Center for Mechanistic Biology and Biotechnology; Univ. of Chicago; Univ. of Texas Medical Branch

    1994-01-01

    Independent Chinese hamster ovary (CHO)-K1 cell mutants at the hypoxanthine-guanine phosphoribosyltransferase (hprt) locus were isolated from untreated, {sup 60}Co {gamma} ray-and {sup 212}Bi {alpha}-exposed cells and the genetic changes underlying the mutation determined by multiplex polymerase chain reaction (PCR)-based exon deletion analysis. In the 71 spontaneous mutants analyzed, 77.5% of the clones showed no change in exon number or size, 15.5% showed a loss of a single exon, 4.2% showed a loss of 2-8 exons, and 2.8% showed loss of all nine hprt exons (total gene deletion). Exposure to 6 Gy of {gamma} rays, which reduced survival levels to 10%, produced a significantly different deletion spectrum that was shifted toward deletions with 45% of the 20 mutants analyzed showing a loss of a single exon and 30% showing a loss of all nine exons. Exposure to 2 Gy {alpha} radiation from 212Bi, a 220Rn daughter, a dose which also reduced survival levels to about 10%, resulted in a deletion spectrum similar to the {gamma}-ray spectrum in that more than 75% of the 49 mutants analyzed were deletions. The {alpha} spectrum, however, was significantly different from both the spontaneous and {gamma} spectra with 55.1% of the {alpha} mutants showing a loss of all nine exons, 10.2% showing loss of a single exon, and 14.3% showing loss of 2-8 exons. Thus, {alpha}-radiation appears to produce larger intragenic deletions than {gamma} radiation. The results suggest that intragenic deletion size should be considered when low- and high linear energy transfer (LET) mutation spectra are compared.

  3. Summer (subarctic) versus winter (subtropic) production affects spinach (Spinacia oleracea L.) leaf bionutrients: vitamins (C, E, Folate, K1, provitamin A), lutein, phenolics, and antioxidants.

    PubMed

    Lester, Gene E; Makus, Donald J; Hodges, D Mark; Jifon, John L

    2013-07-24

    Comparison of spinach (Spinacia oleracea L.) cultivars Lazio and Samish grown during the summer solstice in the subarctic versus the winter solstice in the subtropics provided insight into interactions between production environment (light intensity), cultivar, and leaf age/maturity/position affecting bionutrient concentrations of vitamins (C, E, folate, K1, provitamin A), lutein, phenolics, and antioxidants. Growing spinach during the winter solstice in the subtropics resulted in increased leaf dry matter %, oxidized (dehydro) ascorbic acid (AsA), α- and γ-tocopherol, and total phenols but lower reduced (free) AsA, α-carotene, folate, and antioxidant capacity compared to summer solstice-grown spinac