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Sample records for alt immortalization reveals

  1. A gene expression signature classifying telomerase and ALT immortalization reveals an hTERT regulatory network and suggests a mesenchymal stem cell origin for ALT

    DEFF Research Database (Denmark)

    Lafferty-Whyte, K; Cairney, C J; Will, M B

    2009-01-01

    Telomere length is maintained by two known mechanisms, the activation of telomerase or alternative lengthening of telomeres (ALT). The molecular mechanisms regulating the ALT phenotype are poorly understood and it is unknown how the decision of which pathway to activate is made at the cellular le...

  2. ALT/Space

    Science.gov (United States)

    Rosenfeld, Malke, Ed.; Conarro, Ryan; Upshaw, Allison; Makol, Suzanne; Kelin, Daniel A., II; Redman, Jeff

    2013-01-01

    Stories in the "ALT/Space" section of each issue of "Teaching Artist Journal" illustrate and document a wide variety of topics surrounding the work of teaching artists while simultaneously revealing some larger truths about what it means to be an artist who teaches. This particular section focuses on the process and realities…

  3. Alanine transaminase (ALT) blood test

    Science.gov (United States)

    ... gov/ency/article/003473.htm Alanine transaminase (ALT) blood test To use the sharing features on this page, please enable JavaScript. The alanine transaminase (ALT) blood test measures the level of the enzyme ALT in ...

  4. Analysis of copy number variation in 8,842 Korean individuals reveals 39 genes associated with hepatic biomarkers AST and ALT.

    Science.gov (United States)

    Kim, Hyo Young; Cho, Seoae; Yu, Jeongmi; Sung, Samsun; Kim, Heebal

    2010-08-01

    Biochemical tests such as aspartate aminotransferase (AST) and alanine aminotransferase (ALT) are useful for diagnosing patients with liver disease. In this study, we tested the association between copy number variation and the hepatic biomarkers AST and ALT based on 8,842 samples from population-based cohorts in Korea. We used Affymetrix Genome-Wide Human 5.0 arrays and identified 10,534 CNVs using HelixTree software. Of the CNVs tested using univariate linear regression, 100 CNVs were significant for AST and 16 were significant for ALT (P < 0.05). We identified 39 genes located within the CNV regions. DKK1 and HS3ST3B1 were shown to play roles in heparan sulfate biosynthesis and the Wnt signaling pathway, respectively. NAF1 and NPY1R were associated with glycoprotein processes and neuropeptide Y receptor activity based on GO categories. PTER, SOX14 and TM7SF4 were expressed in liver. DPYS and CTSC were found to be associated with dihydropyrimidinuria and Papillon-Lefevre syndrome phenotypes using OMIM. NPY5R was found to be associated with dyslipidemia using the Genetic Association Database.

  5. Alt. Publicidad Online

    OpenAIRE

    Gómez Benavent, Elena

    2014-01-01

    Treball Final de Grau en Publicitat i Relacions Públiques. Codi: PU0932. Curs: 2013-2014 Alt Publicidad Online es un proyecto de gestión de redes sociales para pequeñas y medianas empresas que apuestan por el mundo online como herramienta esencial para sus negocios. El objetivo principal del trabajo de fin de grado es mostrar la capacidad de las PYMES de hacerse grandes y crecer frente a sus competidores gracias a la calidad y la innovación. Las redes sociales se inician ...

  6. Aerodynamic challenges of ALT

    Science.gov (United States)

    Hooks, I.; Homan, D.; Romere, P. O.

    1985-01-01

    The approach and landing test (ALT) of the Space Shuttle Orbiter presented a number of unique challenges in the area of aerodynamics. The purpose of the ALT program was both to confirm the use of the Boeing 747 as a transport vehicle for ferrying the Orbiter across the country and to demonstrate the flight characteristics of the Orbiter in its approach and landing phase. Concerns for structural fatigue and performance dictated a tailcone be attached to the Orbiter for ferry and for the initial landing tests. The Orbiter with a tailcone attached presented additional challenges to the normal aft sting concept of wind tunnel testing. The landing tests required that the Orbiter be separated from the 747 at approximately 20,000 feet using aerodynamic forces to fly the vehicles apart. The concept required a complex test program to determine the relative effects of the two vehicles on each other. Also of concern, and tested, was the vortex wake created by the 747 and the means for the Orbiter to avoid it following separation.

  7. Exploring the motifs of death and immortality | Maina | Journal of ...

    African Journals Online (AJOL)

    felt threatened by the eventuality of death, inculcating in them a fear so great that all possible strategies are engaged in the search for an avenue that would prepare them for this eventuality. A careful exploration of human activities surrounding the issues of death and immortality reveals an obsession with the expression of ...

  8. The Role of ATRX in the Alternative Lengthening of Telomeres (ALT) Phenotype.

    Science.gov (United States)

    Amorim, João P; Santos, Gustavo; Vinagre, João; Soares, Paula

    2016-09-19

    Telomeres are responsible for protecting chromosome ends in order to prevent the loss of coding DNA. Their maintenance is required for achieving immortality by neoplastic cells and can occur by upregulation of the telomerase enzyme or through a homologous recombination-associated process, the alternative lengthening of telomeres (ALT). The precise mechanisms that govern the activation of ALT or telomerase in tumor cells are not fully understood, although cellular origin may favor one of the other mechanisms that have been found thus far in mutual exclusivity. Specific mutational events influence ALT activation and maintenance: a unifying frequent feature of tumors that acquire this phenotype are the recurrent mutations of the Alpha Thalassemia/Mental Retardation Syndrome X-Linked (ATRX) or Death-Domain Associated Protein (DAXX) genes. This review summarizes the established criteria about this phenotype: its prevalence, theoretical molecular mechanisms and relation with ATRX, DAXX and other proteins (directly or indirectly interacting and resulting in the ALT phenotype).

  9. Dissecting the Immune Response Elicited by WbALT-2, ALT MAP in Clinical Populations and Mouse Model: A Prophylactic Measure Against Lymphatic Filariasis.

    Science.gov (United States)

    Ramanathan, Aparnaa; Immanuel, Christiana; Rao, Donthamsetty Nageswara; Kaliraj, Perumal

    2015-06-01

    Abundant Larval Transcript (ALT) is one of the major groups of immune-dominant proteins produced by filarial worms during their larval stage. The major B-cell and T-cell epitopic domains of the ALT-2 antigen were mapped to develop a multiple antigenic peptide (MAP) prophylactic antigen against lymphatic filariasis. ALT MAP was constructed by solid phase peptide synthesis. The reactivity of whole ALT protein and ALT MAP against clinical sera described a high reactivity of endemic normal sera against ALT MAP compared to WbALT-2 protein. The antibody isotype pattern revealed elevated levels of IgG1 and IgG2 against ALT MAP, followed by IgG3 and IgG4. In this study we also analyzed the immune response pattern elicited by ALT MAP, ALT in mice models, which revealed similar pattern of humoral response, while low T cell proliferation in ALT MAP groups. The low proliferation could be attributed to T/B epitope arrangement on the construct, MHC restriction, and incomplete signal delivery by T cell receptor. The immunodominant epitopes in ALT MAP were found to play a crucial role in inducing high antigen specific proliferation. This revealed the significance of ALT MAP in stimulating innate immunity in offering protective immune response probably through the activation of complement cascade along with stimulation of cellular response. An improved understanding, including the construction of ALT MAP and parasite challenge study in jirds to determine the worm clearance would give a better insight in the characterization ALT MAP construct as a prophylactic vaccine candidate.

  10. The immortal life of Henrietta Lacks

    OpenAIRE

    Duca, Edward; Duca, Edward

    2014-01-01

    Over 60 Best Book of the Year lists, 75 weeks on the New York Best Sellers list, and several prestigious awards, The Immortal life of Henrietta Lacks by Rebecca Skloot is a must read for all. http://www.um.edu.mt/think/the-immortal-life-of-henrietta-lacks/

  11. Immortalization of Werner syndrome and progeria fibroblasts

    International Nuclear Information System (INIS)

    Saito, H.; Moses, R.E.

    1991-01-01

    Human fibroblast cells from two different progeroid syndromes, Werner syndrome (WS) and progeria, were established as immortalized cell lines by transfection with plasmid DNA containing the SV40 early region. The lineage of each immortalized cell line was confirmed by VNTR analysis. Each of the immortalized cell lines maintained its original phenotype of slow growth. DNA repair ability of these cells was also studied by measuring sensitivity to killing by uv or the DNA-damaging drugs methyl methansulfonate, bleomycin, and cis-dichlorodiamine platinum. The results showed that both WS and progeria cells have normal sensitivity to these agents

  12. ALT-114 and ALT-118 Alternative Approaches to NIST ...

    Science.gov (United States)

    In 2016, US EPA approved two separate alternatives (ALT 114 and ALT 118) for the preparation and certification of Hydrogen Chloride (HCl) and Mercury (Hg) cylinder reference gas standards that can serve as EPA Protocol gases where EPA Protocol are required, but unavailable. The alternatives were necessary due to the unavailability of NIST reference materials (SRM, NTRM, CRM or RGM) or VSL reference materials (VSL PRM or VSL CRM), reference materials identified in EPA’s Green Book as necessary to establish the traceability of EPA protocol gases. ALT 114 and ALT 118 provides a pathway for gas vendors to prepare and certify traceable gas cylinder standards for use in certifying Hg and HCl CEMS. In this presentation, EPA will describe the mechanics and requirements of the performance-based approach, provide an update on the availability of these gas standards and also discuss the potential for producing and certifying gas standards for other compounds using this approach. This presentation discusses the importance of NIST-traceable reference gases relative to regulatory source compliance emissions monitoring. Specifically this presentation discusses 2 new approaches for making necessary reference gases available in the absence of NIST reference materials. Moreover, these approaches provide an alternative approach to rapidly make available new reference gases for additional HAPS regulatory compliance emissions measurement and monitoring.

  13. Change alone is eternal, perpetual, immortal : pharmacological immortality in science fiction

    OpenAIRE

    Grech, Victor E.; Vassallo, Clare; Callus, Ivan

    2012-01-01

    Immortality is a common feature in science-fiction (SF). This paper lists the ways in which the pharmacological induction of immortality has been depicted in SF, and the resultant outcomes. Immortality or extreme longevity are often melded with infertility in order to eliminate the overpopulation issues that would inevitably arise. This is only one way in which theoretical utopias which afford life extension become dystopias, cautionary tales that admonish against hubris. In this fashion, SF ...

  14. Følelsen er alt

    DEFF Research Database (Denmark)

    Bonde, Lisbeth

    forrige sekelskifte. Det gælder især de ofte mennesketomme landskabsmalerier med ensomme kastanjetræer, som sukker i efterårslyset, og parkerne med de undertiden flygtige menneskeskikkelser, der som partikler kan forsvinde i vinden. Det er alt sammen skildret med vibrerende, næsten ’sfumatiske’ konturer...

  15. Establishment of immortal normal and ataxia telangiectasia fibroblast cell lines by introduction of the hTERT gene

    Energy Technology Data Exchange (ETDEWEB)

    Nakamura, Hideaki; Fukami, Hiroko; Hayashi, Yuko; Kiyono, Tohru; Ishizaki, Kanji [Aichi Cancer Center, Nagoya (Japan). Research Inst.; Nakatsugawa, Shigekazu; Hamaguchi, Michinari [Nagoya Univ. (Japan). School of Medicine

    2002-06-01

    To establish immortal human cells, we introduced the human catalytic subunit of telomerase (hTERT) gene into skin fibroblast cells obtained from normal and ataxia telangiectasia (AT) individuals of Japanese origin. After hTERT introduction, these cells continue to grow beyond a population doubling number of 200 while maintaining their original radiosensitivity. Inductions of p53, phosphorylation of Serl5 in p53, and induction of p21 by X-ray irradiation in immortal cells derived from normal individual were not affected by the hTERT introduction. Both normal and AT immortal cells exhibited an apparent inhibition of growth as original primary cells when they reached confluence. Karyotype analysis has revealed that they are in a diploid range. These results suggest that cells immortalized by hTERT introduction retain their original characteristics except for immortalization, and that they may be useful for analyzing various effects of radiation on human cells. (author)

  16. Diagnosis and treatment of ALT tumors: is Trabectedin a new therapeutic option?

    Science.gov (United States)

    Pompili, Luca; Leonetti, Carlo; Biroccio, Annamaria; Salvati, Erica

    2017-12-22

    Telomeres are specialized nucleoprotein structures responsible for protecting chromosome ends in order to prevent the loss of genomic information. Telomere maintenance is required for achieving immortality by neoplastic cells. While most cancer cells rely on telomerase re-activation for linear chromosome maintenance and sustained proliferation, a significant population of cancers (10-15%) employs telomerase-independent strategies, collectively referred to as Alternative Lengthening of Telomeres (ALT). ALT mechanisms involve different types of homology-directed telomere recombination and synthesis. These processes are facilitated by loss of the ATRX or DAXX chromatin-remodeling factors and by abnormalities of the telomere nucleoprotein architecture. Although the functional consequences of telomerase and ALT up-regulation are similar in that they both prevent overall telomere shortening in tumors, these telomere maintenance mechanisms (TMMs) differ in several aspects which may account for their differential prognostic significance and response to therapy in various tumor types. Therefore, reliable methods for detecting telomerase activity and ALT are likely to become an important pre-requisite for the use of treatments targeting one or other of these mechanisms. However, the question whether ALT presence can confer sensitivity to rationally designed anti-cancer therapies is still open. Here we review the latest discoveries in terms of mechanisms of ALT activation and maintenance in human tumors, methods for ALT identification in cell lines and human tissues and biomarkers validation. Then, original results on sensitivity to rational based pre-clinical and clinical anti-tumor drugs in ALT vs hTERT positive cells will be presented.

  17. Association of BLM and BRCA1 during Telomere Maintenance in ALT Cells.

    Science.gov (United States)

    Acharya, Samir; Kaul, Zeenia; Gocha, April Sandy; Martinez, Alaina R; Harris, Julia; Parvin, Jeffrey D; Groden, Joanna

    2014-01-01

    Fifteen percent of tumors utilize recombination-based alternative lengthening of telomeres (ALT) to maintain telomeres. The mechanisms underlying ALT are unclear but involve several proteins involved in homologous recombination including the BLM helicase, mutated in Bloom's syndrome, and the BRCA1 tumor suppressor. Cells deficient in either BLM or BRCA1 have phenotypes consistent with telomere dysfunction. Although BLM associates with numerous DNA damage repair proteins including BRCA1 during DNA repair, the functional consequences of BLM-BRCA1 association in telomere maintenance are not completely understood. Our earlier work showed the involvement of BRCA1 in different mechanisms of ALT, and telomere shortening upon loss of BLM in ALT cells. In order to delineate their roles in telomere maintenance, we studied their association in telomere metabolism in cells using ALT. This work shows that BLM and BRCA1 co-localize with RAD50 at telomeres during S- and G2-phases of the cell cycle in immortalized human cells using ALT but not in cells using telomerase to maintain telomeres. Co-immunoprecipitation of BRCA1 and BLM is enhanced in ALT cells at G2. Furthermore, BRCA1 and BLM interact with RAD50 predominantly in S- and G2-phases, respectively. Biochemical assays demonstrate that full-length BRCA1 increases the unwinding rate of BLM three-fold in assays using a DNA substrate that models a forked structure composed of telomeric repeats. Our results suggest that BRCA1 participates in ALT through its interactions with RAD50 and BLM.

  18. Toivo Alt : "Sild tuleb." / Toivo Alt ; interv. Taavi Raudne

    Index Scriptorium Estoniae

    Alt, Toivo, 1957-

    2004-01-01

    Toivo Alt räägib intervjuus marjafirma Saarek tegevusest, Linnuse Kalja kaubamärgi ja Ösel Foods'i aktsiate müügist, erimeelsuste põhjustest endise äripartneri Vjatsheslav Leedoga ning äriideest ehitada mandri ja Muhumaa vahele 2010. aastaks sild. Vt. samas: Kohe näha, et vanad sõbrad; Leedo arvamus

  19. Immortality in view of Maimonides and Spinoza

    Directory of Open Access Journals (Sweden)

    Morteza Shajari

    2014-12-01

    Full Text Available Desire for immortality can be seen as the essential natural impulse. Therefore, different religions and thinkers have attempted to see the issue from different viewpoints. The great Jewish philosopher. Maimonides, due to deep fixation to Judaism, has tried to express their issues to be consistent with the Bible and his own community believes. He, in his discussion of resurrection, believed to three basic steps: The Messiah, the resurrection, and the world hereafter. His standpoint of eternity is dedicated to the hereafter. And we can be immortalized only by acting and teachings in accordance with the Bible and righteousness. Like Maimonides, Spinoza – the other Jewish philosopher - considered the immortality as Ultimate bliss through which the “immutable and eternal love of God" can be achieved. In his opinion, a person reaches this stage, when the lusts and emotions can reasonably be overcome, and also, when the power and anger and contempt and disregard others will respond with love and dignity. Thus, a man can be reached its proper perfection and immortality is reached. The difference between these two philosophers is that Maimonides believes through "actual intellect" -that is Emanation of the active intellect- can be immortalized but, for Spinoza, eternity can be reached through the adequate Ideas.

  20. Network signatures of cellular immortalization in human lymphoblastoid cell lines

    Energy Technology Data Exchange (ETDEWEB)

    Shim, Sung-Mi; Jung, So-Young; Nam, Hye-Young; Kim, Hye-Ryun; Lee, Mee-Hee; Kim, Jun-Woo; Han, Bok-Ghee [National Biobank of Korea, Center for Genome Science, Korea National Institute of Health, Osong 363-951 (Korea, Republic of); Jeon, Jae-Pil, E-mail: jaepiljeon@hanmail.net [Division of Brain Diseases, Center for Biomedical Science, Korea National Institute of Health, Osong 363-951 (Korea, Republic of)

    2013-11-15

    Highlights: •We identified network signatures of LCL immortalization from transcriptomic profiles. •More than 41% of DEGs are possibly regulated by miRNAs in LCLs. •MicroRNA target genes in LCLs are involved in apoptosis and immune-related functions. •This approach is useful to find functional miRNA targets in specific cell conditions. -- Abstract: Human lymphoblastoid cell line (LCL) has been used as an in vitro cell model in genetic and pharmacogenomic studies, as well as a good model for studying gene expression regulatory machinery using integrated genomic analyses. In this study, we aimed to identify biological networks of LCL immortalization from transcriptomic profiles of microRNAs and their target genes in LCLs. We first selected differentially expressed genes (DEGs) and microRNAs (DEmiRs) between early passage LCLs (eLCLs) and terminally differentiated late passage LCLs (tLCLs). The in silico and correlation analysis of these DEGs and DEmiRs revealed that 1098 DEG–DEmiR pairs were found to be positively (n = 591 pairs) or negatively (n = 507 pairs) correlated with each other. More than 41% of DEGs are possibly regulated by miRNAs in LCL immortalizations. The target DEGs of DEmiRs were enriched for cellular functions associated with apoptosis, immune response, cell death, JAK–STAT cascade and lymphocyte activation while non-miRNA target DEGs were over-represented for basic cell metabolisms. The target DEGs correlated negatively with miR-548a-3p and miR-219-5p were significantly associated with protein kinase cascade, and the lymphocyte proliferation and apoptosis, respectively. In addition, the miR-106a and miR-424 clusters located in the X chromosome were enriched in DEmiR–mRNA pairs for LCL immortalization. In this study, the integrated transcriptomic analysis of LCLs could identify functional networks of biologically active microRNAs and their target genes involved in LCL immortalization.

  1. Genome-Wide Transcriptional Reorganization Associated with Senescence-to-Immortality Switch during Human Hepatocellular Carcinogenesis

    Science.gov (United States)

    Konu, Ozlen; Yuzugullu, Haluk; Gursoy-Yuzugullu, Ozge; Ozturk, Nuri; Ozen, Cigdem; Ozdag, Hilal; Erdal, Esra; Karademir, Sedat; Sagol, Ozgul; Mizrak, Dilsa; Bozkaya, Hakan; Ilk, Hakki Gokhan; Ilk, Ozlem; Bilen, Biter; Cetin-Atalay, Rengul; Akar, Nejat; Ozturk, Mehmet

    2013-01-01

    Senescence is a permanent proliferation arrest in response to cell stress such as DNA damage. It contributes strongly to tissue aging and serves as a major barrier against tumor development. Most tumor cells are believed to bypass the senescence barrier (become “immortal”) by inactivating growth control genes such as TP53 and CDKN2A. They also reactivate telomerase reverse transcriptase. Senescence-to-immortality transition is accompanied by major phenotypic and biochemical changes mediated by genome-wide transcriptional modifications. This appears to happen during hepatocellular carcinoma (HCC) development in patients with liver cirrhosis, however, the accompanying transcriptional changes are virtually unknown. We investigated genome-wide transcriptional changes related to the senescence-to-immortality switch during hepatocellular carcinogenesis. Initially, we performed transcriptome analysis of senescent and immortal clones of Huh7 HCC cell line, and identified genes with significant differential expression to establish a senescence-related gene list. Through the analysis of senescence-related gene expression in different liver tissues we showed that cirrhosis and HCC display expression patterns compatible with senescent and immortal phenotypes, respectively; dysplasia being a transitional state. Gene set enrichment analysis revealed that cirrhosis/senescence-associated genes were preferentially expressed in non-tumor tissues, less malignant tumors, and differentiated or senescent cells. In contrast, HCC/immortality genes were up-regulated in tumor tissues, or more malignant tumors and progenitor cells. In HCC tumors and immortal cells genes involved in DNA repair, cell cycle, telomere extension and branched chain amino acid metabolism were up-regulated, whereas genes involved in cell signaling, as well as in drug, lipid, retinoid and glycolytic metabolism were down-regulated. Based on these distinctive gene expression features we developed a 15-gene

  2. The (not so immortal strand hypothesis

    Directory of Open Access Journals (Sweden)

    Cristian Tomasetti

    2015-03-01

    Significance: Utilizing an approach that is fundamentally different from previous efforts to confirm or refute the immortal strand hypothesis, we provide evidence against non-random segregation of DNA during stem cell replication. Our results strongly suggest that parental DNA is passed randomly to stem cell daughters and provides new insight into the mechanism of DNA replication in stem cells.

  3. Immortalization of human myogenic progenitor cell clone retaining multipotentiality

    International Nuclear Information System (INIS)

    Hashimoto, Naohiro; Kiyono, Tohru; Wada, Michiko R.; Shimizu, Shirabe; Yasumoto, Shigeru; Inagawa, Masayo

    2006-01-01

    Human myogenic cells have limited ability to proliferate in culture. Although forced expression of telomerase can immortalize some cell types, telomerase alone delays senescence of human primary cultured myogenic cells, but fails to immortalize them. In contrast, constitutive expression of both telomerase and the E7 gene from human papillomavirus type 16 immortalizes primary human myogenic cells. We have established an immortalized primary human myogenic cell line preserving multipotentiality by ectopic expression of telomerase and E7. The immortalized human myogenic cells exhibit the phenotypic characteristics of their primary parent, including an ability to undergo myogenic, osteogenic, and adipogenic terminal differentiation under appropriate culture conditions. The immortalized cells will be useful for both basic and applied studies aimed at human muscle disorders. Furthermore, immortalization by transduction of telomerase and E7 represents a useful method by which to expand human myogenic cells in vitro without compromising their ability to differentiate

  4. Immortality versus resurrection in the Christian tradition.

    Science.gov (United States)

    Murphy, Nancey

    2011-10-01

    For those in contemporary society who believe in an afterlife, there are a number of views available. The most common may be based on belief in an immortal soul. However, the early Christian account was, instead, bodily resurrection. As Christianity moved throughout the Mediterranean world, apologists and theologians adapted their teaching on human nature and the afterlife to Greek and Roman philosophies. By the time of Augustine (d. 430), the doctrines of body-soul dualism and immortality of the soul were firmly entrenched in Christian teaching. The incorporation of the concept of an immortal soul into Christian accounts of life after death produced a hybrid account. The body dies, the soul (at least of those who were to be saved) travels to heaven. At the end of history, there would be a general resurrection, and the souls would be reunited with their bodies, although the bodies would be in a transformed, indestructible state. This hybrid account of life after death went largely uncontested until the twentieth century. In this essay, I describe this history and argue for a return to the early Christian view of humans as a unity, not a duality, and for belief in resurrection of the body as the appropriate expectation for eternal life. This would not only be truer to Christian sources, but, valuable, I believe, in focusing Christian attention on the need to care for the environment. © 2011 New York Academy of Sciences.

  5. Production of the Allergenic Protein Alt a 1 by Alternaria Isolates from Working Environments

    Directory of Open Access Journals (Sweden)

    Justyna Skóra

    2015-02-01

    Full Text Available The aim of the study was to evaluate the ability of Alternaria isolates from workplaces to produce Alt a 1 allergenic protein, and to analyze whether technical materials (cellulose, compost, leather present within the working environment stimulate or inhibit Alt a 1 production (ELISA test. Studies included identification of the isolated molds by nucleotide sequences analyzing of the ITS1/ITS2 regions, actin, calmodulin and Alt a 1 genes. It has been shown that Alternaria molds are significant part of microbiocenosis in the archive, museum, library, composting plant and tannery (14%–16% frequency in the air. The presence of the gene encoding the Alt a 1 protein has been detected for the strains: Alternaria alternata, A. lini, A. limoniasperae A. nobilis and A. tenuissima. Environmental strains produced Alt a 1 at higher concentrations (1.103–6.528 ng/mL than a ATCC strain (0.551–0.975 ng/mL. It has been shown that the homogenization of the mycelium and the use of ultrafiltration allow a considerable increase of Alt a 1 concentration. Variations in the production of Alt a 1 protein, depend on the strain and extraction methods. These studies revealed no impact of the technical material from the workplaces on the production of Alt a 1 protein.

  6. Production of the Allergenic Protein Alt a 1 by Alternaria Isolates from Working Environments

    Science.gov (United States)

    Skóra, Justyna; Otlewska, Anna; Gutarowska, Beata; Leszczyńska, Joanna; Majak, Iwona; Stępień, Łukasz

    2015-01-01

    The aim of the study was to evaluate the ability of Alternaria isolates from workplaces to produce Alt a 1 allergenic protein, and to analyze whether technical materials (cellulose, compost, leather) present within the working environment stimulate or inhibit Alt a 1 production (ELISA test). Studies included identification of the isolated molds by nucleotide sequences analyzing of the ITS1/ITS2 regions, actin, calmodulin and Alt a 1 genes. It has been shown that Alternaria molds are significant part of microbiocenosis in the archive, museum, library, composting plant and tannery (14%–16% frequency in the air). The presence of the gene encoding the Alt a 1 protein has been detected for the strains: Alternaria alternata, A. lini, A. limoniasperae A. nobilis and A. tenuissima. Environmental strains produced Alt a 1 at higher concentrations (1.103–6.528 ng/mL) than a ATCC strain (0.551–0.975 ng/mL). It has been shown that the homogenization of the mycelium and the use of ultrafiltration allow a considerable increase of Alt a 1 concentration. Variations in the production of Alt a 1 protein, depend on the strain and extraction methods. These studies revealed no impact of the technical material from the workplaces on the production of Alt a 1 protein. PMID:25689994

  7. Production of the allergenic protein Alt a 1 by Alternaria isolates from working environments.

    Science.gov (United States)

    Skóra, Justyna; Otlewska, Anna; Gutarowska, Beata; Leszczyńska, Joanna; Majak, Iwona; Stępień, Łukasz

    2015-02-16

    The aim of the study was to evaluate the ability of Alternaria isolates from workplaces to produce Alt a 1 allergenic protein, and to analyze whether technical materials (cellulose, compost, leather) present within the working environment stimulate or inhibit Alt a 1 production (ELISA test). Studies included identification of the isolated molds by nucleotide sequences analyzing of the ITS1/ITS2 regions, actin, calmodulin and Alt a 1 genes. It has been shown that Alternaria molds are significant part of microbiocenosis in the archive, museum, library, composting plant and tannery (14%-16% frequency in the air). The presence of the gene encoding the Alt a 1 protein has been detected for the strains: Alternaria alternata, A. lini, A. limoniasperae A. nobilis and A. tenuissima. Environmental strains produced Alt a 1 at higher concentrations (1.103-6.528 ng/mL) than a ATCC strain (0.551-0.975 ng/mL). It has been shown that the homogenization of the mycelium and the use of ultrafiltration allow a considerable increase of Alt a 1 concentration. Variations in the production of Alt a 1 protein, depend on the strain and extraction methods. These studies revealed no impact of the technical material from the workplaces on the production of Alt a 1 protein.

  8. ALT Blood Test: MedlinePlus Lab Test Information

    Science.gov (United States)

    ... page: https://medlineplus.gov/labtests/altbloodtest.html ALT Blood Test To use the sharing features on this page, please enable JavaScript. What is an ALT Blood Test? ALT, which stands for alanine transaminase, is an ...

  9. 'Immortal' energy systems and intergenerational justice

    International Nuclear Information System (INIS)

    Weinberg, A.M.

    1985-01-01

    Some critics of our technological society have asserted that we are leaving a legacy of problems for our descendants - in the shape, for example, of CO 2 pollution of the atmosphere and radioactive waste. The author argues that if some of our power generation systems turn out to be near 'immortal', with lives much longer than their book lives, on the contrary, great benefits may be bequeathed to our successors - in fully amortized plant with very low running costs. There are examples in history of similar benefits conferred by dams built hundreds of years ago but which still serve useful purposes today. (author)

  10. [Liver disorders in adults: ALT and AST].

    Science.gov (United States)

    Goorden, Susanna M I; Buffart, Tineke E; Bakker, Annemieke; Buijs, Madelon M

    2013-01-01

    Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) are commonly used biomarkers for liver damage. As well as in liver tissue, AST is also present in cardiac and skeletal muscle and in erythrocytes, making ALT the most specific marker for liver damage. Here, we describe two patients with sustained increases in ALT and AST levels. The first patient is a 79-year-old woman who developed elevated serum transaminases shortly after having a myocardial infarction. The second patient, an obese 40-year-old woman presented with increased ALT and AST levels in the absence of physical symptoms. Notably, her father died of liver cirrhosis without a history of alcohol abuse. Based upon these case reports we discuss the differential diagnostic work-up of elevated serum transaminase levels. Furthermore, we explain monitoring, test performance, reference values and analytical pitfalls of these biomarkers.

  11. ALT-114 and ALT-118 Alternative Approaches to NIST-Traceable Reference Gases

    Science.gov (United States)

    In 2016, US EPA approved two separate alternatives (ALT 114 and ALT 118) for the preparation and certification of Hydrogen Chloride (HCl) and Mercury (Hg) cylinder reference gas standards that can serve as EPA Protocol gases where EPA Protocol are required, but unavailable. The a...

  12. Rule of changes in serum GGT levels and GGT/ALT and AST/ALT ratios in primary hepatic carcinoma patients with different AFP levels.

    Science.gov (United States)

    Yang, Jian-Gong; He, Xiao-Feng; Huang, Bing; Zhang, Hui-Ai; He, Yong-Kang

    2017-12-22

    This study aims to explore the rule of changes in serum GGT activity, as well as GGT/ALT and AST/ALT ratios, in primary hepatic carcinoma (PHC) patients with different alpha-fetal protein (AFP) levels. GGT, AST and ALT were detected in 370 PHC patients with positive HBs-Ag using a automatic biochemical analyzer, and AFP was detected using a Roche E170 modular analytics immunoassay analyzer. GGT level, as well as AST/ALT and GGT/ALT, ratios were compared among PHC patients with different AFP levels. As shown in Table 1, GGT levels were 109.59 ± 111.06, 151.13 ± 190.43, 135.86 ± 107.62, 151.36 ± 176.59 and 172.58 ± 188.84, respectively, in the groups of primary PHC patients with AFP levels of ⩽ 10, 10-100, 100-200, 200-400 and ⩾ 400 ng/ml; and the differences among these groups were not statistically significant (P> 0.05). AST/ALT ratios were 1.55 ± 1.02, 1.30 ± 0.81, 2.02 ± 1.89, 2.12 ± 1.11 and 1.73 ± 1.25, respectively; and the differences among these groups were not statistically significant (P> 0.05). GGT/ALT ratios were 3.43 ± 3.12, 3.57 ± 5.70, 3.57 ± 2.94, 3.89 ± 4.58 and 3.43 ± 3.61, respectively; and the differences among these groups were not statistically significant (P> 0.05). For patients with chronic hepatitis B and cirrhosis after hepatitis B, no matter how AFP level is, when liver function report reveals increased GGT, AST/ALT > 1 and GGT/ALT > 1 (that is, AST > ALT and GGT > ALT), even if AFP is negative, we should also be alert to the existence of PHC.

  13. The conserved mitochondrial gene distribution in relatives of Turritopsis nutricula, an immortal jellyfish

    OpenAIRE

    Devarapalli, Pratap; Kumavath, Ranjith N; Barh, Debmalya; Azevedo, Vasco

    2014-01-01

    Turritopsis nutricula (T. nutricula) is the one of the known reported organisms that can revert its life cycle to the polyp stage even after becoming sexually mature, defining itself as the only immortal organism in the animal kingdom. Therefore, the animal is having prime importance in basic biological, aging, and biomedical researches. However, till date, the genome of this organism has not been sequenced and even there is no molecular phylogenetic study to reveal its close relatives. Here,...

  14. Increased AST/ALT ratio in azotaemic dogs infected with Babesia canis.

    Science.gov (United States)

    Zygner, W; Gójska-Zygner, O; Norbury, L J; Wedrychowicz, H

    2012-01-01

    The AST/ALT ratio was estimated in 182 dogs infected with Babesia canis. Among these dogs 65 had anaemia and 68 were azotaemic. Student's t test was used to compare means of the AST/ALT ratio in anaemic and non-anaemic dogs, and in azotaemic and non-azotaemic dogs (p < 0.05). The differences in AST/ALT ratio between anaemic (1.52 +/- 1.15) and non-anaemic (1.76 +/- 1.34) dogs were statistically insignificant (p = 0.23), however, the comparison of AST/ALT ratio between azotaemic (2.68 +/- 1.52) and non-azotaemic (1.08 +/- 0.53) dogs revealed a significantly higher value of this index in azotaemic dogs (p = 0.00). The present results suggest that kidney injury contributed to increased AST activity in these dogs.

  15. Epitope mapping of Brugia malayi ALT-2 and the development of a multi-epitope vaccine for lymphatic filariasis.

    Science.gov (United States)

    Madhumathi, J; Prince, P R; Rao, D N; Karande, A A; Reddy, M V R; Kaliraj, P

    2017-01-01

    Human lymphatic filariasis is a neglected tropical disease, causing permanent and long-term disability with severe immunopathology. Abundant larval transcript (ALT) plays a crucial role in parasite establishment in the host, due to its multi-faceted ability in host immune regulation. Although ALT protein is a key filarial target, its exact function is yet to be explored. Here, we report epitope mapping and a structural model of Brugia malayi ALT-2, leading to development of a multi-epitope vaccine. Structural analysis revealed that ALT represents unique parasitic defence proteins belonging to a toxin family that carries a 'knottin' fold. ALT-2 has been a favourite vaccine antigen and was protective in filarial models. Due to the immunological significance of ALT-2, we mapped B-cell epitopes systematically and identified two epitope clusters, 1-30 and 89-128. To explore the prophylactic potential of epitope clusters, a recombinant multi-epitopic gene comprising the epitopic domains was engineered and the protective efficacy of recombinant ALT epitope protein (AEP) was tested in the permissive model, Mastomys coucha. AEP elicited potent antibody responses with predominant IgG1 isotype and conferred significantly high protection (74.59%) compared to ALT-2 (61.95%). This proved that these epitopic domains are responsible for the protective efficacy of ALT-2 and engineering protective epitopes as a multi-epitope protein may be a novel vaccine strategy for complex parasitic infections.

  16. Alt a 15 is a new cross-reactive minor allergen of Alternaria alternata.

    Science.gov (United States)

    Gabriel, M F; Postigo, I; Gutiérrez-Rodríguez, A; Suñén, E; Guisantes, J A; Fernández, J; Tomaz, C T; Martínez, J

    2016-02-01

    Alternaria alternata is one of the most common saprophytes worldwide that is clinically and epidemiologically associated with severe asthma. Therefore, the identification and characterization of all A. alternata allergens are of major clinical importance. This study describes a new cross-reactive A. alternata allergen that was officially named Alt a 15 by the official Allergen Nomenclature Subcommittee. The complete coding region for Alt a 15 was amplified using 5' and 3' rapid amplification of cDNA ends and PCR. The recombinant protein was produced in Escherichia coli as a 65-kDa fusion protein, and the protein sequence exhibits high homology with several important fungal allergens. Immunoblotting analyses revealed that IgE antibodies from A. alternata-sensitized patients (n=59) bound to rAlt a 15 with a prevalence of 10.2%. All patients who presented sIgE to rAlt a 15 were apparently poly-sensitized to A. alternata and C. lunata. The extensive cross-reactivity between A. alternata and C. lunata serine proteases was confirmed using immunoblotting inhibition assays. Overall, Alt a 15 is an important new cross-reactive allergen of A. alternata that explains some allergies to A. alternata without Alt a 1 sensitization and initial diagnostic errors for allergies to Alternaria. This molecule may improve the accuracy of the diagnosis, the understanding, and the management of IgE-mediated fungal diseases. Copyright © 2015 Elsevier GmbH. All rights reserved.

  17. Sensitivity and Specificity of Plasma ALT, ALP, and Bile Acids for Hepatitis in Labrador Retrievers.

    Science.gov (United States)

    Dirksen, K; Burgener, I A; Rothuizen, J; van den Ingh, T S G A M; Penning, L C; Spee, B; Fieten, H

    2017-07-01

    Biochemical indicators for diagnosing liver disease are plasma alanine aminotransferase activity (ALT), alkaline phosphatase activity (ALP), and bile acid concentration (BA). To determine the sensitivity and specificity of ALT, ALP, and BA for detecting primary hepatitis (PH) in clinically healthy Labrador retrievers and investigate whether ALT and ALP can discriminate between dogs with PH and nonspecific reactive hepatitis (RH). 191 clinically healthy and 51 clinically ill Labrador retrievers with hepatic histopathology. Retrospective study. Medical records were reviewed for ALT, ALP, preprandial BA, liver histopathology, and hepatic copper concentrations. In 64% (122/191) of the clinically healthy Labrador retrievers, hepatic histology revealed inflammatory infiltrates. This frequency might be biased because part of them was included as first-line relatives of dogs with copper-associated hepatitis. Sensitivity of ALT, ALP, and BA in this population for detecting acute hepatitis was 45, 15, and 15%, respectively. For chronic hepatitis, sensitivity was 71, 35, and 13%, respectively. Specificity of ALT, ALP, and BA was >90% for AH, CH, and RH. When increased liver enzymes were present, median ALT was significantly higher in PH cases (312 U/L, range 38-1,369) compared to RH cases (91 U/L, range 39-139) (P ALT, ALP, and BA for detecting acute and chronic hepatitis in this population was low. More sensitive biomarkers are needed for early detection of liver disease in apparent clinically healthy dogs. Copyright © 2017 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.

  18. Stem cell microRNAs in senescence and immortalization: novel players in cancer therapy.

    Science.gov (United States)

    Leal, Jose A; Feliciano, Andrea; Lleonart, Matilde E

    2013-01-01

    The molecular etiology of malignancy remains one of the most challenging disease processes under scientific investigation; therefore, improved approaches for their treatment are urgently needed. MicroRNAs are highly conserved nonprotein-coding RNAs that regulate gene expression. They are involved in important homeostatic processes, such as cellular proliferation, cell death and development, and affect many diseases, including cancer. High-throughput screenings based on microRNAs related to senescence/immortalization are potential tools for identifying novel proliferative microRNAs that might be involved in carcinogenesis. Recently, a subgroup of highly proliferative microRNAs, which belong to a cluster expressed exclusively in embryonic stem cells and their malignant derivatives (embryonic carcinoma cells), was revealed to play a role in senescence bypass, thereby providing immortalization to human cells. This finding supports the cancer stem cell theory and the relevance of microRNAs in human tumors. This article recapitulates the role of microRNAs that are associated with stem cell properties and their possible link in common pathways related to immortalization and cancer. Ultimately, cancer therapy that is based on the induction of a senescence response is proposed to be highly associated with the loss of stemness properties. Thus, it would be possible to "kill two birds with one stone": along with the inhibition of stemness properties in cancer stem cells, the senescence response could be induced to destroy the cancer stem cell population within a tumor. © 2011 Wiley Periodicals, Inc.

  19. Editorial ALT-J non-electronica

    Directory of Open Access Journals (Sweden)

    Gabriel Jacobs

    1997-12-01

    Full Text Available The ALT Executive Committee has discussed on a number of occasions the question of making ALT-J wholly available in electronic form on our Web site (at present only the editorial and abstracts of papers appear, and even of turning it into a full-blown electronic journal. I have gently but consistently opposed this move, on occasions to the intense irritation of some of my fellow members, and always at the risk of appearing to be a Luddite. Explicitly ('Who, if not we as learning technologists, should be in the forefront of the soon-to-happen electronic-journal revolution?' and implicitly ('It is estimated that within two years, paper-based journals will be considered the dinosaurs of the academic world', I have increasingly come under pressure to accede to an apparently unstoppable, apparently imminent change in the method and form of scholarly publication. I set out here my reasons for my continued resistance.

  20. Ibn Qayyim Al-Jawziyyah and Allameh Tabataba’i on Immortality in Hell

    Directory of Open Access Journals (Sweden)

    Janan Izadi

    2016-09-01

    out from him/her and even enjoys them. The original nature may remain but infidelity is a curtain and constraint (qāsir that does not let the heavenly forms go out of original nature; In this case one can deduce non-immortality in hell according to the philosophical principle “constraint is not permanent.” Allameh replies that in the first case the fiery devil forms that go out of the soul can be perfection in one aspect and can even be consistent and enjoyable. But when truth if revealed in the hereafter and he becomes aware of the harm and consequences of his sins, it will be a great torment for the soul. He will be exactly like an addict who has enjoyed a drug in ignorance but when he becomes aware of the harms, the same joy becomes pain for him. In the second case Allameh agrees that the torment will not be permanent. In explaining the problem of evil both scholars consider evil unrelated to God and believe that God did not predestine torment and torture for His revenge but each scholar has a different explanation. For Ibn Qayyim torment and torture is Divine Action and the essence of evil in hell is not the creation’s purpose and aim in itself but is an object that is foreordained for a desirable result that is heaven. Allameh Tabataba’i, however, believes in the embodiment (tajassum of human actions in life in the hereafter and considers the torments as the result of the actions of sinful people, not the result of the actions of God.

  1. Immortality in the Christian Physicalistic Theology: A Critical Survey

    Directory of Open Access Journals (Sweden)

    Hasan Ahmadizade

    2017-07-01

    Full Text Available Physicalistic Theology is a term that has no exact definition in theologian views. In the 20th century some of Christian thinkers on theology, like Nancy Murphy and Peter van Inwagen, by accepting a Physicalistic approach on human being, tried to analyze the Christian beliefs about human identity and his immortality. This approach today is called Physicalistic Theology. According to this approach, human is not but this physical body itself and so we can simply analyze the immortality problem. In this article we try to by an analytic and descriptive method, analyze the immortality of human according to the view of Physicalistic Theology. We will analyze the most important reasoning of Physicalistic Theology that is: no-interaction between the material and the immaterial, interaction between the person and the body, and the physicalism in Christian beliefs. One of the conclusions of this article is that according to Physicalistic view, the person that at some time has not been in the world, must exists any time to destroyed forever because the Christians believe to things that cannot justify rationally. The problem of immortality is one of these matters. Physicalistic Theology try to prove the immortality based on the miracles and the absolute power of God.

  2. The helicase domain of Polθ counteracts RPA to promote alt-NHEJ.

    Science.gov (United States)

    Mateos-Gomez, Pedro A; Kent, Tatiana; Deng, Sarah K; McDevitt, Shane; Kashkina, Ekaterina; Hoang, Trung M; Pomerantz, Richard T; Sfeir, Agnel

    2017-12-01

    Mammalian polymerase theta (Polθ) is a multifunctional enzyme that promotes error-prone DNA repair by alternative nonhomologous end joining (alt-NHEJ). Here we present structure-function analyses that reveal that, in addition to the polymerase domain, Polθ-helicase activity plays a central role during double-strand break (DSB) repair. Our results show that the helicase domain promotes chromosomal translocations by alt-NHEJ in mouse embryonic stem cells and also suppresses CRISPR-Cas9- mediated gene targeting by homologous recombination (HR). In vitro assays demonstrate that Polθ-helicase activity facilitates the removal of RPA from resected DSBs to allow their annealing and subsequent joining by alt-NHEJ. Consistent with an antagonistic role for RPA during alt-NHEJ, inhibition of RPA1 enhances end joining and suppresses recombination. Taken together, our results reveal that the balance between HR and alt-NHEJ is controlled by opposing activities of Polθ and RPA, providing further insight into the regulation of repair-pathway choice in mammalian cells.

  3. In Vitro Endothelialization Test of Biomaterials Using Immortalized Endothelial Cells.

    Directory of Open Access Journals (Sweden)

    Ken Kono

    Full Text Available Functionalizing biomaterials with peptides or polymers that enhance recruitment of endothelial cells (ECs can reduce blood coagulation and thrombosis. To assess endothelialization of materials in vitro, primary ECs are generally used, although the characteristics of these cells vary among the donors and change with time in culture. Recently, primary cell lines immortalized by transduction of simian vacuolating virus 40 large T antigen or human telomerase reverse transcriptase have been developed. To determine whether immortalized ECs can substitute for primary ECs in material testing, we investigated endothelialization on biocompatible polymers using three lots of primary human umbilical vein endothelial cells (HUVEC and immortalized microvascular ECs, TIME-GFP. Attachment to and growth on polymer surfaces were comparable between cell types, but results were more consistent with TIME-GFP. Our findings indicate that TIME-GFP is more suitable for in vitro endothelialization testing of biomaterials.

  4. Genes involved in immortalization of human mammary cells

    Energy Technology Data Exchange (ETDEWEB)

    Stampfer, Martha R.; Yaswen, Paul

    2001-09-27

    Breast cancer progression is characterized by inappropriate cell growth. Normal cells cease growth after a limited number of cell divisions--a process called cellular senescence-while tumor cells may acquire the ability to proliferate indefinitely (immortality). Inappropriate expression of specific oncogenes in a key cellular signaling pathway (Ras, Raf) can promote tumorigenicity in immortal cells, while causing finite lifespan cells to undergo a rapid senescence-like arrest. We have studied when in the course of transformation of cultured human mammary epithelial cells (HMEC), the response to overexpressed oncogenic Raf changes from being tumor-suppressive to tumor enhancing, and what are the molecular underpinnings of this response. Our data indicate: (1) HMEC acquire the ability to maintain growth in the presence of oncogenic Raf not simply as a consequence of overcoming senescence, but as a result of a newly discovered step in the process of immortal transformation uncovered by our lab, termed conversion. Immortal cells that have not undergone conversion (e.g., cells immortalized by exogenous introduction of the immortalizing enzyme, telomerase) remain growth inhibited. (2) Finite lifespan HMEC growth arrest in response to oncogenic Raf using mediators of growth inhibition that are very different from those used in response to oncogenic Raf by rodent cells and certain other human cell types, including the connective tissue cells from the same breast tissue. While many diverse cell types appear to have in common a tumor-suppressive response to this oncogenic signal, they also have developed multiple mechanisms to elicit this response. Understanding how cancer cells acquire the crucial capacity to be immortal and to abrogate normal tumor-suppressive mechanisms may serve both to increase our understanding of breast cancer progression, and to provide new targets for therapeutic intervention. Our results indicate that normal HMEC have novel means of enforcing a Raf

  5. [Immortal time bias in pharmacoepidemiological studies: definition, solutions and examples].

    Science.gov (United States)

    Faillie, Jean-Luc; Suissa, Samy

    2015-01-01

    Among the observational studies of drug effects in chronic diseases, many of them have found effects that were exaggerated or wrong. Among bias responsible for these errors, the immortal time bias, concerning the definition of exposure and exposure periods, is relevantly important as it usually tends to wrongly attribute a significant benefit to the study drug (or exaggerate a real benefit). In this article, we define the mechanism of immortal time bias, we present possible solutions and illustrate its consequences through examples of pharmacoepidemiological studies of drug effects. © 2014 Société Française de Pharmacologie et de Thérapeutique.

  6. Association of ALT and the metabolic syndrome among Mexican children.

    Science.gov (United States)

    Elizondo-Montemayor, Leticia; Ugalde-Casas, Patricia A; Lam-Franco, Lorena; Bustamante-Careaga, Humberto; Serrano-González, Mónica; Gutiérrez, Norma G; Martínez, Ubaldo

    2014-01-01

    Nonalcoholic fatty liver disease (NAFLD) is emerging as a component of the metabolic syndrome (MetS); Hispanics being particularly predisposed. Alanine aminotransferase (ALT) is considered a marker of NAFLD. The aim of this study was to determine the prevalence and associations between ALT elevations and MetS in normal-weight, overweight and obese Mexican children and adolescents, since data in Mexico is scarce. Body mass index (BMI), waist circumference (WC), percentage body fat, blood pressure, glucose, lipid profiles, ALT and aspartate aminotransferase (AST) were measured in 236, 6-12yo normal-weight, overweight and obese Mexicans from eight public schools. The results showed that elevated ALT (>40 IU/L) was found in 17.7% of the obese and overweight population, with no gender difference. The prevalence of elevated ALT increased linearly across BMI categories (p = 0.001), from 0.0% for the normal-weight group (95%CI 0.0-€“8.0) to 22.4% for the obese one (95%CI 16.2-€“30.2). AST/ALT ratio ALT (p = 0.002), 50% in the elevated ALT group (95%CI 34.1-€“65.9) and 24.1% in the normal ALT one (95%CI 18.1-€“31.3). There was also a strong association between MetS and an AST/ALT ratio ALT (AOR = 7.13). Pearson correlation showed that MetS components were significantly correlated with elevated ALT. Therefore elevated ALT levels were highly prevalent and strongly associated with MetS in Mexican children, it should be screened in overweight and obese children. © 2014 Asian Oceanian Association for the Study of Obesity . Published by Elsevier Ltd. All rights reserved.

  7. TEAM TEACHING IN JAPAN FROM THE PERSPECTIVES OF THE ALTs, THE JTEs, AND THE STUDENTS

    Directory of Open Access Journals (Sweden)

    Andrea Ann Johannes

    2012-07-01

    Full Text Available This case study explores team teaching in Japan from the perspectives of JTEs (Japanese English Teachers, ALTs (Assistant Language Teachers, and students. Special focus is attributed to teachers and students’ perceptions of ALT and JTE roles. To determine the perspectives of all three participants, 112 students, 4 JTES and 2 ALTs from a Japanese high school in Chiba Prefecture were surveyed. Fieldwork was conducted over a ten-day period at the research site in 2009 to collect data in the form of questionnaires, interviews, and class observations. The findings indicate more of a mismatch between students’ perceptions of their teachers’ roles than between teachers. The results also reveal the students prefer a combination of both teachers and find team-taught classes more beneficial to developing their English skills.

  8. ALT1, a Snf2 Family Chromatin Remodeling ATPase, Negatively Regulates Alkaline Tolerance through Enhanced Defense against Oxidative Stress in Rice

    Science.gov (United States)

    Guo, Mingxin; Wang, Ruci; Wang, Juan; Hua, Kai; Wang, Yueming; Liu, Xiaoqiang; Yao, Shanguo

    2014-01-01

    Alkaline salt stress adversely affects rice growth, productivity and grain quality. However, the mechanism underlying this process remains elusive. We characterized here an alkaline tolerant mutant, alt1 in rice. Map-based cloning revealed that alt1 harbors a mutation in a chromatin remodeling ATPase gene. ALT1-RNAi transgenic plants under different genetic background mimicked the alt1 phenotype, exhibiting tolerance to alkaline stress in a transcript dosage-dependent manner. The predicted ALT1 protein belonged to the Ris1 subgroup of the Snf2 family and was localized in the nucleus, and transcription of ALT1 was transiently suppressed after alkaline treatment. Although the absorption of several metal ions maintained well in the mutant under alkaline stress, expression level of the genes involved in metal ions homeostasis was not altered in the alt1 mutant. Classification of differentially expressed abiotic stress related genes, as revealed by microarray analysis, found that the majority (50/78) were involved in ROS production, ROS scavenging, and DNA repair. This finding was further confirmed by that alt1 exhibited lower levels of H2O2 under alkaline stress and tolerance to methyl viologen treatment. Taken together, these results suggest that ALT1 negatively functions in alkaline tolerance mainly through the defense against oxidative damage, and provide a potential two-step strategy for improving the tolerance of rice plants to alkaline stress. PMID:25473841

  9. ELEVATED ALT AND AST IN AN ASYMPTOMATIC PERSON

    OpenAIRE

    KEW ST; LOH KY

    2009-01-01

    -Abnormal liver function test with raised alanine aminotransferase (ALT) and raised aspartate aminotransferase (AST) are commonly seen in primary care setting. -Chronic alcohol consumption, drugs, non-alcoholic steatohepatitis (NASH) and chronic viral hepatitis are common causes associated with raised ALT and AST. -In chronic viral hepatitis, the elevation of liver enzyme may not correlate well with the degree of liver damage. -Non-hepatic causes of raised ALT and AST include polymyositis, a...

  10. Zeitschrift für Alte Gesehichte

    Directory of Open Access Journals (Sweden)

    Pedro Moacyr Campos

    1951-12-01

    Full Text Available HISTORIA — Zeitschrift für Alte Gesehichte, Jahrgang I. 1950, Heft 2. —  Baden-Baden, Verlag für Kunst und Wissenschaft (Primeiro Parágrafo do Artigo O prof. Friedrich Matz, do Instituto Arqueológico da Universidade de Marburg, principia o seu artigo por uma referência à pausa forçada que sofreram as pesquisas arqueológicas na Grécia e na Àsia Menor durante a segunda guerra mundial acentuando a relativa utilidade desta pausa para uma revisão dos dados cronólogicos referentes à pre-história daquelas regiões.

  11. Accelerated life testing (ALT) in microelectronics and photonics: its role, attributes, challenges, pitfalls, and interaction with qualification tests

    Science.gov (United States)

    Suhir, Ephraim

    2002-06-01

    Accelerated life tests (ALTs) are aimed at the revealing and understanding the physics of the expected or occurred failures, i.e. are able to detect the possible failure modes and mechanisms. Another objective of the ALTs is to accumulate representative failure statistics. Adequately designed, carefully conducted, and properly interpreted ALTs provide a consistent basis for obtaining the ultimate information of the reliability of a product - the predicted probability of failure after the given time of service. Such tests can dramatically facilitate the solution to the cost effectiveness and time-to-market problems. ALTs should play an important role in the evaluation, prediction and assurance of the reliability of microelectronics and optoelectronics devices and systems. In the majority of cases, ALTs should be conducted in addition to the qualification tests, which are required by the existing standards. There might be also situations, when ALTs can be (and, probably, should be) used as an effective substitution for such standards, or, at least, as the basis for the improvement of the existing qualification specifications. We describe different types (categories) of accelerated tests, with an emphasis on the role that ALTs should play in the development, design, qualification and manufacturing of microelectronics and photonics products. We discuss the challenges associated with the implementation and use of the ALTs, potential pitfalls (primarily those associated with possible shifts in the mechanisms and modes of failure), and the interaction of the ALTs with other types of accelerated tests. The role of the nondestructive evaluations is also briefly outlined. The case of a laser welded optoelectronic package assembly is used to illustrate the concepts addressed.

  12. Koschei the immortal and anti-aging drugs.

    Science.gov (United States)

    Blagosklonny, M V

    2014-12-04

    In Slavic folklore, Koschei the Immortal was bony, thin and lean. Was his condition caused by severe calorie restriction (CR)? CR deactivates the target of rapamycin pathway and slows down aging. But the life-extending effect of severe CR is limited by starvation. What if Koschei's anti-aging formula included rapamycin? And was rapamycin (or another rapalog) combined with commonly available drugs such as metformin, aspirin, propranolol, angiotensin II receptor blockers and angiotensin-converting enzyme inhibitors.

  13. Homologous plasmid recombination is elevated in immortally transformed cells.

    OpenAIRE

    Finn, G K; Kurz, B W; Cheng, R Z; Shmookler Reis, R J

    1989-01-01

    The levels of intramolecular plasmid recombination, following transfection of a plasmid substrate for homologous recombination into normal and immortally transformed cells, have been examined by two independent assays. In the first assay, recovered plasmid was tested for DNA rearrangements which regenerate a functional neomycin resistance gene from two overlapping fragments. Following transformation of bacteria, frequencies of recombinationlike events were determined from the ratio of neomyci...

  14. Homologous plasmid recombination is elevated in immortally transformed cells.

    Science.gov (United States)

    Finn, G K; Kurz, B W; Cheng, R Z; Shmookler Reis, R J

    1989-09-01

    The levels of intramolecular plasmid recombination, following transfection of a plasmid substrate for homologous recombination into normal and immortally transformed cells, have been examined by two independent assays. In the first assay, recovered plasmid was tested for DNA rearrangements which regenerate a functional neomycin resistance gene from two overlapping fragments. Following transformation of bacteria, frequencies of recombinationlike events were determined from the ratio of neomycin-resistant (recombinant) colonies to ampicillin-resistant colonies (indicating total plasmid recovery). Such events, yielding predominantly deletions between the directly repeated sequences, were substantially more frequent in five immortal cell lines than in any of three normal diploid cell strains tested. Effects of plasmid replication or interaction with T antigen and of bacterially mediated rejoining of linear molecules generated in mammalian cells were excluded by appropriate controls. The second assay used limited coamplification of a control segment of plasmid DNA, and of the predicted recombinant DNA region, primed by two sets of flanking oligonucleotides. Each amplified band was quantitated by reference to a near-linear standard curve generated concurrently, and recombination frequencies were determined from the ratio of recombinant/control DNA regions. The results confirmed that recombinant DNA structures were generated within human cells at direct repeats in the transfected plasmid and were markedly more abundant in an immortal cell line than in the diploid normal cells from which that line was derived.

  15. De Ritis Ratio (AST/ALT) as a Significant Prognostic Factor in Patients With Upper Tract Urothelial Cancer Treated With Surgery.

    Science.gov (United States)

    Lee, Hakmin; Choi, Young Hyo; Sung, Hyun Hwan; Han, Deok Hyun; Jeon, Hwang Gyun; Chang Jeong, Byong; Seo, Seong Il; Jeon, Seong Soo; Lee, Hyun Moo; Choi, Han Yong

    2017-06-01

    We investigated the clinical prognostic value of preoperative De Ritis ratio (aspartate aminotransferase [AST]/alanine aminotransferase [ALT]) on postsurgical survival outcomes in patients with upper tract urothelial cancer (UTUC). We retrospectively analyzed the data of 623 patients who underwent radical nephrouretectomy for UTUC. Multivariate regression tests were performed to identify possible associations between adverse pathologic events and AST/ALT. The risk of postoperative progression and survival were tested using Kaplan-Meier analyses and Cox proportional hazards models. According to the receiver operator characteristic curve of AST/ALT for cancer-specific mortality, patients with AST/ALT value ≥1.5 were regarded as the high AST/ALT group, and the remaining patients formed the low AST/ALT group. In Kaplan-Meier analyses, the high AST/ALT group showed worse progression-free survival (PFS), cancer-specific survival (CSS), and overall survival (all P ALT was associated with higher T stage (hazard ratio [HR], 1.577; 95% confidence interval [CI], 1.077-2.311; P = .033) and higher cellular grade (HR, 1.538; 95% CI, 1.034-2.287; P = .041) in multivariate regression tests. In multivariate Cox analyses, high AST/ALT was revealed as an independent predictor of PFS (HR, 2.335; 95% CI, 1.633-3.340; P ALT was a significant predictor of worse postoperative survival in patients surgically treated for UTUC. Further large prospective studies are needed for better understanding of the prognostic value of preoperative AST/ALT. Copyright © 2016 Elsevier Inc. All rights reserved.

  16. SMARCAL1 Resolves Replication Stress at ALT Telomeres.

    Science.gov (United States)

    Cox, Kelli E; Maréchal, Alexandre; Flynn, Rachel Litman

    2016-02-09

    Cancer cells overcome replicative senescence by exploiting mechanisms of telomere elongation, a process often accomplished by reactivation of the enzyme telomerase. However, a subset of cancer cells lack telomerase activity and rely on the alternative lengthening of telomeres (ALT) pathway, a recombination-based mechanism of telomere elongation. Although the mechanisms regulating ALT are not fully defined, chronic replication stress at telomeres might prime these fragile regions for recombination. Here, we demonstrate that the replication stress response protein SMARCAL1 is a critical regulator of ALT activity. SMARCAL1 associates with ALT telomeres to resolve replication stress and ensure telomere stability. In the absence of SMARCAL1, persistently stalled replication forks at ALT telomeres deteriorate into DNA double-strand breaks promoting the formation of chromosome fusions. Our studies not only define a role for SMARCAL1 in ALT telomere maintenance, but also demonstrate that resolution of replication stress is a crucial step in the ALT mechanism. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

  17. Lower liver cancer risk with antiviral therapy in chronic hepatitis B patients with normal to minimally elevated ALT and no cirrhosis.

    Science.gov (United States)

    Hoang, Joseph K; Yang, Hwai-I; Le, An; Nguyen, Nghia H; Lin, Derek; Vu, Vinh D; Chaung, Kevin; Nguyen, Vincent; Trinh, Huy N; Li, Jiayi; Zhang, Jian Q; Chen, Chien-Jen; Nguyen, Mindie H

    2016-08-01

    For chronic hepatitis B (CHB), alanine aminotransferase (ALT) ≥2 × upper limit of normal (ULN) is often used as a major criteria to initiate treatment in absence of cirrhosis, though patients with lower ALT may not be free from future risk of hepatocellular carcinoma (HCC). We aimed to examine the effect of antiviral therapy on HCC incidence based on ALT levels.We performed a retrospective study on 3665 patients consisting of United States and Taiwanese REVEAL-HBV cohort who were consecutive, treatment-naïve, noncirrhotic CHB patients aged ≥40 years. Patients were categorized by ALT cutoffs (≥2 × ULN vs ALT ALT ALT ALT levels.

  18. Comparison of FIB-4 and APRI in Chinese HBV-infected patients with persistently normal ALT and mildly elevated ALT.

    Science.gov (United States)

    Wang, H; Xue, L; Yan, R; Zhou, Y; Wang, M S; Cheng, M J; Huang, H J

    2013-04-01

    Significant liver disease has been reported in chronic hepatitis B patients with normal alanine aminotransferase (ALT). Liver biopsy (LB) is the current gold standard for assessing hepatic inflammation and fibrosis in patients with chronic HBV. However, associated risks have led to the development of noninvasive models. Their utility in patients with normal ALT is unknown. FIB-4 and aspartate aminotransferase (AST)-to-platelet ratio index (APRI) were calculated for patients with chronic HBV infection undergoing biopsy. The performance of each model and AUROC for predicting significant fibrosis (Scheuer's score ≥ S2) were determined for the entire cohort and stratified by elevated (≥50 U/L) and normal ALT. Two-hundred and thirty-one liver biopsies were included. The number of patient with normal ALT was 140, and 22.1% had significant fibrosis. The AUROC curve for patients with normal ALT was 0.81 for FIB-4 and 0.80 for APRI, compared with 0.71 for FIB-4 and 0.72 for APRI for those with mildly elevated ALT level. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of FIB-4 were 0.63, 0.88, 0.61 and 0.93, for patients with normal ALT; the values for APRI were 0.40, 0.88, 0.33 and 0.93. Both FIB-4 and APRI are useful for identification of those without significant fibrosis. However, because they have poor PPV, LB will continue to be used for assessment of HBV-infected patients with normal ALT and mildly elevated ALT. © 2012 Blackwell Publishing Ltd.

  19. Kui kapital veab alt, siis turvaliselt / Aleksander Tsapov

    Index Scriptorium Estoniae

    Tsapov, Aleksander

    2006-01-01

    Kuraatorinäitus "Kapital (see veab meid alt)" Tallinna Kunstihoones. Kuraator Simon Sheikh. Fia-Stina Sandlundi, Katya Sanderi, Oliver Ressleri, Ashley Hunti ja Susan Kelly&Stephen Mortoni töödest näitusel

  20. PISCES and ALT-II: Juelich PSI papers

    Energy Technology Data Exchange (ETDEWEB)

    Conn, R.W.; Hirooka, Y.; LaBombard, B.; Moyer, R.; Goebel, D.M.; Leung, W.K.; Nygren, R.E.; Corbett, W.J.; Lehmer, R.; Ra, Y.; Tynan, G.; Dippel, K.H.; Finken, K.H.; Hardkte, A.; Kohlhaas, W.; Wolf, G.; Vandenplas, P.; Messian, M.; Van Oost, G.; Weynants, R.; Franconi, E.; Miyahara, A.; Sagara, A.; Gauster, W.B.; Koski, J.A.; McGrath, R.T.; Watkins, J.G.; Malinowski, M.E.

    1988-08-01

    This publication comprises papers from the PISCES and ALT-II Programs at UCLA which were presented at the International Plasma Surface Interactions Meeting held in Juelich, FRG, on May 2-6, 1988. A list of publications from the PISCES and ALT-II contained in this report are: Deuterium pumping and erosion behavior of selected graphite materials under high flux plasma bombardment in PISCES; Erosion and redeposition behavior of selected NET-candidate materials under high-flux hydrogen, deuterium plasma bombardment in PISCES; Presheath profiles in simulated tokamak edge plasmas; Boundary asymmetries and plasma flow to the ALT-II toroidal belt pump limiter; ALT-II toroidal belt pump limiter performance in TEXTOR; and An in-situ spectroscopic erosion yield measurement with applications to sputtering and surface morphology alterations.

  1. PISCES and ALT-II: Juelich PSI papers

    International Nuclear Information System (INIS)

    Conn, R.W.; Hirooka, Y.; LaBombard, B.

    1988-08-01

    This publication comprises papers from the PISCES and ALT-II Programs at UCLA which were presented at the International Plasma Surface Interactions Meeting held in Juelich, FRG, on May 2-6, 1988. A list of publications from the PISCES and ALT-II contained in this report are: Deuterium pumping and erosion behavior of selected graphite materials under high flux plasma bombardment in PISCES; Erosion and redeposition behavior of selected NET-candidate materials under high-flux hydrogen, deuterium plasma bombardment in PISCES; Presheath profiles in simulated tokamak edge plasmas; Boundary asymmetries and plasma flow to the ALT-II toroidal belt pump limiter; ALT-II toroidal belt pump limiter performance in TEXTOR; and An in-situ spectroscopic erosion yield measurement with applications to sputtering and surface morphology alterations

  2. Decreasing ALT donor site morbidity with the keystone flap.

    Science.gov (United States)

    Turin, Sergey Y; Spitz, Jamie A; Alexander, Karina; Ellis, Marco F

    2018-03-09

    The anterolateral thigh (ALT) flap is a workhorse flap for a variety of wounds. The primary disadvantage of ALT flaps is donor site morbidity associated with large skin paddle defects. We present a strategy of managing the donor site with the Keystone flap to avoid skin grafts in cases where primary closure is not possible. A retrospective, multi-institutional review from December 2015 to March 2017 based on a single surgeon's experience was performed. Inclusion criteria were harvest of an ALT flap and closure of the thigh donor site with a keystone flap. Six patients underwent reconstruction of the ALT donor site with a keystone flap. The average width of the ALT skin paddle was 7.8 cm (range 7-8 cm) and the defects could not be closed primarily or with wide undermining. Surface area of the ALT flaps averaged 96.8 cm 2 (range 64-152 cm 2 ). Mean patient BMI was 24.6 (range 16-37). Keystone flap dimensions averaged 9.5 × 17.8 cm. There were no major donor or recipient site complications, and one ×minor complication of delayed wound healing. No patients exhibited decreased range of motion at the hip or knee. Primary closure of the ALT donor site is preferred, but becomes difficult as skin paddle width approaches 8 cm. Managing the ALT flap donor site with a keystone flap is safe, does not appear to have significant morbidity, and can allow the surgeon to avoid the morbidity and secondary donor site associated with skin grafting. © 2018 Wiley Periodicals, Inc.

  3. Establishment and characterization of a telomerase immortalized porcine luteal cells.

    Science.gov (United States)

    Zhang, Liang; Huang, Yong; Wang, Zhenyu; Luo, Xiaomao; Zhang, Hongling; Du, Qian; Chang, Lingling; Zhao, Xiaomin; Tong, Dewen

    2017-05-01

    Luteal cells play a crucial role in pregnancy through secreting progesterone to maintain pregnancy and support of fetus. However, low cellular yields and inability to passage primary porcine luteal cells (PLCs) in vitro limit the luteal cell study. Therefore, developing an immortalized porcine luteal cell line is necessary for studying luteal cells activity and function in different diseases. In this study, primary PLCs were obtained from gilts at day 30 to day 50 of gestation and immortalized by human telomerase reverse transcriptase (hTERT). The porcine corpus luteal cell line (hTERT-PLCs) expressed hTERT gene steady, maintained high hTERT activity and normal karyotype. The phase contrast microscope and transmission electron microscope observation showed primary PLCs and hTERT-PLCs were polygonal and exhibited abundant mitochondria, smooth endoplasmic reticulum and lipid droplets. 3β hydroxysteroid dehydrogenase (3βHSD) and Oil-Red-O staining showed that hTERT-PLCs at passage 30 and 50 were similar to primary PLCs. The hTERT-PLCs expressed steroidogenesis-related proteins, enzymes and receptors, such as steroidogenic acute regulatory protein, P450 cholesterol side-chain cleavage, 3βHSD, 20αHSD, luteinizing hormone receptor, progesterone receptor, prolactin receptor, estrogen receptorα/β, as well as primary PLCs. Consequently, hTERT-PLCs could secret progesterone and exhibited similar responses to luteinizing hormone and prostaglandin F2α as primary PLCs. In addition, the hTERT-PLCs did not show neoplastic transformation or anchorage independent growth. In summary, we developed an immortalized porcine luteal cell line which maintained its originally morphological, biological and functional characteristics. Copyright © 2017 Elsevier Inc. All rights reserved.

  4. ImmunoPET imaging of tissue factor expression in pancreatic cancer with89Zr-Df-ALT-836.

    Science.gov (United States)

    Hernandez, Reinier; England, Christopher G; Yang, Yunan; Valdovinos, Hector F; Liu, Bai; Wong, Hing C; Barnhart, Todd E; Cai, Weibo

    2017-10-28

    Overexpression of tissue factor (TF) has been associated with increased tumor growth, tumor angiogenesis, and metastatic potential in many malignancies, including pancreatic cancer. Additionally, high TF expression was shown to strongly correlate with poor prognoses and decreased survival in pancreatic cancer patients. Herein, we exploited the potential targeting of TF for positron emission tomography (PET) imaging of pancreatic cancer. The TF-targeted tracer was developed through radiolabeling of the anti-human TF monoclonal antibody (ALT-836) with 89 Zr. The tracer was characterized by fluorescence microscopy and flow cytometry assays in BXPC-3 and PANC-1 cells, two pancreatic cancer cell lines with high and low TF expression levels, respectively. Non-invasive PET scans were acquired in tumor-bearing mice injected with 89 Zr-Df-ALT-836. Additionally, ex vivo biodistribution, blocking, and histological studies were performed to establish the affinity and specificity of 89 Zr-Df-ALT-836 for TF in vivo. 89 Zr-labeling of Df-ALT-836 was achieved in high yield and good specific activity. Flow cytometry and microscopy studies revealed no detectable difference in TF-binding affinity between ALT-836 and Df-ALT-836 in vitro. Longitudinal PET scans unveiled a lasting and prominent 89 Zr-Df-ALT-836 uptake in BXPC-3 tumors (peak at 31.5±6.0%ID/g at 48h post-injection; n=3), which was significantly abrogated (2.3±0.5%ID/g at 48h post-injection; n=3) when mice were pre-injected with a blocking dose (50mg/kg) of unlabeled ALT-836. Ex vivo biodistribution data confirmed the accuracy of the PET results, and histological analysis correlated high tumor uptake with in situ TF expression. Taken together, these results attest to the excellent affinity and TF-specificity of 89 Zr-Df-ALT-836. With elevated, persistent, and specific accumulation in TF-positive BXPC-3 tumors, PET imaging using 89 Zr-Df-ALT-836 promises to open new avenues for improving future diagnosis, stratification

  5. HDAC9 regulates the alternative lengthening of telomere (ALT) pathway via the formation of ALT-associated PML bodies.

    Science.gov (United States)

    Jamiruddin, Mohd Raeed; Kaitsuka, Taku; Hakim, Farzana; Fujimura, Atsushi; Wei, Fan-Yan; Saitoh, Hisato; Tomizawa, Kazuhito

    2016-12-02

    Cancer cells overcome cellular senescence by activating the telomere maintenance mechanism, which can be either through telomerase or the alternative lengthening of telomeres (ALT). Being exclusive to cancer cells, targeting ALT is a more promising route for the development of drugs against cancer. The histone deacetylase (HDAC) family plays significant roles in various cellular processes. In addition to the regulation of gene expression, HDACs are also known to directly interact with many proteins. We focused on this family, and found that HDAC9 was up-regulated in ALT-positive cells. In ALT-positive cells treated with HDAC9 siRNA, there was a decrease in the telomere replicative capacity, which was evident from the C-circles assay. Furthermore, the formation of ALT-associated promyelocytic leukemia (PML) nuclear bodies (APBs) was inhibited by HDAC9 knockdown. Based on this study, it is suggested that HDAC9 regulates the formation of APBs and could be a candidate for the target of ALT-cancer therapy. Copyright © 2016 Elsevier Inc. All rights reserved.

  6. [The importance of AST / ALT rate in nonalcoholic steatohepatitis diagnosis].

    Science.gov (United States)

    Zamin, Júnior Idilio; de Mattos, Angelo Alves; Perin, Christiano; Ramos, Gabriel Zatti

    2002-01-01

    There is a histologic similarity between nonalcoholic steatohepatitis and alcoholic liver disease and in some cases differential diagnosis may be difficult, since some patients do not report abusive alcohol consumption. Evaluating the usefulness of setting the rate AST/ALT for the differential diagnosis of nonalcoholic steatohepatitis and alcoholic liver disease. Twenty nine obese patients with nonalcoholic steatohepatitis were compared with 28 patients with alcoholic liver disease. The diagnosis of nonalcoholic steatohepatitis was made after exclusion of other causes of liver disease and by histologic findings of, at least, macrovesicular steatosis and hepatocellular necrosis. In patients with nonalcoholic steatohepatitis the medium AST value was 52.3 +/- 21.2 U/L and ALT of 90.1 +/- 37.9 U/L, being the AST/ALT rate lower than 1 in all patients. In patients with alcoholic liver disease the medium AST value was 140 +/- 82.5 U/L and ALT was 50.6 +/- 40.3 U/L. The rate was higher than 1 in all cases and higher than 2 in 24 (85.7%), being statistically significant when compared with patients with nonalcoholic steatohepatitis. The AST/ALT rate seems to be useful in the differential diagnosis of liver diseases, while lower than 1 is highly suggestive of nonalcoholic steatohepatitis.

  7. Evaluation of the biological activities of the IL-15 superagonist complex, ALT-803, following intravenous versus subcutaneous administration in murine models.

    Science.gov (United States)

    Liu, Bai; Jones, Monica; Kong, Lin; Noel, Terra; Jeng, Emily K; Shi, Sixiang; England, Christopher G; Alter, Sarah; Miller, Jeffrey S; Cai, Weibo; Rhode, Peter R; Wong, Hing C

    2018-02-13

    ALT-803 is a fusion protein complex consisting of an interleukin (IL)-15 superagonist and a dimeric IL-15 receptor alpha sushi domain IgG1 Fc fusion protein. When administered to mice, ALT-803 is capable of inducing natural killer (NK) and CD8 + T cell proliferation and activation, and effectively promoting potent anti-tumor responses. Currently, ALT-803 is in clinical trials for treatment of various solid tumors and hematological malignancies. In the initial phase of these clinical studies, intravenous (iv) injection was used according to the route used in pre-clinical efficacy studies. In order to evaluate the possible advantage of subcutaneous (sc) injection versus iv injection, this study compared the biological activity of the two treatment regimens of ALT-803 in pre-clinical in vivo models. The pharmacokinetics, immune stimulation, and anti-tumor efficacy of iv and sc injection routes of ALT-803 in C57BL/6 mice were compared. The half-life of ALT-803 was 7.5 h for iv versus 7.7 h for sc with the maximal detected serum concentration of ALT-803 to be 3926 ng/ml at 0.5 h time-point following iv injection versus 495 ng/ml at 16 h post sc injection. Biodistribution studies indicated that sc ALT-803, similarly to iv ALT-803 as previously reported, has a greater tissue distribution and longer residence time in lymphoid tissues compared to recombinant IL-15. Notably, ALT-803 when administered either iv or sc induced comparable proliferation and activation of CD8 + T and NK cells and resulted in similar reductions of tumor burden. A toxicity study of mice receiving multiple injections of ALT-803 for 4 weeks by iv or sc routes revealed equivalent immune-related changes. The gradual absorbance into the blood stream and lower maximal blood levels of ALT-803 in sc-injected mice, along with similar anti-tumor efficacy support the administration of ALT-803 by sc injection in patients with various malignancies and infectious diseases. Copyright © 2017 Elsevier

  8. Effects of hTERT immortalization on osteogenic and adipogenic differentiation of dental pulp stem cells

    Directory of Open Access Journals (Sweden)

    Ikbale El-Ayachi

    2016-03-01

    Full Text Available These data relate to the differentiation of human dental pulp stem cells (DPSC and DPSC immortalized by constitutively expressing human telomerase reverse transcriptase (hTERT through both osteogenic and adipogenic lineages (i.e. to make bone producing and fat producing cells from these dental pulp stem cells. The data augment another study to characterize immortalized DPSC for the study of neurogenetic “Characterization of neurons from immortalized dental pulp stem cells for the study of neurogenetic disorders” [1]. Two copies of one typical control cell line (technical replicates were used in this study. The data represent the differentiation of primary DPSC into osteoblast cells approximately 60% more effectively than hTERT immortalized DPSC. Conversely, both primary and immortalized DPSC are poorly differentiated into adipocytes. The mRNA expression levels for both early and late adipogenic and osteogenic gene markers are shown.

  9. ELEVATED ALT AND AST IN AN ASYMPTOMATIC PERSON

    Directory of Open Access Journals (Sweden)

    KEW ST

    2009-01-01

    Full Text Available -Abnormal liver function test with raised alanine aminotransferase (ALT and raised aspartate aminotransferase (AST are commonly seen in primary care setting. -Chronic alcohol consumption, drugs, non-alcoholic steatohepatitis (NASH and chronic viral hepatitis are common causes associated with raised ALT and AST. -In chronic viral hepatitis, the elevation of liver enzyme may not correlate well with the degree of liver damage. -Non-hepatic causes of raised ALT and AST include polymyositis, acute muscles injury, acute myocardial infarction and hypothyroidism. -In the primary care setting, the doctor should obtain a complete history regarding the risk factors for viral hepatitis, substance abuse and request investigations accordingly. -Suspected chronic viral hepatitis and liver cirrhosis are best referred to hepatologist for further management.

  10. Antioxidant and Antifatigue Activities of Polygonatum Alte-lobatum Hayata Rhizomes in Rats

    Directory of Open Access Journals (Sweden)

    Chi-Ting Horng

    2014-11-01

    Full Text Available Polygonatum alte-lobatum Hayata, a rhizomatous perennial herb, belongs to the Liliaceae family and is endemic to Taiwan. We investigated the antioxidant and anti-fatigue activities of P. alte-lobatum in exercised rats. Levels of polyphenols, flavonoids and polysaccharides and 2,2-diphenyl-1-picrylhydrazyl (DPPH free-radical scavenging activity were measured in extracts of P. alte-lobatum (EPA. Sprague-Dawley rats were randomly divided into four groups for 8-week treatment with vehicle (control and low-, medium-, and high-dose EPA (LEPA, MEPA, HEPA; 0, 75, 150, and 375 mg/kg/day, respectively. Exercise performance was evaluated by exhaustive treadmill exercise time and by changes in body composition and biochemical variables at the end of the experiment. EPA contained polyphenols, flavonoids and polysaccharides, with polysaccharide content at least 26 times greater than that of polyphenols and flavonoids. Trend analysis revealed that EPA dose-dependently scavenged DPPH free radicals. EPA treatment dose-dependently increased endurance running time to exhaustion and superoxide dismutase activity and total antioxidant ability of blood. EPA dose-dependently decreased serum urea nitrogen and malondialdehyde levels after exercise. Hepatic glycogen content, an important energy source for exercise, was significantly increased with EPA treatment. EPA could be a potential agent with an anti-fatigue pharmacological function.

  11. ALT-II armor tile design for upgraded TEXTOR operation

    International Nuclear Information System (INIS)

    Newberry, B.L.; McGrath, R.T.; Watson, R.D.; Kohlhaas, W.; Finken, K.H.

    1994-01-01

    The upgrade of the TEXTOR tokamak at KFA Juelich was recently completed. This upgrade extended the TEXTOR pulse length from 5 seconds to 10 seconds. The auxiliary heating was increased to a total of 8.0 MW through a combination of neutral beam injection and radio frequency heating. Originally, the inertially cooled armor tiles of the full toroidal belt Advanced Limiter Test -- II (ALT-II) were designed for a 5-second operation with total heating of 6.0 MW. The upgrade of TEXTOR will increase the energy deposited per pulse onto the ALT-II by about 300%. Consequently, the graphite armor tiles for the ALT-II had to be redesigned to avoid excessively high graphite armor surface temperatures that would lead to unacceptable contamination of the plasma. This redesign took the form of two major changes in the ALT-II armor tile geometry. The first design change was an increase of the armor tile thermal mass, primarily by increasing the radial thickness of each tile from 17 mm to 20 mm. This increase in the radial tile dimension reduces the overall pumping efficiency of the ALT-II pump limiter by about 30%. The reduction in exhaust efficiency is unfortunate, but could be avoided only by active cooling of the ALT-II armor tiles. The active cooling option was too complicated and expensive to be considered at this time. The second design change involved redefining the plasma facing surface of each armor tile in order to fully utilize the entire surface area. The incident charged particle heat flux was distributed uniformly over the armor tile surfaces by carefully matching the radial, poloidal and toroidal curvature of each tile to the plasma flow in the TEXTOR boundary layer. This geometry redefinition complicates the manufacturing of the armor tiles, but results in significant thermal performance gains. In addition to these geometry upgrades, several material options were analyzed and evaluated

  12. ALT-II armor tile design for upgraded TEXTOR operation

    International Nuclear Information System (INIS)

    Newberry, B.L.; McGrath, R.T.; Watson, R.D.

    1994-01-01

    The upgrade of the TEXTOR tokamak at KFA Julich will be completed in the spring of 1994. The upgrade will extend the TEXTOR pulse length from 5 seconds to 10 seconds. The auxiliary heating systems are also scheduled to be upgraded so that eventually a total of 8.0 MW auxiliary heating will be available through a combination of neutral beam injection and radio frequency heating. Originally, the inertially cooled armor tiles on the full toroidal belt Advanced Limiter Test - II (ALT-II) were designed for 5-second operation with a total heating power of 6.0 MW. The upgrade of TEXTOR will increase the energy deposited per pulse onto ALT-II by more than 300%. Consequently, the graphite armor tiles for ALT-II had to be redesigned in order to increase their thermal inertia and, thereby, avoid excessively high graphite armor surface temperatures that would lead to unacceptable contamination of the plasma. The armor tile thermal inertia had been increase primarily by expanding the radial thickness of the tiles from 17 mm to 20 mm. This increase in radial tile dimension will reduce the overall pumping efficiency of the ALT-II pump limiter by about 30%. The final armor tile design was a compromise between increasing the power handling capability and reducing the particle exhaust efficiency of ALT-II. The reduction in exhaust efficiency is unfortunate, but could only be avoided by active cooling of the ALT-II armor tiles. The active cooling option was too complicated and expensive to be considered at this time

  13. ALT-II armor tile design for upgraded TEXTOR operation

    Energy Technology Data Exchange (ETDEWEB)

    Newberry, B.L. [Univ. of Michigan, Ann Arbor, MI (United States); McGrath, R.T.; Watson, R.D. [Sandia National Labs., Albuquerque, NM (United States); Kohlhaas, W.; Finken, K.H. [Kernforschungsanlage Juelich GmbH (Germany). Inst. fuer Plasmaphysik; Noda, N. [National Inst. for Fusion Science, Nagoya (Japan)

    1994-08-01

    The upgrade of the TEXTOR tokamak at KFA Juelich was recently completed. This upgrade extended the TEXTOR pulse length from 5 seconds to 10 seconds. The auxiliary heating was increased to a total of 8.0 MW through a combination of neutral beam injection and radio frequency heating. Originally, the inertially cooled armor tiles of the full toroidal belt Advanced Limiter Test -- II (ALT-II) were designed for a 5-second operation with total heating of 6.0 MW. The upgrade of TEXTOR will increase the energy deposited per pulse onto the ALT-II by about 300%. Consequently, the graphite armor tiles for the ALT-II had to be redesigned to avoid excessively high graphite armor surface temperatures that would lead to unacceptable contamination of the plasma. This redesign took the form of two major changes in the ALT-II armor tile geometry. The first design change was an increase of the armor tile thermal mass, primarily by increasing the radial thickness of each tile from 17 mm to 20 mm. This increase in the radial tile dimension reduces the overall pumping efficiency of the ALT-II pump limiter by about 30%. The reduction in exhaust efficiency is unfortunate, but could be avoided only by active cooling of the ALT-II armor tiles. The active cooling option was too complicated and expensive to be considered at this time. The second design change involved redefining the plasma facing surface of each armor tile in order to fully utilize the entire surface area. The incident charged particle heat flux was distributed uniformly over the armor tile surfaces by carefully matching the radial, poloidal and toroidal curvature of each tile to the plasma flow in the TEXTOR boundary layer. This geometry redefinition complicates the manufacturing of the armor tiles, but results in significant thermal performance gains. In addition to these geometry upgrades, several material options were analyzed and evaluated.

  14. Orbiter 'Enterprise' turns and banks during second ALT

    Science.gov (United States)

    1977-01-01

    The Orbiter 101 'Enterprise' makes a turn and bank maneuver during the second free flight of the Shuttle Apporach and Landing Tests (ALTs) conducted on September 13, 1977 at Dryden Flight Research Center in Southern California. The 'Enterprise' separated form the NASA 747 carrier aircraft and after a five-minute, 28-second unpowered flight landed on a runway below. Astronauts Joe H. Engle, and Richard H. Truly were the crew of the 'Enterprise.' The ALT free flights are designed to verify Orbiter subsonic airworthiness, integrated systems operations and pilot-guided approach and landing capability and satisfying prerequisites to automatic flight control and navigation mode.

  15. Preventive treatment of calcium oxalate crystal deposition with immortal flowers.

    Science.gov (United States)

    Orhan, Nilüfer; Onaran, Metin; Şen, İlker; Işık Gönül, İpek; Aslan, Mustafa

    2015-04-02

    A number of medicinal plants are used for their diuretic, urolithiatic and anti-inflammatory effects on urinary system problems in Turkey and the most common traditional remedy for kidney stones is the tea of immortal flowers. The aim of this study is to evaluate the preventive effect of infusions prepared from capitulums of Helichrysum graveolens (M.Bieb.) Sweet (HG) and Helichrysum stoechas ssp. barellieri (Ten.) Nyman (HS) on formation of kidney stones. Sodium oxalate (Ox-70mg/kg intraperitoneally) was used to induce kidney stones on Wistar albino rats. At the same time, two different doses of the plant extracts (HG: 62.5 and 125mg/kg; HS: 78 and 156mg/kg) were dissolved in the drinking water and administered to animals for 5 days. Potassium citrate was used as positive control in the experiments. During the experiment, water intake, urine volume and body weights of the animals were recorded. At the end of the experiments, liver, kidney and body weights of the animals were determined; biochemical analysis were conducted on urine, blood and plasma samples. Histopathological changes in kidney tissues were examined and statistical analysis were evaluated. HS extract showed the highest preventive effect at 156mg/kg dose (stone formation score: 1.16), whereas a number of kidney stones were maximum in sodium oxalate group (stone formation score: 2.66). Helichrysum extracts decreased urine oxalate and uric acid levels and increased citrate levels significantly. In addition, Helichrysum extracts regulated the negative changes in biochemical and hematological parameters occurred after Ox injection. We conclude that Helichrysum extracts could reduce the formation and growth of kidney stones in Ox-induced urolithiasis and can be beneficial for patients with recurrent stones. In addition, this is the first study on the preventive effect of immortal flowers. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  16. Oxidative DNA damage correlates with cell immortalization and mir-92 expression in hepatocellular carcinoma

    Directory of Open Access Journals (Sweden)

    Romilda Cardin

    2012-05-01

    Full Text Available Abstract Background MicroRNAs expression has been extensively studied in hepatocellular carcinoma but little is known regarding the relationship, if any, with inflammation, production of reactive oxygen species (ROS, host’s repair mechanisms and cell immortalization. This study aimed at assessing the extent of oxidative DNA damage (8-hydroxydeoxyguanosine - 8-OHdG in different phases of the carcinogenetic process, in relation to DNA repair gene polymorphism, telomeric dysfunction and to the expression of several microRNAs, non-coding genes involved in post-transcriptional regulation, cell proliferation, differentiation and death. Methods Tissue samples obtained either at surgery, [neoplastic (HCC and adjacent non-cancerous cirrhotic tissues (NCCT] at percutaneous or laparoscopic biopsy (patients with HCV or HBV-related hepatitis or patients undergoing cholecystectomy were analysed for 8-OHdG (HPLC-ED, OGG1 (a DNA repair gene polymorphism (PCR-RFLP, telomerase activity, telomere length (T/S, by RT-PCR, Taqman microRNA assay and Bad/Bax mRNA (RT-PCR. Fifty-eight samples from 29 HCC patients (obtained in both neoplastic and peritumoral tissues, 22 from chronic hepatitis (CH and 10 controls (cholecystectomy patients - CON were examined. Results Eight-OHdG levels were significantly higher in HCC and NCCT than in CH and CON (p=0.001. Telomerase activity was significantly higher in HCC than in the remaining subgroups (p=0.002; conversely T/S was significantly lower in HCC (p=0.05. MiR-199a-b, -195, -122, -92a and −145 were down-regulated in the majority of HCCs while miR-222 was up-regulated. A positive correlation was observed among 8-OHdG levels, disease stage, telomerase activity, OGG1 polymorphisms and ALT/GGT levels. In HCC, miR-92 expression correlated positively with telomerase activity, 8-OHdG levels and Bad/Bax mRNA. Conclusions The above findings confirm the accumulation, in the progression of chronic liver damage to HCC, of a ROS

  17. Down-regulation of T-STAR, a growth inhibitory protein, after SV40-mediated immortalization.

    Science.gov (United States)

    Kool, J; van Zaane, W; van der Eb, A J; Terleth, C

    2001-11-01

    Normal human cells can undergo a limited number of divisions, whereas transformed cells may have an extended life span and can give rise to immortal cells. To isolate genes involved in the immortalization process, gene expression in SV40-transformed preimmortal human fibroblasts was compared with expression in SV40-transformed immortalized fibroblasts using an mRNA differential display. We found that the growth-inhibitory protein testis-signal transduction and activation of RNA (T-STAR) a homologue of cell-cycle regulator Sam68, is strongly down-regulated in immortalized cells. Overexpression of T-STAR in the SV40-transformed immortalized cells resulted in a strong reduction of colony formation, whereas deletion of the RNA-binding domain of T-STAR abrogated this effect. Down-regulation of testis-signal transduction and activation of RNA (T-STAR) expression is found only in immortal cells isolated after a proliferative crisis accompanied with massive cell death. The strict correlation of down-regulation of T-STAR expression only in those immortal cells that arose after a clear proliferative crisis suggests that the loss of T-STAR might be necessary to bypass crisis.

  18. Establishment and culture optimization of a new type of pituitary immortalized cell line

    Energy Technology Data Exchange (ETDEWEB)

    Kokubu, Yuko [Graduate School of Life and Environmental Sciences, The University of Tsukuba, Tsukuba, Ibaraki 305-8562 (Japan); Asashima, Makoto [Graduate School of Life and Environmental Sciences, The University of Tsukuba, Tsukuba, Ibaraki 305-8562 (Japan); Life Science Center of TARA, The University of Tsukuba, Ibaraki-ken 305-8577 (Japan); Kurisaki, Akira, E-mail: akikuri@hotmail.com [Graduate School of Life and Environmental Sciences, The University of Tsukuba, Tsukuba, Ibaraki 305-8562 (Japan); Biotechnology Research Institute for Drug Discovery, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki 305-8562 (Japan)

    2015-08-07

    The pituitary gland is a center of the endocrine system that controls homeostasis in an organism by secreting various hormones. The glandular anterior pituitary consists of five different cell types, each expressing specific hormones. However, their regulation and the appropriate conditions for their in vitro culture are not well defined. Here, we report the immortalization of mouse pituitary cells by introducing TERT, E6, and E7 transgenes. The immortalized cell lines mainly expressed a thyrotroph-specific thyroid stimulating hormone beta (Tshb). After optimization of the culture conditions, these immortalized cells proliferated and maintained morphological characteristics similar to those of primary pituitary cells under sphere culture conditions in DMEM/F12 medium supplemented with N2, B27, basic FGF, and EGF. These cell lines responded to PKA or PKC pathway activators and induced the expression of Tshb mRNA. Moreover, transplantation of the immortalized cell line into subcutaneous regions and kidney capsules of mice further increased Tshb expression. These results suggest that immortalization of pituitary cells with TERT, E6, and E7 transgenes is a useful method for generating proliferating cells for the in vitro analysis of pituitary regulatory mechanisms. - Highlights: • Mouse pituitary cell lines were immortalized by introducing TERT, E6, and E7. • The immortalized cell lines mainly expressed thyroid stimulating hormone beta. • The cell lines responded to PKA or PKC pathway activators, and induced Tshb.

  19. Reconsidering the Use of Autoregressive Latent Trajectory (ALT) Models

    Science.gov (United States)

    Voelkle, Manuel C.

    2008-01-01

    The simultaneous estimation of autoregressive (simplex) structures and latent trajectories, so called ALT (autoregressive latent trajectory) models, is becoming an increasingly popular approach to the analysis of change. Although historically autoregressive (AR) and latent growth curve (LGC) models have been developed quite independently from each…

  20. Presentation and demonstration at Alt-i-lab 2005

    NARCIS (Netherlands)

    Tattersall, Colin

    2005-01-01

    These documents were used to accompany the Learning Design presentation and demonstration at Alt-i-lab 2005. The slides were shown during a plenary session describing the interoperability demo. The scenario was used to guide the demonstration process, and the UNFOLD handout was used to illustrate

  1. ALT and e-Learning in the UK

    NARCIS (Netherlands)

    Micha Wijngaarden; dr. Pierre Gorissen

    2002-01-01

    The Learning Technology Research Institute (LTRI) and the Association for Learning Technology (ALT) are two organisations within the UK that focus on ICT in the field of learning and teaching. Chapter of report on the Exchange Study Trip 2002, organised by SURF from the 21st till the 26th of April

  2. Astronauts Engle and Truly at breakfast priot to second ALT

    Science.gov (United States)

    1977-01-01

    Astronauts Joe H. Engle, center, commander and Richard H. Truly, left, pilot, enjoy breakfast prior to the second free flight of the Space Shuttle Approach and Landing Tests (ALT), on September 13, 1977, in Trailer 69 at Dryden Flight Research Center in Southern California. Joining the crew at breakfast is training officer Rich Nygren.

  3. Peek ja Lepik soovisid varade aresti alt vabastamist

    Index Scriptorium Estoniae

    2005-01-01

    LHV endine töötaja Oliver Peek väidab, et tema vastu esitatud süüdistused on alusetud ja tõendamata ning nõuab oma varade aresti alt vabastamist. Kristjan Lepingu kohtuasja arutamine lükati edasi 21. detsembrile. Endiselt käib uurimine, kui palju väidetavalt teabevargusega raha teeniti

  4. New pediatric percentiles of liver enzyme serum levels (ALT, AST, GGT): Effects of age, sex, BMI and pubertal stage.

    Science.gov (United States)

    Bussler, Sarah; Vogel, Mandy; Pietzner, Diana; Harms, Kristian; Buzek, Theresa; Penke, Melanie; Händel, Norman; Körner, Antje; Baumann, Ulrich; Kiess, Wieland; Flemming, Gunter

    2017-09-19

    The present study aims to clarify the effects of sex, age, BMI and puberty on transaminase serum levels in children and adolescents and to provide new age- and sex-related percentiles for alanine aminotransferase (ALT), aspartate aminotransferase (AST) and γ-glutamyltransferase (GGT). Venous blood and anthropometric data were collected from 4,126 cases. Excluded were cases of participants with potential hepatotoxic medication, with evidence of potential illness at the time of blood sampling and non-normal BMI (BMI  90 th ). The resulting data (N = 3,131 cases) were used for the calculations of ALT, AST, and GGT percentiles. Age- and sex-related reference intervals were established by using an LMSP-type method. Serum levels of transaminases follow age-specific patterns and relate to the onset of puberty. This observation is more pronounced in girls than in boys. The ALT percentiles showed similar shaped patterns in both sexes. Multivariate regression confirmed significant effects of puberty and BMI-SDS (β = 2.21) on ALT. Surprisingly, AST serum levels were negatively influenced by age (β = -1.42) and BMI-SDS (β = -0.15). The GGT percentiles revealed significant sex-specific differences, correlated positively with age (β = 0.37) and showed significant association with BMI-SDS (β = 1.16). Current reference values of ALT, AST and GGT serum levels were calculated for children between 11 months and 16.0 years, using modern analytical and statistical methods. This study extends the current knowledge about transaminases by revealing influences of age, sex, BMI, and puberty on the serum concentrations of all three parameters and has for these parameters one of the largest sample sizes published so far. This article is protected by copyright. All rights reserved. © 2017 by the American Association for the Study of Liver Diseases.

  5. An assessment of the clinical utility of serum ALT and AST in chronic hepatitis C.

    Science.gov (United States)

    Anderson; Zeng; Rock; Yoshida

    2000-07-01

    Background: Alanine aminotransferase (ALT) is frequently used as the sole biochemical marker for chronic hepatitis C (CHC), however, its value may be normal in cases with active disease. Recently, aspartate aminotransferase (AST) has been suggested as a useful predictor of liver pathology and conflicting results were obtained by using AST/ALT ratio to predict cirrhosis. Aims: To evaluate clinical utility of serum ALT and AST in CHC. Methods: The charts of 133 patients with CHC, whose ALT and AST were simultaneously tested from 1994 to 1996, were reviewed. ALT and AST were analyzed for both the entire cohort of patients and subgroups stratified for histopathology, age, gender, alcohol consumption, and risk factors of transmission. In 53 patients, the AST/ALT ratio was evaluated during interferon treatment. Results: The elevation of ALT significantly correlated with that of AST (r=0.79). The AST/ALT ratio increased with liver histological progression. The ratio >/=1 was predominantly in cirrhotic patients. During treatment the ratio increased. The AST remained elevated in eight of the 33 patients in whom the ALT had returned to normal during and after treatment. Conclusions: Both ALT and AST are useful markers for CHC. However, the AST may elevate alone, suggesting that measuring AST may be useful when the ALT is consistently normal. The AST/ALT ratio varies in different patients but increases with histological progression of fibrosis. An AST/ALT ratio >/=1 is highly suggestive of the presence of cirrhosis.

  6. Transporter mRNA expression in a conditionally immortalized rat small intestine epithelial cell line (TR-SIE).

    Science.gov (United States)

    Hosoya, Ken-ichi; Tomi, Masatoshi; Takayama, Megumi; Komokata, Yuko; Nakai, Daisuke; Tokui, Taro; Nishimura, Kenji; Ueda, Masatsugu; Obinata, Masuo; Hori, Satoko; Ohtsuki, Sumio; Amidon, Gordon L; Terasaki, Tetsuya

    2004-08-01

    Small intestine epithelial cell lines (TR-SIE), which are established from the small intestine of transgenic rats harboring temperature-sensitive simian virus 40 large T-antigen gene (tsA58 Tg rat), were used to characterize the mRNA expression of small intestine transporters. TR-SIE cells had a polygonal morphology and expressed cytokeratin protein and villin mRNA. Although the large T-antigen was strongly expressed at 33 degrees C, this was reduced at 37 and 39 degrees C. Concomitantly, the cell growth was arrested at 37 and 39 degrees C compared with that at 33 degrees C, suggesting that TR-SIE cells are conditionally immortalized cell lines. RT-PCR analysis revealed that TR-SIE cells expressed ABCB1 (mdr1a and mdr1b), ABCB4 (mdr2), ABCC2 (mrp2), ABCC6 (mrp6), ABCG1, ABCG2 (bcrp/mxr), Slc21a7 (Oatp3), Slc15a1 (PepT1), and Slc16a1 (Mct1). Conditionally immortalized rat small intestine epithelial cell lines were established from tsA58 Tg rats and expressed the mRNA of intestinal transporters.

  7. Expectations of immortality: dam safety management into the next millennium

    International Nuclear Information System (INIS)

    Palmer, M.D.

    1999-01-01

    Topics concerning the problems associated with older and aging dams are considered including: what can be done to extent the lifetime of an old dam, the decision to decommission a dam based on a value judgment that the risk of maintaining the dam is too great for society's acceptance, the possibility of change in the level of risk tolerance with time in a technological environment, traditional surveillance methods used by dam owners in the Y2K situation, and the unreality of dam immortality. Trends and means for preserving older dams for their owner's purposes are outlined, as well as their lifetime compared to that of the downstream systems they serve. Despite the fact that we live in a throwaway society, dam owners cannot just leave their dam asset when they are through with using it. Someone has to maintain the dam, or ensure that it is safely decommissioned when the owner is finished with it. On a worldwide scale the available pool of experienced dam engineers is shrinking. This problem needs to be addressed by a shift towards operating and dam safety management skills based on a firm awareness of dam design principles. A shift in society's expectations has occurred such that dam designers and owners must now recognize the impact a dam can have both on its natural and social environments. Because of the increasing emphasis on paying attention to the impacts of people's activities on the planet, engineers more than anyone else must have a significant influence in that direction. 9 refs

  8. Spontaneous Immortalization of Clinically Normal Colon-Derived Fibroblasts from a Familial Adenomatous Polyposis Patient

    Directory of Open Access Journals (Sweden)

    Nicholas R. Forsyth

    2004-05-01

    Full Text Available Normal human diploid cells do not spontaneously immortalize in culture, but instead enter replicative senescence after a finite number of population doublings. Ablation of key checkpoint arrest or cancersuppressor genes, through dominantly inherited germline mutation (p53+/-, Li-Fraumeni or viral oncogene expression (SV40 large T, HPV16/18, E6/E7 can lead to escape from senescence, additional doublings, entrance into crisis phase, where immortal clones emerge at low frequency. In the vast majority of cases, telomerase is reactivated and telomeres are stabilized. Here we describe the spontaneous immortalization of clinically normal fibroblasts derived from colonic stroma of a familial adenomatous polyposis (FAP patient. The preimmortal (C26C and the spontaneously immortalized derivative (C26Ci cells are heterozygous for a characterized germline mutation in exon 15 of the adenomatous polyposis coli gene. Immortalization was accompanied by spontaneous reactivation of endogenous telomerase and establishment of telomeres at presenescent lengths. Normal checkpoint behavior is retained and a diploid karyotype is maintained. These cells provide a valuable new addition to the limited number of spontaneously immortalized human cell types, particularly fibroblast cells, will be useful in experimentally determining the functional pathways in neoplastic development and in the identification of potential molecular targets for cancer chemoprevention.

  9. Rescue excavations on Alt-Laari settlement site, Tartumaa / Anti Lillak, Heiki Valk

    Index Scriptorium Estoniae

    Lillak, Anti

    2009-01-01

    Alt-Laari linnusasula tekkis nähtavasti rooma rauaajal. Kui Alt-Laari linnus oli kasutused arvatavasti I aastatuhandest II aastatuhandeni pKr. siis asula kestis edasi keskajal ning jäeti maha hiljemalt 14. sajandil

  10. Sensitivity and Specificity of Plasma ALT, ALP, and Bile Acids for Hepatitis in Labrador Retrievers

    NARCIS (Netherlands)

    Dirksen, K|info:eu-repo/dai/nl/412424428; Burgener, I A; Rothuizen, J|info:eu-repo/dai/nl/071276033; van den Ingh, T S G A M; Penning, L C|info:eu-repo/dai/nl/110369181; Spee, B|info:eu-repo/dai/nl/304830925; Fieten, H|info:eu-repo/dai/nl/314112596

    2017-01-01

    BACKGROUND: Biochemical indicators for diagnosing liver disease are plasma alanine aminotransferase activity (ALT), alkaline phosphatase activity (ALP), and bile acid concentration (BA). OBJECTIVES: To determine the sensitivity and specificity of ALT, ALP, and BA for detecting primary hepatitis (PH)

  11. The AST/ALT (De-Ritis) ratio

    OpenAIRE

    Rief, Peter; Pichler, Martin; Raggam, Reinhard; Hafner, Franz; Gerger, Armin; Eller, Philipp; Brodmann, Marianne; Gary, Thomas

    2016-01-01

    Abstract The aspartat aminotransferase (AST)/alanin aminotransferase (ALT) (De-Ritis) ratio (AAR) is an easily applicable blood test. An elevated AAR on the one hand has been associated with an increase in nonalcoholic fatty liver disease (NAFLD). NAFLD on the other hand is associated with an increase in cardiovascular disease, all-cause mortality, and diabetes. As the AAR is also elevated in case of muscular damage, we investigated AAR and its association with critical limb ischemia (CLI) in...

  12. Understanding and Targeting the ALT Pathway in Human Breast Cancer

    Science.gov (United States)

    2013-09-01

    both ATRX-deficient ALT cell lines and in the ATRX flox MEFs using methylation-sensitive restriction endonucleases . MspI and HpaII are isochizomers...block digestion by HpaII and restrict its activity to regions farther from the chromosome end, producing longer telomere fragments and an observed...methylation status and the level of T- SCEs41. Furthermore, I observed no change in the pattern of restriction digest after Cre-mediated deletion of ATRX in

  13. Shuttle Orbiter 'Enterprise' lands at Edwards AFB after second ALT

    Science.gov (United States)

    1977-01-01

    The Shuttle Orbiter 101 'Enterprise' lands on the desert at Edwards Air Force Base to conclude a five-minute, 31-second unpowered flight during the second free-flight of the Shuttle Approach and and Landing Test (ALT) series, on September 13, 1977. Two T-38 chase planes remain with the 'Enterprise' for the landing. Astronauts Joe H. Engle, commander, and Richard H. Truly, pilot, were the crewmen for the flight.

  14. Shuttle Orbiter 'Enterprise' lands at Edwards AFB after third ALT

    Science.gov (United States)

    1977-01-01

    The Shuttle Orbiter 101 'Enterprise' approaches touchdown on the runnway at Edwards Air Force Base to conclude a five-minute, 34-second unpowered flight during the third free-flight of the Shuttle Approach and and Landing Test (ALT) series, on September 23, 1977. Three T-38 chase planes follow close by. Astronauts Fred W. Haise,Jr., commander, and C. Gordon Fullerton, pilot, were the crewmen for the flight.

  15. Hepatic ALT isoenzymes are elevated in gluconeogenic conditions including diabetes and suppressed by insulin at the protein level.

    Science.gov (United States)

    Qian, Kun; Zhong, Shao; Xie, Keming; Yu, Daozhan; Yang, Rongze; Gong, Da-Wei

    2015-09-01

    Alanine transaminase (ALT) plays an important role in gluconeogenesis by converting alanine into pyruvate for glucose production. Early studies have shown that ALT activities are upregulated in gluconeogenic conditions and may be implicated in the development of diabetes. ALT consists of two isoforms, ALT1 and ALT2, with distinctive subcellular and tissue distributions. Whether and how they are regulated are largely unknown. By using Western blotting analysis, we measured hepatic ALT isoforms at the protein level in obese and diabetic animals and in Fao hepatoma cells treated with dexamethasone and insulin. In addition, we measured glucose output in Fao cells over-expressing ALT1 and ALT2. Both ALT isoforms in the liver were increased in diabetic Goto-Kakizaki rats and during fasting. However, in ob/ob mice, only ALT2, but not ALT1, protein levels were elevated, and the increase of ALT2 was correlated with that of ALT activity. We further demonstrated that, in vitro, both ALT1 and ALT2 were induced by glucocorticoid dexamethasone, but suppressed by insulin in Fao cells. Finally, we showed that the over-expression of ALT1 and ALT2 in Fao cells directly increased glucose output. We have shown the similarity and difference in the regulation of ALT isoforms in gluconeogenic conditions at the protein level, supporting that ALT isoenzymes play an important role in glucose metabolism and may be implicated the development of insulin resistance and diabetes. Copyright © 2015 John Wiley & Sons, Ltd.

  16. 21 CFR 862.1030 - Alanine amino transferase (ALT/SGPT) test system.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Alanine amino transferase (ALT/SGPT) test system... Test Systems § 862.1030 Alanine amino transferase (ALT/SGPT) test system. (a) Identification. An alanine amino transferase (ALT/SGPT) test system is a device intended to measure the activity of the...

  17. The conserved mitochondrial gene distribution in relatives of Turritopsis nutricula, an immortal jellyfish.

    Science.gov (United States)

    Devarapalli, Pratap; Kumavath, Ranjith N; Barh, Debmalya; Azevedo, Vasco

    2014-01-01

    Turritopsis nutricula (T. nutricula) is the one of the known reported organisms that can revert its life cycle to the polyp stage even after becoming sexually mature, defining itself as the only immortal organism in the animal kingdom. Therefore, the animal is having prime importance in basic biological, aging, and biomedical researches. However, till date, the genome of this organism has not been sequenced and even there is no molecular phylogenetic study to reveal its close relatives. Here, using phylogenetic analysis based on available 16s rRNA gene and protein sequences of Cytochrome oxidase subunit-I (COI or COX1) of T. nutricula, we have predicted the closest relatives of the organism. While we found Nemopsis bachei could be closest organism based on COX1 gene sequence; T. dohrnii may be designated as the closest taxon to T. nutricula based on rRNA. Moreover, we have figured out four species that showed similar root distance based on COX1 protein sequence.

  18. Alternative Liquid Fuels Simulation Model (AltSim).

    Energy Technology Data Exchange (ETDEWEB)

    Williams, Ryan; Baker, Arnold Barry; Drennen, Thomas E.

    2009-12-01

    The Alternative Liquid Fuels Simulation Model (AltSim) is a high-level dynamic simulation model which calculates and compares the production and end use costs, greenhouse gas emissions, and energy balances of several alternative liquid transportation fuels. These fuels include: corn ethanol, cellulosic ethanol from various feedstocks (switchgrass, corn stover, forest residue, and farmed trees), biodiesel, and diesels derived from natural gas (gas to liquid, or GTL), coal (coal to liquid, or CTL), and coal with biomass (CBTL). AltSim allows for comprehensive sensitivity analyses on capital costs, operation and maintenance costs, renewable and fossil fuel feedstock costs, feedstock conversion ratio, financial assumptions, tax credits, CO{sub 2} taxes, and plant capacity factor. This paper summarizes the structure and methodology of AltSim, presents results, and provides a detailed sensitivity analysis. The Energy Independence and Security Act (EISA) of 2007 sets a goal for the increased use of biofuels in the U.S., ultimately reaching 36 billion gallons by 2022. AltSim's base case assumes EPA projected feedstock costs in 2022 (EPA, 2009). For the base case assumptions, AltSim estimates per gallon production costs for the five ethanol feedstocks (corn, switchgrass, corn stover, forest residue, and farmed trees) of $1.86, $2.32, $2.45, $1.52, and $1.91, respectively. The projected production cost of biodiesel is $1.81/gallon. The estimates for CTL without biomass range from $1.36 to $2.22. With biomass, the estimated costs increase, ranging from $2.19 per gallon for the CTL option with 8% biomass to $2.79 per gallon for the CTL option with 30% biomass and carbon capture and sequestration. AltSim compares the greenhouse gas emissions (GHG) associated with both the production and consumption of the various fuels. EISA allows fuels emitting 20% less greenhouse gases (GHG) than conventional gasoline and diesels to qualify as renewable fuels. This allows several of the

  19. Thallium stimulates ethanol production in immortalized hippocampal neurons

    Science.gov (United States)

    2017-01-01

    Lactate and ethanol (EtOH) were determined in cell culture medium (CCM) of immortalized hippocampal neurons (HN9.10e cell line) before and after incubation with Thallium (Tl). This cell line is a reliable, in vitro model of one of the most vulnerable regions of central nervous system. Cells were incubated for 48 h with three different single Tl doses: 1, 10, 100 μg/L (corresponding to 4.9, 49 and 490 nM, respectively). After 48 h, neurons were “reperfused” with fresh CCM every 24/48 h until 7 days after the treatment and the removed CCM was collected and analysed. Confocal microscopy was employed to observe morphological changes. EtOH was determined by head space—solid phase microextraction -gas chromatography -mass spectrometry (HS-SPME-GCMS), lactate by RP-HPLC with UV detection. Tl exposure had significant effects on neuronal growth rate and morphology. The damage degree was dose-dependent. In not exposed cells, EtOH concentration was 0.18 ± 0.013 mM, which represents about 5% of lactate concentration (3.4 ± 0.10 mM). After Tl exposure lactate and EtOH increased. In CCM of 100 and 10 μg/L Tl-treated cells, lactate increased 24 h after reperfusion up to 2 and 3.3 times the control value, respectively. In CCM of 10 and 100 μg/L Tl-treated cells 24 h after reperfusion, EtOH increased up to 0.3 and 0.58 mmol/L. respectively. These results are consistent with significant alterations in energy metabolism, despite the low doses of Tl employed and the relatively short incubation time. PMID:29161327

  20. Thallium stimulates ethanol production in immortalized hippocampal neurons.

    Science.gov (United States)

    Colombaioni, Laura; Onor, Massimo; Benedetti, Edoardo; Bramanti, Emilia

    2017-01-01

    Lactate and ethanol (EtOH) were determined in cell culture medium (CCM) of immortalized hippocampal neurons (HN9.10e cell line) before and after incubation with Thallium (Tl). This cell line is a reliable, in vitro model of one of the most vulnerable regions of central nervous system. Cells were incubated for 48 h with three different single Tl doses: 1, 10, 100 μg/L (corresponding to 4.9, 49 and 490 nM, respectively). After 48 h, neurons were "reperfused" with fresh CCM every 24/48 h until 7 days after the treatment and the removed CCM was collected and analysed. Confocal microscopy was employed to observe morphological changes. EtOH was determined by head space-solid phase microextraction -gas chromatography -mass spectrometry (HS-SPME-GCMS), lactate by RP-HPLC with UV detection. Tl exposure had significant effects on neuronal growth rate and morphology. The damage degree was dose-dependent. In not exposed cells, EtOH concentration was 0.18 ± 0.013 mM, which represents about 5% of lactate concentration (3.4 ± 0.10 mM). After Tl exposure lactate and EtOH increased. In CCM of 100 and 10 μg/L Tl-treated cells, lactate increased 24 h after reperfusion up to 2 and 3.3 times the control value, respectively. In CCM of 10 and 100 μg/L Tl-treated cells 24 h after reperfusion, EtOH increased up to 0.3 and 0.58 mmol/L. respectively. These results are consistent with significant alterations in energy metabolism, despite the low doses of Tl employed and the relatively short incubation time.

  1. Thallium stimulates ethanol production in immortalized hippocampal neurons.

    Directory of Open Access Journals (Sweden)

    Laura Colombaioni

    Full Text Available Lactate and ethanol (EtOH were determined in cell culture medium (CCM of immortalized hippocampal neurons (HN9.10e cell line before and after incubation with Thallium (Tl. This cell line is a reliable, in vitro model of one of the most vulnerable regions of central nervous system. Cells were incubated for 48 h with three different single Tl doses: 1, 10, 100 μg/L (corresponding to 4.9, 49 and 490 nM, respectively. After 48 h, neurons were "reperfused" with fresh CCM every 24/48 h until 7 days after the treatment and the removed CCM was collected and analysed. Confocal microscopy was employed to observe morphological changes. EtOH was determined by head space-solid phase microextraction -gas chromatography -mass spectrometry (HS-SPME-GCMS, lactate by RP-HPLC with UV detection. Tl exposure had significant effects on neuronal growth rate and morphology. The damage degree was dose-dependent. In not exposed cells, EtOH concentration was 0.18 ± 0.013 mM, which represents about 5% of lactate concentration (3.4 ± 0.10 mM. After Tl exposure lactate and EtOH increased. In CCM of 100 and 10 μg/L Tl-treated cells, lactate increased 24 h after reperfusion up to 2 and 3.3 times the control value, respectively. In CCM of 10 and 100 μg/L Tl-treated cells 24 h after reperfusion, EtOH increased up to 0.3 and 0.58 mmol/L. respectively. These results are consistent with significant alterations in energy metabolism, despite the low doses of Tl employed and the relatively short incubation time.

  2. Characterization of Neurons from Immortalized Dental Pulp Stem Cells for the Study of Neurogenetic Disorders

    Science.gov (United States)

    Urraca, Nora; Memon, Rawaha; El-Iyachi, Ikbale; Goorha, Sarita; Valdez, Colleen; Tran, Quynh T.; Scroggs, Reese; Miranda-Carboni, Gustavo A.; Donaldson, Martin; Bridges, Dave; Reiter, Lawrence T.

    2015-01-01

    A major challenge to the study and treatment of neurogenetic syndromes is accessing live neurons for study from affected individuals. Although several sources of stem cells are currently available, acquiring these involve invasive procedures, may be difficult or expensive to generate and are limited in number. Dental pulp stem cells (DPSC) are multipotent stem cells that reside deep the pulp of shed teeth. To investigate the characteristics of DPSC that make them a valuable resource for translational research, we performed a set of viability, senescence, immortalization and gene expression studies on control DPSC and derived neurons. We investigated the basic transport conditions and maximum passage number for primary DPSC. We immortalized control DPSC using human telomerase reverse transcriptase (hTERT) and evaluated neuronal differentiation potential and global gene expression changes by RNA-seq. We show that neurons from immortalized DPSC share morphological and electrophysiological properties with non-immortalized DPSC. We also show that differentiation of DPSC into neurons significantly alters gene expression for 1305 transcripts. Here we show that these changes in gene expression are concurrent with changes in protein levels of the transcriptional repressor REST/NSRF, which is known to be involved in neuronal differentiation. Immortalization significantly altered the expression of 183 genes after neuronal differentiation, 94 of which also changed during differentiation. Our studies indicate that viable DPSC can be obtained from teeth stored for ≥72hrs, these can then be immortalized and still produce functional neurons for in vitro studies, but that constitutive hTERT immortalization is not be the best approach for long term use of patient derived DPSC for the study of disease. PMID:26599327

  3. Immortalized human myotonic dystrophy muscle cell lines to assess therapeutic compounds.

    Science.gov (United States)

    Arandel, Ludovic; Polay Espinoza, Micaela; Matloka, Magdalena; Bazinet, Audrey; De Dea Diniz, Damily; Naouar, Naïra; Rau, Frédérique; Jollet, Arnaud; Edom-Vovard, Frédérique; Mamchaoui, Kamel; Tarnopolsky, Mark; Puymirat, Jack; Battail, Christophe; Boland, Anne; Deleuze, Jean-Francois; Mouly, Vincent; Klein, Arnaud F; Furling, Denis

    2017-04-01

    Myotonic dystrophy type 1 (DM1) and type 2 (DM2) are autosomal dominant neuromuscular diseases caused by microsatellite expansions and belong to the family of RNA-dominant disorders. Availability of cellular models in which the DM mutation is expressed within its natural context is essential to facilitate efforts to identify new therapeutic compounds. Here, we generated immortalized DM1 and DM2 human muscle cell lines that display nuclear RNA aggregates of expanded repeats, a hallmark of myotonic dystrophy. Selected clones of DM1 and DM2 immortalized myoblasts behave as parental primary myoblasts with a reduced fusion capacity of immortalized DM1 myoblasts when compared with control and DM2 cells. Alternative splicing defects were observed in differentiated DM1 muscle cell lines, but not in DM2 lines. Splicing alterations did not result from differentiation delay because similar changes were found in immortalized DM1 transdifferentiated fibroblasts in which myogenic differentiation has been forced by overexpression of MYOD1. As a proof-of-concept, we show that antisense approaches alleviate disease-associated defects, and an RNA-seq analysis confirmed that the vast majority of mis-spliced events in immortalized DM1 muscle cells were affected by antisense treatment, with half of them significantly rescued in treated DM1 cells. Immortalized DM1 muscle cell lines displaying characteristic disease-associated molecular features such as nuclear RNA aggregates and splicing defects can be used as robust readouts for the screening of therapeutic compounds. Therefore, immortalized DM1 and DM2 muscle cell lines represent new models and tools to investigate molecular pathophysiological mechanisms and evaluate the in vitro effects of compounds on RNA toxicity associated with myotonic dystrophy mutations. © 2017. Published by The Company of Biologists Ltd.

  4. Immortalized human myotonic dystrophy muscle cell lines to assess therapeutic compounds

    Directory of Open Access Journals (Sweden)

    Ludovic Arandel

    2017-04-01

    Full Text Available Myotonic dystrophy type 1 (DM1 and type 2 (DM2 are autosomal dominant neuromuscular diseases caused by microsatellite expansions and belong to the family of RNA-dominant disorders. Availability of cellular models in which the DM mutation is expressed within its natural context is essential to facilitate efforts to identify new therapeutic compounds. Here, we generated immortalized DM1 and DM2 human muscle cell lines that display nuclear RNA aggregates of expanded repeats, a hallmark of myotonic dystrophy. Selected clones of DM1 and DM2 immortalized myoblasts behave as parental primary myoblasts with a reduced fusion capacity of immortalized DM1 myoblasts when compared with control and DM2 cells. Alternative splicing defects were observed in differentiated DM1 muscle cell lines, but not in DM2 lines. Splicing alterations did not result from differentiation delay because similar changes were found in immortalized DM1 transdifferentiated fibroblasts in which myogenic differentiation has been forced by overexpression of MYOD1. As a proof-of-concept, we show that antisense approaches alleviate disease-associated defects, and an RNA-seq analysis confirmed that the vast majority of mis-spliced events in immortalized DM1 muscle cells were affected by antisense treatment, with half of them significantly rescued in treated DM1 cells. Immortalized DM1 muscle cell lines displaying characteristic disease-associated molecular features such as nuclear RNA aggregates and splicing defects can be used as robust readouts for the screening of therapeutic compounds. Therefore, immortalized DM1 and DM2 muscle cell lines represent new models and tools to investigate molecular pathophysiological mechanisms and evaluate the in vitro effects of compounds on RNA toxicity associated with myotonic dystrophy mutations.

  5. Immortalized human myotonic dystrophy muscle cell lines to assess therapeutic compounds

    Science.gov (United States)

    Arandel, Ludovic; Polay Espinoza, Micaela; Matloka, Magdalena; Bazinet, Audrey; De Dea Diniz, Damily; Naouar, Naïra; Rau, Frédérique; Jollet, Arnaud; Edom-Vovard, Frédérique; Mamchaoui, Kamel; Tarnopolsky, Mark; Puymirat, Jack; Battail, Christophe; Boland, Anne; Deleuze, Jean-Francois; Mouly, Vincent; Klein, Arnaud F.

    2017-01-01

    ABSTRACT Myotonic dystrophy type 1 (DM1) and type 2 (DM2) are autosomal dominant neuromuscular diseases caused by microsatellite expansions and belong to the family of RNA-dominant disorders. Availability of cellular models in which the DM mutation is expressed within its natural context is essential to facilitate efforts to identify new therapeutic compounds. Here, we generated immortalized DM1 and DM2 human muscle cell lines that display nuclear RNA aggregates of expanded repeats, a hallmark of myotonic dystrophy. Selected clones of DM1 and DM2 immortalized myoblasts behave as parental primary myoblasts with a reduced fusion capacity of immortalized DM1 myoblasts when compared with control and DM2 cells. Alternative splicing defects were observed in differentiated DM1 muscle cell lines, but not in DM2 lines. Splicing alterations did not result from differentiation delay because similar changes were found in immortalized DM1 transdifferentiated fibroblasts in which myogenic differentiation has been forced by overexpression of MYOD1. As a proof-of-concept, we show that antisense approaches alleviate disease-associated defects, and an RNA-seq analysis confirmed that the vast majority of mis-spliced events in immortalized DM1 muscle cells were affected by antisense treatment, with half of them significantly rescued in treated DM1 cells. Immortalized DM1 muscle cell lines displaying characteristic disease-associated molecular features such as nuclear RNA aggregates and splicing defects can be used as robust readouts for the screening of therapeutic compounds. Therefore, immortalized DM1 and DM2 muscle cell lines represent new models and tools to investigate molecular pathophysiological mechanisms and evaluate the in vitro effects of compounds on RNA toxicity associated with myotonic dystrophy mutations. PMID:28188264

  6. Characterization of neurons from immortalized dental pulp stem cells for the study of neurogenetic disorders

    Directory of Open Access Journals (Sweden)

    Nora Urraca

    2015-11-01

    Full Text Available A major challenge to the study and treatment of neurogenetic syndromes is accessing live neurons for study from affected individuals. Although several sources of stem cells are currently available, acquiring these involve invasive procedures, may be difficult or expensive to generate and are limited in number. Dental pulp stem cells (DPSCs are multipotent stem cells that reside deep the pulp of shed teeth. To investigate the characteristics of DPSCs that make them a valuable resource for translational research, we performed a set of viability, senescence, immortalization and gene expression studies on control DPSC and derived neurons. We investigated the basic transport conditions and maximum passage number for primary DPSCs. We immortalized control DPSCs using human telomerase reverse transcriptase (hTERT and evaluated neuronal differentiation potential and global gene expression changes by RNA-seq. We show that neurons from immortalized DPSCs share morphological and electrophysiological properties with non-immortalized DPSCs. We also show that differentiation of DPSCs into neurons significantly alters gene expression for 1305 transcripts. Here we show that these changes in gene expression are concurrent with changes in protein levels of the transcriptional repressor REST/NRSF, which is known to be involved in neuronal differentiation. Immortalization significantly altered the expression of 183 genes after neuronal differentiation, 94 of which also changed during differentiation. Our studies indicate that viable DPSCs can be obtained from teeth stored for ≥72 h, these can then be immortalized and still produce functional neurons for in vitro studies, but that constitutive hTERT immortalization is not be the best approach for long term use of patient derived DPSCs for the study of disease.

  7. Are simple noninvasive scoring systems for fibrosis reliable in patients with NAFLD and normal ALT levels?

    Science.gov (United States)

    McPherson, Stuart; Anstee, Quentin M; Henderson, Elsbeth; Day, Christopher P; Burt, Alastair D

    2013-06-01

    Nonalcoholic fatty liver disease (NAFLD) is common and many affected individuals have normal-range alanine aminotransferase (ALT) levels. There is a need for a robust screening tool to triage individuals with advanced fibrosis for specialist care. The aim of this study was to assess the performance of noninvasive fibrosis tests in patients with biopsy-proven NAFLD and normal levels of ALT. Patients presenting at a fatty liver clinic between 1999 and 2009 were included in the study. Liver biopsies were assessed using the Kleiner score. The aspartate aminotransferase (AST)/ALT ratio, BARD, FIB-4 and NAFLD fibrosis scores were calculated. A total of 305 patients were included [70 with normal ALT levels (women: ALT≤30 IU/l, men: ALT≤45 IU/l) and 235 with elevated levels]. In total, 24% of patients with normal ALT levels and 17% of those with elevated ALT levels had advanced fibrosis (Kleiner stage 3-4). The FIB-4 performed best in identifying advanced fibrosis in patients with normal ALT (area under receiver operating characteristic curve=0.86, 82% sensitivity, 77% specificity and 92% negative predictive value). The sensitivity of the AST/ALT ratio and BARD and NAFLD fibrosis scores for advanced fibrosis was good in patients with normal ALT levels (94, 94 and 82%, respectively), but the specificity was low (44, 26 and 51%, respectively). The FIB-4 yielded best results in patients with elevated ALT levels. Using the FIB-4, 61% of patients with normal ALT levels and 63% of those with elevated ALT levels could avoid liver biopsy to exclude advanced fibrosis. In contrast, AST/ALT ratio and BARD and NAFLD scores would have led to a high proportion of patients with mild disease having to undergo a biopsy. The FIB-4 yielded good results in patients with normal or elevated ALT levels, reliably excluding advanced fibrosis and reducing the need for liver biopsy.

  8. Clinical course of chronic hepatitis B (CHB) presented with normal ALT in Asian American patients.

    Science.gov (United States)

    Nguyen, K; Pan, C; Xia, V; Hu, J; Hu, K-Q

    2015-10-01

    The clinical course for chronic hepatitis B (CHB) patients with normal ALT and with or without minimal histologic activity remains unclear. We assessed frequency, amplitude, disease activities, and associated factors of ALT and/or AST flares in this subpopulation. Forty-seven consecutive treatment naïve Asian patients with CHB were enrolled from two liver clinics between December 2003 and January 2013, who had normal baseline ALT by routine clinical biochemical testing performed 6 weeks before or after the liver biopsy. We defined a flare as elevation of ALT/AST above the upper limit of normal of ALT/AST. The mean follow-up was 37.6 (CI = 12, 88) months, and the mean age at entry into the study was 43.3 (CI = 19, 65); 22/47 (46.8%) were males; 15/45 (33.3%), HBeAg+; 68.1% had stage 0-1 fibrosis; 63.8% had grade 0-1 inflammation. During follow-up, 13/47 (27.7%) cases developed ALT flare at least once in a mean of 13.5 (CI = 2, 43) months after liver biopsy; ALT flare was not associated with baseline ALT level, fibrosis stage, inflammation grade, hepatitis B virus (HBV) DNA load, HBeAg status, HBV genotype, HBV precore and basal core promoter mutations. 11/13 (84/6%) of ALT flares resolved during follow-up. 13/13 (100%) of ALT flares met AASLD treatment criteria, but only 6/13 (46.2%) were on HBV treatment. Serum ALT and/or AST flares occur frequently in CHB carriers who initially presented with normal ALT during pretreatment period. Thus, regular follow-up is warranted despite status of ALT/AST. No clinical factors were found to be associated with ALT flares. © 2015 John Wiley & Sons Ltd.

  9. Liver Injury Assessment by Vetscan VS2 Analyzer and Most Frequently Used ALT/GTP Reagent.

    Science.gov (United States)

    Vatsalya, Vatsalya; Avila, Diana; Frimodig, Jane C; Barve, Shirish S; McClain, Craig J; Gobejishvili, Leila

    Liver injury is estimated by serum alanine aminotransferases (ALT) levels in experimental animal models. Laboratories use various techniques to measure ALT levels including assay reagents and chemistry analyzers. VetScan VS2 (VS2) is widely used in veterinary clinics and research laboratories for highly reproducible, convenient and effective testing. Alternatively, ALT liquid reagent is used by laboratories to estimate liver injury in animal studies. The aim of this study was to perform comparative analyses of data obtained from these two assays in two different animal models. In this study, we used two different mouse models and compared the ALT levels measured using VetScan VS2 chemistry analyzer and ALT liquid reagent. Immunohistochemical analysis of hepatic tissue was also performed to document liver pathology. The first model is a high fat diet feeding model that results in a mild hepatic steatosis (fat accumulation in the liver) without elevation of ALT levels. For a severe liver injury model, we employed a hepatotoxin-induced liver injury model (carbon tetrachloride, CCl4), which leads to the development of hepatic fibrosis and very high ALT levels. VetScan VS2 and ALT reagent gave different values of ALT for all animal groups. However, linear regression analysis showed a significantly high association between ALT levels obtained by VS2 and ALT liquid reagent in a high-fat feeding model with no liver injury. For the CCl 4 induced liver injury model, serum dilution (5 and 10 times) was performed to obtain accurate results with ALT reagent. ALT levels acquired from both techniques showed a close association. Interestingly, this correlation was closer when serum was diluted 5 fold. This study demonstrates that both methods give similar results when evaluating liver injury in animal studies. However, the serum dilution factor is critical for severe liver injury assessment when using ALT reagent and requires some optimization. In this regard, VetScan VS2 is

  10. DSLR Double Star Astrometry Using an Alt-Az Telescope

    Science.gov (United States)

    Frey, Thomas; Haworth, David

    2014-07-01

    The goal of this project was to determine if the double star's angular separation and position angle measurements could be successfully measured with a motor driven, alt-azimuth Dobsonian-mounted Newtonian telescope (without a field rotator), and a digital single-lens reflex (DSLR) camera. Additionally, the project was constrained by using as much existing equipment as much as possible, including an Apple MacBook Pro laptop and a Canon T2i camera. This project was additionally challenging because the first author had no experience with astrophotography.

  11. Two crews for the Shuttle Approach and Landing Tests (ALT)

    Science.gov (United States)

    1976-01-01

    The two crews for the Space Shuttle Approach and Landing Tests (ALT) are photographed at the Rockwell International Space Division's Orbiter assembly facility at Palmdale, California on the day of the rollout of the Shuttle Orbiter 101 'Enterprise' spacecraft. They are, left to right, Astronauts C. Gordon Fullerton, pilot of the first crew; Fred W. Haise Jr., commander of the first crew; Joe H. Engle, commander of the second crew; and Richard H. Truly, pilot of the second crew. The DC-9 size airplane-like Orbiter 101 is in the background.

  12. Establishment of immortalized human erythroid progenitor cell lines able to produce enucleated red blood cells.

    Directory of Open Access Journals (Sweden)

    Ryo Kurita

    Full Text Available Transfusion of red blood cells (RBCs is a standard and indispensable therapy in current clinical practice. In vitro production of RBCs offers a potential means to overcome a shortage of transfusable RBCs in some clinical situations and also to provide a source of cells free from possible infection or contamination by microorganisms. Thus, in vitro production of RBCs may become a standard procedure in the future. We previously reported the successful establishment of immortalized mouse erythroid progenitor cell lines that were able to produce mature RBCs very efficiently. Here, we have developed a reliable protocol for establishing immortalized human erythroid progenitor cell lines that are able to produce enucleated RBCs. These immortalized cell lines produce functional hemoglobin and express erythroid-specific markers, and these markers are upregulated following induction of differentiation in vitro. Most importantly, these immortalized cell lines all produce enucleated RBCs after induction of differentiation in vitro, although the efficiency of producing enucleated RBCs remains to be improved further. To the best of our knowledge, this is the first demonstration of the feasibility of using immortalized human erythroid progenitor cell lines as an ex vivo source for production of enucleated RBCs.

  13. Conditionally immortalized human pancreatic stellate cell lines demonstrate enhanced proliferation and migration in response to IGF-I

    Energy Technology Data Exchange (ETDEWEB)

    Rosendahl, Ann H., E-mail: ann.rosendahl@med.lu.se [Lund University, Department of Clinical Sciences Lund, Division of Surgery, Lund (Sweden); Lund University and Skåne University Hospital, Department of Clinical Sciences Lund, Division of Oncology and Pathology, Lund (Sweden); Gundewar, Chinmay; Said Hilmersson, Katarzyna [Lund University, Department of Clinical Sciences Lund, Division of Surgery, Lund (Sweden); Ni, Lan; Saleem, Moin A. [University of Bristol, School of Clinical Sciences, Children' s Renal Unit and Academic Renal Unit, Bristol (United Kingdom); Andersson, Roland [Lund University, Department of Clinical Sciences Lund, Division of Surgery, Lund (Sweden)

    2015-01-15

    Pancreatic stellate cells (PSCs) play a key role in the dense desmoplastic stroma associated with pancreatic ductal adenocarcinoma. Studies on human PSCs have been minimal due to difficulty in maintaining primary PSC in culture. We have generated the first conditionally immortalized human non-tumor (NPSC) and tumor-derived (TPSC) pancreatic stellate cells via transformation with the temperature-sensitive SV40 large T antigen and human telomerase (hTERT). These cells proliferate at 33°C. After transfer to 37°C, the SV40LT is switched off and the cells regain their primary PSC phenotype and growth characteristics. NPSC contained cytoplasmic vitamin A-storing lipid droplets, while both NPSC and TPSC expressed the characteristic markers αSMA, vimentin, desmin and GFAP. Proteome array analysis revealed that of the 55 evaluated proteins, 27 (49%) were upregulated ≥3-fold in TPSC compared to NPSC, including uPA, pentraxin-3, endoglin and endothelin-1. Two insulin-like growth factor binding proteins (IGFBPs) were inversely expressed. Although discordant IGFBP-2 and IGFBP-3 levels, IGF-I was found to stimulate proliferation of both NPSC and TPSC. Both basal and IGF-I stimulated motility was significantly enhanced in TPSC compared to NPSC. In conclusion, these cells provide a unique resource that will facilitate further study of the active stroma compartment associated with pancreatic cancer. - Highlights: • Generation of human conditionally immortalized human pancreatic stellate cell lines. • Temperature-sensitive SV40LT allows switch to primary PSC phenotype characteristics. • Proteome profiling revealed distinct expression patterns between TPSC and NPSC. • Enhanced IGF-I-stimulated proliferation and motility by TPSC compared to NPSC.

  14. The human CTC1/STN1/TEN1 complex regulates telomere maintenance in ALT cancer cells

    Energy Technology Data Exchange (ETDEWEB)

    Huang, Chenhui; Jia, Pingping; Chastain, Megan; Shiva, Olga; Chai, Weihang, E-mail: wchai@wsu.edu

    2017-06-15

    Maintaining functional telomeres is important for long-term proliferation of cells. About 15% of cancer cells are telomerase-negative and activate the alternative-lengthening of telomeres (ALT) pathway to maintain their telomeres. Recent studies have shown that the human CTC1/STN1/TEN1 complex (CST) plays a multi-faceted role in telomere maintenance in telomerase-expressing cancer cells. However, the role of CST in telomere maintenance in ALT cells is unclear. Here, we report that human CST forms a functional complex localizing in the ALT-associated PML bodies (APBs) in ALT cells throughout the cell cycle. Suppression of CST induces telomere instabilities including telomere fragility and elevates telomeric DNA recombination, leading to telomere dysfunction. In addition, CST deficiency significantly diminishes the abundance of extrachromosomal circular telomere DNA known as C-circles and t-circles. Suppression of CST also results in multinucleation in ALT cells and impairs cell proliferation. Our findings imply that the CST complex plays an important role in regulating telomere maintenance in ALT cells. - Highlights: • CST localizes at telomeres and ALT-associated PML bodies in ALT cells throughout the cell cycle. • CST is important for promoting telomeric DNA replication in ALT cells. • CST deficiency decreases ECTR formation and increases T-SCE. • CST deficiency impairs ALT cell proliferation and results in multinucleation.

  15. Immortalization protocols used in cell culture models of human breast morphogenesis

    DEFF Research Database (Denmark)

    Gudjonsson, T; Villadsen, R; Rønnov-Jessen, L

    2004-01-01

    Defining the key players in normal breast differentiation is instrumental to understanding how morphogenesis becomes defective during breast cancer progression. During the past 2 decades much effort has been devoted to the development of technologies for purification and expansion of primary human...... of the tissue of origin. In recent years, we have sought to establish immortalized primary breast cells, which retain crucial characteristics of their original in situ tissue pattern. This review discusses various approaches to immortalization of breast-derived epithelial and stromal cells and the application...

  16. C60 fullerene localization and membrane interactions in RAW 264.7 immortalized mouse macrophages

    Science.gov (United States)

    Russ, K. A.; Elvati, P.; Parsonage, T. L.; Dews, A.; Jarvis, J. A.; Ray, M.; Schneider, B.; Smith, P. J. S.; Williamson, P. T. F.; Violi, A.; Philbert, M. A.

    2016-02-01

    There continues to be a significant increase in the number and complexity of hydrophobic nanomaterials that are engineered for a variety of commercial purposes making human exposure a significant health concern. This study uses a combination of biophysical, biochemical and computational methods to probe potential mechanisms for uptake of C60 nanoparticles into various compartments of living immune cells. Cultures of RAW 264.7 immortalized murine macrophage were used as a canonical model of immune-competent cells that are likely to provide the first line of defense following inhalation. Modes of entry studied were endocytosis/pinocytosis and passive permeation of cellular membranes. The evidence suggests marginal uptake of C60 clusters is achieved through endocytosis/pinocytosis, and that passive diffusion into membranes provides a significant source of biologically-available nanomaterial. Computational modeling of both a single molecule and a small cluster of fullerenes predicts that low concentrations of fullerenes enter the membrane individually and produce limited perturbation; however, at higher concentrations the clusters in the membrane causes deformation of the membrane. These findings are bolstered by nuclear magnetic resonance (NMR) of model membranes that reveal deformation of the cell membrane upon exposure to high concentrations of fullerenes. The atomistic and NMR models fail to explain escape of the particle out of biological membranes, but are limited to idealized systems that do not completely recapitulate the complexity of cell membranes. The surprising contribution of passive modes of cellular entry provides new avenues for toxicological research that go beyond the pharmacological inhibition of bulk transport systems such as pinocytosis.There continues to be a significant increase in the number and complexity of hydrophobic nanomaterials that are engineered for a variety of commercial purposes making human exposure a significant health concern

  17. ALT-II toroidal belt pump limiter performance in TEXTOR

    Energy Technology Data Exchange (ETDEWEB)

    Goebel, D.M.; Conn, R.W.; Corbett, W.J.; Moyer, R.; Dippel, K.H.; Finken, K.H.; Hardtke, A.; Kohlhaas, W.; Wolf, G.; Gauster, W.B.

    1989-04-01

    The Advanced Limiter Test (ALT-II) is a toroidal belt pump limiter in the TEXTOR tokamak. ALT-II is composed of 8 blade segments which form an axisymmetric toroidal belt of 3.4 m/sup 2/ exposed surface area, located on the outside of the torus at 45/sup 0/ below the horizontal midplane. Ohmic plasma operation with ALT-II as the main limiter is characterized by a line-averaged density range of 5x10/sup 12/ to 5.5x10/sup 13/ cm/sup -3/ at B/sub T/=2 T and I/sub p/=340 kA, Z/sub eff/=1.1 to 2 and typically 40 to 95% of the power radiated depending on the plasma density. ICRH heating of the plasma with up to 2.6 MW of incident power has been achieved, which modifies the scrape-off layer (SOL) and the pump limiter performance. The recycling coefficient in TEXTOR is normally close to one, but helium RG conditioning and baking of the limiter at 400/sup 0/C is found to lower the recycling coefficient to 0.8 for the order of 10 shots. Measurements by arrays of probes in the SOL and thermocouples in the limiter tiles indicate the flow to the limiter is toroidally symmetric and poloidally asymmetric. The asymmetries result in different power and particle fluxes to the ion and electron drift sides of the limiter. The density and power scrape-off lengths are on the order of 1 cm and significantly longer on the outside of the torus. In spite of the flow asymmetry favoring the ion drift side near the tangency point, the longer e-folding lengths on the electron side in the SOL result in equal or higher particle collection by the electron side. The probe arrays indicate that during ohmic heating a total of 15 to 20% of the core efflux is incident on the neutralizer plates located in scoops beneath the blades. More particles are collected during ICRH auxiliary heating due to changes in the SOL profiles and shorter particle confinement times. (Abstract Truncated)

  18. Human papillomavirus type 59 immortalized keratinocytes express late viral proteins and infectious virus after calcium stimulation

    International Nuclear Information System (INIS)

    Lehr, Elizabeth E.; Qadadri, Brahim; Brown, Calla R.; Brown, Darron R.

    2003-01-01

    Human papillomavirus type 59 (HPV 59) is an oncogenic type related to HPV 18. HPV 59 was recently propagated in the athymic mouse xenograft system. A continuous keratinocyte cell line infected with HPV 59 was created from a foreskin xenograft grown in an athymic mouse. Cells were cultured beyond passage 50. The cells were highly pleomorphic, containing numerous abnormally shaped nuclei and mitotic figures. HPV 59 sequences were detected in the cells by DNA in situ hybridization in a diffuse nuclear distribution. Southern blots were consistent with an episomal state of HPV 59 DNA at approximately 50 copies per cell. Analysis of the cells using a PCR/reverse blot strip assay, which amplifies a portion of the L1 open reading frame, was strongly positive. Differentiation of cells in monolayers was induced by growth in F medium containing 2 mM calcium chloride for 10 days. Cells were harvested as a single tissue-like sheet, and histologic analysis revealed a four-to-six cell-thick layer. Transcripts encoding involucrin, a cornified envelope protein, and the E1-circumflexE4 and E1-circumflexE4-circumflexL1 viral transcripts were detected after several days of growth in F medium containing 2 mM calcium chloride. The E1-circumflexE4 and L1 proteins were detected by immunohistochemical analysis, and virus particles were seen in electron micrographs in a subset of differentiated cells. An extract of differentiated cells was prepared by vigorous sonication and was used to infect foreskin fragments. These fragments were implanted into athymic mice. HPV 59 was detected in the foreskin xenografts removed 4 months later by DNA in situ hybridization and PCR/reverse blot assay. Thus, the complete viral growth cycle, including production on infectious virus, was demonstrated in the HPV 59 immortalized cells grown in a simple culture system

  19. Effects of magnesium isoglycyrrhizinate on AST, ALT, and serum levels of Th1 cytokines in patients with allo-HSCT.

    Science.gov (United States)

    Lv, Jinglong; Xiao, Qing; Chen, Yongping; Fan, Xuegong; Liu, Xin; Liu, Fen; Luo, Guoping; Zhang, Bangshuo; Wang, Sheng

    2017-05-01

    This study aimed to investigate the protective effects of magnesium isoglycyrrhizinate (MGL) on aspartate aminotransferase (AST), alanine aminotransferase (ALT), and serum levels of T helper 1 (Th1) cytokines in patients with allogeneic hematopoietic stem cell transplantation (allo-HSCT). The study included 42 patients prepared for allo-HSCT, who were divided equally into MGL and reduced glutathione groups. The ALT and AST levels were detected 1day before pretreatment and transplantation, and 7, 14, and 21days after transplantation. The total days and times of fever, treatment time of patients in the laminar flow room, white blood cell (WBC) count, platelet (PTL) implantation time, and success rate of transplantation were recorded. The serum levels of Th1/Th2 cytokines were detected. MGL had a significant protective effect on AST 1day before transplantation and 7, 14, and 21days after transplantation, while ALT had a statistical difference only 7days after transplantation. MGL could shorten the duration of fever during transplantation and advance the WBC and PTL implantation time. Significant differences in Th1-like cytokines (P0.05) were found in the MGL group compared with the control group. MGL had significant protective effects on AST after transplantation. MGL could reduce the duration of fever during transplantation, help the reconstruction and recovery of WBCs and PTLs, and regulate Th1 cytokines, revealing its protective effects on hepatic transaminases and graft versus host disease in allo-HSCT patients. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. Elevated Alt and Ast in an Asymptomatic Person: What the primary care doctor should do?

    OpenAIRE

    Yin, Loh Keng; Tong, Kew Siang

    2009-01-01

    Abnormal liver function test with raised alanine aminotransferase (ALT) and raised aspartate aminotransferase (AST) are commonly seen in primary care setting.Chronic alcohol consumption, drugs, non-alcoholic steatohepatitis (NASH) and chronic viral hepatitis are common causes associated with raised ALT and AST.In chronic viral hepatitis, the elevation of liver enzyme may not correlate well with the degree of liver damage.Non-hepatic causes of raised ALT and AST include polymyositis, acute mus...

  1. Diorganosilacetylene-alt-diorganosilvinylene polymers and a process of preparation

    Science.gov (United States)

    Barton, Thomas J.; Ijadi-Maghsoodi, Sina; Pang, Yi

    1995-10-10

    The present invention provides linear organosilicon polymers including acetylene and vinylene moieties, and a process for their preparation. These diorganosilacetylene-alt-diorganosilvinylene linear polymers can be represented by the formula: --[--(R.sup.1)(R.sup.2)Si--C.tbd.C--(R.sup.3)(R.sup.4)Si--CH.dbd.CH--].sub .n --, wherein n.gtoreq.2; and each R.sup.1, R.sup.2, R.sup.3, and R.sup.4 is independently selected from the group consisting of hydrogen, halogen, alkyl, alkenyl, aryl, and aralkyl radicals. The polymers are soluble in organic solvents, air stable, and can be pulled into fibers or cast into films. They can be thermally converted into silicon carbide ceramic materials.

  2. Teaching from "The Immortal Life of Henrietta Lacks": Student Perspectives on Health Disparities and Medical Ethics

    Science.gov (United States)

    Dimaano, Christian; Spigner, Clarence

    2017-01-01

    Objective: "The Immortal Life of Henrietta Lacks" by Rebecca Skloot is an award-winning biography engaging its readers on important topics ranging from race, science and ethics to the social determinants of health. However, the multiple pedagogic impacts of this book on the public health classroom setting have yet to be comprehensively…

  3. Differential In Vitro Immortalization Capacity of Eleven, Probable High-Risk Human Papillomavirus Types

    NARCIS (Netherlands)

    Schutze, Denise M.; Snijders, Peter J. F.; Bosch, Leontien; Kramer, Duco; Meijer, Chris J. L. M.; Steenbergen, Renske D. M.

    Epidemiological studies identified 12 high-risk HPV (hrHPV) types and 8 probable/possible hrHPV types that display different cancer risks. Functional studies on transforming properties of hrHPV are mainly limited to HPV16 and -18, which induce immortalization of human foreskin keratinocytes (HFKs)

  4. Functional analysis of a novel human serotonin transporter gene promoter in immortalized raphe cells

    DEFF Research Database (Denmark)

    Mortensen, O V; Thomassen, M; Larsen, M B

    1999-01-01

    were found to possess the additional 379 bp fragment. The integrity of the promoter was furthermore confirmed by genomic Southern blotting. The promoter activity was analyzed by reporter gene assays in neuronal and non-neuronal serotonergic cell lines. In immortalized serotonergic raphe neurons, RN46A...

  5. Tissue distribution and engraftment of human mesenchymal stem cells immortalized by human telomerase reverse transcriptase gene

    DEFF Research Database (Denmark)

    Bentzon, J.F.; Stenderup, K.; Hansen, F.D.

    2005-01-01

    Engraftment of mesenchymal stem cells (MSC) in peripheral tissues for replenishing of local stem cell function has been proposed as a therapeutic approach to degenerative diseases. We have previously reported the development of an immortalized human telomerase reverse transcriptase transduced MSC...

  6. Establishment of an immortalized mouse dermal papilla cell strain with optimized culture strategy

    Directory of Open Access Journals (Sweden)

    Haiying Guo

    2018-01-01

    Full Text Available Dermal papilla (DP plays important roles in hair follicle regeneration. Long-term culture of mouse DP cells can provide enough cells for research and application of DP cells. We optimized the culture strategy for DP cells from three dimensions: stepwise dissection, collagen I coating, and optimized culture medium. Based on the optimized culture strategy, we immortalized primary DP cells with SV40 large T antigen, and established several immortalized DP cell strains. By comparing molecular expression and morphologic characteristics with primary DP cells, we found one cell strain named iDP6 was similar with primary DP cells. Further identifications illustrate that iDP6 expresses FGF7 and α-SMA, and has activity of alkaline phosphatase. During the process of characterization of immortalized DP cell strains, we also found that cells in DP were heterogeneous. We successfully optimized culture strategy for DP cells, and established an immortalized DP cell strain suitable for research and application of DP cells.

  7. Immortality of Prejudice in Striving Ubuntu: Case Studies of Community Managed Schools in Nepal

    Science.gov (United States)

    Rajbhandari, Mani Man Singh; Rajbhandari, Smriti

    2016-01-01

    The immortality of prejudice after the school management transfer has not been judged. This makes communities to take responsibility for schools further by compelling the government to mandate amendments of Community Managed Schools (CMS) Directives. The purpose was to explore the CMS enduring Ubuntu against immorality of prejudice, through…

  8. Human papilloma virus DNAs immortalize normal human mammary epithelial cells and reduce their growth factor requirements

    Energy Technology Data Exchange (ETDEWEB)

    Band, V.; Zajchowski, D.; Kulesa, V.; Sager, R. (Dana-Farber Cancer Institute, Boston, MA (USA))

    1990-01-01

    Human papilloma virus (HPV) types 16 and 18 are most commonly associated with cervical carcinoma in patients and induce immortalization of human keratinocytes in culture. HPV has not been associated with breast cancer. This report describes the immortalization of normal human mammary epithelial cells (76N) by plasmid pHPV18 or pHPV16, each containing the linearized viral genome. Transfectants were grown continuously for more than 60 passages, whereas 76N cells senesce after 18-20 passages. The transfectants also differ from 76N cells in cloning in a completely defined medium called D2 and growing a minimally supplemented defined medium (D3) containing epidermal growth factor. All transfectant tested contain integrated HPV DNA, express HPV RNA, and produce HPV E7 protein. HPV transfectants do not form tumors in a nude mouse assay. It is concluded that products of the HPV genome induce immortalization of human breast epithelial cells and reduce their growth factor requirements. This result raises the possibility that HPV might be involved in breast cancer. Furthermore, other tissue-specific primary epithelial cells that are presently difficult to grown and investigate may also be immortalized by HPV.

  9. Histone H3 Lysine 79 Methyltransferase Dot1 Is Required for Immortalization by MLL Oncogenes

    Science.gov (United States)

    Chang, Ming-Jin; Wu, Hongyu; Achille, Nicholas J.; Reisenauer, Mary Rose; Chou, Chau-Wen; Zeleznik-Le, Nancy J.; Hemenway, Charles S.; Zhang, Wenzheng

    2011-01-01

    Chimeric oncoproteins resulting from fusion of MLL to a wide variety of partnering proteins cause biologically distinctive and clinically aggressive acute leukemias. However, the mechanism of MLL-mediated leukemic transformation is not fully understood. Dot1, the only known histone H3 lysine 79 (H3K79) methyltransferase, has been shown to interact with multiple MLL fusion partners including AF9, ENL, AF10, and AF17. In this study, we utilize a conditional Dot1l deletion model to investigate the role of Dot1 in hematopoietic progenitor cell immortalization by MLL fusion proteins. Western blot and mass spectrometry show that Dot1-deficient cells are depleted of the global H3K79 methylation mark. We find that loss of Dot1 activity attenuates cell viability and colony formation potential of cells immortalized by MLL oncoproteins but not by the leukemic oncoprotein E2a-Pbx1. Although this effect is most pronounced for MLL-AF9, we find that Dot1 contributes to the viability of cells immortalized by other MLL oncoproteins that are not known to directly recruit Dot1. Cells immortalized by MLL fusions also show increased apoptosis, suggesting the involvement of Dot1 in survival pathways. In summary, our data point to a pivotal requirement for Dot1 in MLL fusion protein–mediated leukemogenesis and implicate Dot1 as a potential therapeutic target. PMID:21159644

  10. Computational study of pH-dependent oligomerization and ligand binding in Alt a 1, a highly allergenic protein with a unique fold.

    Science.gov (United States)

    Garrido-Arandia, María; Bretones, Jorge; Gómez-Casado, Cristina; Cubells, Nuria; Díaz-Perales, Araceli; Pacios, Luis F

    2016-05-01

    Alt a 1 is a highly allergenic protein from Alternaria fungi responsible for several respiratory diseases. Its crystal structure revealed a unique β-barrel fold that defines a new family exclusive to fungi and forms a symmetrical dimer in a butterfly-like shape as well as tetramers. Its biological function is as yet unknown but its localization in cell wall of Alternaria spores and its interactions in the onset of allergy reactions point to a function to transport ligands. However, at odds with binding features in β-barrel proteins, monomeric Alt a 1 seems unable to harbor ligands because the barrel is too narrow. Tetrameric Alt a 1 is able to bind the flavonoid quercetin, yet the stability of the aggregate and the own ligand binding are pH-dependent. At pH 6.5, which Alt a 1 would meet when secreted by spores in bronchial epithelium, tetramer-quercetin complex is stable. At pH 5.5, which Alt a 1 would meet in apoplast when infecting plants, the complex breaks down. By means of a combined computational study that includes docking calculations, empirical pKa estimates, Poisson-Boltzmann electrostatic potentials, and Molecular Dynamics simulations, we identified a putative binding site at the dimeric interface between subunits in tetramer. We propose an explanation on the pH-dependence of both oligomerization states and protein-ligand affinity of Alt a 1 in terms of electrostatic variations associated to distinct protonation states at different pHs. The uniqueness of this singular protein can thus be tracked in the combination of all these features.

  11. The People Paradox: Self-Esteem Striving, Immortality Ideologies, and Human Response to Climate Change

    Directory of Open Access Journals (Sweden)

    Janis L. Dickinson

    2009-06-01

    Full Text Available In 1973, Ernest Becker, a cultural anthropologist cross-trained in philosophy, sociology, and psychiatry, invoked consciousness of self and the inevitability of death as the primary sources of human anxiety and repression. He proposed that the psychological basis of cooperation, competition, and emotional and mental health is a tendency to hold tightly to anxiety-buffering cultural world views or "immortality projects" that serve as the basis for self-esteem and meaning. Although he focused mainly on social and political outcomes like war, torture, and genocide, he was increasingly aware that materialism, denial of nature, and immortality-striving efforts to control, rather than sanctify, the natural world were problems whose severity was increasing. In this paper I review Becker's ideas and suggest ways in which they illuminate human response to global climate change. Because immortality projects range from belief in technology and materialism to reverence for nature or belief in a celestial god, they act both as barriers to and facilitators of sustainable practices. I propose that Becker's cross-disciplinary "science of man," and the predictions it generates for proximate-level determinants of social behavior, add significantly to our understanding of and potential for managing the people paradox, i.e., that the very things that bring us symbolic immortality often conflict with our prospects for survival. Analysis of immortality projects as one of the proximate barriers to addressing climate change is both cautionary and hopeful, providing insights that should be included in the cross-disciplinary quest to uncover new pathways toward rational, social change.

  12. Can AST/ALT ratio indicate recovery after acute paracetamol poisoning?

    Science.gov (United States)

    McGovern, Allison J; Vitkovitsky, Irena V; Jones, Daniel L; Mullins, Michael E

    2015-03-01

    Paracetamol (acetaminophen or APAP) is the most common pharmaceutical exposure in the US. Elevations in aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels indicate hepatic toxicity. AST and ALT levels rise in similar proportions but later decline at different rates, with AST falling more rapidly than ALT. To determine whether the AST/ALT ratio can indicate that a patient has passed the time of peak AST concentration. We retrospectively identified cases of patients hospitalized for acute APAP poisoning by querying the pharmacy database of all patients treated with acetylcysteine (NAC) from January 1, 2001 to March 19, 2013. We included all patients with severe APAP poisoning, defined as AST or ALT greater than 1000 IU/L. Patients who were given NAC for other indications, those without APAP poisoning, and those receiving liver transplantation were excluded. We then recorded paired AST and ALT concentrations from each patient's hospital course. We classified each pair as clearly post-peak or not, and calculated the AST/ALT ratio for each pair of values. We compared different thresholds of AST/ALT ratio in increments of 0.1 to find the optimal value that reliably indicated resolving transaminases. We identified 1820 patients who received NAC during the study period. Of these, 333 received NAC for suspected poisoning by APAP. After excluding patients without severe APAP poisoning, other diagnoses explaining transaminase elevations, and patients who underwent liver transplantation, we had 37 evaluable patients with 343 evaluable pairs of AST and ALT concentrations. An AST/ALT ratio less than or equal to 0.4 was 99% sensitive for identifying patients with resolving transaminases. An AST/ALT ratio less than or equal to 0.4 following severe hepatoxicity from paracetamol poisoning appears to be highly predictive of recovery in patients treated with NAC. This has potential to be an indicator of safe discontinuation of NAC treatment.

  13. A block in lineage differentiation of immortal human mammary stem / progenitor cells by ectopically-expressed oncogenes

    Directory of Open Access Journals (Sweden)

    Xiangshan Zhao

    2011-01-01

    Full Text Available Introduction: Emerging evidence suggests a direct role of cancer stem cells (CSCs in the development of breast cancer. In vitro cellular models that recapitulate properties of CSCs are therefore highly desirable. We have previously shown that normal human mammary epithelial cells (hMECs immortalized with human telomerase reverse transcriptase (hTERT possess properties of mammary stem / progenitor cells. Materials and Methods: In the present study, we used this cell system to test the idea that other known hMEC-immortalizing oncogenes (RhoA, HPVE6, HPVE7, p53 mutant, and treatment with g-radiation, share with hTERT, the ability to maintain mammary stem / progenitor cells. Results: The results presented here demonstrate that similar to hMECs immortalized with hTERT, all hMEC cell lines immortalized using various oncogenic strategies express stem / progenitor cell markers. Furthermore, analyses using 2D and 3D culture assays demonstrate that all the immortal cell lines retain their ability to self-renew and to differentiate along the luminal lineage. Remarkably, the stem / progenitor cell lines generated using various oncogenic strategies exhibit a block in differentiation along the myoepithelial lineage, a trait that is retained on hTERT-immortalized stem / progenitors. The inability to differentiate along the myoepithelial lineage could be induced by ectopic mutant p53 expression in hTERT-immortalized hMEC. Conclusions: Our studies demonstrate that stem / progenitor cell characteristics of hMECs are maintained upon immortalization by using various cancer-relevant oncogenic strategies. Oncogene-immortalized hMECs show a block in their ability to differentiate along the myoepithelial lineage. Abrogation of the myoepithelial differentiation potential by a number of distinct oncogenic insults suggests a potential explanation for the predominance of luminal and rarity of myoepithelial breast cancers.

  14. The Genetic Architecture of Liver Enzyme Levels: GGT, ALT and AST

    NARCIS (Netherlands)

    van Beek, J.H.D.A.; de Moor, M.H.M.; de Geus, E.J.C.; Lubke, G.H.; Vink, J.M.; Willemsen, G.; Boomsma, D.I.

    2013-01-01

    High levels of liver enzymes GGT, ALT and AST are predictive of disease and all-cause mortality and can reflect liver injury, fatty liver and/or oxidative stress. Variation in GGT, ALT and AST levels is heritable. Moderation of the heritability of these liver enzymes by age and sex has not often

  15. Progressive liver functional impairment is associated with an increase in AST/ALT ratio.

    Science.gov (United States)

    Giannini, E; Botta, F; Fasoli, A; Ceppa, P; Risso, D; Lantieri, P B; Celle, G; Testa, R

    1999-06-01

    The ratio of serum aspartate aminotransferase to alanine aminotransferase (AST/ALT ratio) has been proposed as a noninvasive method of assessing liver fibrosis and cirrhosis. Our aims were to confirm the usefulness of the AST/ALT ratio in diagnosing cirrhosis noninvasively as well as to verify the existence of a relationship between the ratio and liver functional impairment. In all, 348 patients (177 with chronic hepatitis, 171 with cirrhosis) were retrospectively evaluated and the AST/ALT ratio was related to monoethyl glycine xylidide (MEGX) formation. Moreover, in a subgroup of 54 patients we analyzed the relationships among the AST/ALT ratio and indocyanine green clearance and half-life. The AST/ALT ratio was able to separate patients with mild fibrosis from those with severe fibrosis and cirrhosis. The AST/ALT ratio, MEGX, prothrombin activity, and platelet count were selected by multivariate analysis as variables associated with cirrhosis. The AST/ALT ratio showed significant correlations both with MEGX formation and with indocyanine green clearance and half-life. The alterations of indocyanine green kinetics, which depend upon liver blood flow and uptake, were likely due to progressive fibrosis. These findings might partially explain the increase in the AST/ALT ratio as disease progresses.

  16. Preliminary design analysis of the ALT-II limiter for TEXTOR

    International Nuclear Information System (INIS)

    Koski, J.A.; Boyd, R.D.; Kempka, S.M.; Romig, A.D. Jr.; Smith, M.F.; Watson, R.D.; Whitley, J.B.; Conn, R.W.; Grotz, S.P.

    1983-01-01

    Installation of a large toroidal belt pump limiter, Advanced Limiter Test II (ALT-II), on the TEXTOR tokamak at Juelich, FRG is anticipated for early 1986. This paper discusses the preliminary mechanical design and materials considerations undertaken as part of the feasibility study phase for ALT-II

  17. Telomerase suppresses formation of ALT-associated single-stranded telomeric C-circles.

    Science.gov (United States)

    Plantinga, Matthew J; Pascarelli, Kara M; Merkel, Anna S; Lazar, Alexander J; von Mehren, Margaret; Lev, Dina; Broccoli, Dominique

    2013-06-01

    Telomere maintenance is an essential characteristic of cancer cells, most commonly achieved by activation of telomerase. Telomeres can also be maintained by a recombination-based mechanism, alternative lengthening of telomeres (ALT). Cells using ALT are characterized by the presence of ALT-associated promyelocytic leukemia (PML) bodies (APB), long, heterogeneously sized telomeres, extrachromosomal telomeric circular DNA, and elevated telomeric recombination. Consistent with other reports, we found that liposarcomas containing APBs, but lacking telomerase expression, always contained C-rich circles (C-circles), and these C-circles were never present in the absence of APBs, indicating a tight link between these features in ALT cells. However, a rare subgroup of tumors showing evidence of telomere maintenance by both telomerase and ALT did not contain C-circles. To test the hypothesis that telomerase expression disrupts the tight link between APBs and C-circles, we used ALT cell lines that were engineered to express telomerase. Introduction of telomerase activity in these ALT cells resulted in, on average, shorter telomeres with retention of APBs. However, at high passage, the level of C-circles was significantly reduced, which was paralleled by a switch from C-strand overhangs to G-strand overhangs. We propose that by extending critically short telomeres in these cells, telomerase is disrupting a key step in the ALT pathway necessary for production and/or maintenance of C-circles. ©2013 AACR.

  18. Switch telomerase to ALT mechanism by inducing telomeric DNA damages and dysfunction of ATRX and DAXX.

    Science.gov (United States)

    Hu, Yang; Shi, Guang; Zhang, Laichen; Li, Feng; Jiang, Yuanling; Jiang, Shuai; Ma, Wenbin; Zhao, Yong; Songyang, Zhou; Huang, Junjiu

    2016-08-31

    Activation of telomerase or alternative lengthening of telomeres (ALT) is necessary for tumours to escape from dysfunctional telomere-mediated senescence. Anti-telomerase drugs might be effective in suppressing tumour growth in approximately 85-90% of telomerase-positive cancer cells. However, there are still chances for these cells to bypass drug treatment after switching to the ALT mechanism to maintain their telomere integrity. But the mechanism underlying this switch is unknown. In this study, we used telomerase-positive cancer cells (HTC75) to discover the mechanism of the telomerase-ALT switch by inducing telomere-specific DNA damage, alpha-thalassemia X-linked syndrome protein (ATRX) knockdown and deletion of death associated protein (DAXX). Surprisingly, two important ALT hallmarks in the ALT-like HTC75 cells were observed after treatments: ALT-associated promyelocytic leukaemia bodies (APBs) and extrachromosomal circular DNA of telomeric repeats. Moreover, knocking out hTERT by utilizing the CRISPR/Cas9 technique led to telomere elongation in a telomerase-independent manner in ALT-like HTC75 cells. In summary, this is the first report to show that inducing telomeric DNA damage, disrupting the ATRX/DAXX complex and inhibiting telomerase activity in telomerase-positive cancer cells lead to the ALT switch.

  19. The genetic architecture of liver enzyme levels: GGT, ALT and AST

    NARCIS (Netherlands)

    Beek, J.H.D.H. van; Moor, M.H.M. de; Geus, E.J.C. de; Lubke, G.H.; Vink, J.M.; Willemsen, G.; Boomsma, D.I.

    2013-01-01

    %High levels of liver enzymes GGT, ALT and AST are predictive of disease and all-cause mortality and can reflect liver injury, fatty liver and/or oxidative stress. Variation in GGT, ALT and AST levels is heritable. Moderation of the heritability of these liver enzymes by age and sex has not often

  20. Editorial: a new direction for ALT-J

    Directory of Open Access Journals (Sweden)

    Gráinne Conole

    2003-12-01

    Full Text Available The Association for Learning Technology celebrated its tenth anniversary this year and it can now be said that learning technology is a recognized research field, with a growing body of researchers and associated conferences and journals. Over the past two years as an editorial team we have undertaken a review of the position of ALT-J in relation to other journals in the area and considered the nature and scope of the papers we publish. We continue to expand our list of referees and have provided them with more detailed review forms to help guide them when considering recommendations on submissions and to encourage detailed, critical (and hopefully! helpful feedback to authors. We would like to take this opportunity to thank all the referees for their involvement and commitment to the journal and for their continued work on refereeing papers. As part of the review process we will be moving to a new publisher, Taylor and Francis, commencing with Volume 12; this is, in part, driven by a desire to increase our international profile and readership, as well as seeking to have a better online presence including electronic access to all papers. We are, after all, a research journal on learning technologies! We are looking forward to taking the journal forward with Taylor and Francis but would also like to take this opportunity to thank our current publisher, University of Wales Press, for all their support and professional work over the years; it has been very good working with them.

  1. Carrier-bound Alt a 1 peptides without allergenic activity for vaccination against Alternaria alternata allergy

    Science.gov (United States)

    Twaroch, T. E.; Focke, M.; Fleischmann, K.; Balic, N.; Lupinek, C.; Blatt, K.; Ferrara, R.; Mari, A.; Ebner, C.; Valent, P.; Spitzauer, S.; Swoboda, I.; Valenta, R.

    2017-01-01

    Background The mould Alternaria alternata is a major elicitor of allergic asthma. Diagnosis and specific immunotherapy (SIT) of Alternaria allergy are often limited by the insufficient quality of natural mould extracts. Objective To investigate whether recombinant Alt a 1 can be used for reliable diagnosis of Alternaria alternata allergy and to develop a safe, non-allergenic vaccine for SIT of Alternaria allergy. Methods The qualitative sensitization profile of 80 Alternaria-allergic patients from Austria and Italy was investigated using an allergen micro-array and the amount of Alternaria-specific IgE directed to rAlt a 1 was quantified by ImmunoCAP measurements. Peptides spanning regions of predicted high surface accessibility of Alt a 1 were synthesized and tested for IgE reactivity and allergenic activity, using sera and basophils from allergic patients. Carrier-bound peptides were studied for their ability to induce IgG antibodies in rabbits which recognize Alt a 1 and inhibit allergic patients’ IgE reactivity to Alt a 1. Results rAlt a 1 allowed diagnosis of Alternaria allergy in all tested patients, bound the vast majority (i.e. >95%) of Alternaria-specific IgE and elicited basophil activation already at a concentration of 0.1 ng/mL. Four non-allergenic peptides were synthesized which, after coupling to the carrier protein keyhole limpet hemocyanin, induced Alt a 1-specific IgG and inhibited allergic patients’ IgE binding to Alt a 1. Conclusions and clinical relevance rAlt a 1 is a highly allergenic molecule allowing sensitive diagnosis of Alternaria allergy. Carrier-bound non-allergenic Alt a 1 peptides are candidates for safe SIT of Alternaria allergy. PMID:22909168

  2. Development of a PCR-based tool for detecting immunologically relevant Alt a 1 and Alt a 1 homologue coding sequences.

    Science.gov (United States)

    Gabriel, M F; Postigo, I; Gutiérrez-Rodríguez, A; Suñén, E; Tomaz, C T; Martínez, J

    2015-08-01

    Alt a 1 has been recognized as the most important allergen produced by the Pleosporaceae family and is a risk factor for asthma development and/or exacerbation. The aim of this study was to develop a detection tool that is highly specific for species that produced airborne Alt a 1. Based on the highly conserved internal nucleotide region of the several Alt a 1 sequences that are available in GenBank, a pair of primers (Alta1CF/Alta1CR) was designed. A set of primers used by other authors for the production of recombinant Alt a 1 (A21F/A21R) was also tested. The molecular analyses were based on the polymerase chain reaction (PCR) amplification and sequencing of the cDNA that was obtained from thirteen common fungal species. The PCR system that utilized Alta1CF/Alta1CR was highly specific, sensitive, and was able to detect an amplicon of approximately 180 bp from Alt a 1 and Alt a 1-like encoding genes from A. alternata, A. tenuissima, A. infectoria, U. botrytis, and S. botryosum. In contrast, the A21F/A21R primers were specific for the very close taxonomically related species A. alternata and A. tenuissima. Thus, this rapid, sensitive, and specific detection tool can be used to assess Alt a 1 exposure levels and to inform the implementation of the appropriate public health measures. © The Author 2015. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  3. Regulation of Telomere Homeostasis during Epstein-Barr virus Infection and Immortalization.

    Science.gov (United States)

    Kamranvar, Siamak A; Masucci, Maria G

    2017-08-09

    The acquisition of unlimited proliferative potential is dependent on the activation of mechanisms for telomere maintenance, which counteracts telomere shortening and the consequent triggering of the DNA damage response, cell cycle arrest, and apoptosis. The capacity of Epstein Barr virus (EBV) to infect B-lymphocytes in vitro and transform the infected cells into autonomously proliferating immortal cell lines underlies the association of this human gamma-herpesvirus with a broad variety of lymphoid and epithelial cell malignancies. Current evidence suggests that both telomerase-dependent and -independent pathways of telomere elongation are activated in the infected cells during the early and late phases of virus-induced immortalization. Here we review the interaction of EBV with different components of the telomere maintenance machinery and the mechanisms by which the virus regulates telomere homeostasis in proliferating cells. We also discuss how these viral strategies may contribute to malignant transformation.

  4. The AST/ALT (De-Ritis) ratio

    Science.gov (United States)

    Rief, Peter; Pichler, Martin; Raggam, Reinhard; Hafner, Franz; Gerger, Armin; Eller, Philipp; Brodmann, Marianne; Gary, Thomas

    2016-01-01

    Abstract The aspartat aminotransferase (AST)/alanin aminotransferase (ALT) (De-Ritis) ratio (AAR) is an easily applicable blood test. An elevated AAR on the one hand has been associated with an increase in nonalcoholic fatty liver disease (NAFLD). NAFLD on the other hand is associated with an increase in cardiovascular disease, all-cause mortality, and diabetes. As the AAR is also elevated in case of muscular damage, we investigated AAR and its association with critical limb ischemia (CLI) in peripheral arterial occlusive disease (PAOD) patients. In our cross-sectional study, we included 1782 PAOD patients treated at our institution from 2005 to 2010. Patients with chronic alcohol consumption (>20 g/day) were excluded. AAR was calculated and the cohort was categorized into tertiles according to the AAR. An optimal cut-off value for the continuous AAR was calculated by applying a receiver operating curve analysis to discriminate between CLI and non-CLI. In our cohort, occurrence of CLI significantly increased with an elevation in AAR. As an optimal cut-off value, an AAR of 1.67 (sensitivity 34.1%, specificity 81.0%) was identified. Two groups were categorized, 1st group containing 1385 patients (AAR  1.67). CLI was more frequent in AAR > 1.67 patients (166 [41.9%]) compared to AAR  1.67 was associated with an odds ratio (OR) of 2.0 (95% confidence interval [CI] 1.7–2.3) for CLI even after adjustment for other well-established vascular risk factors. An increased AAR is significantly associated with patients at high risk for CLI and other cardiovascular endpoints. The AAR is a broadly available and cheap marker, which might be useful to highlight patients at high risk for vascular endpoints. PMID:27310963

  5. The People Paradox: Self-Esteem Striving, Immortality Ideologies, and Human Response to Climate Change

    OpenAIRE

    Janis L. Dickinson

    2009-01-01

    In 1973, Ernest Becker, a cultural anthropologist cross-trained in philosophy, sociology, and psychiatry, invoked consciousness of self and the inevitability of death as the primary sources of human anxiety and repression. He proposed that the psychological basis of cooperation, competition, and emotional and mental health is a tendency to hold tightly to anxiety-buffering cultural world views or "immortality projects" that serve as the basis for self-esteem and meaning. Although he focused m...

  6. Immortalized human myotonic dystrophy muscle cell lines to assess therapeutic compounds

    OpenAIRE

    Arandel, Ludovic; Polay Espinoza, Micaela; Matloka, Magdalena; Bazinet, Audrey; De Dea Diniz, Damily; Naouar, Na?ra; Rau, Fr?d?rique; Jollet, Arnaud; Edom-Vovard, Fr?d?rique; Mamchaoui, Kamel; Tarnopolsky, Mark; Puymirat, Jack; Battail, Christophe; Boland, Anne; Deleuze, Jean-Francois

    2017-01-01

    International audience; Myotonic dystrophy type 1 (DM1) and type 2 (DM2) are autosomal dominant neuromuscular diseases caused by microsatellite expansions and belong to the family of RNA-dominant disorders. Availability of cellular models in which the DM mutation is expressed within its natural context is essential to facilitate efforts to identify new therapeutic compounds. Here, we generated immortalized DM1 and DM2 human muscle cell lines that display nuclear RNA aggregates of expanded rep...

  7. Derivation and Osmotolerance Characterization of Three Immortalized Tilapia (Oreochromis mossambicus) Cell Lines

    Science.gov (United States)

    Gardell, Alison M.; Qin, Qin; Rice, Robert H.; Li, Johnathan; Kültz, Dietmar

    2014-01-01

    Fish cell cultures are becoming more widely used models for investigating molecular mechanisms of physiological response to environmental challenge. In this study, we derived two immortalized Mozambique tilapia (Oreochromis mossambicus) cell lines from brain (OmB) and lip epithelium (OmL), and compared them to a previously immortalized bulbus arteriosus (TmB) cell line. The OmB and OmL cell lines were generated without or with Rho-associated kinase (ROCK) inhibitor/3T3 feeder layer supplementation. Although both approaches were successful, ROCK inhibitor/feeder layer supplementation was found to offer the advantages of selecting for epithelial-like cell type and decreasing time to immortalization. After immortalization (≥ passage 5), we characterized the proteomes of the newly derived cell lines (OmB and OmL) using LCMS and identified several unique cell markers for each line. Subsequently, osmotolerance for each of the three cell lines following acute exposure to elevated sodium chloride was evaluated. The acute maximum osmotolerance of these tilapia cell lines (>700 mOsm/kg) was markedly higher than that of any other known vertebrate cell line, but was significantly higher in the epithelial-like OmL cell line. To validate the physiological relevance of these tilapia cell lines, we quantified the effects of acute hyperosmotic challenge (450 mOsm/kg and 700 mOsm/kg) on the transcriptional regulation of two enzymes involved in biosynthesis of the compatible organic osmolyte, myo-inositol. Both enzymes were found to be robustly upregulated in all three tilapia cell lines. Therefore, the newly established tilapia cells lines represent valuable tools for studying molecular mechanisms involved in the osmotic stress response of euryhaline fish. PMID:24797371

  8. Derivation and osmotolerance characterization of three immortalized tilapia (Oreochromis mossambicus cell lines.

    Directory of Open Access Journals (Sweden)

    Alison M Gardell

    Full Text Available Fish cell cultures are becoming more widely used models for investigating molecular mechanisms of physiological response to environmental challenge. In this study, we derived two immortalized Mozambique tilapia (Oreochromis mossambicus cell lines from brain (OmB and lip epithelium (OmL, and compared them to a previously immortalized bulbus arteriosus (TmB cell line. The OmB and OmL cell lines were generated without or with Rho-associated kinase (ROCK inhibitor/3T3 feeder layer supplementation. Although both approaches were successful, ROCK inhibitor/feeder layer supplementation was found to offer the advantages of selecting for epithelial-like cell type and decreasing time to immortalization. After immortalization (≥ passage 5, we characterized the proteomes of the newly derived cell lines (OmB and OmL using LCMS and identified several unique cell markers for each line. Subsequently, osmotolerance for each of the three cell lines following acute exposure to elevated sodium chloride was evaluated. The acute maximum osmotolerance of these tilapia cell lines (>700 mOsm/kg was markedly higher than that of any other known vertebrate cell line, but was significantly higher in the epithelial-like OmL cell line. To validate the physiological relevance of these tilapia cell lines, we quantified the effects of acute hyperosmotic challenge (450 mOsm/kg and 700 mOsm/kg on the transcriptional regulation of two enzymes involved in biosynthesis of the compatible organic osmolyte, myo-inositol. Both enzymes were found to be robustly upregulated in all three tilapia cell lines. Therefore, the newly established tilapia cells lines represent valuable tools for studying molecular mechanisms involved in the osmotic stress response of euryhaline fish.

  9. Generation and characterization of a spontaneously immortalized endothelial cell line from mice microcirculation.

    Science.gov (United States)

    Loiola, Rodrigo A; Torres, Tathiany C; Aburaya, Carla M; Landgraf, Maristella A; Landgraf, Richardt G; Bosco Pesquero, João; Fernandes, Liliam

    2013-05-01

    Endothelial cells from microvasculature are directly involved in a large number of vascular diseases; however, culture of these cells is problematic, since most methodologies employ proteolytic enzymes or mechanical techniques, leading to cell damage and contamination of endothelial cultures with other cellular types. Besides, primary cultured cells have a short life span in vitro and undergo replicative senescence after 3-4 passages, limiting long-term studies. In the present work we report the generation of a spontaneously immortalized endothelial culture obtained from mice pulmonary capillaries. Firstly, primary (third passage) and immortalized (100th) cultures were established. Further, monoclonal populations were obtained by serial dilutions from immortalized cultures. Cells were analyzed according to: (1) morphological appearance, (2) expression of specific endothelial markers by fluorescent staining [von Willebrand Factor (vWF), endothelial nitric oxide synthase (eNOS), angiotensin converting enzyme (ACE) and Ulex europaeus (UEA-1)] and by flow cytometry (endoglin, VE-cadherin and VCAM-1), and (3) release of nitric oxide (NO), assessed by the specific fluorescent dye DAF-2 DA, and prostacyclin (PGI2), quantified by enzyme immune assay. In both cultures cells grew in monolayers and presented cobblestone appearance at confluence. Positive staining for vWF, eNOS, ACE and UEA-1 was detected in cloned as well as in early-passage cultured cells. Similarly, cultures presented equal expressions of endoglin, VE-cadherin and VCAM-1. Values of NO and PGI2 levels did not differ between cultures. From these results we confirm that the described spontaneously immortalized endothelial cell line is capable of unlimited growth and retains typical morphological and functional properties exhibited by primary cultured cells. Therefore, the endothelial cell line described in the present study can become a suitable tool in the field of endothelium research and can be useful for

  10. Characterisation of a flavonoid ligand of the fungal protein Alt a 1.

    Science.gov (United States)

    Garrido-Arandia, María; Silva-Navas, Javier; Ramírez-Castillejo, Carmen; Cubells-Baeza, Nuria; Gómez-Casado, Cristina; Barber, Domingo; Pozo, Juan C; Melendi, Pablo G; Pacios, Luis F; Díaz-Perales, Araceli

    2016-09-16

    Spores of pathogenic fungi are virtually ubiquitous and cause human disease and severe losses in crops. The endophytic fungi Alternaria species produce host-selective phytotoxins. Alt a 1 is a strongly allergenic protein found in A. alternata that causes severe asthma. Despite the well-established pathogenicity of Alt a 1, the molecular mechanisms underlying its action and physiological function remain largely unknown. To gain insight into the role played by this protein in the pathogenicity of the fungus, we studied production of Alt a 1 and its activity in spores. We found that Alt a 1 accumulates inside spores and that its release with a ligand is pH-dependent, with optimum production in the 5.0-6.5 interval. The Alt a 1 ligand was identified as a methylated flavonoid that inhibits plant root growth and detoxifies reactive oxygen species. We also found that Alt a 1 changes its oligomerization state depending on the pH of the surrounding medium and that these changes facilitate the release of the ligand. Based on these results, we propose that release of Alt a 1 should be a pathogenic target in approaches used to block plant defenses and consequently to favor fungal entry into the plant.

  11. IL-15 super-agonist (ALT-803) enhances natural killer (NK) cell function against ovarian cancer.

    Science.gov (United States)

    Felices, M; Chu, S; Kodal, B; Bendzick, L; Ryan, C; Lenvik, A J; Boylan, K L M; Wong, H C; Skubitz, A P N; Miller, J S; Geller, M A

    2017-06-01

    Natural killer (NK) cells represent a powerful immunotherapeutic target as they lyse tumors directly, do not require differentiation, and can elicit potent inflammatory responses. The objective of these studies was to use an IL-15 super-agonist complex, ALT-803 (Altor BioScience Corporation), to enhance the function of both normal and ovarian cancer patient derived NK cells by increasing cytotoxicity and cytokine production. NK cell function from normal donor peripheral blood mononuclear cells (PBMCs) and ovarian cancer patient ascites was assessed using flow cytometry and chromium release assays ±ALT-803 stimulation. To evaluate the ability of ALT-803 to enhance NK cell function in vivo against ovarian cancer, we used a MA148-luc ovarian cancer NOD scid gamma (NSG) xenogeneic mouse model with transferred human NK cells. ALT-803 potently enhanced functionality of NK cells against all ovarian cancer cell lines with significant increases seen in CD107a, IFNγ and TNFα expression depending on target cell line. Function was also rescued in NK cells derived from ovarian cancer patient ascites. Finally, only animals treated with intraperitoneal ALT-803 displayed an NK dependent significant decrease in tumor. ALT-803 enhances NK cell cytotoxicity against ovarian cancer in vitro and in vivo and is able to rescue functionality of NK cells derived from ovarian cancer patient ascites. These findings suggest that ALT-803 has the potential to enhance NK cell-based immunotherapeutic approaches for the treatment of ovarian cancer. Copyright © 2017 Elsevier Inc. All rights reserved.

  12. Pattern of AST and ALT changes in Relation to Hemolysis in sickle cell Disease

    Directory of Open Access Journals (Sweden)

    K. Nsiah

    2011-01-01

    Full Text Available Background Elevated aminotransferase levels are commonly associated with compromised hepatic integrity from various insults. In sickle cell disease, aspartate transaminase (AST is also released via intravascular hemolysis. This study was done to determine the pattern of changes in AST and alanine transaminase (ALT, in particular the AST:ALT ratio, and to relate these to the hemolytic state, which we consider to be more important than hepatic and cardiac dysfunction in some individuals with sickle cell disease. Methods Serum aminotransferase levels were measured in 330 subjects with sickle cell disease, as well as hemoglobin, reticulocytes, and lactate dehydrogenase. The AST:ALT ratio was designated as a hemolytic marker, and simple and multivariate regression analyses were carried out between this ratio and other hemolytic markers. Results Mean AST and ALT levels were 48.24 % 27.78 and 26.48 % 22.73 U/L, respectively. However, for 49 subjects without sickle cell disease, mean AST and ALT levels were the same, ie, 23.0 U/L. In the subjects with sickle cell disease, the increases in AST levels were far higher than for ALT, supporting its release via intravascular hemolysis. In 95.8% of the subjects with sickle cell disease, the AST:ALT ratio was > 1, but our results did not suggest overt malfunctioning of the liver and heart in the majority of subjects. Conclusion Regression analyses support the use of the AST:ALT ratio as a hemolytic marker, because it has an inverse association with the hemoglobin level. Whether in steady state or in crisis, provided hepatic and cardiac integrity has not been compromised, subjects with sickle cell disease would have higher AST levels due to the hemolytic nature of the condition. This is the first report highlighting the AST:ALT ratio in sickle cell disease.

  13. Some features of alt texts associated with images in Web pages

    Directory of Open Access Journals (Sweden)

    Timothy C. Craven

    2006-01-01

    Full Text Available Introduction. This paper extends a series on summaries of Web objects, in this case, the alt attribute of image files. Method. Data were logged from 1894 pages from Yahoo!'s random page service and 4703 pages from the Google directory; an img tag was extracted randomly from each where present; its alt attribute, if any, was recorded; and the header for the corresponding image file was retrieved if possible. Analysis. Associations were measured between image type and use of null alt values, image type and image file size, image file size and alt text length, and alt text length and number of images on the page. Results. 16.6% and 17.3% of pages respectively showed no img elements. Of 1579 and 3888 img tags randomly selected from the remainder, 47.7% and 49.4% had alt texts, of which 26.3% and 27.5% were null. Of the 1316 and 3384 images for which headers could be retrieved, 71.2% and 74.2% were GIF, 28.1% and 20.5%, JPEG; and 0.8% and 0.8% PNG. GIF images were more commonly assigned null alt texts than JPEG images, and GIF files tended to be shorter than JPEG files. Weak positive correlations were observed between image file size and alt text length, except for JPEG files in the Yahoo! set. Alt texts for images from pages containing more images tended to be slightly shorter. Conclusion. Possible explanations for the results include GIF files' being more suited to decorative images and the likelihood that many images on image-rich pages are content-poor.

  14. The prevalence and clinical correlates of elevated ALT levels in an urban Chinatown community.

    Science.gov (United States)

    Cotler, S J; Dhamija, M K; Luc, B J; Siqueira, F; Bartram, A H; Layden, T J; Wong, S S

    2010-02-01

    Alanine aminotransferase (ALT) levels are a primary consideration in treatment decisions regarding hepatitis B virus (HBV) infection. Levels can be elevated as a result of HBV-related liver injury or to other causes. The aim of this study was to assess the background prevalence and clinical correlates of elevated ALT levels in Chinese immigrants who were HBsAg negative. Subjects consisted of adults seen in an urban Chinatown Internal Medicine practice between 1, 2006 and 12, 2007. Clinical information was collected retrospectively. An elevated ALT level was defined as >19 U/L for women and >30 U/L for men. The primary analysis focused on 2457 persons who were HBsAg negative. Their mean age was 55 +/- 18 years, 64% were female and 36% were obese. All subjects were ethnically Chinese and 96% were born in Asia. Thirty percent had an elevated ALT level. In univariate analysis, factors associated with elevated ALT included older age (P < 0.001), female gender (P = 0.001), obesity (P < 0.001), diabetes (P < 0.001), hypercholesterolemia (P < 0.001) and hypertension (P < 0.001). In multivariate analysis BMI, diabetes, female gender and hypercholesterolemia were significantly associated with elevated ALT level. An additional analysis focused on a group of patients who were HBsAg positive, HBeAg negative and had HBV-DNA levels <1000 copies/mL. Fifty percent had elevated ALT levels. In conclusion, elevated ALT levels are common among Chinese immigrants without HBV (30%) and are associated with features of the metabolic syndrome. Liver biopsy should be performed in selected patients with HBV to distinguish the cause of ALT elevation before initiating antiviral therapy.

  15. Baseline ALT levels as a marker of glycemic response to treatment with GLP-1 receptor agonists.

    Science.gov (United States)

    Gimeno-Orna, Jose A; Verdes-Sanz, Guayente; Borau-Maorad, Laura; Campos-Fernández, Julia; Lardiés-Sánchez, Beatriz; Monreal-Villanueva, Marta

    2016-04-01

    This study aimed to assess if ALT levels, as a marker of non-alcoholic fatty liver disease, may predict HbA1c response to treatment with GLP-1 receptor agonists (GLP-1 RAs). A retrospective, longitudinal, analytical study was conducted including patients with type 2 diabetes mellitus continuously treated with GLP-1 agonists (85% with liraglutide) for one year. Patients were divided into two groups according to baseline ALT levels, with 24 U/L (the median of the distribution) as the cut-off point. The dependent variable was HbA1c change (one-year follow-up minus baseline). The predictive value of ALT levels above 24 U/L and ALT change was analyzed using multivariate linear regression adjusted to age, gender, diabetes duration, type and dose of GLP-1 RA, baseline HbA1c, baseline body mass index (BMI), and change in BMI. A total of 117 patients (48% females) aged 58.6 (SD 9.6) years were enrolled into the study. Treatment was associated with a change in ALT of -4.3 U/L (p=0.041) and a change in HbA1c of -1.1% (pALT (-9.25 vs 0.46 U/L; p=0.002) were significantly higher in patients with ALT levels above the median. In the multivariate analysis, both ALT>24 U/L (b=-0.74; 95%CI: -1.31 to -0.18; p=0.011) and ALT change (b=0.028; 95%CI: 0.010 to 0.046; p=0.003), were significant response predictors. Elevated baseline transaminase values and decreased transaminase levels during follow-up are associated to a favorable glycemic response to GLP-1 RAs. Copyright © 2016 SEEN. Published by Elsevier España, S.L.U. All rights reserved.

  16. A Cisplatin Derivative Tetra-Pt(bpy) as an Oncotherapeutic Agent for Targeting ALT Cancer.

    Science.gov (United States)

    Zheng, Xiao-Hui; Nie, Xin; Fang, Yiming; Zhang, Zepeng; Xiao, Yingnan; Mao, Zongwan; Liu, Haiying; Ren, Jian; Wang, Feng; Xia, Lixin; Huang, Junjiu; Zhao, Yong

    2017-10-01

    In approximately 15% of human cancers, telomere length is maintained independently of telomerase by the homologous recombination (HR)-mediated alternative lengthening of telomeres (ALT) pathway. Whether the ALT pathway can be exploited for therapeutic treatment remains unknown. The purpose of this study is to develop oncotherapeutic agent to target ALT cancers. Surface plasmon resonance assay, antibody to G-quadruplex, and fluorescence in situ hybridization (FISH) were used to discover Tetra-Pt(bpy), a cisplatin derivative that specifically targets telomeric G-quadruplex. We used immunofluorescence, FISH, C-circle assay, and chromosome orientation FISH to evaluate the inhibitory effect of Tetra-Pt(bpy) on ALT activity in human ALT cancers. The shortening of telomere length induced by Tetra-Pt(bpy) was determined by telomere restriction fragment or Q-FISH. Cell destination after Tetra-Pt(bpy) treatment was determined by β-gal staining or apoptosis assay. Nude mice (n = 4 per group) were injected with U2OS cells to evaluate the effects of Tetra-Pt(bpy) on tumor growth. All statistical tests were two-sided. Tetra-Pt(bpy) inhibits the strand invasion/annealing step of telomeric homologous recombination by selectively converting telomeric ssDNA to a G-quadruplex. ALT-cells treated with Tetra-Pt(bpy) show fewer ALT-associated promyelocytic leukemia bodies (untreated: mean±SD = 5.9±0.2 vs treated: mean±SD = 3.1±0.1, P ALT-cell xenograft tumors in mice (untreated: mean±SD = 57.1±3.7 mm 3 vs treated: mean±SD = 19.0±3.2 mm 3 , P ALT cancer cells. © The Author 2017. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  17. Comparison of the Superagonist Complex, ALT-803, to IL15 as Cancer Immunotherapeutics in Animal Models.

    Science.gov (United States)

    Rhode, Peter R; Egan, Jack O; Xu, Wenxin; Hong, Hao; Webb, Gabriela M; Chen, Xiaoyue; Liu, Bai; Zhu, Xiaoyun; Wen, Jinghai; You, Lijing; Kong, Lin; Edwards, Ana C; Han, Kaiping; Shi, Sixiang; Alter, Sarah; Sacha, Jonah B; Jeng, Emily K; Cai, Weibo; Wong, Hing C

    2016-01-01

    IL15, a potent stimulant of CD8(+) T cells and natural killer (NK) cells, is a promising cancer immunotherapeutic. ALT-803 is a complex of an IL15 superagonist mutant and a dimeric IL15 receptor αSu/Fc fusion protein that was found to exhibit enhanced biologic activity in vivo, with a substantially longer serum half-life than recombinant IL15. A single intravenous dose of ALT-803, but not IL15, eliminated well-established tumors and prolonged survival of mice bearing multiple myeloma. In this study, we extended these findings to demonstrate the superior antitumor activity of ALT-803 over IL15 in mice bearing subcutaneous B16F10 melanoma tumors and CT26 colon carcinoma metastases. Tissue biodistribution studies in mice also showed much greater retention of ALT-803 in the lymphoid organs compared with IL15, consistent with its highly potent immunostimulatory and antitumor activities in vivo. Weekly dosing with 1 mg/kg ALT-803 in C57BL/6 mice was well tolerated, yet capable of increasing peripheral blood lymphocyte, neutrophil, and monocyte counts by >8-fold. ALT-803 dose-dependent stimulation of immune cell infiltration into the lymphoid organs was also observed. Similarly, cynomolgus monkeys treated weekly with ALT-803 showed dose-dependent increases of peripheral blood lymphocyte counts, including NK, CD4(+), and CD8(+) memory T-cell subsets. In vitro studies demonstrated ALT-803-mediated stimulation of mouse and human immune cell proliferation and IFNγ production without inducing a broad-based release of other proinflammatory cytokines (i.e., cytokine storm). Based on these results, a weekly dosing regimen of ALT-803 has been implemented in multiple clinical studies to evaluate the dose required for effective immune cell stimulation in humans. ©2015 American Association for Cancer Research.

  18. Hopelessly mortal: The role of mortality salience, immortality and trait self-esteem in personal hope.

    Science.gov (United States)

    Wisman, Arnaud; Heflick, Nathan A

    2016-08-01

    Do people lose hope when thinking about death? Based on Terror Management Theory, we predicted that thoughts of death (i.e., mortality salience) would reduce personal hope for people low, but not high, in self-esteem, and that this reduction in hope would be ameliorated by promises of immortality. In Studies 1 and 2, mortality salience reduced personal hope for people low in self-esteem, but not for people high in self-esteem. In Study 3, mortality salience reduced hope for people low in self-esteem when they read an argument that there is no afterlife, but not when they read "evidence" supporting life after death. In Study 4, this effect was replicated with an essay affirming scientific medical advances that promise immortality. Together, these findings uniquely demonstrate that thoughts of mortality interact with trait self-esteem to cause changes in personal hope, and that literal immortality beliefs can aid psychological adjustment when thinking about death. Implications for understanding personal hope, trait self-esteem, afterlife beliefs and terror management are discussed.

  19. Immune activation induces immortalization of HTLV-1 LTR-Tax transgenic CD4+ T cells.

    Science.gov (United States)

    Swaims, Alison Y; Khani, Francesca; Zhang, Yingyu; Roberts, Arthur I; Devadas, Satish; Shi, Yufang; Rabson, Arnold B

    2010-10-21

    Infection with the human T-cell leukemia virus-1 (HTLV-1) results in a variety of diseases including adult T-cell leukemia/lymphoma (ATL). Although the pathogenesis of these disorders is poorly understood, it involves complex interactions with the host immune system. Activation of infected T cells may play an important role in disease pathogenesis through induction of the oncogenic HTLV-1 Tax transactivator protein. To test this hypothesis, we employed transgenic mice in which Tax is regulated by the HTLV-1 LTR. T-cell receptor stimulation of LTR-Tax CD4(+) T cells induced Tax expression, hyper-proliferation, and immortalization in culture. The transition to cellular immortalization was accompanied by markedly increased expression of the antiapoptotic gene, mcl-1, previously implicated as important in T-cell survival. Immortalized cells exhibited a CD4(+)CD25(+)CD3(-) phenotype commonly observed in ATL. Engraftment of activated LTR-Tax CD4(+) T cells into NOD/Shi-scid/IL-2Rγ null mice resulted in a leukemia-like phenotype with expansion and tissue infiltration of Tax(+), CD4(+) lymphocytes. We suggest that immune activation of infected CD4(+) T cells plays an important role in the induction of Tax expression, T-cell proliferation, and pathogenesis of ATL in HTLV-1-infected individuals.

  20. Immortalization of chicken preadipocytes by retroviral transduction of chicken TERT and TR.

    Directory of Open Access Journals (Sweden)

    Wei Wang

    Full Text Available The chicken is an important agricultural animal and model for developmental biology, immunology and virology. Excess fat accumulation continues to be a serious problem for the chicken industry. However, chicken adipogenesis and obesity have not been well investigated, because no chicken preadipocyte cell lines have been generated thus far. Here, we successfully generated two immortalized chicken preadipocyte cell lines through transduction of either chicken telomerase reverse transcriptase (chTERT alone or in combination with chicken telomerase RNA (chTR. Both of these cell lines have survived >100 population doublings in vitro, display high telomerase activity and have no sign of replicative senescence. Similar to primary chicken preadipocytes, these two cell lines display a fibroblast-like morphology, retain the capacity to differentiate into adipocytes, and do not display any signs of malignant transformation. Isoenzyme analysis and PCR-based analysis confirmed that these two cell lines are of chicken origin and are free from inter-species contamination. To our knowledge, this is the first report demonstrating the generation of immortal chicken cells by introduction of chTERT and chTR. Our established chicken preadipocyte cell lines show great promise as an in vitro model for the investigation of chicken adipogenesis, lipid metabolism, and obesity and its related diseases, and our results also provide clues for immortalizing other avian cell types.

  1. The Genetic Architecture of Liver Enzyme Levels: GGT, ALT and AST

    OpenAIRE

    van Beek, Jenny H. D. A.; de Moor, Marleen H. M.; de Geus, Eco J. C.; Lubke, Gitta H.; Vink, Jacqueline M.; Willemsen, Gonneke; Boomsma, Dorret I.

    2013-01-01

    High levels of liver enzymes GGT, ALT and AST are predictive of disease and all-cause mortality and can reflect liver injury, fatty liver and/or oxidative stress. Variation in GGT, ALT and AST levels is heritable. Moderation of the heritability of these liver enzymes by age and sex has not often been explored, and it is not clear to what extent non-additive genetic and shared environmental factors may play a role. To examine the genetic architecture of GGT, ALT and AST, plasma levels were ass...

  2. Post-test navigation data analysis techniques for the shuttle ALT

    Science.gov (United States)

    1975-01-01

    Postflight test analysis data processing techniques for shuttle approach and landing tests (ALT) navigation data are defined. Postfight test processor requirements are described along with operational and design requirements, data input requirements, and software test requirements. The postflight test data processing is described based on the natural test sequence: quick-look analysis, postflight navigation processing, and error isolation processing. Emphasis is placed on the tradeoffs that must remain open and subject to analysis until final definition is achieved in the shuttle data processing system and the overall ALT plan. A development plan for the implementation of the ALT postflight test navigation data processing system is presented. Conclusions are presented.

  3. First-in-human Phase 1 Clinical Study of the IL-15 Superagonist Complex ALT-803 to Treat Relapse after Transplantation.

    Science.gov (United States)

    Romee, Rizwan; Cooley, Sarah; Berrien-Elliott, Melissa M; Westervelt, Peter; Verneris, Michael R; Wagner, John E; Weisdorf, Daniel J; Blazar, Bruce R; Ustun, Celalettin; DeFor, Todd E; Vivek, Sithara; Peck, Lindsey; DiPersio, John F; Cashen, Amanda F; Kyllo, Rachel; Musiek, Amy; Schaffer, András; Anadkat, Milan J; Rosman, Ilana; Miller, Daniel; Egan, Jack O; Jeng, Emily K; Rock, Amy; Wong, Hing C; Fehniger, Todd A; Miller, Jeffrey S

    2018-02-20

    New therapies for patients with hematologic malignancies who relapse after allogeneic hematopoietic cell transplantation (allo-HCT) are needed. Interleukin (IL)-15 is a cytokine that stimulates CD8 + T cell and NK cell anti-tumor responses, and we hypothesized this cytokine may augment anti-leukemia/lymphoma immunity in vivo. To test this, we performed a first-in-human multi-center phase 1 trial (ClinicalTrials.gov NCT01885897) of the IL-15 superagonist complex ALT-803 in patients who relapsed >60 days after allo-HCT. ALT-803 was administered to 33 patients via the intravenous (IV) or subcutaneous (SQ) routes once weekly for 4 doses (dose levels of 1, 3, 6, and 10 mcg/kg). ALT-803 was well-tolerated, and no dose-limiting toxicities or treatment-emergent graft-versus-host-disease (GVHD) requiring systemic therapy were observed in this clinical setting. Adverse events following IV administration included constitutional symptoms temporally related to increased serum IL-6 and IFN-γ. To mitigate these effects the subcutaneous (SQ) route was tested. SQ delivery resulted in self-limited injection site rashes infiltrated with lymphocytes without acute constitutional symptoms. PK analysis revealed a prolonged (>96 hours) serum concentrations following SQ, but not IV injection. ALT-803 stimulated activation, proliferation, and expansion of NK cells and CD8 + T cells without increasing regulatory T cells. Responses were observed in 19% of evaluable patients, including one complete remission lasting 7 months. Thus, ALT-803 is a safe, well-tolerated agent that significantly increased NK and CD8 + T cell number and function. This immunostimulatory IL-15 superagonist warrants further investigation to augment anti-tumor immunity alone and combined with other immunotherapies. Copyright © 2018 American Society of Hematology.

  4. Immortal time bias in drug safety cohort studies: spontaneous abortion following nonsteroidal antiinflammatory drug exposure.

    Science.gov (United States)

    Daniel, Sharon; Koren, Gideon; Lunenfeld, Eitan; Levy, Amalia

    2015-03-01

    Experimental research of drug safety in pregnancy is generally not feasible because of ethical issues. Therefore, most of the information about drug safety in general and teratogenicity in particular is obtained through observational studies, which require careful methodologic design to obtain unbiased results. Immortal time bias occurs when some cases do not "survive" sufficient time in the study, and as such, they have reduced chances of being defined as "exposed" simply because the durations of their follow-ups were shorter. For example, studies that examine the risk for spontaneous abortions in women exposed to a drug during pregnancy are susceptible to immortal time bias because the chance of drug exposure increases the longer a pregnancy lasts. Therefore, the drug tested may falsely be found protective against the outcome tested. The objective of the current study was to illustrate the extent of immortal time bias using a cohort study of pregnancies assessing the risk for spontaneous abortions following nonsteroidal antiinflammatory drug exposure. We assembled 3 databases containing data on spontaneous abortions, births and drug dispensions to create the present study's cohort. The risk for spontaneous abortion was assessed using 2 statistical analysis methods that were compared for 2 definitions of exposure (dichotomous, exposed vs unexposed, regular Cox regression vs Cox regression with time-varying exposure). Significant differences were found in the risk for spontaneous abortions between the 2 statistical methods, both for groups and for most specific nonsteroidal antiinflammatory drugs (nonselective Cox inhibitors - hazard ratio, 0.70; 95% confidence interval, 0.61-0.94 vs hazard ratio, 1.10; 95% confidence interval, 0.99-1.22 for dichotomous vs time-varying exposure analyses, respectively). Furthermore, a significant correlation was found between the median misclassified immortal time for each drug and the extent of the bias. Immortal time bias can

  5. Pre-ABoVE: Ground-penetrating Radar Measurements of ALT on the Alaska North Slope

    Data.gov (United States)

    National Aeronautics and Space Administration — This data set includes estimates of permafrost Active Layer Thickness (ALT; cm), and calculated uncertainties, derived using a ground-penetrating radar (GPR) system...

  6. Alte Harfe. Vollständige Sammlung alter estnischer Volkslieder : Vorrede / Jakob Hurt

    Index Scriptorium Estoniae

    Hurt, Jakob, 1839-1907

    2005-01-01

    Eessõna rmt. : Hurt, Jakob. Vana kannel. 1. kogu : täieline kogu vanu eesti rahvalaulusid = Alte Harfe : vollständige Sammlung alter estnischen Volkslieder. Tartu, 1875-1886. - (Eesti Kirjameeste Seltsi toimetised ; 3)

  7. Pattern of AST and ALT changes in Relation to Hemolysis in sickle cell Disease

    OpenAIRE

    K. Nsiah; V.P. Dzogbefia; D. Ansong; A. Osei Akoto; H. Boateng; D. Ocloo

    2011-01-01

    Background Elevated aminotransferase levels are commonly associated with compromised hepatic integrity from various insults. In sickle cell disease, aspartate transaminase (AST) is also released via intravascular hemolysis. This study was done to determine the pattern of changes in AST and alanine transaminase (ALT), in particular the AST:ALT ratio, and to relate these to the hemolytic state, which we consider to be more important than hepatic and cardiac dysfunction in some individuals with ...

  8. Aspartate Aminotransferase (AST/GOT) and Alanine Aminotransferase (ALT/GPT) Detection Techniques

    OpenAIRE

    Huang, Xing-Jiu; Choi, Yang-Kyu; Im, Hyung-Soon; Yarimaga, Oktay; Yoon, Euisik; Kim, Hak-Sung

    2006-01-01

    The levels of aspartate aminotransferase (AST/GOT) and alanine aminotransferase (ALT/GPT) in serum can help people diagnose body tissues especially the heart and the liver are injured or not. This article provides a comprehensive review of research activities that concentrate on AST/GOT and ALT/GPT detection techniques due to their clinical importance. The detection techniques include colorimetric, spectrophotometric, chemiluminescence, chromatography, fluorescence and UV absorbance, radioche...

  9. The ASCUS/LSIL Triage Study for Cervical Cancer (ALTS) | Division of Cancer Prevention

    Science.gov (United States)

    ALTS was a clinical trial to find the best way to help women and their doctors decide what to do about the mildly abnormal and very common Pap test results known as ASCUS and LSIL. | ALTS was a clinical trial to find the best way to help women and their doctors decide what to do about the mildly abnormal and very common Pap test results known as ASCUS and LSIL.

  10. Association of MBOAT7 gene variant with plasma ALT levels in children: the PANIC study.

    Science.gov (United States)

    Viitasalo, Anna; Eloranta, Aino-Maija; Atalay, Mustafa; Romeo, Stefano; Pihlajamäki, Jussi; Lakka, Timo A

    2016-11-01

    We studied for the first time among children differences in plasma alanine aminotransferase (ALT) among genotypes of the rs641738 polymorphism in the MBOAT7 gene that has been associated with increased risk of nonalcoholic fatty liver disease among adults. We also investigated the associations of a genetic risk score combining information from the MBOAT7, PNPLA3, and TM6SF2 polymorphisms with plasma ALT. We performed a 2-y follow-up study in 467 Caucasian children aged 6-9 y, genotyped the MBOAT7, PNPLA3, and TM6SF2 polymorphisms, calculated a genetic risk score from these polymorphisms (scored 0-3) and assessed plasma ALT. Children carrying the T allele of the MBOAT7 polymorphism had 7% higher plasma ALT at baseline (17.8 vs. 19.1 U/l, P = 0.022) and 10% higher plasma ALT at 2-y follow-up (18.0 vs. 19.7 U/l, P = 0.022) than the noncarriers. A higher genetic risk score was associated with higher plasma ALT at baseline (17.5, 18.5, 19.2, and 22.8 U/l, P = 0.008 for linear trend) and 2-y follow-up (18.2, 18.9, 18.9, and 32.8 U/l, P = 0.017 for linear trend). Children carrying the T allele of the MBOAT7 polymorphism had higher plasma ALT than the noncarriers. Children with the MBOAT7, PNPLA3, and TM6SF2 variants had the highest plasma ALT.

  11. "Control-alt-delete": rebooting solutions for the E-waste problem.

    Science.gov (United States)

    Li, Jinhui; Zeng, Xianlai; Chen, Mengjun; Ogunseitan, Oladele A; Stevels, Ab

    2015-06-16

    A number of efforts have been launched to solve the global electronic waste (e-waste) problem. The efficiency of e-waste recycling is subject to variable national legislation, technical capacity, consumer participation, and even detoxification. E-waste management activities result in procedural irregularities and risk disparities across national boundaries. We review these variables to reveal opportunities for research and policy to reduce the risks from accumulating e-waste and ineffective recycling. Full regulation and consumer participation should be controlled and reinforced to improve local e-waste system. Aiming at standardizing best practice, we alter and identify modular recycling process and infrastructure in eco-industrial parks that will be expectantly effective in countries and regions to handle the similar e-waste stream. Toxicity can be deleted through material substitution and detoxification during the life cycle of electronics. Based on the idea of "Control-Alt-Delete", four patterns of the way forward for global e-waste recycling are proposed to meet a variety of local situations.

  12. High AST/ALT ratio may indicate advanced alcoholic liver disease rather than heavy drinking.

    Science.gov (United States)

    Nyblom, H; Berggren, U; Balldin, J; Olsson, R

    2004-01-01

    To assess the place of AST/ALT ratio (the ratio of serum aspartate aminotransferase to serum alanine aminotransferase) as a diagnostic marker in medical populations. Laboratory tests were viewed retrospectively in three groups of patients: 313 patients with alcohol dependence, consecutively admitted to an alcohol and drug treatment unit for treatment of withdrawal (W) symptoms, 78 patients with alcohol abuse or dependence consecutively admitted to surgical or medical wards with various primary somatic (S) diagnoses (e.g. respiratory, gastrointestinal and metabolic), and 48 consecutive patients with alcohol abuse or dependence admitted to surgical or medical wards for treatment of alcohol-related liver cirrhosis and its complications (C). Comparison between groups was made of the pattern of patients' AST/ALT ratios using, for Groups S and C, laboratory data from patients' first admission for their condition. There was a significant rise in the AST/ALT ratio from the W to the S patients, and from the S to the C patients. In the W group, the ratio was or = 2. In the C group, 69% had a ratio > or = 2, and 8% a ratio < or = 1.0. The mean ratio was midway in the S group. In the C group, there was a progressive decline in aspartate (AST/ALT) ratios after admission. Most patients with high alcohol consumption but without severe liver disease do not have an AST/ALT ratio above 1. High AST/ALT ratio suggests advanced alcoholic liver disease.

  13. The human CTC1/STN1/TEN1 complex regulates telomere maintenance in ALT cancer cells.

    Science.gov (United States)

    Huang, Chenhui; Jia, Pingping; Chastain, Megan; Shiva, Olga; Chai, Weihang

    2017-06-15

    Maintaining functional telomeres is important for long-term proliferation of cells. About 15% of cancer cells are telomerase-negative and activate the alternative-lengthening of telomeres (ALT) pathway to maintain their telomeres. Recent studies have shown that the human CTC1/STN1/TEN1 complex (CST) plays a multi-faceted role in telomere maintenance in telomerase-expressing cancer cells. However, the role of CST in telomere maintenance in ALT cells is unclear. Here, we report that human CST forms a functional complex localizing in the ALT-associated PML bodies (APBs) in ALT cells throughout the cell cycle. Suppression of CST induces telomere instabilities including telomere fragility and elevates telomeric DNA recombination, leading to telomere dysfunction. In addition, CST deficiency significantly diminishes the abundance of extrachromosomal circular telomere DNA known as C-circles and t-circles. Suppression of CST also results in multinucleation in ALT cells and impairs cell proliferation. Our findings imply that the CST complex plays an important role in regulating telomere maintenance in ALT cells. Copyright © 2017 Elsevier Inc. All rights reserved.

  14. The Effect of Serum Vitamin D on Serum ALT Levels in Healthy Individuals.

    Science.gov (United States)

    Shehata, Elaine; Qayyum, Rehan

    2016-10-01

    Several studies have examined the relationship between vitamin D (VD) and liver disease but none have explored this relationship in adults with normal liver enzymes. Our aim was to explore an independent association of VD with alanine aminotransferase (ALT) in a large sample of the US adults with liver enzymes in normal range (≤39 U/L). We used the continuous National Health and Nutrition Examination Survey from 2001 to 2006. We excluded individuals with serum ALT>39 U/L. We built linear regression models to estimate unadjusted and adjusted (age, sex, race, diabetes, hypertension, alcohol use, smoking, and body mass index) effect sizes, taking into account the complex probability survey design. Of the 12,155 participants, 6635 (54.6%) were women, mean±SD age was 49.9±19.4 years, VD was 21.9±9.2 ng/mL, and ALT was 20.9±6.9 U/L. In unadjusted analysis, VD was significantly associated with serum ALT (0.02 U/L/ng/mL of VD, P=0.007). After adjustment for confounders, VD remained statistically significantly associated with serum ALT levels (0.04 U/L, PALT than those in the lowest quartile (unadjusted difference=0.98 U/L, PALT after excluding individuals with suspected active liver injury (ALT>39 U/L). The underlying mechanisms for this association are not known and needs further study.

  15. Microsomal Triglyceride Transfer Protein Inhibition Induces Endoplasmic Reticulum Stress and Increases Gene Transcription via Ire1α/cJun to Enhance Plasma ALT/AST*

    Science.gov (United States)

    Josekutty, Joby; Iqbal, Jahangir; Iwawaki, Takao; Kohno, Kenji; Hussain, M. Mahmood

    2013-01-01

    Microsomal triglyceride transfer protein (MTP) is a target to reduce plasma lipids because of its indispensable role in triglyceride-rich lipoprotein biosynthesis. MTP inhibition in Western diet fed mice decreased plasma triglycerides/cholesterol, whereas increasing plasma alanine/aspartate aminotransferases (ALT/AST) and hepatic triglycerides/free cholesterol. Free cholesterol accumulated in the endoplasmic reticulum (ER) and mitochondria resulting in ER and oxidative stresses. Mechanistic studies revealed that MTP inhibition increased transcription of the GPT/GOT1 genes through up-regulation of the IRE1α/cJun pathway leading to increased synthesis and release of ALT1/AST1. Thus, transcriptional up-regulation of GPT/GOT1 genes is a major mechanism, in response to ER stress, elevating plasma transaminases. Increases in plasma and tissue transaminases might represent a normal response to stress for survival. PMID:23532846

  16. Microsomal triglyceride transfer protein inhibition induces endoplasmic reticulum stress and increases gene transcription via Ire1α/cJun to enhance plasma ALT/AST.

    Science.gov (United States)

    Josekutty, Joby; Iqbal, Jahangir; Iwawaki, Takao; Kohno, Kenji; Hussain, M Mahmood

    2013-05-17

    Microsomal triglyceride transfer protein (MTP) is a target to reduce plasma lipids because of its indispensable role in triglyceride-rich lipoprotein biosynthesis. MTP inhibition in Western diet fed mice decreased plasma triglycerides/cholesterol, whereas increasing plasma alanine/aspartate aminotransferases (ALT/AST) and hepatic triglycerides/free cholesterol. Free cholesterol accumulated in the endoplasmic reticulum (ER) and mitochondria resulting in ER and oxidative stresses. Mechanistic studies revealed that MTP inhibition increased transcription of the GPT/GOT1 genes through up-regulation of the IRE1α/cJun pathway leading to increased synthesis and release of ALT1/AST1. Thus, transcriptional up-regulation of GPT/GOT1 genes is a major mechanism, in response to ER stress, elevating plasma transaminases. Increases in plasma and tissue transaminases might represent a normal response to stress for survival.

  17. Generation of an immortalized mesenchymal stem cell line producing a secreted biosensor protein for glucose monitoring.

    Directory of Open Access Journals (Sweden)

    Evangelia K Siska

    Full Text Available Diabetes is a chronic disease characterized by high levels of blood glucose. Diabetic patients should normalize these levels in order to avoid short and long term clinical complications. Presently, blood glucose monitoring is dependent on frequent finger pricking and enzyme based systems that analyze the drawn blood. Continuous blood glucose monitors are already on market but suffer from technical problems, inaccuracy and short operation time. A novel approach for continuous glucose monitoring is the development of implantable cell-based biosensors that emit light signals corresponding to glucose concentrations. Such devices use genetically modified cells expressing chimeric genes with glucose binding properties. MSCs are good candidates as carrier cells, as they can be genetically engineered and expanded into large numbers. They also possess immunomodulatory properties that, by reducing local inflammation, may assist long operation time. Here, we generated a novel immortalized human MSC line co-expressing hTERT and a secreted glucose biosensor transgene using the Sleeping Beauty transposon technology. Genetically modified hMSCs retained their mesenchymal characteristics. Stable transgene expression was validated biochemically. Increased activity of hTERT was accompanied by elevated and constant level of stem cell pluripotency markers and subsequently, by MSC immortalization. Furthermore, these cells efficiently suppressed PBMC proliferation in MLR transwell assays, indicating that they possess immunomodulatory properties. Finally, biosensor protein produced by MSCs was used to quantify glucose in cell-free assays. Our results indicate that our immortalized MSCs are suitable for measuring glucose concentrations in a physiological range. Thus, they are appropriate for incorporation into a cell-based, immune-privileged, glucose-monitoring medical device.

  18. Immortalized myeloid suppressor cells trigger apoptosis in antigen-activated T lymphocytes.

    Science.gov (United States)

    Apolloni, E; Bronte, V; Mazzoni, A; Serafini, P; Cabrelle, A; Segal, D M; Young, H A; Zanovello, P

    2000-12-15

    We described a generalized suppression of CTL anamnestic responses that occurred in mice bearing large tumor nodules or immunized with powerful recombinant viral immunogens. Immune suppression entirely depended on GM-CSF-driven accumulation of CD11b(+)/Gr-1(+) myeloid suppressor cells (MSC) in secondary lymphoid organs. To further investigate the nature and properties of MSC, we immortalized CD11b(+)/Gr-1(+) cells isolated from the spleens of immunosuppressed mice, using a retrovirus encoding the v-myc and v-raf oncogenes. Immortalized cells expressed monocyte/macrophage markers (CD11b, F4/80, CD86, CD11c), but they differed from previously characterized macrophage lines in their capacities to inhibit T lymphocyte activation. Two MSC lines, MSC-1 and MSC-2, were selected based upon their abilities to inhibit Ag-specific proliferative and functional CTL responses. MSC-1 line was constitutively inhibitory, while suppressive functions of MSC-2 line were stimulated by exposure to the cytokine IL-4. Both MSC lines triggered the apoptotic cascade in Ag-activated T lymphocytes by a mechanism requiring cell-cell contact. Some well-known membrane molecules involved in the activation of apoptotic pathways (e.g., TNF-related apoptosis-inducing ligand, Fas ligand, TNF-alpha) were ruled out as candidate effectors for the suppression mechanism. The immortalized myeloid lines represent a novel, useful tool to shed light on the molecules involved in the differentiation of myeloid-related suppressors as well as in the inhibitory pathway they use to control T lymphocyte activation.

  19. Characterization of immortalized human mammary epithelial cell line HMEC 2.6.

    Science.gov (United States)

    Joshi, Pooja S; Modur, Vishnu; Cheng, JiMing; Robinson, Kathy; Rao, Krishna

    2017-10-01

    Primary human mammary epithelial cells have a limited life span which makes it difficult to study them in vitro for most purposes. To overcome this problem, we have developed a cell line that was immortalized using defined genetic elements, and we have characterized this immortalized non-tumorigenic human mammary epithelial cell line to establish it as a potential model system. human mammary epithelial cells were obtained from a healthy individual undergoing reduction mammoplasty at SIU School of Medicine. The cells were transduced with CDK4R24C followed by transduction with human telomerase reverse transcriptase. Post all manipulation, the cells displayed a normal cell cycle phase distribution and were near diploid in nature, which was confirmed by flow cytometry and karyotyping. In vitro studies showed that the cells were anchorage dependent and were non-invasive in nature. The cell line expressed basal epithelial markers such as cytokeratin 7, CD10, and p63 and was negative for the expression of estrogen receptor and progesterone receptor. Upon G-band karyotyping, the cell line displayed the presence of a few cytogenic abnormalities, including trisomy 20 and trisomy 7, which are also commonly present in other immortalized mammary cell lines. Furthermore, the benign nature of these cells was confirmed by multiple in vitro and in vivo experiments. Therefore, we think that this cell line could serve as a good model to understand the molecular mechanisms involved in the development and progression of breast cancer and to also assess the effect of novel therapeutics on human mammary epithelial cells.

  20. [Differential liver histopathological features of chronic HBV infection patients with normal and mildly elevated serum ALT].

    Science.gov (United States)

    Ying, Ruo-su; Yang, Zhan; Chen, Yan-yu; Yang, Ke-li; Xiao, Yan-hua; Wu, Ling-jie; Fan, Hui-min

    2012-08-01

    To study the liver histopathological features that are distinctive between chronic hepatitis B virus (HBV) infection patients who have normal serum alanine aminotransferase (ALT)/asparatate aminotransferase (AST) and those with mildly elevated serum ALT/AST. One-hundred-and-thrity-four chronic HBV infection patients with normal serum ALT/AST and 165 chronic HBV infection patients with mildly elevated serum ALT/AST were included in the study. Liver biopsies were performed and used to assess the histological changes by hematoxylin-eosin and reticular fiber staining; mild to severe scoring for inflammation was made as grade G0-G4 and for fibrosis stage as S0-S4. HBV DNA levels were detected by fluorescent quantitative PCR. HBV serological markers were examined by chemiluminescence. The mildly elevated serum ALT/AST group had more male patients than the normal serum ALT/AST group. In the normal serum ALT/AST group, 50.0% (67/134) of the patients had moderate histological changes and only 3.0% (4/134) had severe changes (G3-4 and/or S3-4). In the mildly elevated ALT/AST group, 65.7% (174/265) of patients had moderate histological changes and 16.2% (43/265) had severe changes (G3-4 and/or S3-4). Hepatic inflammation and fibrosis were significantly more severe in the mildly elevated serum ALT/AST group than in the normal ALT/AST group (x2 = 26.386, P less than 0.01; x2 = 15.299, P less than 0.01). In the normal ALT/AST group, the severity of inflammation and fibrosis were positively correlated with age (rs = 0.620, P less than 0.01; rs = 0.347, P less than 0.01). In the mildly elevated ALT/AST group, the severity of inflammation and fibrosis were negatively correlated with age (rs = -0.807, P less than 0.01; rs = -0.557, P less than 0.01). In both groups, the severity of inflammation and fibrosis were negatively correlated with HBV DNA levels (rs = -0.215, P less than 0.01, rs = -0.527, P less than 0.01, rs = -0.951, P less than 0.01; rs = -0.715, P less than 0.01) and

  1. Use of the landmark method to address immortal person-time bias in comparative effectiveness research: a simulation study.

    Science.gov (United States)

    Mi, Xiaojuan; Hammill, Bradley G; Curtis, Lesley H; Lai, Edward Chia-Cheng; Setoguchi, Soko

    2016-11-20

    Observational comparative effectiveness and safety studies are often subject to immortal person-time, a period of follow-up during which outcomes cannot occur because of the treatment definition. Common approaches, like excluding immortal time from the analysis or naïvely including immortal time in the analysis, are known to result in biased estimates of treatment effect. Other approaches, such as the Mantel-Byar and landmark methods, have been proposed to handle immortal time. Little is known about the performance of the landmark method in different scenarios. We conducted extensive Monte Carlo simulations to assess the performance of the landmark method compared with other methods in settings that reflect realistic scenarios. We considered four landmark times for the landmark method. We found that the Mantel-Byar method provided unbiased estimates in all scenarios, whereas the exclusion and naïve methods resulted in substantial bias when the hazard of the event was constant or decreased over time. The landmark method performed well in correcting immortal person-time bias in all scenarios when the treatment effect was small, and provided unbiased estimates when there was no treatment effect. The bias associated with the landmark method tended to be small when the treatment rate was higher in the early follow-up period than it was later. These findings were confirmed in a case study of chronic obstructive pulmonary disease. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  2. Characterization of lipid metabolism in insulin-sensitive adipocytes differentiated from immortalized human mesenchymal stem cells

    DEFF Research Database (Denmark)

    Prawitt, Janne; Niemeier, Andreas; Kassem, Moustapha

    2008-01-01

    There is a great demand for cell models to study human adipocyte function. Here we describe the adipogenic differentiation of a telomerase-immortalized human mesenchymal stem cell line (hMSC-Tert) that maintains numerous features of terminally differentiated adipocytes even after prolonged...... receptor (VLDLR, LRP1) and apolipoprotein E expression during differentiation. Despite this increased expression, the receptor-mediated endocytosis of lipoproteins was decreased in differentiated adipocytes, suggesting that these proteins may have an additional function in adipose tissue beyond lipoprotein...

  3. Reference Intervals of Total Bilirubin, ALT, AST, and Creatinine in Healthy Elderly Chinese

    Science.gov (United States)

    Zhang, Guo-ming; Xia, Yong-jie; Guo, Xu-xiao; Zhu, Bao-lin; Zhang, Gao-ming; Ma, Xiao-bo; Yu, Hong; Wang, Hong-jian; Wang, Guang-sheng; Yang, Li; Zhou, Ye-ting

    2014-01-01

    Background The aim of this study was to establish the reference intervals (RIs) of total bilirubin (TBIL), alanine aminotransferase (ALT), aspartate transaminase (AST), and creatinine (CREA) for apparently healthy elderly (Han ethnicity) in Shuyang, China. Material/Methods A total of 54 912 blood specimens from elderly residents age 65–104 years were collected by standard procedures in Shuyang county of Jiangsu province. TBIL, ALT, AST, and CREA for each participant were determined by automatic biochemical analyzer. Distribution and differences of TBIL, ALT, AST, and CREA were analyzed and compared between the elderly of the same age of different sexes and different ages of the same sex. RIs of TBIL, ALT, AST, and CREA were compared with the current RIs. The RIs and 95% confidence intervals were calculated using nonparametric method (2.5th–97.5th percentiles) according to the guideline of the Clinical and Laboratory Standards Institute. Results RIs established for the healthy elderly include: TBIL 7.8~30.6 μmol/L for males and 7.3~26.1 μmol/L for females; ALT 8.7~47.3 U/L for males and 8.4~45.2 U/L for females; AST 15.7~46.9 U/L for males and 15.1~46.2 U/L for females; and CREA 45.1~100.9 μmol/L for males and 38.7~85.0 μmol/L for females. Reference intervals of TBIL, ALT, AST, and CREA for male elderly were higher than those of females, and values of CREA increased with increasing age. Conclusions We have established a panel of locally relevant RIs. It is necessary to establish scientific and reasonable RIs of TBIL, ALT, AST, and CREA for the healthy elderly in our region, which will provide a reference for clinicians and inspection officers. PMID:25272068

  4. The genetic architecture of liver enzyme levels: GGT, ALT and AST.

    Science.gov (United States)

    van Beek, Jenny H D A; de Moor, Marleen H M; de Geus, Eco J C; Lubke, Gitta H; Vink, Jacqueline M; Willemsen, Gonneke; Boomsma, Dorret I

    2013-07-01

    High levels of liver enzymes GGT, ALT and AST are predictive of disease and all-cause mortality and can reflect liver injury, fatty liver and/or oxidative stress. Variation in GGT, ALT and AST levels is heritable. Moderation of the heritability of these liver enzymes by age and sex has not often been explored, and it is not clear to what extent non-additive genetic and shared environmental factors may play a role. To examine the genetic architecture of GGT, ALT and AST, plasma levels were assessed in a large sample of twins, their siblings, parents and spouses (N = 8,371; age range 18-90). For GGT and ALT, but not for AST, genetic structural equation modeling showed evidence for quantitative sex differences in the genetic architecture. There was no evidence for qualitative sex differences, i.e. the same genes were expressed in males and females. Both additive and non-additive genetic factors were important for GGT in females (total heritability h(2) 60 %) and AST in both sexes (total h(2) 43 %). The heritability of GGT in males and ALT for both sexes was due to additive effects only (GGT males 30 %; ALT males 40 %, females 22 %). Evidence emerged for shared environmental factors influencing GGT in the male offspring generation (variance explained 28 %). Thus, the same genes influence liver enzyme levels across sex and age, but their relative contribution to the variation in GGT and ALT differs in males and females and for GGT across age. Given adequate sample sizes these results suggest that genome-wide association studies may result in the detection of new susceptibility loci for liver enzyme levels when pooling results over sex and age.

  5. Significant fibrosis is not rare in Chinese chronic hepatitis B patients with persistent normal ALT.

    Science.gov (United States)

    Liao, Baolin; Wang, Zhanhui; Lin, Siwei; Xu, Ying; Yi, Junqing; Xu, Min; Huang, Zuxiong; Zhou, Ying; Zhang, Fuchun; Hou, Jinlin

    2013-01-01

    Limited studies have been done on chronic hepatitis B (CHB) patients defined according to the latest Asian-Pacific Association for the Study of the Liver guideline with liver histology by a large sample size. We retrospectively evaluated liver histological characteristics on a cohort of consecutive treatment-naive CHB patients with persistent normal alanine aminotransferase (PNALT) or elevated ALT from May 2005 to October 2011. Histological assessment was based on the Metavir scoring system, significant abnormality was defined as necroinflammation grade ≥A2 and/or fibrosis stage ≥F2. A total of 675 CHB patients were recruited, including 516 HBeAg-positive and 159 HBeAg-negative patients. In HBeAg-positive patients, significant fibrosis was found 49.4% (42/85) in PNALT, 69.8% (88/126) in ALT 1-2×upper limit normal (ULN) and 81.6% (249/305) in ALT>2×ULN group, respectively. In HBeAg-negative patients, significant fibrosis was found 30.9% (17/55) in PNALT, 73.3% (33/45) in ALT 1-2×ULN and 94.9% (56/59) in ALT>2×ULN group, respectively. HBeAg-positive patients with PNALT over 30 years old had a higher frequency of significant fibrosis than those under 30 years old (87.5% vs. 45.5%, P = 0.058). Multivariate logistic regression analysis indicated increasing age (P = 0.012), higher aspartate aminotransferase (AST) (P AST (P ALT. Higher AST was associated with significant necroinflammation in HBeAg-negative patients with elevated ALT (P = 0.009). Significant fibrosis is not rare in Chinese CHB patients with PNALT, especially HBeAg-positive patients over 30 years old.

  6. Reference intervals for total bilirubin, ALT, AST and creatinine in healthy Chinese elderly.

    Science.gov (United States)

    Zhang, Guo-ming; Xia, Yong-jie; Guo, Xu-xiao; Zhu, Bao-lin; Zhang, Gao-ming; Ma, Xiao-bo; Yu, Hong; Wang, Hong-jian; Wang, Guang-sheng; Yang, Li; Zhou, Ye-ting

    2014-10-01

    The aim of this study was to establish the reference intervals (RIs) of total bilirubin (TBIL), alanine aminotransferase (ALT), aspartate transaminase (AST), and creatinine (CREA) for apparently healthy elderly (Han ethnicity) in Shuyang, China. A total of 54 912 blood specimens from elderly residents age 65-104 years were collected by standard procedures in Shuyang county of Jiangsu province. TBIL, ALT, AST, and CREA for each participant were determined by automatic biochemical analyzer. Distribution and differences of TBIL, ALT, AST, and CREA were analyzed and compared between the elderly of the same age of different sexes and different ages of the same sex. RIs of TBIL, ALT, AST, and CREA were compared with the current RIs. The RIs and 95% confidence intervals were calculated using nonparametric method (2.5th-97.5th percentiles) according to the guideline of the Clinical and Laboratory Standards Institute. RIs established for the healthy elderly include: TBIL 7.8~30.6 µmol/L for males and 7.3~26.1 µmol/L for females; ALT 8.7~47.3 U/L for males and 8.4~45.2 U/L for females; AST 15.7~46.9 U/L for males and 15.1~46.2 U/L for females; and CREA 45.1~100.9 µmol/L for males and 38.7~85.0 µmol/L for females. Reference intervals of TBIL, ALT, AST, and CREA for male elderly were higher than those of females, and values of CREA increased with increasing age. We have established a panel of locally relevant RIs. It is necessary to establish scientific and reasonable RIs of TBIL, ALT, AST, and CREA for the healthy elderly in our region, which will provide a reference for clinicians and inspection officers.

  7. Attempting immortality

    International Nuclear Information System (INIS)

    Krull, W.

    1995-01-01

    The world's population of research reactors is growing old. Many have been adapted to serve new purposes over their lives, from testing materials for nuclear power programmes and supporting neutron physics experiments, to colouring gemstones, doping silicon and generating medical isotopes. In the first article of this survey of research reactor issues, Wilfried Krull from GKSS in Germany describes the effects on a reactor of supporting these changes in application as ''design ageing'' . Managing this and other symptoms of ageing to extend plant life is a key task for operators, and Krull discusses the efforts being made internationally to handle them. Eventually, terminal decline of one vital component can determine when a reactor has to be shutdown for refurbishment. For BR2 in Belgium, it was the beryllium matrix. Edgar Koonen from SCK-CEN explains work being done to replace it and extend reactor life for around 15 years. The Triga at the University of Austria's Atomic Institute has already been running for over 30 years and Helmuth Bock, of the Institute, credits its good health to thorough routine maintenance procedures and custom-made tools for the work. The HANARO reactor in Korea achieved first criticality in February this year - one of the few new research reactors to have started up in recent years. Seong-yun Kim of KAERI describes its design and uses, including medical isotope production. About 80% of medical Mo-99 comes from the 38-year-old NRU in Canada. Russell Ball from B and W Nuclear Environmental Services describes a conceptual reactor for dedicated Mo-99 production in the last article of the survey. (UK)

  8. Differential Evolutionary Selection and Natural Evolvability Observed in ALT Proteins of Human Filarial Parasites.

    Science.gov (United States)

    Devoe, Neil C; Corbett, Ian J; Barker, Linsey; Chang, Robert; Gudis, Polyxeni; Mullen, Nathan; Perez, Kailey; Raposo, Hugo; Scholz, John; May, Meghan

    2016-01-01

    The abundant larval transcript (ALT-2) protein is present in all members of the Filarioidea, and has been reported as a potential candidate antigen for a subunit vaccine against lymphatic filariasis. To assess the potential for vaccine escape or heterologous protection, we examined the evolutionary selection acting on ALT-2. The ratios of nonsynonymous (K(a)) to synonymous (K(s)) mutation frequencies (ω) were calculated for the alt-2 genes of the lymphatic filariasis agents Brugia malayi and Wuchereria bancrofti and the agents of river blindness and African eyeworm disease Onchocerca volvulus and Loa loa. Two distinct Bayesian models of sequence evolution showed that ALT-2 of W. bancrofti and L. loa were under significant (PALT-2 of B. malayi and O. volvulus were under neutral to stabilizing selection. Diversifying selection as measured by ω values was notably strongest on the region of ALT-2 encoding the signal peptide of L. loa and was elevated in the variable acidic domain of L. loa and W. bancrofti. Phylogenetic analysis indicated that the ALT-2 consensus sequences formed three clades: the first consisting of B. malayi, the second consisting of W. bancrofti, and the third containing both O. volvulus and L. loa. ALT-2 selection was therefore not predictable by phylogeny or pathology, as the two species parasitizing the eye were selected differently, as were the two species parasitizing the lymphatic system. The most immunogenic regions of L. loa and W. bancrofti ALT-2 sequence as modeled by antigenicity prediction analysis did not correspond with elevated levels of diversifying selection, and were not selected differently than predicted antigenic epitopes in B. malayi and O. volvulus. Measurements of ALT-2 evolvability made by χ2 analysis between alleles that were stable (O. volvulus and B. malayi) and those that were under diversifying selection (W. bancrofti and L. loa) indicated significant (PALT-2 of the four organisms examined, and the described

  9. AST/ALT ratio predicts cirrhosis in patients with chronic hepatitis C virus infection.

    Science.gov (United States)

    Sheth, S G; Flamm, S L; Gordon, F D; Chopra, S

    1998-01-01

    A liver biopsy is necessary to grade and stage chronic hepatitis C virus (HCV) infection. In a previous study of patients with nonalcoholic liver disease, an aspartate aminotransferase (AST) to alanine aminotransferase (ALT) ratio >1 suggested cirrhosis. We sought to examine the value of the AST/ALT ratio in distinguishing cirrhotic patients with chronic HCV infection from noncirrhotic patients and to correlate the ratio with the grade and stage of hepatitis and other biochemical indices. We retrospectively studied 139 patients with chronic HCV infection. Routine biochemical indices were determined, and the histological grade of necroinflammatory activity and the stage of fibrosis of the liver biopsy specimens were scored. The mean AST/ALT ratio in the cirrhotic patients (n = 47) was higher than in the noncirrhotic patients (n = 92) (1.06 +/- 0.06 vs 0.60 +/- 0.09; p or =1 had 100% specificity and positive predictive value in distinguishing cirrhotic from noncirrhotic patients, with a 53.2% sensitivity and 80.7% negative predictive value. The ratio correlated positively with the stage of fibrosis but not with the grade of activity or other biochemical indices. Of the cirrhotic patients, 17% had no clinical or biochemical features suggestive of chronic liver disease except for an AST/ALT ratio > or =1. The AST/ALT ratio is a dependable marker of fibrosis stage and cirrhosis in patients with chronic HCV infection.

  10. The AST/ALT ratio as an indicator of cirrhosis in patients with PBC.

    Science.gov (United States)

    Nyblom, Helena; Björnsson, Einar; Simrén, Magnus; Aldenborg, Frank; Almer, Sven; Olsson, Rolf

    2006-09-01

    A non-invasive, simple and non-expensive test to predict cirrhosis would be highly desirable. The aspartate aminotransferase/alanine aminotransferase (AST/ALT) ratio has been proven to be such an indicator of cirrhosis in alcoholic liver disease, hepatitis C. To test whether the AST/ALT ratio is a marker of cirrhosis also in patients with primary biliary cirrhosis (PBC). The study consisted of 160 patients. In 126 patients, we had clinical and laboratory data at the time of diagnosis and follow-up with outcome: liver-related death, liver transplantation and survival. In 121 patients, we had laboratory data and liver histology. We found that the AST/ALT ratio was significantly higher in cirrhotic patients than in non-cirrhotic patients. A high AST/ALT ratio was significantly associated with esophageal varices and ascites. In a multivariate analysis, bilirubin and ALP were predictors of poor prognosis. The AST/ALT ratio seems to be of clinical value as a hint to the diagnosis of cirrhosis in patients with PBC but not as a prognostic factor.

  11. Coverage of supraumbilical abdominal wall defects: The tunnelled-pedicled ALT technique.

    Science.gov (United States)

    Fernandez-Alvarez, Jose-Alberto; Barrera-Pulido, Fernando; Lagares-Borrego, Araceli; Narros-Gimenez, Rocio; Gacto-Sanchez, Purificación; Gomez-Cia, Tomas

    2017-02-01

    Abdominal wall defects are a challenge for reconstructive surgeons. Although the utility of anterolateral thigh perforator (ALT) flap has been well established for lower abdominal wall reconstruction, pedicled ALT flap is usually not considered for supraumbilical defects in the most recent algorithms. The purpose of this paper is to report the results of a tunneled pedicled ALT flap for reconstruction of supraumbilical defect from a series of patients. From July 2009 to September2014, six patients underwent delayed abdominal wall coverage using pedicled ALT flaps and reinforcement with polypropylene meshes. Defects occurred after surgical complications and abdominal trauma. Flaps were tunneled beneath the rectus femoris and sartorius muscles to increase the pedicle length. The size of the skin islands ranged from 22-29 × 10-14 cm. All flaps survived and the healing of the wounds was successful. Partial dehiscence of donor site occurred in one patient, and small wound dehiscence due to minimal distal necrosis was observed in another patient. No functional problems were reported in donor site, and no complications occurred in 6-68 months of follow-up. The tunneled pedicled ALT flap may provide a reliable alternative method for abdominal wall reconstruction, including supraumbilical defects. © 2015 Wiley Periodicals, Inc. Microsurgery 37:119-127, 2017. © 2015 Wiley Periodicals, Inc.

  12. Immortal time bias: a frequently unrecognized threat to validity in the evaluation of postoperative radiotherapy.

    Science.gov (United States)

    Park, Henry S; Gross, Cary P; Makarov, Danil V; Yu, James B

    2012-08-01

    To evaluate the influence of immortal time bias on observational cohort studies of postoperative radiotherapy (PORT) and the effectiveness of sequential landmark analysis to account for this bias. First, we reviewed previous studies of the Surveillance, Epidemiology, and End Results (SEER) database to determine how frequently this bias was considered. Second, we used SEER to select three tumor types (glioblastoma multiforme, Stage IA-IVM0 gastric adenocarcinoma, and Stage II-III rectal carcinoma) for which prospective trials demonstrated an improvement in survival associated with PORT. For each tumor type, we calculated conditional survivals and adjusted hazard ratios of PORT vs. postoperative observation cohorts while restricting the sample at sequential monthly landmarks. Sixty-two percent of previous SEER publications evaluating PORT failed to use a landmark analysis. As expected, delivery of PORT for all three tumor types was associated with improved survival, with the largest associated benefit favoring PORT when all patients were included regardless of survival. Preselecting a cohort with a longer minimum survival sequentially diminished the apparent benefit of PORT. Although the majority of previous SEER articles do not correct for it, immortal time bias leads to altered estimates of PORT effectiveness, which are very sensitive to landmark selection. We suggest the routine use of sequential landmark analysis to account for this bias. Copyright © 2012 Elsevier Inc. All rights reserved.

  13. Immortal Time Bias: A Frequently Unrecognized Threat to Validity in the Evaluation of Postoperative Radiotherapy

    International Nuclear Information System (INIS)

    Park, Henry S.; Gross, Cary P.; Makarov, Danil V.; Yu, James B.

    2012-01-01

    Purpose: To evaluate the influence of immortal time bias on observational cohort studies of postoperative radiotherapy (PORT) and the effectiveness of sequential landmark analysis to account for this bias. Methods and Materials: First, we reviewed previous studies of the Surveillance, Epidemiology, and End Results (SEER) database to determine how frequently this bias was considered. Second, we used SEER to select three tumor types (glioblastoma multiforme, Stage IA–IVM0 gastric adenocarcinoma, and Stage II–III rectal carcinoma) for which prospective trials demonstrated an improvement in survival associated with PORT. For each tumor type, we calculated conditional survivals and adjusted hazard ratios of PORT vs. postoperative observation cohorts while restricting the sample at sequential monthly landmarks. Results: Sixty-two percent of previous SEER publications evaluating PORT failed to use a landmark analysis. As expected, delivery of PORT for all three tumor types was associated with improved survival, with the largest associated benefit favoring PORT when all patients were included regardless of survival. Preselecting a cohort with a longer minimum survival sequentially diminished the apparent benefit of PORT. Conclusions: Although the majority of previous SEER articles do not correct for it, immortal time bias leads to altered estimates of PORT effectiveness, which are very sensitive to landmark selection. We suggest the routine use of sequential landmark analysis to account for this bias.

  14. Immortal Time Bias: A Frequently Unrecognized Threat to Validity in the Evaluation of Postoperative Radiotherapy

    Energy Technology Data Exchange (ETDEWEB)

    Park, Henry S. [Department of Therapeutic Radiology, Yale University School of Medicine, New Haven, CT (United States); Gross, Cary P. [Cancer Outcomes Policy and Effectiveness Research Center, Yale University School of Medicine, New Haven, CT (United States); Makarov, Danil V. [Department of Urology, New York University School of Medicine, New York, NY (United States); Yu, James B., E-mail: james.b.yu@yale.edu [Department of Therapeutic Radiology, Yale University School of Medicine, New Haven, CT (United States); Cancer Outcomes Policy and Effectiveness Research Center, Yale University School of Medicine, New Haven, CT (United States)

    2012-08-01

    Purpose: To evaluate the influence of immortal time bias on observational cohort studies of postoperative radiotherapy (PORT) and the effectiveness of sequential landmark analysis to account for this bias. Methods and Materials: First, we reviewed previous studies of the Surveillance, Epidemiology, and End Results (SEER) database to determine how frequently this bias was considered. Second, we used SEER to select three tumor types (glioblastoma multiforme, Stage IA-IVM0 gastric adenocarcinoma, and Stage II-III rectal carcinoma) for which prospective trials demonstrated an improvement in survival associated with PORT. For each tumor type, we calculated conditional survivals and adjusted hazard ratios of PORT vs. postoperative observation cohorts while restricting the sample at sequential monthly landmarks. Results: Sixty-two percent of previous SEER publications evaluating PORT failed to use a landmark analysis. As expected, delivery of PORT for all three tumor types was associated with improved survival, with the largest associated benefit favoring PORT when all patients were included regardless of survival. Preselecting a cohort with a longer minimum survival sequentially diminished the apparent benefit of PORT. Conclusions: Although the majority of previous SEER articles do not correct for it, immortal time bias leads to altered estimates of PORT effectiveness, which are very sensitive to landmark selection. We suggest the routine use of sequential landmark analysis to account for this bias.

  15. Grafting amino drugs to poly(styrene-alt-maleic anhydride) as a potential method for drug release

    Energy Technology Data Exchange (ETDEWEB)

    Khazaei, Ardeshir; Saednia, Shahnaz; Saien, Javad; Abbasi, Fatemeh, E-mail: Khazaei_1326@yahoo.com, E-mail: ssaednia@gmail.com [Faculty of Chemistry, Bu-Ali Sina University, Hamedan (Iran, Islamic Republic of); Kazem-Rostami, Masoud [Young Researchers Club and Elite, Takestan Branch, Islamic Azad University, Takestan (Iran, Islamic Republic of); Sadeghpour, Mahdieh [Department of Chemistry, Takestan Branch, Islamic Azad University, Takestan (Iran, Islamic Republic of); Borazjani, Maryam Kiani [Faculty of Science, Department of Chemistry, Bushehr Payame Noor University (PNU), Bushehr (Iran, Islamic Republic of)

    2013-07-15

    Drug delivery systems based on polymer-drug conjugates give an improved treatment with lower toxicity or side effects and be used for the treatment of different diseases. Conjugates of biodegradable poly(styrene-alt-maleic anhydride) (PSMA), with a therapeutic agents such as amantadine hydrochloride, amlodipine, gabapentin, zonisamide and mesalamine, were afforded by the formation of the amide bonds of the amino drugs that reacted with the PSMA anhydride groups. The amounts of covalently conjugated drugs were determined by a {sup 1}H NMR spectroscopic method, and the in vitro release rate in buffer solution (pH 1.3) was studied at body temperature 37 Degree-Sign C. In kinetic studies, different dissolution models were examined to obtain drug release data and the collected data were well-fitted to the Korsmeyer-Peppas equation, revealing a dominant Fickian diffusion mechanism for drug release under the in vitro conditions. (author)

  16. Elevated Alt and Ast in an Asymptomatic Person: What the primary care doctor should do?

    Science.gov (United States)

    Yin, Loh Keng; Tong, Kew Siang

    2009-01-01

    Abnormal liver function test with raised alanine aminotransferase (ALT) and raised aspartate aminotransferase (AST) are commonly seen in primary care setting.Chronic alcohol consumption, drugs, non-alcoholic steatohepatitis (NASH) and chronic viral hepatitis are common causes associated with raised ALT and AST.In chronic viral hepatitis, the elevation of liver enzyme may not correlate well with the degree of liver damage.Non-hepatic causes of raised ALT and AST include polymyositis, acute muscles injury, acute myocardial infarction and hypothyroidism.In the primary care setting, the doctor should obtain a complete history regarding the risk factors for viral hepatitis, substance abuse and request investigations accordingly.Suspected chronic viral hepatitis and liver cirrhosis are best referred to hepatologist for further management.

  17. Alt a 1 from Alternaria interacts with PR5 thaumatin-like proteins.

    Science.gov (United States)

    Gómez-Casado, Cristina; Murua-García, Amaya; Garrido-Arandia, María; González-Melendi, Pablo; Sánchez-Monge, Rosa; Barber, Domingo; Pacios, Luis F; Díaz-Perales, Araceli

    2014-05-02

    Alt a 1 is a protein found in Alternaria alternata spores related to virulence and pathogenicity and considered to be responsible for chronic asthma in children. We found that spores of Alternaria inoculated on the outer surface of kiwifruits did not develop hyphae. Nevertheless, the expression of Alt a 1 gene was upregulated, and the protein was detected in the pulp where it co-localized with kiwi PR5. Pull-down assays demonstrated experimentally that the two proteins interact in such a way that Alt a 1 inhibits the enzymatic activity of PR5. These results are relevant not only for plant defense, but also for human health as patients with chronic asthma could suffer from an allergic reaction when they eat fruit contaminated with Alternaria. Copyright © 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

  18. Environmental Stress Induces Trinucleotide Repeat Mutagenesis in Human Cells by Alt-Nonhomologous End Joining Repair.

    Science.gov (United States)

    Chatterjee, Nimrat; Lin, Yunfu; Yotnda, Patricia; Wilson, John H

    2016-07-31

    Multiple pathways modulate the dynamic mutability of trinucleotide repeats (TNRs), which are implicated in neurodegenerative disease and evolution. Recently, we reported that environmental stresses induce TNR mutagenesis via stress responses and rereplication, with more than 50% of mutants carrying deletions or insertions-molecular signatures of DNA double-strand break repair. We now show that knockdown of alt-nonhomologous end joining (alt-NHEJ) components-XRCC1, LIG3, and PARP1-suppresses stress-induced TNR mutagenesis, in contrast to the components of homologous recombination and NHEJ, which have no effect. Thus, alt-NHEJ, which contributes to genetic mutability in cancer cells, also plays a novel role in environmental stress-induced TNR mutagenesis. Published by Elsevier Ltd.

  19. Comparison of histologic characteristics of Chinese chronic hepatitis B patients with persistently normal or mildly elevated ALT.

    Science.gov (United States)

    Wang, Hong; Xue, Li; Yan, Rong; Zhou, Yin; Wang, Ming-Shan; Cheng, Mei-Juan; Hai-Jun Huang

    2013-01-01

    Liver disease can develop in chronic hepatitis B (CHB) patients with normal or mildly elevated alanine aminotransferase (ALT) who seldom undergo liver biopsy. We aimed to determine histologic characteristics of a large cohort of Chinese CHB patients undergoing liver biopsy and to evaluate the utility of ALT and HBV DNA values at the time of biopsy in predicting liver disease in this population. This prospective study enrolled 230 treatment-naïve patients with persistently normal or mildly elevated ALT. All patients had a liver biopsy. ALT, aspartate aminotransferase (AST), and HBV DNA levels were some of the other parameters measured. Using Scheuer's classification, significant histology was defined as stage ≧2 fibrosis and/or stage 1 fibrosis plus≧ grade 2 inflammation. Liver disease was observed in 34.4% and 61.8% of patients with normal ALT and mildly elevated ALT, respectively. Patients with mildly elevated ALT levels had significantly more events, including liver disease, elevated AST, and moderate to severe inflammation and liver fibrosis, than patients with normal ALT (all P≤0.005). A total of 107 patients (46.5%) had liver disease and 123 (53.5%) did not. PLT and ALT were significantly associated with liver disease (both PALT, lower platelet count and HBV DNA ALT while gender and HBV DNA levels were significantly associated with liver disease in patients with mildly elevated ALT. Assessing liver damage via biopsy in patients with normal or mildly elevated ALT may help to identify those who would benefit from antiviral therapy.

  20. Risk of alanine transferase (ALT) elevation in patients with rheumatoid arthritis treated with methotrexate in a DAS-steered strategy.

    Science.gov (United States)

    Dirven, L; Klarenbeek, N B; van den Broek, M; van Groenendael, J H L M; de Sonnaville, P B J; Kerstens, P J S M; Huizinga, T W J; Dijkmans, B A C; Lems, W F; Allaart, C F

    2013-05-01

    To determine incidence of increased levels of alanine transferase (ALT) >2× upper limit of normal (ULN) in patients receiving methotrexate (MTX), treated according to a dynamic strategy, and to identify predictors of ALT of >2× ULN. Data of 508 recent-onset rheumatoid arthritis (RA) patients from the BeSt study, randomized to initial monotherapy or combination therapy, were used. Treatment was dynamic, aiming at a disease activity score = ≤ 2.4. ALT was measured every three months. With logistic regression analyses, baseline variables predictive of first ALT of >2× ULN were identified and the association between use of concomitant antirheumatic drugs, the actual and cumulative dose of MTX and ALT of >2× ULN was determined. In total, 498 patients ever initiated MTX, with a total duration on MTX of 1,416 patient-years. In 89 patients, a first incidence of ALT of >2× ULN occurred. Incidence rate was 6.3 per 100 patient-years and cumulative incidence 18 %. ACPA positivity and baseline ALT of >1× ULN were independent predictors of later ALT of >2× ULN (OR 1.8 (95 % CI, 1.1-3.1) and OR 3.1 (95 % CI, 1.6-6.2), respectively). Smoking showed a trend (OR 1.6 (95 % CI, 0.98-2.7)). Mean MTX dosage over time was higher in patients with an ALT of >2× ULN. Patients who did not have an ALT of >2× ULN used more concomitant disease-modifying antirheumatic drugs and longer. In RA patients treated with MTX according to a dynamic strategy resembling daily clinical practice, incidence of increased ALT of >2× ULN was lower than previously reported, and also without treatment adjustments, persistence was rare. The recommendations for ALT monitoring may be reevaluated.

  1. FANCM, BRCA1, and BLM cooperatively resolve the replication stress at the ALT telomeres.

    Science.gov (United States)

    Pan, Xiaolei; Drosopoulos, William C; Sethi, Louisa; Madireddy, Advaitha; Schildkraut, Carl L; Zhang, Dong

    2017-07-18

    In the mammalian genome, certain genomic loci/regions pose greater challenges to the DNA replication machinery (i.e., the replisome) than others. Such known genomic loci/regions include centromeres, common fragile sites, subtelomeres, and telomeres. However, the detailed mechanism of how mammalian cells cope with the replication stress at these loci/regions is largely unknown. Here we show that depletion of FANCM, or of one of its obligatory binding partners, FAAP24, MHF1, and MHF2, induces replication stress primarily at the telomeres of cells that use the alternative lengthening of telomeres (ALT) pathway as their telomere maintenance mechanism. Using the telomere-specific single-molecule analysis of replicated DNA technique, we found that depletion of FANCM dramatically reduces the replication efficiency at ALT telomeres. We further show that FANCM, BRCA1, and BLM are actively recruited to the ALT telomeres that are experiencing replication stress and that the recruitment of BRCA1 and BLM to these damaged telomeres is interdependent and is regulated by both ATR and Chk1. Mechanistically, we demonstrated that, in FANCM-depleted ALT cells, BRCA1 and BLM help to resolve the telomeric replication stress by stimulating DNA end resection and homologous recombination (HR). Consistent with their roles in resolving the replication stress induced by FANCM deficiency, simultaneous depletion of BLM and FANCM, or of BRCA1 and FANCM, leads to increased micronuclei formation and synthetic lethality in ALT cells. We propose that these synthetic lethal interactions can be explored for targeting the ALT cancers.

  2. Choline transport via choline transporter-like protein 1 in conditionally immortalized rat syncytiotrophoblast cell lines TR-TBT.

    Science.gov (United States)

    Lee, N-Y; Choi, H-M; Kang, Y-S

    2009-04-01

    Choline is an essential nutrient for phospholipids and acetylcholine biosynthesis in normal development of fetus. In the present study, we investigated the functional characteristics of choline transport system and inhibitory effect of cationic drugs on choline transport in rat conditionally immortalized syncytiotrophoblast cell line (TR-TBT). Choline transport was weakly Na(+) dependent and significantly influenced by extracellular pH and by membrane depolarization. The transport process of choline is saturable with Michaelis-Menten constants (K(m)) of 68microM and 130microM in TR-TBT 18d-1 and TR-TBT 18d-2 respectively. Choline uptake in the cells was inhibited by unlabeled choline and hemicholinium-3 as well as various organic cations including guanidine, amiloride and acetylcholine. However, the prototypical organic cation tetraethylammonium and cimetidine showed very little inhibitory effect of choline uptake in TR-TBT cells. RT-PCR revealed that choline transporter-like protein 1 (CTL1) and organic cation transporter 2 (OCT2) are expressed in TR-TBT cells. The transport properties of choline in TR-TBT cells were similar or identical to that of CTL1 but not OCT2. CTL1 was also detected in human placenta. In addition, several cationic drugs such as diphenhydramine and verapamil competitively inhibited choline uptake in TR-TBT 18d-1 with K(i) of 115microM and 55microM, respectively. Our results suggest that choline transport system, which has intermediate affinity and weakly Na(+) dependent, in TR-TBT seems to occur through a CTL1 and this system may have relevance with the uptake of pharmacologically important organic cation drugs.

  3. Persistent Exposure to Porphyromonas gingivalis Promotes Proliferative and Invasion Capabilities, and Tumorigenic Properties of Human Immortalized Oral Epithelial Cells.

    Science.gov (United States)

    Geng, Fengxue; Liu, Junchao; Guo, Yan; Li, Chen; Wang, Hongyang; Wang, Hongyan; Zhao, Haijiao; Pan, Yaping

    2017-01-01

    Recent epidemiological studies revealed a significant association between oral squamous cell carcinoma (OSCC) and Porphyromonas gingivalis , a major pathogen of periodontal disease. As a keystone pathogen of periodontitis, P. gingivalis is known not only to damage local periodontal tissues, but also to evade the host immune system and eventually affect systemic health. However, its role in OSCC has yet to be defined. To explore the underlying effect of chronic P. gingivalis infection on OSCC and to identify relevant biomarkers as promising targets for therapy and prevention, we established a novel model by exposing human immortalized oral epithelial cells (HIOECs) to P. gingivalis at a low multiplicity of infection (MOI) for 5-23 weeks. The P. gingivalis infected HIOECs were monitored for tumor biological alteration by proliferation, wound healing, transwell invasion, and gelatin zymography assays. Microarray and proteomic analyses were performed on HIOECs infected with P. gingivalis for 15 weeks, and some selected data were validated by quantitative real-time PCR and (or) western blot on cells infected for 15 and 23 weeks. Persistent exposure to P. gingivalis caused cell morphological changes, increased proliferation ability with higher S phase fraction in the cell cycle, and promoted cell migratory and invasive properties. In combining results of bioinformatics analyses and validation assays, tumor-related genes such as NNMT, FLI1, GAS6, lncRNA CCAT1, PDCD1LG2, and CD274 may be considered as the key regulators in tumor-like transformation in response to long-time exposure of P. gingivalis . In addition, some useful clinical biomarkers and novel proteins were also presented. In conclusion, P. gingivalis could promote tumorigenic properties of HIOECs, indicating that chronic P. gingivalis infection may be considered as a potential risk factor for oral cancer. The key regulators detected from the present model might be used in monitoring the development of OSCC with

  4. Balancing Benefits and Risks of Immortal Data: Participants' Views of Open Consent in the Personal Genome Project.

    Science.gov (United States)

    Zarate, Oscar A; Brody, Julia Green; Brown, Phil; Ramirez-Andreotta, Mónica D; Perovich, Laura; Matz, Jacob

    2016-01-01

    An individual's health, genetic, or environmental-exposure data, placed in an online repository, creates a valuable shared resource that can accelerate biomedical research and even open opportunities for crowd-sourcing discoveries by members of the public. But these data become "immortalized" in ways that may create lasting risk as well as benefit. Once shared on the Internet, the data are difficult or impossible to redact, and identities may be revealed by a process called data linkage, in which online data sets are matched to each other. Reidentification (re-ID), the process of associating an individual's name with data that were considered deidentified, poses risks such as insurance or employment discrimination, social stigma, and breach of the promises often made in informed-consent documents. At the same time, re-ID poses risks to researchers and indeed to the future of science, should re-ID end up undermining the trust and participation of potential research participants. The ethical challenges of online data sharing are heightened as so-called big data becomes an increasingly important research tool and driver of new research structures. Big data is shifting research to include large numbers of researchers and institutions as well as large numbers of participants providing diverse types of data, so the participants' consent relationship is no longer with a person or even a research institution. In addition, consent is further transformed because big data analysis often begins with descriptive inquiry and generation of a hypothesis, and the research questions cannot be clearly defined at the outset and may be unforeseeable over the long term. In this article, we consider how expanded data sharing poses new challenges, illustrated by genomics and the transition to new models of consent. We draw on the experiences of participants in an open data platform-the Personal Genome Project-to allow study participants to contribute their voices to inform ethical consent

  5. Aspartate Aminotransferase (AST/GOT and Alanine Aminotransferase (ALT/GPT Detection Techniques

    Directory of Open Access Journals (Sweden)

    Hak-Sung Kim

    2006-07-01

    Full Text Available The levels of aspartate aminotransferase (AST/GOT and alanineaminotransferase (ALT/GPT in serum can help people diagnose body tissues especially theheart and the liver are injured or not. This article provides a comprehensive review ofresearch activities that concentrate on AST/GOT and ALT/GPT detection techniques due totheir clinical importance. The detection techniques include colorimetric, spectrophotometric,chemiluminescence, chromatography, fluorescence and UV absorbance, radiochemical, andelectrochemical techniques. We devote the most attention on experimental principle. Insome methods a few representative devices and important conclusions are presented.

  6. Aspartate Aminotransferase (AST/GOT) and Alanine Aminotransferase (ALT/GPT) Detection Techniques

    Science.gov (United States)

    Huang, Xing-Jiu; Choi, Yang-Kyu; Im, Hyung-Soon; Yarimaga, Oktay; Yoon, Euisik; Kim, Hak-Sung

    2006-01-01

    The levels of aspartate aminotransferase (AST/GOT) and alanine aminotransferase (ALT/GPT) in serum can help people diagnose body tissues especially the heart and the liver are injured or not. This article provides a comprehensive review of research activities that concentrate on AST/GOT and ALT/GPT detection techniques due to their clinical importance. The detection techniques include colorimetric, spectrophotometric, chemiluminescence, chromatography, fluorescence and UV absorbance, radiochemical, and electrochemical techniques. We devote the most attention on experimental principle. In some methods a few representative devices and important conclusions are presented.

  7. ALT-803 Transiently Reduces Simian Immunodeficiency Virus Replication in the Absence of Antiretroviral Treatment.

    Science.gov (United States)

    Ellis-Connell, Amy L; Balgeman, Alexis J; Zarbock, Katie R; Barry, Gabrielle; Weiler, Andrea; Egan, Jack O; Jeng, Emily K; Friedrich, Thomas; Miller, Jeffrey S; Haase, Ashley T; Schacker, Timothy W; Wong, Hing C; Rakasz, Eva; O'Connor, Shelby L

    2018-02-01

    Developing biological interventions to control human immunodeficiency virus (HIV) replication in the absence of antiretroviral therapy (ART) could contribute to the development of a functional cure. As a potential alternative to ART, the interleukin-15 (IL-15) superagonist ALT-803 has been shown to boost the number and function of HIV-specific CD8 + T and NK cell populations in vitro Four simian immunodeficiency virus (SIV)-positive rhesus macaques, three of whom possessed major histocompatibility complex alleles associated with control of SIV and all of whom had received SIV vaccine vectors that had the potential to elicit CD8 + T cell responses, were given ALT-803 in three treatment cycles. The first and second cycles of treatment were separated by 2 weeks, while the third cycle was administered after a 29-week break. ALT-803 transiently elevated the total CD8 + effector and central memory T cell and NK cell populations in peripheral blood, while viral loads transiently decreased by ∼2 logs in all animals. Virus suppression was not sustained as T cells became less responsive to ALT-803 and waned in numbers. No effect on viral loads was observed in the second cycle of ALT-803, concurrent with downregulation of the IL-2/15 common γC and β chain receptors on both CD8 + T cells and NK cells. Furthermore, populations of immunosuppressive T cells increased during the second cycle of ALT-803 treatment. During the third treatment cycle, responsiveness to ALT-803 was restored. CD8 + T cells and NK cells increased again 3- to 5-fold, and viral loads transiently decreased again by 1 to 2 logs. IMPORTANCE Overall, our data show that ALT-803 has the potential to be used as an immunomodulatory agent to elicit effective immune control of HIV/SIV replication. We identify mechanisms to explain why virus control is transient, so that this model can be used to define a clinically appropriate treatment regimen. Copyright © 2018 American Society for Microbiology.

  8. A spontaneously immortalized Schwann cell line from aldose reductase-deficient mice as a useful tool for studying polyol pathway and aldehyde metabolism.

    Science.gov (United States)

    Niimi, Naoko; Yako, Hideji; Takaku, Shizuka; Kato, Hiroshi; Matsumoto, Takafumi; Nishito, Yasumasa; Watabe, Kazuhiko; Ogasawara, Saori; Mizukami, Hiroki; Yagihashi, Soroku; Chung, Sookja K; Sango, Kazunori

    2018-03-01

    The increased glucose flux into the polyol pathway via aldose reductase (AR) is recognized as a major contributing factor for the pathogenesis of diabetic neuropathy, whereas little is known about the functional significance of AR in the peripheral nervous system. Spontaneously immortalized Schwann cell lines established from long-term cultures of AR-deficient and normal C57BL/6 mouse dorsal root ganglia and peripheral nerves can be useful tools for studying the physiological and pathological roles of AR. These cell lines, designated as immortalized knockout AR Schwann cells 1 (IKARS1) and 1970C3, respectively, demonstrated distinctive Schwann cell phenotypes, such as spindle-shaped morphology and immunoreactivity to S100, p75 neurotrophin receptor, and vimentin, and extracellular release of neurotrophic factors. Conditioned media obtained from these cells promoted neuronal survival and neurite outgrowth of cultured adult mouse dorsal root ganglia neurons. Microarray and real-time RT-PCR analyses revealed significantly down-regulated mRNA expression of polyol pathway-related enzymes, sorbitol dehydrogenase and ketohexokinase, in IKARS1 cells compared with those in 1970C3 cells. In contrast, significantly up-regulated mRNA expression of aldo-keto reductases (AKR1B7 and AKR1B8) and aldehyde dehydrogenases (ALDH1L2, ALDH5A1, and ALDH7A1) was detected in IKARS1 cells compared with 1970C3 cells. Exposure to reactive aldehydes (3-deoxyglucosone, methylglyoxal, and 4-hydroxynonenal) significantly up-regulated the mRNA expression of AKR1B7 and AKR1B8 in IKARS1 cells, but not in 1970C3 cells. Because no significant differences in viability between these two cell lines after exposure to these aldehydes were observed, it can be assumed that the aldehyde detoxification is taken over by AKR1B7 and AKR1B8 in the absence of AR. © 2017 International Society for Neurochemistry.

  9. The E7 protein of the cottontail rabbit papillomavirus immortalizes normal rabbit keratinocytes and reduces pRb levels, while E6 cooperates in immortalization but neither degrades p53 nor binds E6AP

    International Nuclear Information System (INIS)

    Ganzenmueller, Tina; Matthaei, Markus; Muench, Peter; Scheible, Michael; Iftner, Angelika; Hiller, Thomas; Leiprecht, Natalie; Probst, Sonja; Stubenrauch, Frank; Iftner, Thomas

    2008-01-01

    Human papillomaviruses (HPVs) cause cervical cancer and are associated with the development of non-melanoma skin cancer. A suitable animal model for papillomavirus-associated skin carcinogenesis is the infection of domestic rabbits with the cottontail rabbit papillomavirus (CRPV). As the immortalizing activity of CRPV genes in the natural target cells remains unknown, we investigated the properties of CRPV E6 and E7 in rabbit keratinocytes (RK) and their influence on the cell cycle. Interestingly, CRPV E7 immortalized RK after a cellular crisis but showed no such activity in human keratinocytes. Co-expressed CRPV E6 prevented cellular crisis. The HPV16 or CRPV E7 protein reduced rabbit pRb levels thereby causing rabbit p19 ARF induction and accumulation of p53 without affecting cellular proliferation. Both CRPV E6 proteins failed to degrade rabbit p53 in vitro or to bind E6AP; however, p53 was still inducible by mitomycin C. In summary, CRPV E7 immortalizes rabbit keratinocytes in a species-specific manner and E6 contributes to immortalization without directly affecting p53

  10. Development of Public Immortal Mapping Populations, Molecular Markers and Linkage Maps for Rapid Cycling Brassica rapa and B. oleracea

    Science.gov (United States)

    In this study we describe public immortal mapping populations of self-compatible lines, molecular markers, and linkage maps for Brassica rapa and B. oleracea. We propose that these resources are valuable reference tools for the Brassica community. The B. rapa population consists of 150 recombinant...

  11. A long-term hepatitis B viremia model generated by transplanting nontumorigenic immortalized human hepatocytes in Rag-2-deficient mice

    NARCIS (Netherlands)

    Parashar, B; Moshage, H; Tanaka, KE; Engelhardt, D; Rabbani, E; Roy-Chowdhury, N; Roy-Chowdhury, J

    Development of new therapies for human hepatitis B virus infection (HBV) would be greatly facilitated by the availability of a suitable small-animal model for HBV virus production in vivo. To develop a murine model for HBV production, we established an immortalized, cloned liver cell line by

  12. Stepwise DNA Methylation Changes Are Linked to Escape from Defined Proliferation Barriers and Mammary Epithelial Cell Immortalization

    Energy Technology Data Exchange (ETDEWEB)

    Novak, Petr; Jensen, Taylor J.; Garbe, James C.; Stampfer, Martha R.; Futscher, Bernard W.

    2009-04-20

    The timing and progression of DNA methylation changes during carcinogenesis are not completely understood. To develop a timeline of aberrant DNA methylation events during malignant transformation, we analyzed genome-wide DNA methylation patterns in an isogenic human mammary epithelial cell (HMEC) culture model of transformation. To acquire immortality and malignancy, the cultured finite lifespan HMEC must overcome two distinct proliferation barriers. The first barrier, stasis, is mediated by the retinoblastoma protein and can be overcome by loss of p16(INK4A) expression. HMEC that escape stasis and continue to proliferate become genomically unstable before encountering a second more stringent proliferation barrier, telomere dysfunction due to telomere attrition. Rare cells that acquire telomerase expression may escape this barrier, become immortal, and develop further malignant properties. Our analysis of HMEC transitioning from finite lifespan to malignantly transformed showed that aberrant DNA methylation changes occur in a stepwise fashion early in the transformation process. The first aberrant DNA methylation step coincides with overcoming stasis, and results in few to hundreds of changes, depending on how stasis was overcome. A second step coincides with immortalization and results in hundreds of additional DNA methylation changes regardless of the immortalization pathway. A majority of these DNA methylation changes are also found in malignant breast cancer cells. These results show that large-scale epigenetic remodeling occurs in the earliest steps of mammary carcinogenesis, temporally links DNA methylation changes and overcoming cellular proliferation barriers, and provides a bank of potential epigenetic biomarkers that mayprove useful in breast cancer risk assessment.

  13. Transcriptome profile and cytogenetic analysis of immortalized neuronally restricted progenitor cells derived from the porcine olfactory bulb

    Science.gov (United States)

    Recently, we established and phenotypically characterized an immortalized porcine olfactory bulb neuroblast cell line, OBGF400 (Uebing-Czipura et al., 2008). To facilitate the future application of these cells in studies of neurological dysfunction and neuronal replacement therapies, a comprehensive...

  14. Influence of gender, BMI, and ethnicity on serum ALT levels of healthy students of a medical school

    International Nuclear Information System (INIS)

    Bilal, M.; Tariq, A.; Khan, S.; Quratulain, A.; Tariq, A.; Shahid, M.F.; Khan, M.W.; Naveed, A.K.

    2011-01-01

    Background: Alanine Aminotransferase (ALT) is an enzyme found in Liver and indicates injury to Hepatocytes. It is influenced by various factors. The objectives of this study were to identify the correlates of ALT activity among healthy medical students of Army Medical College, National University of Sciences and Technology, aged 18-22 years. This was to establish the mean ALT levels of the students and compare them with those in various parts of the world and observe various correlations that exist and factors that may influence ALT levels. Methods: This population included 143 volunteer students (93 men and 50 women) selected on the basis of negative answers to a detailed medical questionnaire including past medical history, drug and alcohol consumption, on the absence of clinical signs of liver disease, on the negativity of serological testing for Hepatitis B and C virus. Results: The mean ALT level of the entire population was 28.7 IU/L. A major sex-difference in ALT value was observed, the mean ALT value being higher in men than in women (32.1+- 21.7 vs. 22.6 +- 9.7 IU/L, p<0.004). According to WHO criteria for Asians, normal BMI was taken from 18.5-23.0 Kg/m/sup 2/. There was a positive significant correlation between serum ALT level and BMI (p<0.002). ALT level strongly correlates with body mass index and gender. There was no significant variation in ALT levels among Punjabis and Sindhis, Balochis, Pathans, and Kashmiris. Conclusion: We suggest the need of taking into account these parameters in a clinical interpretation of ALT level. (author)

  15. Relationships between alanine aminotransferase (ALT), visceral adipose tissue (AT) and metabolic risk factors in a middle-aged Japanese population.

    Science.gov (United States)

    Oka, Rie; Yagi, Kunimasa; Nakanishi, Chiaki; Konno, Tetsuo; Kawashiri, Masa-aki; Hayashi, Kenshi; Nohara, Atsushi; Inazu, Akihiro; Yamagishi, Masakazu

    2014-01-01

    The commonly observed relationship between increased visceral adiposity and metabolic abnormalities may be partly mediated by a concomitant increase in liver fat content. We evaluated the independent association between the level of alanine aminotransferase (ALT) as a surrogate marker of the liver fat content and the incidence of metabolic abnormalities after adjusting for the amount of visceral adipose tissue (AT). The subjects included 1,118 Japanese individuals (44% women) who underwent computed tomography to assess the amount of visceral AT on medical checkups. Cross-sectional associations between the serum ALT, visceral AT and metabolic risk factors were examined. The ALT level and visceral AT were found to show a significant correlation(r=0.41 in men and r=0.36 in women, p<0.001). In a multivariable linear regression analysis, the ALT level and visceral AT were found to be independently associated with blood pressure in men and triglycerides and 2-hour post-challenge glucose in both genders(p<0.01), whereas only visceral AT was found to be associated with HDL-cholesterol(p<0.01). When the participants were classified into four subgroups based on the 75th percentiles of ALT and visceral AT, the low ALT/high-visceral AT group, but not the high-ALT/low-visceral AT group, had a significantly higher odds ratio for low HDL-cholesterol among both genders(p<0.05) and for hypertriglyceridemia in men only(p<0.05). Meanwhile, the high-ALT/low-visceral AT group, but ot the low-ALT/high-visceral AT group, had a significantly higher odds ratio for IGT among women(p<0.05). Although the ALT level and visceral AT were found to be independently associated with most metabolic risk factors, visceral AT had a dominant association with dyslipidemia in both genders, while the ALT level appeared to have a closer association with IGT in women.

  16. The use of hTERT-immortalized cells in tissue engineering

    DEFF Research Database (Denmark)

    Kassem, Moustapha; Abdallah, Basem; Yu, Zentao

    2004-01-01

    The use of human telomerase reverse transcriptase (hTERT)-immortalized cells in tissue engineering protocols is a potentially important application of telomere biology. Several human cell types have been created that overexpress the hTERT gene with enhanced telomerase activity, extended life span...... and maintained or even improved functional activities. Furthermore, some studies have employed the telomerized cells in tissue engineering protocols with very good results. However, high telomerase activity allows extensive cell proliferation that may be associated with genomic instability and risk for cell...... transformation. Thus, safety issues should be studied carefully before using the telomerized tissues in the clinic. Alternatively, the development of conditional or intermittent telomerase activation protocols is needed....

  17. Immortalization and characterization of mouse floxed Bmp2/4 osteoblasts

    Energy Technology Data Exchange (ETDEWEB)

    Wu, Li-An [Department of Pediatric Dentistry, The University of Texas Health Science Center at San Antonio, TX (United States); Department of Pediatric Dentistry, School of Stomatology, The Fourth Military Medical University, Xi-an (China); Yuan, Guohua; Yang, Guobin [Department of Pediatric Dentistry, The University of Texas Health Science Center at San Antonio, TX (United States); Key Laboratory of Oral Biomedical Engineering Ministry of Education, Wuhan (China); Ortiz-Gonzalez, Iris [Department of Pediatric Dentistry, The University of Texas Health Science Center at San Antonio, TX (United States); Yang, Wuchen; Cui, Yong [Department of Periodontics, Dental School, The University of Texas Health Science Center at San Antonio, TX (United States); MacDougall, Mary [Department of Oral/Maxillofacial Surgery, University of Alabama, Birmingham, AL (United States); Donly, Kevin J. [Department of Pediatric Dentistry, The University of Texas Health Science Center at San Antonio, TX (United States); Harris, Stephen [Department of Periodontics, Dental School, The University of Texas Health Science Center at San Antonio, TX (United States); Chen, Shuo, E-mail: chens0@uthscsa.edu [Department of Pediatric Dentistry, The University of Texas Health Science Center at San Antonio, TX (United States)

    2009-08-14

    Generation of a floxed Bmp2/4 osteoblast cell line is a valuable tool for studying the modulatory effects of Bmp2 and Bmp4 on osteoblast differentiation as well as relevant molecular events. In this study, primary floxed Bmp2/4 mouse osteoblasts were cultured and transfected with simian virus 40 large T-antigen. Transfection was verified by polymerase chain reaction (PCR) and immunohistochemistry. To examine the characteristics of the transfected cells, morphology, proliferation and mineralization were analyzed, expression of cell-specific genes including Runx2, ATF4, Dlx3, Osx, dentin matrix protein 1, bone sialoprotein, osteopontin, osteocalcin, osteonectin and collagen type I was detected. These results show that transfected floxed Bmp2/4 osteoblasts bypassed senescence with a higher proliferation rate, but retain the genotypic and phenotypic characteristics similar to the primary cells. Thus, we for the first time demonstrate the establishment of an immortalized mouse floxed Bmp2/4 osteoblast cell line.

  18. Immortalized Adult Rodent Schwann Cells as In Vitro Models to Study Diabetic Neuropathy

    Directory of Open Access Journals (Sweden)

    Kazunori Sango

    2011-01-01

    Full Text Available We have established spontaneously immortalized Schwann cell lines from normal adult mice and rats and murine disease models. One of the normal mouse cell lines, IMS32, possesses some biological properties of mature Schwann cells and high proliferative activities. The IMS32 cells under hyperglycemic and/or hyperlipidemic conditions have been utilized to investigate the pathogenesis of diabetic neuropathy, especially the polyol pathway hyperactivity, glycation, increased oxidative stress, and reduced synthesis of neurotrophic factors. In addition to the mouse cell lines, our current study focuses on the characterization of a normal rat cell line, IFRS1, under normal and high glucose conditions. These Schwann cell lines can be valuable tools for exploring the detailed mechanisms leading to diabetic neuropathy and novel therapeutic approaches against that condition.

  19. Recovery of important physiological functions in 3D culture of immortal hepatocytes

    DEFF Research Database (Denmark)

    Wrzesinski, Krzysztof; Fey, S. J.

    2011-01-01

    differentiated to a much greater degree and showed many characteristics seen in liver tissue. These results suggest that growth in 2D preferentially supports proliferation whereas growth in 3D preferentially supports cellular differentiation. References: Griffith LG, Swartz MA. (2006): Capturing complex 3D...... to grow human liver cells in ‘3 dimensional’ cultures so that they behave very similar to the liver in our bodies. By growing the immortal hepatocytes in specially designed bioreactors they form small pieces of ‘pseudotissue’ which exhibit several of the functions seen in the adult liver. We have grown...... and glycogen granules. In other words, the cells grown in the classical 2D growth conditions were proliferating rapidly and showing little sign of the differentiated phenotype that hepatocytes in liver tissue show. Conversely the same cells grown as spheroids were proliferating very slowly but were...

  20. Immortalization and characterization of mouse floxed Bmp2/4 osteoblasts

    International Nuclear Information System (INIS)

    Wu, Li-An; Yuan, Guohua; Yang, Guobin; Ortiz-Gonzalez, Iris; Yang, Wuchen; Cui, Yong; MacDougall, Mary; Donly, Kevin J.; Harris, Stephen; Chen, Shuo

    2009-01-01

    Generation of a floxed Bmp2/4 osteoblast cell line is a valuable tool for studying the modulatory effects of Bmp2 and Bmp4 on osteoblast differentiation as well as relevant molecular events. In this study, primary floxed Bmp2/4 mouse osteoblasts were cultured and transfected with simian virus 40 large T-antigen. Transfection was verified by polymerase chain reaction (PCR) and immunohistochemistry. To examine the characteristics of the transfected cells, morphology, proliferation and mineralization were analyzed, expression of cell-specific genes including Runx2, ATF4, Dlx3, Osx, dentin matrix protein 1, bone sialoprotein, osteopontin, osteocalcin, osteonectin and collagen type I was detected. These results show that transfected floxed Bmp2/4 osteoblasts bypassed senescence with a higher proliferation rate, but retain the genotypic and phenotypic characteristics similar to the primary cells. Thus, we for the first time demonstrate the establishment of an immortalized mouse floxed Bmp2/4 osteoblast cell line.

  1. On the Trajectories of the Predetermined ALT Model: What Are We Really Modeling?

    Science.gov (United States)

    Jongerling, Joran; Hamaker, Ellen L.

    2011-01-01

    This article shows that the mean and covariance structure of the predetermined autoregressive latent trajectory (ALT) model are very flexible. As a result, the shape of the modeled growth curve can be quite different from what one might expect at first glance. This is illustrated with several numerical examples that show that, for example, a…

  2. Effect of Ramadan fasting on alanine transferase (ALT in nonalcoholic fatty liver disease (NAFLD

    Directory of Open Access Journals (Sweden)

    Hojjatolah Rahimi

    2017-09-01

    Full Text Available Background: The effects of Ramadan fasting on NAFLD are unknown and there are very limited studies have done in this area. Some nutritional and behavioral changes of fasting people in Ramadan can affect NAFLD. These include nutrition with high fat and calories, altering in weight and sleep and low physical activity. We decided to evaluate the effects of these changes on one of the important indicators of deterioration of NAFLD, ALT.Methods: Sixty patients with fatty liver disease performed two consecutive ALT exams before and after Ramadan month of whom finally 34 were fasting and 26 nonfasting. After collecting data they were divided in two groups of fasting and nonfasting and compared using SPSS software.Results: Mean ALT change from before to after Ramadan was higher and positive in fasting (+7.38±8.47 IU/L compared to nonfastng patients that was negative (-0.12±10.15 IU/L (P=0.002 and this change was mainly in patients who had fasted 21-30 days.Conclusion: Ramadan fasting may increase ALT. It is needed to perform more studies in patients with NAFLD during Ramadan fasting with larger sample size and in various conditions especially weight loss with patients’ education for observing dietary regimen.

  3. Preparation of Poly(MA-alt-α-olefin-C6,8,12,18)

    Indian Academy of Sciences (India)

    form solution by methanol. Poly(MA-alt-1-octadecene). (C18) was supplied from Aldrich with Mn = 30000–. 50000 g mol−1. All other solvents and reagents were of analytical grade and used without purifi- cation. γ-Aminopropyltrimethoxysilane (APTS) and tetraethoxysilane (tetraethyl orthosilicate) (TEOS) were obtained ...

  4. Laser Ablative Structural Modification of Poly(ethylene-alt-maleic anhydride)

    Czech Academy of Sciences Publication Activity Database

    Pola, Josef; Kupčík, Jaroslav; Durani, S. M. A.; Khavaja, E. E.; Masoudi, H. M.; Bastl, Zdeněk; Šubrt, Jan

    2003-01-01

    Roč. 15, č. 20 (2003), s. 3887-3893 ISSN 0897-4756 Institutional research plan: CEZ:AV0Z4032918; CEZ:AV0Z4072921 Keywords : laser ablation * poly(ethylene- alt -maleic)anhydride Subject RIV: CH - Nuclear ; Quantum Chemistry Impact factor: 4.374, year: 2003

  5. Astronauts Engle and Truly egress Shuttle Orbiter 101 after fourth ALT

    Science.gov (United States)

    1977-01-01

    Astronauts Joe H. Engle, left, commander and Richard H. Truly, pilot, egress the Shuttle Orbiter 101 'Enterprise' following completion of the fourth Approach and Landing Test (ALT) free flight at Dryden Flight Research Center (DFRC) in Southern California. The landed the craft in the desert at Edwards Air Force Base following a two-minute, 34-second unpowered mission.

  6. Astronauts Haise and Fullerton in cockpit of Orbiter 101 prior to fifth ALT

    Science.gov (United States)

    1977-01-01

    Astronauts Fred W. Haise Jr., commander, left, and C. Gordon Fullerton in the cockpit of the Space shuttle Orbiter 101 'Enterprise' prior to the fifth and final free flight in the Approach and Landing Test (ALT) series, from Dryden Flight Research Center (DFRC).

  7. Cu (II), Zn (II) andMn (II) complexes of poly (methyl vinyl ether-alt ...

    Indian Academy of Sciences (India)

    MVE-alt-MA)) with Zn(II), Mn(II) and Cu(II) ions were synthesized from the reaction of the aqueous solution of copolymer and metal(II) chlorides at different temperatures ranging from 25° to 40°C. Elemental analysis of themetal-polymer complexes ...

  8. Measurement and purification of Alanine aminotransferase (ALT enzyme activity in patients with celiac disease

    Directory of Open Access Journals (Sweden)

    Taghreed U. Mohammed

    2017-09-01

    Full Text Available Celiac disease (CD is the most common genetically - based disease in correlation with food intolerance. The aim of this study is to measure the activity of ALT enzyme and purify enzyme from sera women with celiac disease. Alanine aminotransferase (ALT activity has been assayed in (30 women serum samples with celiac disease, age range between (20-40 year and (30 serum of healthy women as control group, age range between (22-38 year. In the present study, the mean value of ALT activity was significantly higher in patients with celiac disease than healthy group (p<0.01. The ALT enzyme was partial purified from sera women with celiac disease by dialysis, gel filtration using Sephadex G- 50 and ion exchange chromatography using DEAE- cellulose A-50 . The results showed a single peak by using gel filtration and the activity reached 31-15 U/L .Two isoenzymes were obtained by using ion exchange chromatography and the purity degree of isoenzymse (I, II were (5.7 and (5.53 fold respectively

  9. Generation of human cortical neurons from a new immortal fetal neural stem cell line

    International Nuclear Information System (INIS)

    Cacci, E.; Villa, A.; Parmar, M.; Cavallaro, M.; Mandahl, N.; Lindvall, O.; Martinez-Serrano, A.; Kokaia, Z.

    2007-01-01

    Isolation and expansion of neural stem cells (NSCs) of human origin are crucial for successful development of cell therapy approaches in neurodegenerative diseases. Different epigenetic and genetic immortalization strategies have been established for long-term maintenance and expansion of these cells in vitro. Here we report the generation of a new, clonal NSC (hc-NSC) line, derived from human fetal cortical tissue, based on v-myc immortalization. Using immunocytochemistry, we show that these cells retain the characteristics of NSCs after more than 50 passages. Under proliferation conditions, when supplemented with epidermal and basic fibroblast growth factors, the hc-NSCs expressed neural stem/progenitor cell markers like nestin, vimentin and Sox2. When growth factors were withdrawn, proliferation and expression of v-myc and telomerase were dramatically reduced, and the hc-NSCs differentiated into glia and neurons (mostly glutamatergic and GABAergic, as well as tyrosine hydroxylase-positive, presumably dopaminergic neurons). RT-PCR analysis showed that the hc-NSCs retained expression of Pax6, Emx2 and Neurogenin2, which are genes associated with regionalization and cell commitment in cortical precursors during brain development. Our data indicate that this hc-NSC line could be useful for exploring the potential of human NSCs to replace dead or damaged cortical cells in animal models of acute and chronic neurodegenerative diseases. Taking advantage of its clonality and homogeneity, this cell line will also be a valuable experimental tool to study the regulatory role of intrinsic and extrinsic factors in human NSC biology

  10. #AltPlanets: Exploring the Exoplanet Catalogue with Neural Networks

    Science.gov (United States)

    Laneuville, M.; Tasker, E. J.; Guttenberg, N.

    2017-12-01

    The launch of Kepler in 2009 brought the number of known exoplanets into the thousands, in a growth explosion that shows no sign of abating. While the data available for individual planets is presently typically restricted to orbital and bulk properties, the quantity of data points allows the potential for meaningful statistical analysis. It is not clear how planet mass, radius, orbital path, stellar properties and neighbouring planets influence one another, therefore it seems inevitable that patterns will be missed simply due to the difficulty of including so many dimensions. Even simple trends may be overlooked if they fall outside our expectation of planet formation; a strong risk in a field where new discoveries have destroyed theories from the first observations of hot Jupiters. A possible way forward is to take advantage of the capabilities of neural network autoencoders. The idea of such algorithms is to learn a representation (encoding) of the data in a lower dimension space, without a priori knowledge about links between the elements. This encoding space can then be used to discover the strongest correlations in the original dataset.The key point is that trends identified by a neural network are independent of any previous analysis and pre-conceived ideas about physical processes. Results can reveal new relationships between planet properties and verify existing trends. We applied this concept to study data from the NASA Exoplanet Archive and while we have begun to explore the potential use of neural networks for exoplanet data, there are many possible extensions. For example, the network can produce a large number of 'alternative planets' whose statistics should match the current distribution. This larger dataset could highlight gaps in the parameter space or indicate observations are missing particular regimes. This could guide instrument proposals towards objects liable to yield the most information.

  11. Telomere Length Maintenance in Cancer: At the Crossroad between Telomerase and Alternative Lengthening of Telomeres (ALT).

    Science.gov (United States)

    De Vitis, Marco; Berardinelli, Francesco; Sgura, Antonella

    2018-02-18

    Eukaryotic cells undergo continuous telomere shortening as a consequence of multiple rounds of replications. During tumorigenesis, cells have to acquire telomere DNA maintenance mechanisms (TMMs) in order to counteract telomere shortening, to preserve telomeres from DNA damage repair systems and to avoid telomere-mediated senescence and/or apoptosis. For this reason, telomere maintenance is an essential step in cancer progression. Most human tumors maintain their telomeres expressing telomerase, whereas a lower but significant proportion activates the alternative lengthening of telomeres (ALT) pathway. However, evidence about the coexistence of ALT and telomerase has been found both in vivo in the same cancer populations and in vitro in engineered cellular models, making the distinction between telomerase- and ALT-positive tumors elusive. Indeed, after the development of drugs able to target telomerase, the capability for some cancer cells to escape death, switching from telomerase to ALT, was highlighted. Unfortunately, to date, the mechanism underlying the possible switching or the coexistence of telomerase and ALT within the same cell or populations is not completely understood and different factors could be involved. In recent years, different studies have tried to shed light on the complex regulation network that controls the transition between the two TMMs, suggesting a role for embryonic cancer origin, epigenetic modifications, and specific genes activation-both in vivo and in vitro. In this review, we examine recent findings about the cancer-associated differential activation of the two known TMMs and the possible factors implicated in this process. Furthermore, some studies on cancers are also described that did not display any TMM.

  12. A prospective study on the causes of notably raised alanine aminotransferase (ALT).

    Science.gov (United States)

    Björnsson, Helgi Kristinn; Olafsson, Sigurdur; Bergmann, Ottar M; Björnsson, Einar S

    2016-01-01

    High levels of alanine aminotransferase (ALT) can be a marker of severe liver disease with variable aetiologies and prognosis. Very few prospective studies have been undertaken on the aetiology and prognosis of patients with high ALT levels. No population-based prospective study has systematically evaluated drug-induced liver injury (DILI) among these patients. The objective was to determine the aetiology and prognosis of patients with high ALT. In a catchment area of 160,000 inhabitants, a population-based prospective study identified all adult patients with serum level of ALT >500 U/L during a 12-month period. All underwent thorough diagnostic work-up and follow-up. In suspected DILI, causality was assessed with Roussel Uclaf Causality Assessment Method. A total of 155 patients were identified with ALT >500 U/L, 12 children and one with ALT of non-liver-related origin, leaving 142 patients for the analysis: 73 (51%) males, median age 52 (IQR 36-68, range 19-89 years). The most common causes were choledocholithiasis 48/142 (34%), ischaemic hepatitis 26 (18%), viral hepatitis 16 (11%) and DILI 15 (11%), hepatobiliary malignancy (n = 6), surgery/interventions (n = 8) and other aetiologies (n = 23). No specific aetiology was found in 6% of cases. In the total study cohort 99 (70%) required hospitalisation, 78 (55%) had jaundice and 22 (16%) died, liver-related death in 10%, 35% in IH and 7% in DILI. The most common cause of notably high ALT was choledocholithiasis. Ischaemic hepatitis was a common aetiology with approximately 35% liver-related mortality. Viral hepatitis and DILI were important aetiologies among these patients.

  13. Bile acid sequestration by cholestyramine mitigates FGFR4 inhibition-induced ALT elevation.

    Science.gov (United States)

    Schadt, Heiko S; Wolf, Armin; Mahl, Joerg Andreas; Wuersch, Kuno; Couttet, Philippe; Schwald, Marianne; Fischer, Audrey; Lienard, Mathilde; Emotte, Corinne; Teng, Chi-Hse; Skuba, Elizabeth; Richardson, Terrilyn A; Manenti, Luigi; Weiss, Andreas; Graus Porta, Diana; Fairhurst, Robin A; Kullak-Ublick, Gerd A; Chibout, Salah-Dine; Pognan, Francois; Kluwe, William; Kinyamu-Akunda, Jacqueline

    2018-02-08

    The FGF19-FGFR4-βKlotho (KLB) pathway plays an important role in the regulation of bile acid (BA) homeostasis. Aberrant activation of this pathway has been described in the development and progression of a subset of liver cancers including hepatocellular carcinoma (HCC), establishing FGFR4 as an attractive therapeutic target for such solid tumors. FGF401 is a highly selective FGFR4 kinase inhibitor being developed for HCC, currently in Phase I/II clinical studies. In preclinical studies in mice and dogs, oral administration of FGF401 led to induction of Cyp7a1, elevation of its peripheral marker 7alpha-hydroxy-4-cholesten-3-one (C4), increased BA pool size, decreased serum cholesterol and diarrhea in dogs. FGF401 was also associated with increases of serum aminotransferases, primarily alanine aminotransferase (ALT), in the absence of any observable adverse histopathological findings in the liver, or in any other organs. We hypothesized that the increase in ALT could be secondary to increased BAs and conducted an investigative study in dogs with FGF401 and co-administration of the BA sequestrant cholestyramine (CHO). CHO prevented and reversed FGF401-related increases in ALT in dogs in parallel to its ability to reduce BAs in the circulation. Correlation analysis showed that FGF401-mediated increases in ALT strongly correlated with increases in taurolithocholic acid (TLCA) and taurodeoxycholic acid (TDCA), the major secondary BAs in dog plasma, indicating a mechanistic link between ALT elevation and changes in BA pool hydrophobicity. Thus, CHO may offer the potential to mitigate elevations in serum aminotransferases in human subjects that are caused by targeted FGFR4 inhibition and elevated intracellular BA levels. © The Author 2018. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  14. Establishing reference intervals for ALT, AST, UR, Cr, and UA in apparently healthy Chinese adolescents.

    Science.gov (United States)

    Li, Ying; Mussa, Ahmed Ebrahim; Tang, Aiguo; Xiang, Zhongyuan; Mo, Ximing

    2018-03-01

    The current child-specific reference intervals (RIs) are inadequate or even unavailable for many analyses in China. Many of the RIs used in Chinese laboratories were derived from Chinese adult standards or from foreign studies. The aim of this study was to establish specific RIs for alanine aminotransferase (ALT), aspartate aminotransferase (AST), urea (UR), creatinine (Cr) and uric acid (UA) for apparently healthy Chinese adolescents. Overall, 1682 apparently healthy adolescents were enrolled. Serum ALT, AST, UR, Cr and UA were measured by an ARCHITECT C-8000 automated chemistry analyzer. The 2.5th and 97.5th percentile RIs were determined using non-parametric methods. The established reference intervals for ALT, AST, UR, CR and UA were 7.5-42.8 U/L, 12.8-40.2 U/L, 3.12-6.38 mmol/L, 42.7-91.2 μmol/L, and 180.2-409.6 μmol/L in boys and 6.5-32.8 U/L, 10.4-32.5 U/L, 3.05-6.47 mmol/L, 40.2-88.8 μmol/L and 176.5-394.0 μmol/L in girls, respectively. The median and upper and lower limits for the RIs of ALT, AST, Cr and UA were higher in boys than they were in girls (P ALT, AST, UR, Cr and UA that are defined specifically for Chinese adolescents and are appropriate for universal use among Chinese laboratories. Copyright © 2018 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

  15. Association between ALT level and the rate of cardio/cerebrovascular events in HIV-positive individuals

    DEFF Research Database (Denmark)

    Sabin, Caroline A; Ryom, Lene; Kovari, Helen

    2013-01-01

    An inverse association between serum alanine aminotransferase (ALT) levels and the risk of myocardial infarction (MI) has been reported in the general population. We investigated associations between ALT levels and the risk of various cardiovascular and cerebrovascular outcomes in a large cohort...

  16. Extrachromosomal telomere repeat DNA is linked to ALT development via cGAS-STING DNA sensing pathway.

    Science.gov (United States)

    Chen, Yi-An; Shen, Yi-Ling; Hsia, Hsuan-Yu; Tiang, Yee-Peng; Sung, Tzu-Ling; Chen, Liuh-Yow

    2017-12-01

    Extrachromosomal telomere repeat (ECTR) DNA is unique to cancer cells that maintain telomeres through the alternative lengthening of telomeres (ALT) pathway, but the role of ECTRs in ALT development remains elusive. We found that induction of ECTRs in normal human fibroblasts activated the cGAS-STING-TBK1-IRF3 signaling axis to trigger IFNβ production and a type I interferon response, resulting in cell-proliferation defects. In contrast, ALT cancer cells are commonly defective in sensing cytosolic DNA. We found that STING expression was inhibited in ALT cancer cell lines and transformed ALT cells. Notably, the ALT suppressors histone H3.3 and the ATRX-Daxx histone chaperone complex were also required to activate the DNA-sensing pathway. Collectively, our data suggest that the loss of the cGAS-STING pathway may be required to evade ECTR-induced anti-proliferation effects and permit ALT development, and this requirement may be exploited for treatments specific to cancers utilizing the ALT pathway.

  17. Interaction and joint effect of ALT and chronic liver disease on liver cancer in type 2 diabetes patients.

    Science.gov (United States)

    Li, Tsai-Chung; Li, Chia-Ing; Liu, Chiu-Shong; Lin, Pao-Hsuan; Lin, Wen-Yuan; Lin, Chih-Hsueh; Yang, Sing-Yu; Chiang, Jen-Huai; Lin, Cheng-Chieh

    2017-11-28

    This study examined whether serum alanine transaminase (ALT) and chronic liver diseases were interactively, jointly, or independently associated with hepatocellular carcinoma (HCC) risk in type 2 diabetic patients. A retrospective cohort study was conducted in 46,369 Chinese type 2 diabetic patients, aged 30 and older, in National Diabetes Care Management Program in 2002-2004. These data were analyzed by multivariate Cox proportional hazards models. Mean follow-up period was 8.20 years. Multivariate-adjusted hazard ratios of HCC were 2.85 (95% confidence interval, CI: 2.45-3.31), 3.80 (3.04-4.76), and 3.89 (3.08-4.91) for patients with a level of ALT 40-80, 80-120, and >120 U/L, respectively, compared with patients with a level of ALT ALT ≥ 40 U/L and alcoholic liver damage, nonalcoholic fatty liver disease, liver cirrhosis, hepatitis B virus and hepatitis C virus infection, or any one of these chronic liver diseases compared with patients with ALT level ALT level with liver cirrhosis and HBV. Results suggest significant effect modification and joint associations of ALT ≥ 40 U/L and chronic liver diseases. Diabetes care should provide lifestyle or treatment interventions to manage ALT level, liver cirrhosis and hepatitis B virus infection for reducing burden of HCC.

  18. Risk of alanine transferase (ALT) elevation in patients with rheumatoid arthritis treated with methotrexate in a DAS-steered strategy

    NARCIS (Netherlands)

    Dirven, L.; Klarenbeek, N.B.; van den Broek, M.; Groenendael, J.H.L.M.; de Sonnaville, P.B.J.; Kerstens, P.J.S.M.; Huizinga, T.W.J.; Dijkmans, B.A.C.; Lems, W.F.; Allaart, C.F.

    2013-01-01

    Objective: To determine incidence of increased levels of alanine transferase (ALT) >2× upper limit of normal (ULN) in patients receiving methotrexate (MTX), treated according to a dynamic strategy, and to identify predictors of ALT of >2× ULN. Methods: Data of 508 recent-onset rheumatoid arthritis

  19. Low ALT blood levels predict long-term all-cause mortality among adults. A historical prospective cohort study.

    Science.gov (United States)

    Ramaty, E; Maor, E; Peltz-Sinvani, N; Brom, A; Grinfeld, A; Kivity, S; Segev, S; Sidi, Y; Kessler, T; Sela, B A; Segal, G

    2014-12-01

    Increased blood levels of alanine amino transferase (ALT, also known as SGPT; serum glutamic pyruvic transaminase) serve as a marker of liver injury by various mechanisms. Less is known about the clinical implications associated with low-normal ALT levels. Previous studies showed low ALT levels to be associated with poor long-term outcomes among elderlies, serving as a biomarker for increased incidence of frailty and subsequent risk of mortality. However, it has not been determined yet whether low-normal ALT values might be predictive of frailty and mortality in younger, middle-aged adults. We conducted a historical prospective cohort analysis. A total of 23,506 adults with ALT levels within the normal range, at the mean age of 48 ± 11 years, participating in an annual screening program for preventive medicine, were followed-up for a median period of 8.5 years during which 638 died. Low-normal ALT values (serum ALT activity ALT values may serve as an independent predictive marker for increased long-term mortality in middle-aged adults. Copyright © 2014 European Federation of Internal Medicine. Published by Elsevier B.V. All rights reserved.

  20. Mutational analysis of the activator of late transcription, Alt , in the lactococcal bacteriophage TP901-1

    DEFF Research Database (Denmark)

    Pedersen, Margit; Hammer, Karin

    2007-01-01

    An activator protein, Alt, synthesized during the early state of lytic infection is required for transcription of the late operon in the lactococcal phage TP901-1. In order to identify amino acid residues in the Alt protein required for activation of the TP901-1 late promoter, Plate, hydroxylamin...

  1. Life and cancer without telomerase: ALT and other strategies for making sure ends (don't) meet.

    Science.gov (United States)

    Apte, Manasi S; Cooper, Julia Promisel

    2017-02-01

    While most cancer cells rely on telomerase expression/re-activation for linear chromosome maintenance and sustained proliferation, a significant population of cancers (10-15%) employs telomerase-independent strategies, collectively dubbed Alternative Lengthening of Telomeres (ALT). Most ALT cells relax the usual role of telomeres as inhibitors of local homologous recombination while maintaining the ability of telomeres to prohibit local non-homologous end joining reactions. Here we review current concepts surrounding how ALT telomeres achieve this new balance via alterations in chromatin landscape, DNA damage repair processes and handling of telomeric transcription. We also discuss telomerase independent end maintenance strategies utilized by other organisms, including fruitflies and yeasts, to draw parallels and contrasts and highlight additional modes, beyond ALT, that may be available to telomerase-minus cancers. We conclude by commenting on promises and challenges in the development of effective anti-ALT cancer therapies.

  2. Mre11 and Blm-Dependent Formation of ALT-Like Telomeres in Ku-Deficient Ustilago maydis.

    Directory of Open Access Journals (Sweden)

    Eun Young Yu

    2015-10-01

    Full Text Available A subset of human cancer cells uses a specialized, aberrant recombination pathway known as ALT to maintain telomeres, which in these cells are characterized by complex aberrations including length heterogeneity, high levels of unpaired C-strand, and accumulation of extra-chromosomal telomere repeats (ECTR. These phenotypes have not been recapitulated in any standard budding or fission yeast mutant. We found that eliminating Ku70 or Ku80 in the yeast-like fungus Ustilago maydis results initially in all the characteristic telomere aberrations of ALT cancer cells, including C-circles, a highly specific marker of ALT. Subsequently the ku mutants experience permanent G2 cell cycle arrest, accompanied by loss of telomere repeats from chromosome ends and even more drastic accumulation of very short ECTRs (vsECTRs. The deletion of atr1 or chk1 rescued the lethality of the ku mutant, and "trapped" the telomere aberrations in the early ALT-like stage. Telomere abnormalities are telomerase-independent, but dramatically suppressed by deletion of mre11 or blm, suggesting major roles for these factors in the induction of the ALT pathway. In contrast, removal of other DNA damage response and repair factors such as Rad51 has disparate effects on the ALT phenotypes, suggesting that these factors process ALT intermediates or products. Notably, the antagonism of Ku and Mre11 in the induction of ALT is reminiscent of their roles in DSB resection, in which Blm is also known to play a key role. We suggest that an aberrant resection reaction may constitute an early trigger for ALT telomeres, and that the outcomes of ALT are distinct from DSB because of the unique telomere nucleoprotein structure.

  3. Mre11 and Blm-Dependent Formation of ALT-Like Telomeres in Ku-Deficient Ustilago maydis.

    Science.gov (United States)

    Yu, Eun Young; Pérez-Martín, José; Holloman, William K; Lue, Neal F

    2015-10-01

    A subset of human cancer cells uses a specialized, aberrant recombination pathway known as ALT to maintain telomeres, which in these cells are characterized by complex aberrations including length heterogeneity, high levels of unpaired C-strand, and accumulation of extra-chromosomal telomere repeats (ECTR). These phenotypes have not been recapitulated in any standard budding or fission yeast mutant. We found that eliminating Ku70 or Ku80 in the yeast-like fungus Ustilago maydis results initially in all the characteristic telomere aberrations of ALT cancer cells, including C-circles, a highly specific marker of ALT. Subsequently the ku mutants experience permanent G2 cell cycle arrest, accompanied by loss of telomere repeats from chromosome ends and even more drastic accumulation of very short ECTRs (vsECTRs). The deletion of atr1 or chk1 rescued the lethality of the ku mutant, and "trapped" the telomere aberrations in the early ALT-like stage. Telomere abnormalities are telomerase-independent, but dramatically suppressed by deletion of mre11 or blm, suggesting major roles for these factors in the induction of the ALT pathway. In contrast, removal of other DNA damage response and repair factors such as Rad51 has disparate effects on the ALT phenotypes, suggesting that these factors process ALT intermediates or products. Notably, the antagonism of Ku and Mre11 in the induction of ALT is reminiscent of their roles in DSB resection, in which Blm is also known to play a key role. We suggest that an aberrant resection reaction may constitute an early trigger for ALT telomeres, and that the outcomes of ALT are distinct from DSB because of the unique telomere nucleoprotein structure.

  4. Patrika - alt

    Indian Academy of Sciences (India)

    brilliant printers

    1215–1315 Lecture Presentations. Murali Sastry: New methods for the synthesis and assembly of nanomaterials ..... cellular respiration; cytochrome c oxidase from the soil bacterium Paracoccus denitrificans, the yeast cytochrome ...... antibiotics were worked out. Rao was thus a unique combination of a forest chemist and a ...

  5. Patrika - alt

    Indian Academy of Sciences (India)

    brilliant printers

    end he also linked up to recent work on nanostructured materials approaching the ..... designs. Today typically 50–70% of the design cycle time is spent in verifying correctness. While simulation remains a predominant form of validation, exhaustive simulation .... application of genomic information in basic biology, diagnosis.

  6. Biomaterial properties evaluation of poly(vinyl acetate- alt-maleic anhydride)/chitosan nanocapsules

    Science.gov (United States)

    Raţă, Delia Mihaela; Popa, Marcel; Chailan, Jean-François; Zamfir, Carmen Lăcrămioara; Peptu, Cătălina Anişoara

    2014-08-01

    Nanocapsules with diameter around 100 nm based on a natural polymer (chitosan) and a synthetic polymer poly(vinyl acetate- alt-maleic anhydride) [poly(MAVA)] by interfacial condensation method were prepared. The present study proposes a new type of biocompatible nanocapsules based on poly(vinyl acetate- alt-maleic anhydride-chitosan) (MCS) able to become a reliable support for inclusion and release of drugs. The spherical shape of the nanocapsules was evidenced by scanning electron microscopy. Nanocapsules presented a good Norfloxacin loading and release capacity. Haemocompatibility tests have demonstrated that the nanocapsules present a low toxicity and a good compatibility with sanguine medium. The biocompatibility properties of the nanocapsules after their intraperitoneal administration in rats were evidenced by histopathological examination of different organs (brain, liver, kidney, and lung). The results are encouraging and the nanocapsules can be used as controlled drug delivery systems.

  7. The telomere-associated homeobox-containing protein TAH1/HMBOX1 participates in telomere maintenance in ALT cells.

    Science.gov (United States)

    Feng, Xuyang; Luo, Zhenhua; Jiang, Shuai; Li, Feng; Han, Xin; Hu, Yang; Wang, Dan; Zhao, Yong; Ma, Wenbin; Liu, Dan; Huang, Junjiu; Songyang, Zhou

    2013-09-01

    The majority of cancer cells rely on elevated telomerase expression and activity for rapid growth and proliferation. Telomerase-negative cancer cells, by contrast, often employ the alternative lengthening of telomeres (ALT) pathway to maintain telomeres. ALT cells are characterized by long and dynamic telomeres and the presence of ALT-associated promyelocytic leukemia (PML) bodies (APBs). Previous work has shown the importance of APBs to the ALT pathway, but their formation and precise role remain unclear. Here, we demonstrate that a homeobox-containing protein known as HMBOX1 can directly bind telomeric double-stranded DNA and associate with PML nuclear bodies. Hence, we renamed this protein TAH1 for telomere-associated homeobox-containing protein 1. TAH1 knockdown significantly reduced the number of APBs and led to an increase in DNA damage response signals at telomeres. Importantly, TAH1 inhibition also notably reduced the presence of telomere C-circles, indicating altered ALT activity. Our findings point to TAH1 as a novel link between pathways that regulate DNA damage responses, PML nuclear bodies, and telomere homeostasis in ALT cells, and provide insight into how ALT cells may achieve sustained growth and proliferation independent of the telomerase.

  8. Spermine-alt-poly(ethylene glycol) polyspermine as a safe and efficient aerosol gene carrier for lung cancer therapy.

    Science.gov (United States)

    Kim, You-Kyoung; Cho, Chong-Su; Cho, Myung-Haing; Jiang, Hu-Lin

    2014-07-01

    The clinical success of gene therapy critically depends upon the safety and efficiency of delivery system used. Although polyethylenimine (PEI) has been commonly used as an efficient cationic polymeric gene carrier due to its high transfection efficiency, its cytotoxicity and nondegradability limit the polymer's therapeutic applications in clinical trials. In this study, biocompatible polyspermine based on spermine (SPE) and poly(ethylene glycol) (PEG) diacrylate (SPE-alt-PEG) was synthesized using a Michael-type addition reaction, and its ability as an alternative gene carrier for lung cancer therapy was evaluated. SPE-alt-PEG polyspermine was complexed with plasmid DNA, and the resulting complexes were characterized by particle size and surface charge by dynamic light scattering, complex formation and DNA protection ability by gel retardation, and complex shape by energy-filtering transmission electron microscopy. The SPE-alt-PEG copolymer showed low cytotoxicity, and SPE-alt-PEG/DNA complexes showed efficacious transfection efficiency compared with 25 kDa PEI (PEI 25K). Also SPE-alt-PEG/GFP complexes were efficiently transferred into the lungs after aerosol administration without toxicity, and delivery of Pdcd4 gene as a therapeutic gene with SPE-alt-PEG polyspermine greatly reduced tumor size as well as tumor numbers in K-ras(LA1) lung cancer model mice compared relative to the effect observed for PEI 25K. These results suggest that SPE-alt-PEG has potential as a gene carrier for lung cancer gene therapy. © 2013 Wiley Periodicals, Inc.

  9. Low ALT Levels Independently Associated with 22-Year All-Cause Mortality Among Coronary Heart Disease Patients.

    Science.gov (United States)

    Peltz-Sinvani, N; Klempfner, R; Ramaty, E; Sela, B A; Goldenberg, I; Segal, G

    2016-02-01

    Low alanine aminotransferase (ALT) blood levels are known to be associated with frailty and increased risk of long-term mortality in certain populations. However, the contribution of this marker to long-term outcome has not been assessed in patients with chronic coronary heart disease. The aim of the current study was to assess the association between low ALT values and long-term, 22.8-year, all-cause mortality in this population. We examined the association of low ALT (ALT group compared with patients with higher ALT levels (65.6 % vs. 58.4 %; log-rank p ALT is independently associated with 11 % greater long-term (22.8 years) mortality risk [HR 1.11 (95 % confidence interval: 1.03-1.19; adjusted p ALT levels are associated with increased long-term mortality among middle-aged patients with stable coronary heart disease. This association remained statistically significant after adjustment for other well-established risk factors for mortality in this population.

  10. RNaseH1 regulates TERRA-telomeric DNA hybrids and telomere maintenance in ALT tumour cells.

    Science.gov (United States)

    Arora, Rajika; Lee, Yongwoo; Wischnewski, Harry; Brun, Catherine M; Schwarz, Tobias; Azzalin, Claus M

    2014-10-21

    A fraction of cancer cells maintain telomeres through the telomerase-independent, 'Alternative Lengthening of Telomeres' (ALT) pathway. ALT relies on homologous recombination (HR) between telomeric sequences; yet, what makes ALT telomeres recombinogenic remains unclear. Here we show that the RNA endonuclease RNaseH1 regulates the levels of RNA-DNA hybrids between telomeric DNA and the long noncoding RNA TERRA, and is a key mediator of telomere maintenance in ALT cells. RNaseH1 associated to telomeres specifically in ALT cells and its depletion led to telomeric hybrid accumulation, exposure of single-stranded telomeric DNA, activation of replication protein A at telomeres and abrupt telomere excision. Conversely, overexpression of RNaseH1 weakened the recombinogenic nature of ALT telomeres and led to telomere shortening. Altering cellular RNaseH1 levels did not perturb telomere homoeostasis in telomerase-positive cells. RNaseH1 maintains regulated levels of telomeric RNA-DNA hybrids at ALT telomeres to trigger HR without compromising telomere integrity too severely.

  11. Factors underlying the association of body mass index with serum ALT in Chinese hypertensive adults without known hepatic diseases.

    Science.gov (United States)

    Zhang, Yan; Qin, Xian-hui; Li, Jian-ping; Cui, Yi-min; Liu, Ze-yuan; Zhao, Zhi-gang; Ge, Jun-bo; Guan, De-ming; Hu, Jian; Wang, Yan-ni; Zhang, Fu-min; Xu, Xin; Xu, Xi-ping; Huo, Yong

    2013-08-01

    High body mass index (BMI) is considered as the most important risk factor for elevated serum alanine aminotransferase (ALT) concentration. This study examined an array of factors, including waist circumference (WC) and folate deficiency, which may mediate the association of BMI with serum ALT concentration in Chinese hypertensive adults without known hepatic diseases. A multicenter, cross-sectional study was carried out. A total of 378 patients with mild or moderate hypertension and without known hepatic diseases were recruited from five hospitals in Harbin, Shanghai, Beijing, Xi'an, and Nanjing. Of the 360 hypertensive patients with complete data in our final analysis, 13.6% had high ALT concentrations (>40 IU/L). Factors including BMI, WC, triglyceride level, and folate concentration were associated with ALT concentration in univariate analysis. Consistently higher prevalence rates of elevated ALT were observed in subjects with lower folate concentrations (≥12 vs. ALT concentration disappeared (P=0.802 in males and 0.369 in females), while WC in females (PALT concentration. This multicenter study demonstrated that WC and low folate concentration were important factors underlying the association between BMI and ALT concentrations in Chinese hypertensive adults without known hepatic diseases.

  12. RNaseH1 regulates TERRA-telomeric DNA hybrids and telomere maintenance in ALT tumour cells

    Science.gov (United States)

    Arora, Rajika; Lee, Yongwoo; Wischnewski, Harry; Brun, Catherine M.; Schwarz, Tobias; Azzalin, Claus M.

    2014-01-01

    A fraction of cancer cells maintain telomeres through the telomerase-independent, ‘Alternative Lengthening of Telomeres’ (ALT) pathway. ALT relies on homologous recombination (HR) between telomeric sequences; yet, what makes ALT telomeres recombinogenic remains unclear. Here we show that the RNA endonuclease RNaseH1 regulates the levels of RNA–DNA hybrids between telomeric DNA and the long noncoding RNA TERRA, and is a key mediator of telomere maintenance in ALT cells. RNaseH1 associated to telomeres specifically in ALT cells and its depletion led to telomeric hybrid accumulation, exposure of single-stranded telomeric DNA, activation of replication protein A at telomeres and abrupt telomere excision. Conversely, overexpression of RNaseH1 weakened the recombinogenic nature of ALT telomeres and led to telomere shortening. Altering cellular RNaseH1 levels did not perturb telomere homoeostasis in telomerase-positive cells. RNaseH1 maintains regulated levels of telomeric RNA–DNA hybrids at ALT telomeres to trigger HR without compromising telomere integrity too severely. PMID:25330849

  13. Two members of the first crew of the Shuttle Approach and Landing Tests (ALT)

    Science.gov (United States)

    1976-01-01

    The two members of the first crew for the Space Shuttle Approach and Landing Tests (ALT) are photographed at the Rockwell International Space Division's Orbiter assembly facility at Palmdale, California on the day of the rollout of the Shuttle Orbiter 101 'Enterprise' spacecraft. They are Astronauts Fred W. Haise Jr. (left), commander; and C. Gordon Fullerton, pilot. The DC-9 size airplane-like Orbiter 101 is in the background.

  14. Liver stiffness becomes stable in patients with chronic hepatitis C three months after ALT normalization due to antiviral therapy

    Directory of Open Access Journals (Sweden)

    CHEN Feikai

    2013-10-01

    Full Text Available ObjectiveTo investigate the time for liver stiffness measurement (LSM to become stable in chronic hepatitis C (CHC patients with elevated alanine aminotransferase (ALT levels after ALT normalization due to antiviral therapy. MethodsCHC patients who sought initial treatment at Peking University People′s Hospital were screened for elevated ALT levels from May 2011. Liver stiffness was determined by FibroScan. A total of 29 patients had been included in the study by September 2012, who were followed up regularly after antiviral treatment. ALT tests were repeated every four weeks and LSM every eight weeks until their medians did not change significantly. Comparisons of matched data at two adjacent time points were made with the non-parametric Wilcoxon test, while multiple comparisons of repeated measurements were performed using Bonferroni correction. Correlation between two variables was analyzed with the Spearman rank test. ResultsPatients were followed up until 24 weeks after antiviral treatment, and 24 patients were included in analysis. The median ALT levels were 64, 26, 21, 20, and 22 U/L at baseline and 4, 8, 12, and 24 weeks, respectively (P= 0.000, 0.006, 0.337, and 0.109 for comparisons between two adjacent values. ALT decreased significantly below 1 ULN at 4 weeks after antiviral therapy and stabilized at 8 weeks. The median LSM values were 8.7, 7.8, 6.8, and 6.7 kPa at baseline and 8, 16, and 24 weeks, respectively (P= 0.009, 0.001, and 0188 for comparisons between two adjacent values. LSM decreased significantly within 16 weeks after antiviral therapy and stabilized afterwards. LSM stabilized 12 weeks after ALT normalization. ConclusionLSM becomes stable in CHC patients with elevated ALT levels three months after ALT normalization due to antiviral therapy.

  15. AST/ALT ratio > or = 1 is not diagnostic of cirrhosis in patients with chronic hepatitis C.

    Science.gov (United States)

    Reedy, D W; Loo, A T; Levine, R A

    1998-09-01

    Medical guidelines for interferon-alpha2a or -alpha2b (IFN-alpha) treatment of chronic hepatitis C virus (HCV) infection depend upon baseline liver histology. A better long-term response to IFN-alpha therapy correlates with less inflammation and absence of cirrhosis. It has been suggested that the presence of cirrhosis in patients with chronic hepatitis C virus infection may be predicted based on an AST/ALT ratio > or = 1. This study was designed to determine if the presence of cirrhosis can be predicted in patients with chronic HCV infection by such a ratio. Seventy-seven patients, including 23 cirrhotics, with chronic HCV infection were studied. Serum ALT, AST, and HCV-RNA levels and hepatic activity index (HAI), reflecting histologic inflammation in all liver biopsies, were assessed. AST/ALT ratios and mean ALT, AST, and HCV-RNA were determined for both cirrhotic and noncirrhotic patients. HAI was correlated with ALT, AST, and HCV-RNA levels, the latter determined by quantitative RT-PCR. The likelihood ratio (LR) and positive predictive value of an AST/ALT ratio > or = 1 for cirrhosis was 7.3 and only 77%, respectively. In cirrhotics vs noncirrhotics, there were no significant differences between mean serum ALT (149 +/- 28 vs 176 +/- 17 units/liter), AST (139 +/- 28 vs 102 +/- 8 units/liter), or HCV-RNA levels (589,160 +/- 147,053 vs 543,915 +/- 75,497 copies/ml), respectively. There was a significant, but clinically weak, correlation between serum ALT and HAI (r = 0.234), and none between HAI and either serum AST or HCV-RNA levels. Our results support the need for a liver biopsy prior to treatment of chronic HCV infection, since the AST/ALT ratio fails to predict accurately the presence of cirrhosis.

  16. Clinical presentation and disease phases of chronic hepatitis B using conventional versus modified ALT criteria in Asian Americans.

    Science.gov (United States)

    Hsu, Yu-Nan; Pan, Calvin Q; Abbasi, Ali; Xia, Victor; Bansal, Raghav; Hu, Ke-Qin

    2014-04-01

    The modified normal alanine aminotransferase (ALT) value (i.e., males ALT criteria could be better associated with disease phases in a cohort of Asian Americans (AsAm) with CHB. This two-center retrospective study evaluated 198 non-treated AsAm with CHB and a mean follow-up of 21 months. Both conventional and modified ALT criteria were used to determine the differences and clinical value using modified ALT criteria in classifying CHB phases. Among HBeAg (+) patients (29.3 %), HBV pre-core (PC) and basal core promoter (BCP) mutations were detected in 24.4 % and 31.3 %, respectively. Using baseline conventional ALT criteria, 97/153 (63.4 %) patients could be categorized into CHB phases 1 to 4, whereas 56/153 (36.6 %) were indeterminate. Using the modified ALT criteria, 43 (28.1 %) patients had phase changes of which 31/43 (72.1 %) were moved from phase 1 and indeterminate to phases 2 and 4, more active CHB phases. In 13/31 of these patients with liver biopsy, 6 (19.4 %) reported stage 2-4 fibrosis and 10 (32.3 %) reported grade 1-2 inflammation. Using modified ALT criteria to evaluate 48/153 patients with full data at baseline and the end of 1-year follow-up, we observed that 19/48 (39.6 %) changed their CHB phases; 5/48 (10.4 %) moved from phases 1 and 3 to phases 2 and 4; 2/48 (4.2 %) remained in the active phases; 10/48 (20.8 %) became indeterminate. HBV PC and BCP mutations were detectable in 24.4 % and 31.3 % of HBeAg (+) AsAm patients, respectively. Compared with conventional ALT criteria, modified ALT criteria is more sensitive in identifying CHB patients in active phases.

  17. Comparison of the super agonist complex, ALT-803, to IL-15 as cancer immunotherapeutics in animal models

    Science.gov (United States)

    Xu, Wenxin; Hong, Hao; Webb, Gabriela M.; Chen, Xiaoyue; Liu, Bai; Zhu, Xiaoyun; Wen, Jinghai; You, Lijing; Kong, Lin; Edwards, Ana C.; Han, Kaiping; Shi, Sixiang; Alter, Sarah; Sacha, Jonah B.; Jeng, Emily K.; Cai, Weibo; Wong, Hing C.

    2015-01-01

    Interleukin-15 (IL-15), a potent stimulant of CD8+ T and NK cells, is a promising cancer immunotherapeutic. ALT-803 is a complex of an IL-15 superagonist mutant and a dimeric IL-15 receptor αSu/Fc fusion protein that was found to exhibit enhanced biologic activity in vivo with a substantially longer serum half-life than recombinant IL-15. A single intravenous dose of ALT-803, but not IL-15, eliminated well-established tumors and prolonged survival of mice bearing multiple myeloma. In this study, we extended these findings to demonstrate the superior antitumor activity of ALT-803 over IL-15 in mice bearing subcutaneous B16F10 melanoma tumors and CT26 colon carcinoma metastases. Tissue biodistribution studies in mice also showed much greater retention of ALT-803 in the lymphoid organs compared to IL-15, consistent with its highly potent immunostimulatory and antitumor activities in vivo. Weekly dosing with 1 mg/kg ALT-803 in C57BL/6 mice was well-tolerated, yet capable of increasing peripheral blood lymphocyte, neutrophil and monocyte counts by >8-fold. ALT-803 dose-dependent stimulation of immune cell infiltration into the lymphoid organs was also observed. Similarly, cynomolgus monkeys treated weekly with ALT-803 showed dose-dependent increases of peripheral blood lymphocyte counts, including NK, CD4+, and CD8+ memory T cell subsets. In vitro studies demonstrated ALT-803-mediated stimulation of mouse and human immune cell proliferation and IFN-γ production without inducing a broad-based release of other proinflammatory cytokines (i.e., cytokine storm). Based on these results, a weekly dosing regimen of ALT-803 has been implemented in multiple clinical studies to evaluate the dose required for effective immune cell stimulation in humans. PMID:26511282

  18. Twist-3 fragmentation effects for ALT in light hadron production from proton-proton collisions

    Science.gov (United States)

    Koike, Y.; Pitonyak, D.; Takagi, Y.; Yoshida, S.

    2016-01-01

    We compute the contribution from the twist-3 fragmentation function for light hadron production in collisions between transversely and longitudinally polarized protons, i.e., p↑ p → → h X, which can cause a double-spin asymmetry (DSA) ALT. This is a naïve T-even twist-3 observable that we analyze in collinear factorization using both Feynman gauge and lightcone gauge as well as give a general proof of color gauge invariance. So far only twist-3 effects in the transversely polarized proton have been studied for ALT in p↑ p → → h X. However, there are indications that the naïve T-odd transverse single-spin asymmetry (SSA) AN in p↑ p → h X is dominated not by such distribution effects but rather by a fragmentation mechanism. Therefore, one may expect similarly that the fragmentation contribution is important for ALT. Given possible plans at RHIC to measure this observable, it is timely to provide a calculation of this term.

  19. Elevation of ALT to AST ratio in patients with enteroviral myocarditis.

    Science.gov (United States)

    Kanda, T; Kobayashi, I; Suzuki, T; Murata, K; Radio, S J; McManus, B M

    1995-01-01

    Enteroviral myocarditis is often a relatively benign condition in adults. Physicians, therefore, may not always record detailed clinical and laboratory data in such patients. As such, they may not recognize viral involvement in organs beyond the heart. The purpose of this study was to examine the hepatic involvement of enteroviral peri-myocarditis and to compare the other diseases with congestive heart failure. We analyzed 18 patients (ages 15-64) who were diagnosed as having enteroviral myocarditis (n = 16) or pericarditis (n = 2). Serology was positive for coxsackie viruses in 11 patients and echoviruses in six patients. A diagnosis of hepatic involvement was made by the following laboratory data: rising levels of alanine amino transferase (ALT), aspartate amino transferase (AST) and exceeded serum ALT compared with AST levels. A ratio of ALT/AST more than 1.0 was greatly frequent in patients with peri-myocarditis (72%; 13/18) compared with acute myocardial infarction (0%; 0/10) and idiopathic dilated cardiomyopathy (3%; 3/10). In summary, hepatic involvement in the setting of acute enteroviral peri-myocarditis may be considerably more common in adults than previously suspected. The recognition of hepatic involvement in association with enteroviral peri-myocarditis may allow improvement of diagnostic sensitivity and alter approaches to treatments of acute viral myocarditis.

  20. Detection of TTV in peripheral blood cells from patients with altered ALT and AST levels.

    Science.gov (United States)

    de Oliveira, Jaqueline Carvalho; Nasser, Thiago Franco; Oda, Julie Massayo Maeda; Aoki, Mateus Nóbrega; Carneiro, Juliana Laino do Val; Barbosa, Décio Sabbatini; Reiche, Edna Maria Vissoci; Watanabe, Maria Angelica Ehara

    2008-04-01

    This work analyzes the prevalence of TTV DNA in peripheral blood cells from patients with hepatic alterations and healthy blood donors and measures levels of sodium, potassium, urea, creatinine, phosphatase alkaline, total and direct bilirubin, gamma glutamyl transferase (GGT), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in certain randomly selected patients. DNA samples from 111 individuals were evaluated. They were divided into two groups, "A" (study) and "B" (control), including 54 patients with liver enzyme alterations (ALT/AST) presenting non-B-non-C hepatitis and 57 blood donors, respectively. TTV DNA was determined by nested PCR. Certain products of the second-round PCR were sequenced. Serum biochemical assay was performed and disclosed TTV in 31.48% (17/54) of patients in group A and 5.26% (3/57) in the control group B. TTV prevalence was significantly higher in patients with liver disease than in healthy donors. In group A, sodium, potassium, urea, creatinine, phosphatase alkaline, total and direct bilirubin, gamma glutamyl transferase (GGT), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were analyzed in certain randomly selected patients and no significant difference in biochemical levels (p>0.05) was found when TTV infected and noninfected individuals were compared. Knowledge related to TTV has rapidly increased, but many fundamental aspects remain unclear. This led us to question the role of TTV and doubt remains as to whether or not it is just a commensal virus. Further studies are necessary to confirm and extend these findings.

  1. Establishment and characterization of a spontaneously immortalized trophoblast cell line (HPT-8) and its hepatitis B virus-expressing clone.

    Science.gov (United States)

    Zhang, Lei; Zhang, Weilu; Shao, Chen; Zhang, Jingxia; Men, Ke; Shao, Zhongjun; Yan, Yongping; Xu, Dezhong

    2011-08-01

    Most trophoblast cell lines currently available to study vertical transmission of hepatitis B virus (HBV) are immortalized by viral transformation. Our goal was to establish and characterize a spontaneously immortalized human first-trimester trophoblast cell line and its HBV-expressing clone. Chorionic villi of Asian human first-trimester placentae were digested with trypsin and collagenase I to obtain the primary trophoblast cell culture. A spontaneously immortalized trophoblast cell line (HPT-8) was analyzed by scanning and transmission electron microscopy, cell cycle analysis, immunohistochemistry and immunofluorescence. HPT-8 cells were stably transfected with the adr subtype of HBV (HPT-8-HBV) and characterized by PCR and enzyme-linked immunosorbent assay. We obtained a clonal derivative of a spontaneously immortalized primary cell clone (HPT-8). HPT-8 cells were epithelioid and polygonal, and formed multinucleate, giant cells. They exhibited microvilli, distinct desmosomes between adjacent cells, abundant endoplasm, lipid inclusions and glycogen granules, which are all characteristic of cytotrophoblasts. HPT-8 cells expressed cytokeratin 7, cytokeratin 18, vimentin, cluster of differentiation antigen 9, epidermal growth factor receptor, stromal cell-derived factor 1 and placental alkaline phosphatase. They secreted prolactin, estradiol, progesterone and hCG, and were positive for HLA-G, a marker of extravillous trophoblasts. HPT-8-HBV cells were positive for HBV relaxed-circular, covalently closed circular DNA and pre-S sequence. HPT-8-HBV cells also produced and secreted HBV surface antigen and HBV e antigen. We established a trophoblast cell line, HPT-8 and its HBV-expressing clone which could be valuable in exploring the mechanism of HBV viral integration in human trophoblasts during intrauterine infection.

  2. Immortalization of Fetal Bovine Colon Epithelial Cells by Expression of Human Cyclin D1, Mutant Cyclin Dependent Kinase 4, and Telomerase Reverse Transcriptase: An In Vitro Model for Bacterial Infection.

    Directory of Open Access Journals (Sweden)

    Kengo Kuroda

    Full Text Available Cattle are the economically important animals in human society. They are essential for the production of livestock products such as milk and meats. The production efficiency of livestock products is negatively impacted by infection with zoonotic pathogens. To prevent and control infectious diseases, it is important to understand the interaction between cattle tissue and pathogenic bacteria. In this study, we established an in vitro infection model of an immortalized bovine colon-derived epithelial cell line by transducing the cells with lentiviral vectors containing genes encoding cell cycle regulators cyclin D1, mutant cyclin dependent kinase 4 (CDK4, and human telomerase reverse transcriptase (TERT. The established cell line showed continuous cell proliferation, expression of epithelial markers, and an intact karyotype, indicating that the cells maintained their original nature as colon-derived epithelium. Furthermore, we exposed the established cell line to two strains of Salmonella enterica and EHEC. Interestingly, S. Typhimurium showed higher affinity for the established cell line and invaded the cytoplasm than S. Enteritidis. Quantitative RT-PCR revealed that gene expression of Toll-like receptor 1 (TLR1, TLR 2 and TLR 3, whereas TLR 4, 5 and 6 were not detectable in established cells. Our established immortalized colon-derived epithelial cell should be a useful tool for studies evaluating the molecular mechanisms underlying bacterial infection.

  3. Immortalization of Fetal Bovine Colon Epithelial Cells by Expression of Human Cyclin D1, Mutant Cyclin Dependent Kinase 4, and Telomerase Reverse Transcriptase: An In Vitro Model for Bacterial Infection.

    Science.gov (United States)

    Kuroda, Kengo; Kiyono, Tohru; Isogai, Emiko; Masuda, Mizuki; Narita, Moe; Okuno, Katsuya; Koyanagi, Yukako; Fukuda, Tomokazu

    2015-01-01

    Cattle are the economically important animals in human society. They are essential for the production of livestock products such as milk and meats. The production efficiency of livestock products is negatively impacted by infection with zoonotic pathogens. To prevent and control infectious diseases, it is important to understand the interaction between cattle tissue and pathogenic bacteria. In this study, we established an in vitro infection model of an immortalized bovine colon-derived epithelial cell line by transducing the cells with lentiviral vectors containing genes encoding cell cycle regulators cyclin D1, mutant cyclin dependent kinase 4 (CDK4), and human telomerase reverse transcriptase (TERT). The established cell line showed continuous cell proliferation, expression of epithelial markers, and an intact karyotype, indicating that the cells maintained their original nature as colon-derived epithelium. Furthermore, we exposed the established cell line to two strains of Salmonella enterica and EHEC. Interestingly, S. Typhimurium showed higher affinity for the established cell line and invaded the cytoplasm than S. Enteritidis. Quantitative RT-PCR revealed that gene expression of Toll-like receptor 1 (TLR1), TLR 2 and TLR 3, whereas TLR 4, 5 and 6 were not detectable in established cells. Our established immortalized colon-derived epithelial cell should be a useful tool for studies evaluating the molecular mechanisms underlying bacterial infection.

  4. Selling space colonization and immortality: A psychosocial, anthropological critique of the rush to colonize Mars

    Science.gov (United States)

    Slobodian, Rayna Elizabeth

    2015-08-01

    Extensive media coverage regarding the proposal to send four people to Mars by 2025 has exploded recently. Private enterprise has taken the reins to venture into space, which has typically only been reserved for government agencies. I argue, that with this new direction comes less regulation, raising questions regarding the ethics of sending people into outer space to colonize Mars within a decade. Marketers selling colonization to the public include perspectives such as biological drives, species survival, inclusiveness and utopian ideals. I challenge these narratives by suggesting that much of our desire to colonize space within the next decade is motivated by ego, money and romanticism. More specifically, I will examine the roles that fear and stories of immortality play within selling space and how those stories are marketed. I am passionate about space and hope that one day humanity will colonize other worlds, but the rush to settle is dangerous and careless. I assert that humanity should first gain more experience and knowledge before colonizing outer space, using this research to mitigate the risk to astronauts and proceed with careful consideration for the lives of potential astronauts.

  5. Mapping of imprinted quantitative trait loci using immortalized F2 populations.

    Directory of Open Access Journals (Sweden)

    Yongxian Wen

    Full Text Available Mapping of imprinted quantitative trait loci (iQTLs is helpful for understanding the effects of genomic imprinting on complex traits in animals and plants. At present, the experimental designs and corresponding statistical methods having been proposed for iQTL mapping are all based on temporary populations including F2 and BC1, which can be used only once and suffer some other shortcomings respectively. In this paper, we propose a framework for iQTL mapping, including methods of interval mapping (IM and composite interval mapping (CIM based on conventional low-density genetic maps and point mapping (PM and composite point mapping (CPM based on ultrahigh-density genetic maps, using an immortalized F2 (imF2 population generated by random crosses between recombinant inbred lines or doubled haploid lines. We demonstrate by simulations that imF2 populations are very desirable and the proposed statistical methods (especially CIM and CPM are very powerful for iQTL mapping, with which the imprinting effects as well as the additive and dominance effects of iQTLs can be unbiasedly estimated.

  6. Myelinating cocultures of rodent stem cell line-derived neurons and immortalized Schwann cells.

    Science.gov (United States)

    Ishii, Tomohiro; Kawakami, Emiko; Endo, Kentaro; Misawa, Hidemi; Watabe, Kazuhiko

    2017-10-01

    Myelination is one of the most remarkable biological events in the neuron-glia interactions for the development of the mammalian nervous system. To elucidate molecular mechanisms of cell-to-cell interactions in myelin synthesis in vitro, establishment of the myelinating system in cocultures of continuous neuronal and glial cell lines are desirable. In the present study, we performed co-culture experiments using rat neural stem cell-derived neurons or mouse embryonic stem (ES) cell-derived motoneurons with immortalized rat IFRS1 Schwann cells to establish myelinating cultures between these cell lines. Differentiated neurons derived from an adult rat neural stem cell line 1464R or motoneurons derived from a mouse ES cell line NCH4.3, were mixed with IFRS1 Schwann cells, plated, and maintained in serum-free F12 medium with B27 supplement, ascorbic acid, and glial cell line-derived neurotrophic factor. Myelin formation was demonstrated by electron microscopy at 4 weeks in cocultures of 1464R-derived neurons or NCH4.3-derived motoneurons with IFRS1 Schwann cells. These in vitro coculture systems utilizing the rodent stable stem and Schwann cell lines can be useful in studies of peripheral nerve development and regeneration. © 2017 Japanese Society of Neuropathology.

  7. Traceless Targeting and Isolation of Gene-Edited Immortalized Keratinocytes from Epidermolysis Bullosa Simplex Patients

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    Magomet Aushev

    2017-09-01

    Full Text Available Epidermolysis bullosa simplex (EBS is a blistering skin disease caused by dominant-negative mutations in either KRT5 or KRT14, resulting in impairment of keratin filament structure and epidermal fragility. Currently, nearly 200 mutations distributed across the entire length of these genes are known to cause EBS. Genome editing using programmable nucleases enables the development of ex vivo gene therapies for dominant-negative genetic diseases. A clinically feasible strategy involves the disruption of the mutant allele while leaving the wild-type allele unaffected. Our aim was to develop a traceless approach to efficiently disrupt KRT5 alleles using TALENs displaying unbiased monoallelic disruption events and devise a strategy that allows for subsequent screening and isolation of correctly modified keratinocyte clones without the need for selection markers. Here we report on TALENs that efficiently disrupt the KRT5 locus in immortalized patient-derived EBS keratinocytes. Inactivation of the mutant allele using a TALEN working at sub-optimal levels resulted in restoration of intermediate filament architecture. This approach can be used for the functional inactivation of any mutant keratin allele regardless of the position of the mutation within the gene and is furthermore applicable to the treatment of other inherited skin disorders.

  8. Dendritic cells derived from HOXB4-immortalized hematopoietic bone marrow cells.

    Science.gov (United States)

    Baru, Abdul Mannan; Krishnaswamy, Jayendra Kumar; Rathinasamy, Anchana; Scherr, Michaela; Eder, Matthias; Behrens, Georg M N

    2011-11-01

    Dendritic cells (DCs) are essential for the generation and modulation of cell-mediated adaptive immunity against infections. DC-based vaccination involves transplantation of ex vivo-generated DCs loaded with antigen in vitro, but remains limited by the number of autologous or allogeneic cells. While in vitro expansion and differentiation of hematopoietic stem cells (HSCs) into DCs seems to be the most viable alternative to overcome this problem, the complexity of HSC expansion in vitro has posed significant limitations for clinical application. We immortalized lineage-depleted murine hematopoietic bone marrow (lin(-)BM) cells with HOXB4, and differentiated them into CD11c(+)MHCII(+) DCs. These cells showed the typical DC phenotype and upregulated surface expression of co-stimulatory molecules on stimulation with various toll-like receptor ligands. These DCs efficiently presented exogenous antigen to T-cells via major histocompatibility complex (MHC) I and II and viral antigen on infection. Finally, they showed migratory capacity and were able to generate antigen-specific primed T-cells in vivo. In summary, we provide evidence that HOXB4-transduced lin(-)BM cells can serve as a viable means of generating fully functional DCs for scientific and therapeutic applications.

  9. On the acoustic wave sensor response to immortalized hypothalamic neurons at the device-liquid interface

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    Shilin Cheung

    2016-12-01

    Full Text Available The response of a thickness shear mode biosensor to immortalized murine hypothalamic neurons (mHypoE-38 and -46 cells under a variety of conditions and stimuli is discussed. Cellular studies which lead to the production of detectable neuronal responses include neuronal deposition, adhesion and proliferation, alteration in the extent of specific cell-surface interactions, actin filament and microtubule cytoskeletal disruptions, effects of cell depolarization, inhibition of the Na+-K+ pump via ouabain, effects of neuronal synchronization and the effects ligand-receptor interaction (glucagon. In the presence of cells, fs shifts are largely influenced by the damping of the TSM resonator. The formation of cell-surface interactions and hence the increase in coupling and acoustic energy dissipation can be modeled as an additional resistor in the BVD model. Further sensor and cellular changes can be obtained by negating the effects of damping from fs via the use of Rm and θmax. Keywords: Acoustic wave sensor, Hypothalamic neurons, Neuron cell-surface interaction

  10. Biodentine induces immortalized murine pulp cell differentiation into odontoblast-like cells and stimulates biomineralization.

    Science.gov (United States)

    Zanini, Marjorie; Sautier, Jean Michel; Berdal, Ariane; Simon, Stéphane

    2012-09-01

    Biodentine (Septodont, Saint Maur des Faussés, France), a new tricalcium silicate-based cement, has recently been commercialized and advertised as a bioactive material. Its clinical application and physical properties have been widely described, but, so far, its bioactivity and biological effect on pulp cells have not been clearly shown. Thus, the aim of this study was to evaluate the biological effect of Biodentine on immortalized murine pulp cells (OD-21). OD-21 cells were cultured with or without Biodentine. Cell proliferation was determined by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) colorimetric assay after 2, 3, and 5 days of stimulation. The expression of several biomolecular markers was analyzed to screen differentiation pathways, both on a gene level with Real-time reverse transcription polymerase chain reaction and on a protein level by measuring alkaline phosphatase activity. Alizarin red staining was used to assess and quantify biomineralization. The expression patterns of several genes confirmed the differentiation of OD-21 cells into odontoblasts during the period of cell culture. Our results suggest that Biodentine is bioactive because it increased OD-21 cell proliferation and biomineralization in comparison with controls. Because of its bioactivity, Biodentine can be considered as a suitable material for clinical indications of dentin-pulp complex regeneration, such as direct pulp capping. Copyright © 2012 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  11. An elasto-visco-plastic model for immortal foams or emulsions.

    Science.gov (United States)

    Bénito, S; Bruneau, C-H; Colin, T; Gay, C; Molino, F

    2008-03-01

    A variety of complex fluids consists in soft, round objects (foams, emulsions, assemblies of copolymer micelles or of multilamellar vesicles--also known as onions). Their dense packing induces a slight deviation from their preferred circular or spherical shape. As a frustrated assembly of interacting bodies, such a material evolves from one conformation to another through a succession of discrete, topological events driven by finite external forces. As a result, the material exhibits a finite yield threshold. The individual objects usually evolve spontaneously (colloidal diffusion, object coalescence, molecular diffusion), and the material properties under low or vanishing stress may alter with time, a phenomenon known as aging. We neglect such effects to address the simpler behaviour of (uncommon) immortal fluids: we construct a minimal, fully tensorial, rheological model, equivalent to the (scalar) Bingham model. Importantly, the model consistently describes the ability of such soft materials to deform substantially in the elastic regime (be it compressible or not) before they undergo (incompressible) plastic creep--or viscous flow under even higher stresses.

  12. Generation and characterization of the first immortalized alpaca cell line suitable for diagnostic and immunization studies.

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    Valentina Franceschi

    Full Text Available Raising of alpacas as exotic livestock for wool and meat production and as companion animals is growing in importance in the United States, Europe and Australia. Furthermore the alpaca, as well as the rest of the camelids, possesses the peculiarity of producing single-chain antibodies from which nanobodies can be generated. Nanobodies, due to their structural simplicity and reduced size, are very versatile in terms of manipulation and bio-therapeutic exploitation. In fact the biotech companies involved in nanobody production and application continue to grow in number and size. Hence, the development of reagents and tools to assist in the further growth of this new scientific and entrepreneurial reality is becoming a necessity. These are needed mainly to address alpaca disease diagnosis and prophylaxis, and to develop alpaca immunization strategies for nanobody generation. For instance an immortalized alpaca cell line would be extremely valuable. In the present work the first stabilized alpaca cell line from alpaca skin stromal cells (ASSCs was generated and characterized. This cell line was shown to be suitable for replication of viruses bovine herpesvirus-1, bovine viral diarrhea virus and caprine herpesvirus-1 and the endocellular parasite Neospora caninum. Moreover ASSCs were easy to transfect and transduce by several methods. These two latter characteristics are extremely useful when recombinant antigens need to be produced in a host homologous system. This work could be considered as a starting point for the expansion of the biotechnologies linked to alpaca farming and industry.

  13. A lysosomal switch triggers proteostasis renewal in the immortal C. elegans germ lineage.

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    Bohnert, K Adam; Kenyon, Cynthia

    2017-11-30

    Although individuals age and die with time, an animal species can continue indefinitely, because of its immortal germ-cell lineage. How the germline avoids transmitting damage from one generation to the next remains a fundamental question in biology. Here we identify a lysosomal switch that enhances germline proteostasis before fertilization. We find that Caenorhabditis elegans oocytes whose maturation is arrested by the absence of sperm exhibit hallmarks of proteostasis collapse, including protein aggregation. Remarkably, sperm-secreted hormones re-establish oocyte proteostasis once fertilization becomes imminent. Key to this restoration is activation of the vacuolar H + -ATPase (V-ATPase), a proton pump that acidifies lysosomes. Sperm stimulate V-ATPase activity in oocytes by signalling the degradation of GLD-1, a translational repressor that blocks V-ATPase synthesis. Activated lysosomes, in turn, promote a metabolic shift that mobilizes protein aggregates for degradation, and reset proteostasis by enveloping and clearing the aggregates. Lysosome acidification also occurs during Xenopus oocyte maturation; thus, a lysosomal switch that enhances oocyte proteostasis in anticipation of fertilization may be conserved in other species.

  14. Parabens enable suspension growth of MCF-10A immortalized, non-transformed human breast epithelial cells.

    Science.gov (United States)

    Khanna, Sugandha; Darbre, Philippa D

    2013-05-01

    Parabens (alkyl esters of p-hydroxybenzoic acid) are used extensively as preservatives in consumer products, and intact esters have been measured in several human tissues. Concerns of a potential link between parabens and breast cancer have been raised, but mechanistic studies have centred on their oestrogenic activity and little attention has been paid to any carcinogenic properties. In the present study, we report that parabens can induce anchorage-independent growth of MCF-10A immortalized but non-transformed human breast epithelial cells, a property closely related to transformation and a predictor of tumour growth in vivo. In semi-solid methocel suspension culture, MCF-10A cells produced very few colonies and only of a small size but the addition of 5 × 10(-4) M methylparaben, 10(-5) M n-propylparaben or 10(-5) M n-butylparaben resulted in a greater number of colonies per dish (P paraben concentrations in human breast tissue samples from 40 mastectomies (Barr et al., 2012) showed that 22/40 of the patients had at least one of the parabens at the site of the primary tumour at or above these concentrations. To our knowledge, this is the first study to report that parabens can induce a transformed phenotype in human breast epithelial cells in vitro, and further investigation is now justified into a potential link between parabens and breast carcinogenesis. Copyright © 2012 John Wiley & Sons, Ltd.

  15. Characterization of cloned cells from an immortalized fetal pulmonary type II cell line

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    Henderson, R.F.; Waide, J.J.; Lechner, J.F.

    1995-12-01

    A cultured cell line that maintained expression of pulmonary type II cell markers of differentiation would be advantageous to generate a large number of homogenous cells in which to study the biochemical functions of type II cells. Type II epithelial cells are the source of pulmonary surfactant and a cell of origin for pulmonary adenomas. Last year our laboratory reported the induction of expression of two phenotypic markers of pulmonary type II cells (alkaline phosphatase activity and surfactant lipid synthesis) in cultured fetal rat lung epithelial (FRLE) cells, a spontaneously immortalized cell line of fetal rat lung type II cell origin. Subsequently, the induction of the ability to synthesize surfactant lipid became difficult to repeat. We hypothesized that the cell line was heterogenuous and some cells were more like type II cells than others. The purpose of this study was to test this hypothesis and to obtain a cultured cell line with type II cell phenotypic markers by cloning several FRLE cells and characterizing them for phenotypic markers of type II cells (alkaline phosphatase activity and presence of surfactant lipids). Thirty cloned cell lines were analyzed for induced alkaline phosphatase activity (on x-axis) and for percent of phospholipids that were disaturated (i.e., surfactant).

  16. Transformation of SV40-immortalized human uroepithelial cells by 3-methylcholanthrene increases IFN- and Large T Antigen-induced transcripts

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    Easton Marilyn J

    2010-02-01

    Full Text Available Abstract Background Simian Virus 40 (SV40 immortalization followed by treatment of cells with 3-methylcholanthrene (3-MC has been used to elicit tumors in athymic mice. 3-MC carcinogenesis has been thoroughly studied, however gene-level interactions between 3-MC and SV40 that could have produced the observed tumors have not been explored. The commercially-available human uroepithelial cell lines were either SV40-immortalized (HUC or SV40-immortalized and then 3-MC-transformed (HUC-TC. Results To characterize the SV40 - 3MC interaction, we compared human gene expression in these cell lines using a human cancer array and confirmed selected changes by RT-PCR. Many viral Large T Antigen (Tag expression-related changes occurred in HUC-TC, and it is concluded that SV40 and 3-MC may act synergistically to transform cells. Changes noted in IFP 9-27, 2'-5' OAS, IF 56, MxA and MxAB were typical of those that occur in response to viral exposure and are part of the innate immune response. Because interferon is crucial to innate immune host defenses and many gene changes were interferon-related, we explored cellular growth responses to exogenous IFN-γ and found that treatment impeded growth in tumor, but not immortalized HUC on days 4 - 7. Cellular metabolism however, was inhibited in both cell types. We conclude that IFN-γ metabolic responses were functional in both cell lines, but IFN-γ anti-proliferative responses functioned only in tumor cells. Conclusions Synergism of SV40 with 3-MC or other environmental carcinogens may be of concern as SV40 is now endemic in 2-5.9% of the U.S. population. In addition, SV40-immortalization is a generally-accepted method used in many research materials, but the possibility of off-target effects in studies carried out using these cells has not been considered. We hope that our work will stimulate further study of this important phenomenon.

  17. Individual differences in plasma ALT, AST and GGT: contributions of genetic and environmental factors, including alcohol consumption.

    Science.gov (United States)

    Whitfield, J B; Martin, N G

    1985-01-01

    The causes of individuality of the plasma enzymes alanine aminotransferase (ALT; EC 2.6.1.2), aspartate aminotransferase (AST; EC 2.6.1.1) and gamma-glutamyl transferase (GGT; EC 2.3.2.2) were investigated in a study of 206 pairs of twins. Between-person variance was greater in men than women, while within-person variation was similar in both sexes. Plasma ALT and AST levels were affected by genetic factors, while GGT was affected by some environmental factor shared by co-twins. In the men, alcohol intake had a significant but small effect on all three enzyme levels, and since alcohol consumption was highly heritable, this appeared as a genetic influence on enzyme activities. The major factors involved in the observed correlations between these enzymes were a non-shared environmental factor other than alcohol affecting ALT, AST and GGT, and a genetic factor affecting only ALT and AST.

  18. H/sub. cap alpha. / monitors for the Advanced Limiter Test-II (ALT-II) on TEXTOR

    Energy Technology Data Exchange (ETDEWEB)

    Uckan, T.

    1988-05-01

    The H/sub ..cap alpha../ monitor system installed on TEXTOR in conjunction with the Advanced Limiter Test-II (ALT-II) toroidal belt pump limiter is introduced. The monitors are used to study edge particle fluxes and recycling, correlations to confinement properties of core and edge plasma with ALT-II, and also high-power auxiliary heating (/approximately/5 MW) during long-pulse (/approximately/4-s) operation of TEXTOR. A model of the edge particle flux based on the H/sub ..cap alpha../ measurements is presented. The ALT-II experiments are to be carried out in various phases. Here we discuss the results obtained from the monitor system during the initial phase of operations following the installation of the ALT-II limiter, with ohmic heating only. 15 refs., 21 figs.

  19. Incremental Predictive Value of Serum AST-to-ALT Ratio for Incident Metabolic Syndrome: The ARIRANG Study.

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    Dhananjay Yadav

    Full Text Available The ratio of aspartate aminotransferase (AST to alanine aminotransferase (ALT is of great interest as a possible novel marker of metabolic syndrome. However, longitudinal studies emphasizing the incremental predictive value of the AST-to-ALT ratio in diagnosing individuals at higher risk of developing metabolic syndrome are very scarce. Therefore, our study aimed to evaluate the AST-to-ALT ratio as an incremental predictor of new onset metabolic syndrome in a population-based cohort study.The population-based cohort study included 2276 adults (903 men and 1373 women aged 40-70 years, who participated from 2005-2008 (baseline without metabolic syndrome and were followed up from 2008-2011. Metabolic syndrome was defined according to the harmonized definition of metabolic syndrome. Serum concentrations of AST and ALT were determined by enzymatic methods.During an average follow-up period of 2.6-years, 395 individuals (17.4% developed metabolic syndrome. In a multivariable adjusted model, the odds ratio (95% confidence interval for new onset of metabolic syndrome, comparing the fourth quartile to the first quartile of the AST-to-ALT ratio, was 0.598 (0.422-0.853. The AST-to-ALT ratio also improved the area under the receiver operating characteristic curve (AUC for predicting new cases of metabolic syndrome (0.715 vs. 0.732, P = 0.004. The net reclassification improvement of prediction models including the AST-to-ALT ratio was 0.23 (95% CI: 0.124-0.337, P<0.001, and the integrated discrimination improvement was 0.0094 (95% CI: 0.0046-0.0143, P<0.001.The AST-to-ALT ratio independently predicted the future development of metabolic syndrome and had incremental predictive value for incident metabolic syndrome.

  20. Correlation between Aminotransferase Ratio (AST/ALT and Other Biochemical Parameters in Chronic Liver Disease of Viral Origin

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    Shah Md Fazlul Karim

    2015-03-01

    Full Text Available Background: In recent years the ratio of aspartate aminotransferase (AST to alanine aminotransferase (ALT in patients of chronic liver disease (CLD of various origins has gained much attention. This variable is readily available, easy to interpret, and inexpensive and the clinical utility of the AST/ALT ratio in the diagnostic workup of patients with CLD is quite promising. Objective: The present study was designed to find out the link between aminotransferase (AST/ALT ratio with commonly measured biochemical parameters of liver function tests in CLD of viral origin. Materials and method: This cross sectional study was carried out in the department of Biochemistry, Sir Salimullah Medical College, Dhaka, Bangladesh. Forty four biopsy proven diagnosed subjects of chronic viral hepatitis without cirrhosis of both sex were selected purposively. With aseptic precaution 5 mL venous blood was collected from each subject and common liver function tests (serum AST, ALT, AST/ALT ratio, alkaline phosphatase, total bilirubin, serum total protein, serum albumin, serum globulin, serum albumin/globulin ratio, prothrombin time and viral serology (HBsAg, Anti HDV antibody, Anti HCV antibody were performed. Data were analyzed by SPSS version 19 for Windows. Pearson’s correlation test was done to determine association between AST/ALT with other biochemical parameters. Results: Mean(±SD age of the study subjects was 32.55±10.55 years (range 20-50 years with 48 (77.7% male and 14 (22.6% female subjects. Pearson’s correlation test was done between AST to ALT ratio with other biochemical parameters and prothrombin time showed significant positive correlation (p <0.01. Conclusion: In our study we found significant positive correlation between AST/ALT with prothrombin time in CLD subjects without cirrhosis.

  1. Curcumin reverses benzidine-induced epithelial-mesenchymal transition via suppression of ERK5/AP-1 in SV-40 immortalized human urothelial cells.

    Science.gov (United States)

    Liu, Zhiqi; Liu, Jie; Zhao, Li; Geng, Hao; Ma, Jiaxing; Zhang, Zhiqiang; Yu, Dexin; Zhong, Caiyun

    2017-04-01

    Overexposure to benzidine has been manifested as an important cause of bladder cancer. However, the molecular mechanism of benzidine-induced malignancy is still insufficiently interpreted. Epithelial-mesenchymal transition (EMT) is a crucial pathophysiological process in embryonic development as well as initiation and development of epithelium-originated malignant tumors. The role of extracellular regulated protein kinase 5 (ERK5) in benzidine-meditated bladder cancer development has not been explored. In the present study, we explored the role of ERK5/AP-1 pathway in benzidine-induced EMT in human normal urothelial cells and the intervention effect of curcumin on bezidine-induced EMT. We found that benzidine-induced EMT in SV-40 immortalized human urothelial cells (SV-HUC-1) at low concentrations. We detected that ERK5/AP-1 pathway was notably activated. Specific ERK5 inhibitor, XMD8-92 was applied to determine the role of ERK5 in benzidine-induced EMT. Results indicated that XMD8-92 reversed the EMT process. Furthermore, curcumin effectively attenuated benzidine-induced urocystic EMT by suppressing ERK5/AP-1 pathway. In conclusion, the present study revealed the positive role of ERK5/AP-1 in benzidine-provoked urocystic EMT and the curcumin promising use in bladder cancer prevention and intervention via ERK5/AP-1 pathway.

  2. An assessment of racial differences in the upper limits of normal ALT levels in children and the effect of obesity on elevated values.

    Science.gov (United States)

    Kliethermes, S; Ma, M; Purtell, C; Balasubramanian, N; Gonzalez, B; Layden, T J; Cotler, S J

    2017-10-01

    Childhood obesity is a risk factor for non-alcoholic fatty liver disease and poses important public health issues for children. Racial differences in alanine aminotransferase (ALT) levels among children have not been described. This study aimed to identify racial differences in upper limit normal (ULN) ALT levels and evaluate the effect of obesity on elevated levels in children without other metabolic risk factors. National Health and Nutrition Examination Surveys and clinical data from the Loyola University Health System were used to determine ULN ALT by race and gender. Quantile regression was used to evaluate the impact of obesity on elevated ALT and to identify potential risk factors for ALT above the ULN. Upper limit normal (ULN) ALT was approximately 28.0 and 21.0-24.0 U/L for boys and girls, respectively. No significant difference in ULN ALT across race was observed. Obesity was significantly associated with elevated ALT; obese children with elevated ALT had values 10 U/L higher than normal-weight children. Racial differences in ALT levels among adults are not evident in children. Obesity, in the absence of metabolic risk factors and other causes of liver disease, is associated with elevated ALT, providing evidence against the concept of healthy obesity in children. © 2016 World Obesity Federation.

  3. [SOUND CONTACTS] Transcendence and immortality in Busoni’s "Doktor Faust"

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    Antonella DiGiulio

    2015-04-01

    Full Text Available The categorization of the many musical and literary works inspired by the Faust legend is based on their two possible conclusions: Faust ends up in hell or is redeemed and attaints heaven. Busoni’s work is often categorized in terms of the redemptive form. I will analyze a third possibility: during the last part of the nineteenth century, philosophers conceptualized that man’s feelings of unhappiness came from his confinement into an eternal recurrence and that life doesn’t end with the death. Scholars tend to ignore the links between Doktor Faust and these philosophical thoughts because of Busoni’s explicit denial of such intentions. And yet Busoni wrote that an opera is the highest form of artistic expression, where music gives words to the unspoken. Therefore I will argue that Busoni characterizes Doktor Faust as a hybridization of the new philosophical theories with literature and music. Busoni’s depiction of the main character in this opera may not be intentional, but the composer summed up all the qualities of Nietzsche’s Übermensch, D’Annunzio’s Superuomo, and Pascoli’s fanciullino in Faust. Faust discovers his own transcendent power, which is independent from the material universe and yet belongs to nature’s laws that are beyond God and the evil. Doktor Faust’s attainment of immortality does not pray for divine redemption and wins out over everything: in this opera, Faust is neither redeemed nor damned. Faust becomes aware that death is nothing but the means that lead to rebirth: “ich, Faust, ein ewiger Wille”.

  4. Functional Characteristics of Reversibly Immortalized Hepatic Progenitor Cells Derived from Mouse Embryonic Liver

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    Yang Bi

    2014-10-01

    Full Text Available Background/Aims: Liver is a vital organ and retains its regeneration capability throughout adulthood, which requires contributions from different cell populations, including liver precursors and intrahepatic stem cells. To overcome the mortality of hepatic progenitors (iHPs in vitro, we aim to establish reversibly immortalized hepatic progenitor cells from mouse embryonic liver. Methods and Results: Using retroviral system to stably express SV40 T antigen flanked with Cre/LoxP sites, we establish a repertoire of iHP clones with varied differentiation potential. The iHP cells maintain long-term proliferative activity and express varied levels of progenitor markers (Pou5f1/Oct4 and Dlk and hepatocyte markers (AFP, Alb and ApoB. Five representative iHP clones express hepatic/pancreatic transcription factors HNF3α/Foxa1, HNF3β/Foxa2, and HNF4α/MODY1. Dexamethasone is shown to promote the expression of hepatocyte markers AFP and TAT, along with ICG-uptake and glycogen storage functions in the iHP clones. Cre-mediated removal of SV40 T antigen reverses the proliferative activity of iHP cells. When iHP cells are subcutaneously implanted in athymic nude mice, no tumor formation is observed for up to 8 weeks. Conclusions: We demonstrate that the established iHP cells are stable, reversible, and non-tumorigenic hepatic progenitor-like cells, which should be valuable for studying liver organogenesis, metabolic regulations, and hepatic lineage-specific differentiation.

  5. Arsenic compromises conducting airway epithelial barrier properties in primary mouse and immortalized human cell cultures.

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    Cara L Sherwood

    Full Text Available Arsenic is a lung toxicant that can lead to respiratory illness through inhalation and ingestion, although the most common exposure is through contaminated drinking water. Lung effects reported from arsenic exposure include lung cancer and obstructive lung disease, as well as reductions in lung function and immune response. As part of their role in innate immune function, airway epithelial cells provide a barrier that protects underlying tissue from inhaled particulates, pathogens, and toxicants frequently found in inspired air. We evaluated the effects of a five-day exposure to environmentally relevant levels of arsenic {<4μM [~300 μg/L (ppb] as NaAsO2} on airway epithelial barrier function and structure. In a primary mouse tracheal epithelial (MTE cell model we found that both micromolar (3.9 μM and submicromolar (0.8 μM arsenic concentrations reduced transepithelial resistance, a measure of barrier function. Immunofluorescent staining of arsenic-treated MTE cells showed altered patterns of localization of the transmembrane tight junction proteins claudin (Cl Cl-1, Cl-4, Cl-7 and occludin at cell-cell contacts when compared with untreated controls. To better quantify arsenic-induced changes in tight junction transmembrane proteins we conducted arsenic exposure experiments with an immortalized human bronchial epithelial cell line (16HBE14o-. We found that arsenic exposure significantly increased the protein expression of Cl-4 and occludin as well as the mRNA levels of Cl-4 and Cl-7 in these cells. Additionally, arsenic exposure resulted in altered phosphorylation of occludin. In summary, exposure to environmentally relevant levels of arsenic can alter both the function and structure of airway epithelial barrier constituents. These changes likely contribute to the observed arsenic-induced loss in basic innate immune defense and increased infection in the airway.

  6. [Effects of polydatin on ALT, AST, TNF-alpha, and COX-2 in sepsis model mice].

    Science.gov (United States)

    Li, Xiao-Hui; Wu, Meng-Jiao; Zhang, Li-Na; Zheng, Jia-Jia; Zhang, Li; Wan, Jing-Yuan

    2013-02-01

    To investigate the protective effects of polydatin on sepsis-induced acute liver injury (ALI) in mice, and to preliminarily study its mechanisms. The sepsis model was established using cecal ligation and puncture (CLP).A sham-operation control group was also set up. Polydatin (50, 100, and 300 mg/kg, respectively) was administrated to mice 1 h before CLP. The survival and liver injury were evaluated subsequently per 6 h after CLP. The survived mice were scarified 24 h later. The serum and the liver tissue sample were collected. The serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected by colorimetric method. The content of tumor necrosis factor-alpha (TNF-alpha) was assayed by ELISA. The cyclooxygenase-2 (COX-2) expression in the liver tissue was detected by Western blot. The pathological changes of the hepatic tissue were analyzed by hematoxylin and eosin stain. The mortality of mice reached as high as 50% at 24 h after CLP. The biochemical indices and the pathological changes of the liver tissue showed obvious lesion. The success rate of modeling was 90%. Compared with the sham-operation control group, the serum ALT,AST activity, the TNF-alpha content, and the hepatic COX-2 protein expression markedly increased in the CLP group (P < 0.01). Polydatin improved the sepsis-induced mortality dose-dependently, inhibited increased ALT, AST activity and TNF-alpha, decreased the hepatic COX-2 protein expression, and attenuated the pathological injury of the liver (P < 0.05). Polydatin could effectively protect sepsis-induced ALI, which might be achieved possibly through inhibiting serum TNF-alpha production and hepatic COX-2 expression.

  7. AGE-breaker ALT-711 plus insulin could restore erectile function in streptozocin-induced type 1 diabetic rats.

    Science.gov (United States)

    Wang, Lin; Tian, Wenjie; Uwais, Zaid; Li, Guangyong; Li, Huixi; Guan, Ruili; Gao, Zhezhu; Xin, Zhongcheng

    2014-06-01

    The interaction between advanced glycation end-products (AGEs) and its receptors for AGEs (RAGEs) elicits oxidative stress and mediates the development of erectile dysfunction (ED). ALT-711, an AGE cross-link breaker, has the therapeutic potential for ED but has been less intensively investigated. The aim of this study was to investigate the effects of an AGEs breaker 3-phenacyl-4,5-dimethylthiazolium chloride (ALT-711) plus insulin on erectile function in streptozocin (STZ)-induced type 1 diabetic rats. Fifty 8-week-old Sprague-Dawley rats were randomly distributed into five groups: normal control (C), diabetic (D), insulin-treated diabetic (D + I), ALT-711-treated diabetic (D + ALT-711) and insulin plus ALT-711-treated diabetic (D + I + ALT-711) rats. Diabetes was induced by a single intraperitoneal injection of STZ. Eight weeks after induction of diabetes, ALT-711 was administered by intraperitoneal injection. Two to six units of intermediate-acting insulin were utilized to achieve normal levels of glycemic control. After treatment for 6 weeks, erectile function was determined via measurement of intracavernous pressures (ICPs) following electrostimulation of the cavernous nerve. The deposition of AGEs, expression of RAGEs, superoxide dismutase activity, and lipid peroxidation were measured. We also evaluated penile histological changes such as smooth muscle contents, endothelial cells contents, and apoptotic activity. The main outcome measures were the ratio of ICP/mean arterial pressure (MAP), penile endothelial cells, smooth muscle cells, neuronal nitric oxide synthase, AGE and RAGE expression, malondialdehyde concentration, SOD activity, and apoptosis index. Diabetic rats demonstrated significantly reduced ICP/MAP ratio, penile endothelial cells, smooth muscles cells, increased AGEs and RAGE expression, and increased apoptosis. Insulin and ALT-711 monotherapy partially restored erectile function and histological changes. However, the

  8. Incremental Predictive Value of Serum AST-to-ALT Ratio for Incident Metabolic Syndrome: The ARIRANG Study

    Science.gov (United States)

    Ahn, Song Vogue; Baik, Soon Koo; Cho, Youn zoo; Koh, Sang Baek; Huh, Ji Hye; Chang, Yoosoo; Sung, Ki-Chul; Kim, Jang Young

    2016-01-01

    Aims The ratio of aspartate aminotransferase (AST) to alanine aminotransferase (ALT) is of great interest as a possible novel marker of metabolic syndrome. However, longitudinal studies emphasizing the incremental predictive value of the AST-to-ALT ratio in diagnosing individuals at higher risk of developing metabolic syndrome are very scarce. Therefore, our study aimed to evaluate the AST-to-ALT ratio as an incremental predictor of new onset metabolic syndrome in a population-based cohort study. Material and Methods The population-based cohort study included 2276 adults (903 men and 1373 women) aged 40–70 years, who participated from 2005–2008 (baseline) without metabolic syndrome and were followed up from 2008–2011. Metabolic syndrome was defined according to the harmonized definition of metabolic syndrome. Serum concentrations of AST and ALT were determined by enzymatic methods. Results During an average follow-up period of 2.6-years, 395 individuals (17.4%) developed metabolic syndrome. In a multivariable adjusted model, the odds ratio (95% confidence interval) for new onset of metabolic syndrome, comparing the fourth quartile to the first quartile of the AST-to-ALT ratio, was 0.598 (0.422–0.853). The AST-to-ALT ratio also improved the area under the receiver operating characteristic curve (AUC) for predicting new cases of metabolic syndrome (0.715 vs. 0.732, P = 0.004). The net reclassification improvement of prediction models including the AST-to-ALT ratio was 0.23 (95% CI: 0.124–0.337, Pmetabolic syndrome and had incremental predictive value for incident metabolic syndrome. PMID:27560931

  9. Incremental Predictive Value of Serum AST-to-ALT Ratio for Incident Metabolic Syndrome: The ARIRANG Study.

    Science.gov (United States)

    Yadav, Dhananjay; Choi, Eunhee; Ahn, Song Vogue; Baik, Soon Koo; Cho, Youn Zoo; Koh, Sang Baek; Huh, Ji Hye; Chang, Yoosoo; Sung, Ki-Chul; Kim, Jang Young

    2016-01-01

    The ratio of aspartate aminotransferase (AST) to alanine aminotransferase (ALT) is of great interest as a possible novel marker of metabolic syndrome. However, longitudinal studies emphasizing the incremental predictive value of the AST-to-ALT ratio in diagnosing individuals at higher risk of developing metabolic syndrome are very scarce. Therefore, our study aimed to evaluate the AST-to-ALT ratio as an incremental predictor of new onset metabolic syndrome in a population-based cohort study. The population-based cohort study included 2276 adults (903 men and 1373 women) aged 40-70 years, who participated from 2005-2008 (baseline) without metabolic syndrome and were followed up from 2008-2011. Metabolic syndrome was defined according to the harmonized definition of metabolic syndrome. Serum concentrations of AST and ALT were determined by enzymatic methods. During an average follow-up period of 2.6-years, 395 individuals (17.4%) developed metabolic syndrome. In a multivariable adjusted model, the odds ratio (95% confidence interval) for new onset of metabolic syndrome, comparing the fourth quartile to the first quartile of the AST-to-ALT ratio, was 0.598 (0.422-0.853). The AST-to-ALT ratio also improved the area under the receiver operating characteristic curve (AUC) for predicting new cases of metabolic syndrome (0.715 vs. 0.732, P = 0.004). The net reclassification improvement of prediction models including the AST-to-ALT ratio was 0.23 (95% CI: 0.124-0.337, Pmetabolic syndrome and had incremental predictive value for incident metabolic syndrome.

  10. The major Alternaria alternata allergen, Alt a 1: A reliable and specific marker of fungal contamination in citrus fruits.

    Science.gov (United States)

    Gabriel, M F; Uriel, N; Teifoori, F; Postigo, I; Suñén, E; Martínez, J

    2017-09-18

    The ubiquitously present spores of Alternaria alternata can spoil a wide variety of foodstuffs, including a variety of fruits belonging to the Citrus genus. The major allergenic protein of A. alternata, Alt a 1, is a species-specific molecular marker that has been strongly associated with allergenicity and phytopathogenicity of this fungal species. This study aimed to evaluate the potential of the detection of Alt a 1 as a reliable indicator of A. alternata contamination in citrus fruits. To accomplish this aim, sixty oranges were artificially infected with a spore suspension of A. alternata. Internal fruit material was collected at different incubation times (one, two and three weeks after the fungal inoculation) and used for both total RNA extraction and protein extraction. Alt a 1 detection was then performed by polymerase chain reaction (PCR) amplification using Alt a 1 specific primers and by enzyme-linked immunosorbent assay (ELISA). The experimental model presented in this work was effective to simulate the typical Alternaria black rot phenotype and its progression. Although both PCR and ELISA techniques have been successfully carried out for detecting Alt a 1 allergen in A. alternata infected oranges, the PCR method was found to be more sensitive than ELISA. Nevertheless, ELISA results were highly valuable to demonstrate that considerable amounts of Alt a 1 are produced during A. alternata fruit infection process, corroborating the recently proposed hypothesis that this protein plays a role in the pathogenicity and virulence of Alternaria species. Such evidence suggests that the detection of Alt a 1 by PCR-based assay may be used as a specific indicator of the presence of pathogenic and allergenic fungal species, A. alternata, in fruits. This knowledge can be employed to control the fungal infection and mitigate agricultural losses as well as human exposure to A. alternata allergens and toxins. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Self-reported faster eating associated with higher ALT activity in middle-aged, apparently healthy Japanese women.

    Science.gov (United States)

    Mochizuki, Kazuki; Hariya, Natsuyo; Miyauchi, Rie; Misaki, Yasumi; Ichikawa, Yoko; Goda, Toshinao

    2014-01-01

    Faster eating and elevated circulating activity of alanine aminotransferase (ALT), a marker for liver injury, are risk factors for the development of obesity and type 2 diabetes mellitus, and their complications. The aim of this study was to examine the association between self-reported eating rate and circulating ALT activity in apparently healthy middle-aged Japanese women. We conducted a cross-sectional study of 900 apparently healthy women ages 40 to 64 y (mean ± SD, 53.1 ± 7.1 y) who participated in health check-ups in Japan. We analyzed their clinical serum parameters and lifestyle factors, including self-reported eating rate. Associations between liver injury markers (ALT, γ-glutamyl transpeptidase [GTP], and aspartate aminotransferase [AST]), other clinical parameters and lifestyle factors were analyzed using Tukey's multiple range test following analysis of variance and analysis of covariance for three groups, divided by self-reported eating rates. The associations between self-reported faster eating and ALT activity and lifestyle factors were analyzed by multivariate logistic regression analyses. ALT activity, but not γ-GTP or AST activities, was higher in participants who reported relatively fast/very fast eating than in those who reported medium eating after adjusting for age, alcohol intake, energy intake, smoking, and physical activity. The odds ratio of eating rate for ALT activity in T3 (18-128 U/L) compared with T1 (3-12 U/L) was 1.67 (P ALT activity is positively associated with faster eating, but is dependent on BMI in middle-aged, apparently healthy Japanese women. Copyright © 2014 Elsevier Inc. All rights reserved.

  12. "Almha La Vengadora": Protagonista del indigenismo de neovanguardia alteño

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    Maria Irina Soto-Mejia

    2016-03-01

    Este artículo discute la obra de Crispín Portugal (1976-2007, un escritor alteño, en el contexto de la lucha contra la pobreza y el racismo que enfrentan a las ciudades de El Alto y La Paz, y del movimiento en contra del mercado  que comenzó en Argentina produciendo libros con tapas de cartón compradas a los cartoneros. Portugal y otros escritores indígenas iniciaron un proyecto editorial llamado Yerba Mala Cartonera, que cuenta con su propio blog, para diseminar la literatura boliviana que no cuenta con otros medios de difusión. A través de la revisión de un cuento de Portugal (Almha La Vengadora que describe a una chola luchadora de El Alto cuyo nombre en el cuadrilátero es “la vengadora”, este trabajo plantea la configuración de un indigenismo alteño de neovanguardia equiparable al vanguardismo de Gamaliel Churata.  Con este propósito, presta especial atención a El Alto –la ciudad indígena más grande de Latinoamérica- situándolo como escenario clave para el desarrollo de una neovanguardia que se alimenta del neoliberalismo fallido para configurar una propuesta de modernidad que recupera el pasado indígena aymara para inventar otras formas de futuro.

  13. [The effect of blood AST, ALT and lactate after short and middle distance exercise training].

    Science.gov (United States)

    Chuang, C C; Chen, W C; Lee, S Y; Wang, K T

    1996-09-01

    Twenty-four nursing college students aged 16 and 17 years were selected as research subjects and divided into two groups. Group A comprised 12 individuals who were trained for short distance running (5km/day) over a four-week period, while group B was trained for middle distance running (7km/day) during the same period. Blood AST (aspartate aminotransferase), ALT (alanine aminotransferase) and lactate were performed at rest (before training) and after exercise every week. After both short and middle distance exercise training, the lactate values after 1-3 week(s) training period were persistently higher than those before training and the differences between then are significant. However, the lactate value after 4 weeks training period is lower than that after the third week training. There are significant differences between the AST values after 1-4 week(s) training period and those before exercise in short distance exercise training. There are no significant differences between the AST values after training and those before exercise in middle distance exercise training. The ALT values after 1-4 week(s) training period were lower than those before exercise in short and middle distance exercise training. In conclusion, after 4 weeks training, the lactate and AST values can't reduce to those before training in middle distance exercise training, and the lactate value in short distance exercise training is the same as former. Further investigation is needed.

  14. Production and Functional Characterization of Murine Osteoclasts Differentiated from ER-Hoxb8-Immortalized Myeloid Progenitor Cells.

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    Frank Zach

    Full Text Available In vitro differentiation into functional osteoclasts is routinely achieved by incubation of embryonic stem cells, induced pluripotent stem cells, or primary as well as cryopreserved spleen and bone marrow-derived cells with soluble receptor activator of nuclear factor kappa-B ligand and macrophage colony-stimulating factor. Additionally, osteoclasts can be derived from co-cultures with osteoblasts or by direct administration of soluble receptor activator of nuclear factor kappa-B ligand to RAW 264.7 macrophage lineage cells. However, despite their benefits for osteoclast-associated research, these different methods have several drawbacks with respect to differentiation yields, time and animal consumption, storage life of progenitor cells or the limited potential for genetic manipulation of osteoclast precursors. In the present study, we therefore established a novel protocol for the differentiation of osteoclasts from murine ER-Hoxb8-immortalized myeloid stem cells. We isolated and immortalized bone marrow cells from wild type and genetically manipulated mouse lines, optimized protocols for osteoclast differentiation and compared these cells to osteoclasts derived from conventional sources. In vitro generated ER-Hoxb8 osteoclasts displayed typical osteoclast characteristics such as multi-nucleation, tartrate-resistant acid phosphatase staining of supernatants and cells, F-actin ring formation and bone resorption activity. Furthermore, the osteoclast differentiation time course was traced on a gene expression level. Increased expression of osteoclast-specific genes and decreased expression of stem cell marker genes during differentiation of osteoclasts from ER-Hoxb8-immortalized myeloid progenitor cells were detected by gene array and confirmed by semi-quantitative and quantitative RT-PCR approaches. In summary, we established a novel method for the quantitative production of murine bona fide osteoclasts from ER-Hoxb8 stem cells generated from

  15. Accumulation and altered localization of telomere-associated protein TRF2 in immortally transformed and tumor-derived human breast cells

    Energy Technology Data Exchange (ETDEWEB)

    Nijjar, Tarlochan; Bassett, Ekaterina; Garbe, James; Takenaka, Yasuhiro; Stampfer, Martha R.; Gilley, David; Yaswen, Paul

    2004-12-23

    We have used cultured human mammary epithelial cells (HMEC) and breast tumor-derived lines to gain information on defects that occur during breast cancer progression. HMEC immortalized by a variety of agents (the chemical carcinogen benzo(a)pyrene, oncogenes c-myc and ZNF217, and/or dominant negative p53 genetic suppressor element GSE22) displayed marked up regulation (10-15 fold) of the telomere binding protein, TRF2. Up-regulation of TRF2 protein was apparently due to differences in post-transcriptional regulation, as mRNA levels remained comparable in finite life span and immortal HMEC. TRF2 protein was not up-regulated by the oncogenic agents alone in the absence of immortalization, nor by expression of exogenously introduced hTERT genes. We found TRF2 levels to be at least 2-fold higher than in control cells in 11/15 breast tumor cell lines, suggesting that elevated TRF2 levels are a frequent occurrence during the transformation of breast tumor cells in vivo. The dispersed distribution of TRF2 throughout the nuclei in some immortalized and tumor-derived cells indicated that not all the TRF2 was associated with telomeres in these cells. The process responsible for accumulation of TRF2 in immortalized HMEC and breast tumor-derived cell lines may promote tumorigenesis by contributing to the cells ability to maintain an indefinite life span.

  16. Characterization of global transcription profile of normal and HPV-immortalized keratinocytes and their response to TNF treatment

    Directory of Open Access Journals (Sweden)

    Colo Anna

    2008-06-01

    Full Text Available Abstract Background Persistent infection by high risk HPV types (e.g. HPV-16, -18, -31, and -45 is the main risk factor for development of cervical intraepithelial neoplasia and cervical cancer. Tumor necrosis factor (TNF is a key mediator of epithelial cell inflammatory response and exerts a potent cytostatic effect on normal or HPV16, but not on HPV18 immortalized keratinocytes. Moreover, several cervical carcinoma-derived cell lines are resistant to TNF anti-proliferative effect suggesting that the acquisition of TNF-resistance may constitute an important step in HPV-mediated carcinogenesis. In the present study, we compared the gene expression profiles of normal and HPV16 or 18 immortalized human keratinocytes before and after treatment with TNF for 3 or 60 hours. Methods In this study, we determined the transcriptional changes 3 and 60 hours after TNF treatment of normal, HPV16 and HPV18 immortalized keratinocytes by microarray analysis. The expression pattern of two genes observed by microarray was confirmed by Northern Blot. NF-κB activation was also determined by electrophoretic mobility shift assay (EMSA using specific oligonucleotides and nuclear protein extracts. Results We observed the differential expression of a common set of genes in two TNF-sensitive cell lines that differs from those modulated in TNF-resistant ones. This information was used to define genes whose differential expression could be associated with the differential response to TNF, such as: KLK7 (kallikrein 7, SOD2 (superoxide dismutase 2, 100P (S100 calcium binding protein P, PI3 (protease inhibitor 3, skin-derived, CSTA (cystatin A, RARRES1 (retinoic acid receptor responder 1, and LXN (latexin. The differential expression of the KLK7 and SOD2 transcripts was confirmed by Northern blot. Moreover, we observed that SOD2 expression correlates with the differential NF-κB activation exhibited by TNF-sensitive and TNF-resistant cells. Conclusion This is the first in

  17. AKTIVITAS ASPARTAT AMINOTRANSFERASE (AST DAN ALANIN TRANSAMINASE (ALT PADA MONYET EKOR PANJANG (Macaca fascicularis OBESITAS DI PURA LUHUR ULUWATU, BALI

    Directory of Open Access Journals (Sweden)

    Ayu Paramita Lestari

    2016-08-01

    Full Text Available Penelitian obsevasional-deskriptif dengan pendekatan cross-sectional telah dilakukan untuk mengetahui aktivitas aspartat aminotransferase (AST dan alanin transaminase (ALT pada monyet ekor panjang (Macaca fascicularis obesitas yang hidup liar di kawasan Pura Luhur Uluwatu, Bali. Sebanyak 16 ekor monyet ekor panjang berhasil dibius menggunakan ketamine dosis 10 mg/kg berat badan dicampur dengan premedikasi xylasin dosis 1-2 mg/kg berat badan. Sampel yang digunakan adalah serum darah monyet obesitas yang diambil dari monyet dalam keadaan terbius. Dari 16 ekor monyet, 12 ekor tergolong obesitas berdasarkan Indeks Massa Tubuh dan Berat Badan. Aktivitas aspartat aminotransferase (AST dan alanin transaminase (ALT ditentukan menggunakan mesin automatic chemistry analyzer (Indiko-Thermo Scientific. Hasil penelitian menunjukkan nilai AST bervariasi dari 43-88 U/L dan ALT 26-77 U/L dengan rataan 69,4 ± 14,9 U/L. Dari hasil penelitian dapat disimpulkan bahwa nilai AST dan ALT monyet obesitas cenderung lebih tinggi dari nilai AST dan ALT normal.

  18. [Aspartate aminotransferase (AST) to alanine aminotransferase (ALT) ratio in health surveillance of workers exposed to vinyl chloride monomer: preliminary results].

    Science.gov (United States)

    Di Lorenzo, L; Corfiati, M; Bulfaro, D; Liuzzi, G; Zenzola, M; Soleo, L

    2003-01-01

    An AST/ALT ratio > 1 is predictive of liver fibrosis and cirrhosis in patients with chronic hepatitis C virus infection. The aim of this case-control study is to assess AST/ALT ratio in 150 workers exposed to VCM (E) from the beginning of the 1960s to the end of the 1990s. The non-exposed group (NE) consists in 150 male workers employed in the production of a food industry. At least since 1983 exposed subjects worked at VCM environmental concentrations 1 significantly higher than non-exposed. The mean AST/ALT ratio results significantly higher in the exposed group, also after stratification for alcohol consumption. In exposed workers who consume alcoholic beverages and are operating since before 1983 AST/ALT ratio is significantly and positively influenced only by the working age until 1983. If these results will be confirmed, AST/ALT ratio could be proposed to be included in the periodic medical surveillance of VCM workers.

  19. Cdk-dependent phosphorylation regulates TRF1 recruitment to PML bodies and promotes C-circle production in ALT cells.

    Science.gov (United States)

    Wilson, Florence R; Ho, Angus; Walker, John R; Zhu, Xu-Dong

    2016-07-01

    TRF1, a duplex telomeric DNA binding protein, is implicated in homologous-recombination-based alternative lengthening of telomeres, known as ALT. However, how TRF1 promotes ALT activity has yet to be fully characterized. Here we report that Cdk-dependent TRF1 phosphorylation on T371 acts as a switch to create a pool of TRF1, referred to as (pT371)TRF1, which is recruited to ALT-associated PML bodies (APBs) in S and G2 phases independently of its binding to telomeric DNA. We find that phosphorylation of T371 is essential for APB formation and C-circle production, both of which are hallmarks of ALT. We show that the interaction of (pT371)TRF1 with APBs is dependent upon ATM and homologous-recombination-promoting factors Mre11 and BRCA1. In addition, (pT371)TRF1 interaction with APBs is sensitive to transcription inhibition, which also reduces DNA damage at telomeres. Furthermore, overexpression of RNaseH1 impairs (pT371)TRF1 recruitment to APBs in the presence of campothecin, an inhibitor that prevents topoisomerase I from resolving RNA-DNA hybrids. These results suggest that transcription-associated DNA damage, perhaps arising from processing RNA-DNA hybrids at telomeres, triggers (pT371)TRF1 recruitment to APBs to facilitate ALT activity. © 2016. Published by The Company of Biologists Ltd.

  20. Ephedra alte (Joint Pine: An Invasive, Problematic Weedy Species in Forestry and Fruit Tree Orchards in Jordan

    Directory of Open Access Journals (Sweden)

    Jamal R. Qasem

    2012-01-01

    Full Text Available A field survey was carried out to record plant species climbed by Ephedra alte in certain parts of Jordan during 2008–2010. Forty species of shrubs, ornamental, fruit, and forest trees belonging to 24 plant families suffered from the climbing habit of E. alte. Growth of host plants was adversely affected by E. alte growth that extended over their vegetation. In addition to its possible competition for water and nutrients, the extensive growth it forms over host species prevents photosynthesis, smothers growth and makes plants die underneath the extensive cover. However, E. alte did not climb all plant species, indicating a host preference range. Damaged fruit trees included Amygdalus communis, Citrus aurantifolia, Ficus carica, Olea europaea, Opuntia ficus-indica, and Punica granatum. Forestry species that were adversely affected included Acacia cyanophylla, Ceratonia siliqua, Crataegus azarolus, Cupressus sempervirens, Pinus halepensis, Pistacia atlantica, Pistacia palaestina, Quercus coccifera, Quercus infectoria, Retama raetam, Rhamnus palaestina, Rhus tripartita, and Zizyphus spina-christi. Woody ornamentals attacked were Ailanthus altissima, Hedera helix, Jasminum fruticans, Jasminum grandiflorum, Nerium oleander, and Pyracantha coccinea. Results indicated that E. alte is a strong competitive for light and can completely smother plants supporting its growth. A. communis, F. carica, R. palaestina, and C. azarolus were most frequently attacked.

  1. Altération des sulfures des granulats dans les chaussées

    Science.gov (United States)

    Jigorel, A.; Jauberthie, R.

    2002-07-01

    Sulfides present in hornsfeld aggregates, used in light pavanent construction in contact with humid air alter rapidly. Crystallisation of sulfates at the interface of bitumastic material and sub-base creates serious problems (intumescence and crazing) that can lead to a total reconstruction of the project (roads, sidewalks, sports areas, etc). The sulfides in the aggregates and the sulfates produced due to alteration are studied by SEM and XRD. The results show that the intensity of this phenomenon is linked to the nature and the crystallinity of the sulfides. The evolution of the sulfates formed during this alteration process is slow and complex. In new pavements (3 years) the sulfates have a pulverised appearance and consist mostly of epsomite, associated with pickeringite and halotrichite. In older pavements (20 years) the sulfates form a fibrous concretion consisting of pickeringite and small quantities of halotrichite. Les sulfures présents dans les granulats élaborés à partir de cornéen nes s'altèrent rapidement dans les chaussées légères en présence d'air humide. La cristallisation des sulfates à l'interface enrobé-couche de fondation crée des désordres si importants (intumescences, faiençage) qu'il est bien souvent nécessaire d'assurer la réfection totale des ouvrages (routes, trottoirs, plateaux sportifs...). Les sulfures des granulats et les sulfates issus du processus d'altération ont été étudiés par diffractométrie X et examen su Microscope Electronique à Balayage équipé de microanalyse X. Les résultats montrent que !intensité des désordres est liée à la nature et à la cristallinité des sulfures. Les sulfates formés évoluent su cours du processus d'altération qui est long et complexe. Dans les chaussées récentes (3 ans) ils ont un aspect pulvérulent et sont constitués d'epsomite dominante associée à de la pickeringite et à de l'halotrichite. Dans les chaussées plus anciennes (20 ans) ils forment des concr

  2. Molecular characterization of the t(3;9) associated with immortalization in the MCF10A cell line.

    Science.gov (United States)

    Cowell, John K; LaDuca, Jeffrey; Rossi, Michael R; Burkhardt, Tania; Nowak, Norma J; Matsui, Sei-ichi

    2005-11-01

    The t(3;9)(p14;p21) in the MCF10A human mammary gland epithelial cell line was the single cytogenetic event that accompanied the transition from primary culture to immortalized cell line, suggesting that it is related to the development of the immortalization phenotype. To study the molecular consequences of the breakpoints in this rearrangement, we used a combination of fluorescence in situ hybridization (FISH) and array comparative genomic hybridization (CGH). The 3p14 translocation breakpoint occurs within BAC RP11-795e22, which accommodates only the TAFA1 gene, a novel cysteine-rich secreted protein thought to be involved in cytokine signaling. TAFA1 is expressed in normal breast tissue, not in MCF10A, and shows differential expression in a range of breast cancer cell lines. The 9p translocation breakpoint results in a deletion of approximately 4 megabases on the derivative chromosome 9, which includes the CDKN2A (p16) gene. Array CGH and FISH analysis demonstrated that BAC 149i22, which contains the CDKN2A/B genes, is also deleted specifically on the apparently normal copy of chromosome 9, making MCF10A null for the p16/p15 genes. The exact extent of gains and losses of chromosome regions resulting from rearrangements involving chromosomes 1q, 5q, and 8q have also been characterized using the BAC arrays.

  3. Reversible immortalization of Nestin-positive precursor cells from pancreas and differentiation into insulin-secreting cells

    Energy Technology Data Exchange (ETDEWEB)

    Wei, Pei; Li, Li; Qi, Hui [The Clinical Medical Research Center, The Second Clinical Medical College (Shenzhen People' s Hospital), Jinan University, 518020 Shenzhen (China); Zhou, Han-xin [Department of General Surgery, First Hospital (Shenzhen Second People' s Hospital) of Shenzhen University, 518020 Shenzhen (China); Deng, Chun-yan [The Clinical Medical Research Center, The Second Clinical Medical College (Shenzhen People' s Hospital), Jinan University, 518020 Shenzhen (China); Li, Fu-rong, E-mail: frli62@yahoo.com [The Clinical Medical Research Center, The Second Clinical Medical College (Shenzhen People' s Hospital), Jinan University, 518020 Shenzhen (China); Shenzhen Institution of Gerontology, 518020 Shenzhen (China)

    2012-02-10

    Highlights: Black-Right-Pointing-Pointer The NPPCs from mouse pancreas were isolated. Black-Right-Pointing-Pointer Tet-on system for SV40 large in NPPCs was used to get RINPPCs. Black-Right-Pointing-Pointer The RINPPCs can undergo at least 80 population doublings without senescence. Black-Right-Pointing-Pointer The RINPPCs can be induced to differentiate into insulin-producing cells. Black-Right-Pointing-Pointer The combination of GLP-1 and sodium butyrate promoted the differentiation process. -- Abstract: Pancreatic stem cells or progenitor cells posses the ability of directed differentiation into pancreatic {beta} cells. However, these cells usually have limited proliferative capacity and finite lifespan in vitro. In the present study, Nestin-positive progenitor cells (NPPCs) from mouse pancreas that expressed the pancreatic stem cells or progenitor cell marker Nestin were isolated to obtain a sufficient number of differentiated pancreatic {beta} cells. Tet-on system for SV40 large T-antigen expression in NPPCs was used to achieve reversible immortalization. The reversible immortal Nestin-positive progenitor cells (RINPPCs) can undergo at least 80 population doublings without senescence in vitro while maintaining their biological and genetic characteristics. RINPPCs can be efficiently induced to differentiate into insulin-producing cells that contain a combination of glucagon-like peptide-1 (GLP-1) and sodium butyrate. The results of the present study can be used to explore transplantation therapy of type I diabetes mellitus.

  4. Influence of gender on the ratio of serum aspartate aminotransferase (AST) to alanine aminotransferase (ALT) in patients with and without hyperbilirubinemia.

    Science.gov (United States)

    Mera, Jorge R; Dickson, Beverly; Feldman, Mark

    2008-03-01

    The serum asparate aminotransferase (AST)/alanine aminotransferase (ALT) ratio is widely used in the differential diagnosis of icteric and non-icteric hepatic disorders. Our objective was to determine whether there are gender related-differences in the serum AST/ALT ratio. We used sera from 3,618 unselected patients sent to our laboratory for an automated chemistry panel, which included measurements of AST, ALT, alkaline phosphatase, and total bilirubin. Effects of gender on serum AST, ALT, and AST/ALT were examined in different age groups. Among patients with normal total serum bilirubin concentrations, serum AST and ALT concentrations were significantly lower in the females than in the males (P < 0.0001). However, the serum AST/ALT ratio was higher in the females than the males (median values of 0.90 and 0.81, respectively; P < 0.0001). AST and ALT were also lower in the 54 hyperbilirubinemic females than in the 102 hyperbilirubinemic males. Serum AST/ALT ratios were considerably higher in these 156 hyperbilirubinemic patients than in the normobilirubinemic group, with median ratios of 1.09 in females and 0.92 in males (P = 0.02). Significantly higher serum AST/ALT ratios in females were first evident in the 3rd age decade and remained significantly higher than ratios in males through the 8th decade. We conclude that serum AST/ALT ratios are higher in women than men. When clinicians utilize serum AST/ALT ratios to assess the etiology or chronicity of liver disease, the patient's gender also should be taken into consideration.

  5. Predictive value of ALT levels for non-alcoholic steatohepatitis (NASH) and advanced fibrosis in non-alcoholic fatty liver disease (NAFLD).

    Science.gov (United States)

    Verma, Siddharth; Jensen, Donald; Hart, John; Mohanty, Smruti R

    2013-10-01

    Non-alcoholic fatty liver disease (NAFLD) patients with elevated serum alanine aminotransferase (ALT) generally undergo a liver biopsy to evaluate for possible non-alcoholic steatohepatitis (NASH) or advanced fibrosis. However, patients with normal ALT could also have advanced stages of NAFLD. To determine ALT value that will accurately predict NASH and advanced fibrosis using area under the receiver operating characteristics curve (AUROC) analysis. Demographic, clinical and laboratory data of an ethnically diverse cohort of biopsy proven NAFLD patients were retrospectively analysed under univariate and multivariate analyses. Liver biopsies were scored using NASH clinical research network (NASH CRN) system. AUROC were performed for NAFLD Activity Score ≥5 (NASH) and fibrosis score ≥2 (advanced fibrosis). Two hundred and twenty-two patients were analysed. Fifty six (23%) had normal ALT. There was no difference in the rate of advanced fibrosis between normal and elevated ALT (26.8% vs. 18.1%, P = 0.19). However, significantly lower percentage of normal ALT group had NASH compared with elevated ALT group (10.7% vs. 28.9%, P ALT group had NASH or advanced fibrosis, whereas 53% of elevated ALT had no NASH or advanced fibrosis. Higher ALT values correlated with higher specificity, but lower sensitivity for both NASH and advanced fibrosis. AUROC for ALT level correlating NASH and advanced fibrosis were 0.62 and 0.46 respectively. There is no optimal ALT level to predict NASH and advanced fibrosis. Metabolic risk factors should be evaluated to select patients for a liver biopsy to confirm NASH and advanced fibrosis. © 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  6. Incontri di Fisica delle Alte Energie Italian Meeting on High Energy Physics Napoli

    CERN Document Server

    Carlino, Gianpaolo; Merola, Leonardo; Paolucci, Pierluigi; Ricciardi, Giulia; IFAE 2007

    2008-01-01

    This book collects the Proceedings of the Workshop "Incontri di Fisica delle Alte Energie (IFAE) 2007, Napoli, 11-13 April 2007". This is the sixth edition of a series of meetings on fundamental research in particle physics and was attended by about 160 researchers. Presentations, both theoretical and experimental, addressed the status of Physics of the Standard Model and beyond, Flavour phyisc, Neutrino and Astroparticle physics, new technology in high energy physics. Special emphasis was given to the expectations of the forthcoming Large Hadron Collider, due in operation at the end of 2007. The venue of plenary sessions interleaved with parallel ones allowed for a rich exchange of ideas, presented in these Proceedings, that form a coherent picture of the findings and of the open questions in this extremely challenging cultural field. The venue of plenary sessions interleaved with parallel ones allowed for a rich exchange of ideas, presented in these Proceedings, that form a coherent picture of the findings ...

  7. Twist-3 effect from the longitudinally polarized proton for ALT in hadron production from pp collisions

    Science.gov (United States)

    Koike, Yuji; Pitonyak, Daniel; Yoshida, Shinsuke

    2016-08-01

    We compute the contribution from the longitudinally polarized proton to the twist-3 double-spin asymmetry ALT in inclusive (light) hadron production from proton-proton collisions, i.e., p↑ p → → h X. We show that using the relevant QCD equation-of-motion relation and Lorentz invariance relation allows one to eliminate the twist-3 quark-gluon correlator (associated with the longitudinally polarized proton) in favor of one-variable twist-3 quark distributions and the (twist-2) transversity parton density. Including this result with the twist-3 pieces associated with the transversely polarized proton and unpolarized final-state hadron (which have already been calculated in the literature), we now have the complete leading-order cross section for this process.

  8. De l’Illusion de l’altérité

    Directory of Open Access Journals (Sweden)

    Buata Malela

    2008-02-01

    Full Text Available Dans L’Illusion de l’altérité, Bernard Mouralis nous propose une synthèse significative des études littéraires francophones d’Afrique. Son ouvrage rassemble près de quarante années de recherches et de réflexions sur ce qu’il appelle les littératures « négro-africaines ». Il offre une vision claire des progressions multiples de ces recherches, ce qui contribue à faire de ce livre un outil de référence pour se familiariser avec certaines thématiques et questions liées aux études littéraires d’A...

  9. Twist-3 effect from the longitudinally polarized proton for ALT in hadron production from pp collisions

    Directory of Open Access Journals (Sweden)

    Yuji Koike

    2016-08-01

    Full Text Available We compute the contribution from the longitudinally polarized proton to the twist-3 double-spin asymmetry ALT in inclusive (light hadron production from proton–proton collisions, i.e., p↑p→→hX. We show that using the relevant QCD equation-of-motion relation and Lorentz invariance relation allows one to eliminate the twist-3 quark-gluon correlator (associated with the longitudinally polarized proton in favor of one-variable twist-3 quark distributions and the (twist-2 transversity parton density. Including this result with the twist-3 pieces associated with the transversely polarized proton and unpolarized final-state hadron (which have already been calculated in the literature, we now have the complete leading-order cross section for this process.

  10. Diorganosilacetylene-alt-diorganosilvinylene polymers and a process densifying porous silicon-carbide bodies

    Science.gov (United States)

    Barton, Thomas J.; Ijadi-Maghsoodi, Sina; Pang, Yi

    1994-05-17

    The present invention provides linear organosilicon polymers including acetylene and vinylene moieties, and a process for their preparation. These diorganosilacetylene-alt-diorganosilvinylene linear polymers can be represented by the formula: --[--(R.sup.1)(R.sup.2)Si--C.tbd.C--(R.sup.3)(R.sup.4)Si--CH=CH--].sub.n-- , wherein n.gtoreq.2; and each R.sup.1, R.sup.2, R.sup.3, and R.sup.4 is independently selected from the group consisting of hydrogen, halogen, alkyl, alkenyl, aryl, and aralkyl radicals. The polymers are soluble in organic solvents, air stable, and can be pulled into fibers or cast into films. They can be thermally converted into silicon carbide ceramic materials.

  11. 2017-04-28_W88 ALT 370 Program Overview(OUO).

    Energy Technology Data Exchange (ETDEWEB)

    Daniels, Vonceil

    2017-04-01

    All major program milestones have been met and the program is executing within budget. The ALT 370 program achieved Phase 6.4 authorization in February of this year. Five component Final Design Reviews (FDRs) have been completed, indicating progress in finalizing the design and development phase of the program. A series of ground-based qualification activities have demonstrated that designs are meeting functional requirements. The first fully functional flight test, FCET-53, demonstrated end-to-end performance in normal flight environments in February. Similarly, groundbased nuclear safety and hostile environments testing indicates that the design meets requirements in these stringent environments. The first in a series of hostile blast tests was successfully conducted in April.

  12. Comparing the characteristics of highly cited titles and highly alted titles

    Energy Technology Data Exchange (ETDEWEB)

    Didegah, F.; Bowman, T.D.; Bowman, S.; Hartley, J.

    2016-07-01

    This study examines differences in the types of titles for articles that show high altmetric activity (highly alted articles) versus highly cited articles. This work expands on previous research on document titles in combination with a grounded theory approach to develop a codebook in which articles were manually coded based on 11 characteristics. The results show that there are differences and similarities in titles across many of the examined characteristics; highly cited titles and highly mentioned titles on Wikipedia have some similar characteristics such as they have the the highest percentage of substantive words; in addition, there are no or very few titles referencing outside or with humor/lightness on both platforms. Twitter and Facebook also showed some similarities having the highest percentage of humorous/light titles and lowest percentage of substantive words in their titles. (Author)

  13. Segmental dynamics of polyethylene-alt-propylene studied by NMR spin echo techniques

    Science.gov (United States)

    Lozovoi, A.; Mattea, C.; Hofmann, M.; Saalwaechter, K.; Fatkullin, N.; Stapf, S.

    2017-06-01

    Segmental dynamics of a highly entangled melt of linear polyethylene-alt-propylene with a molecular weight of 200 kDa was studied with a novel proton nuclear magnetic resonance (NMR) approach based upon 1H → 2H isotope dilution as applied to a solid-echo build-up function ISE(t), which is constructed from the NMR spin echo signals arising from the Hahn echo (HE) and two variations of the solid-echo pulse sequence. The isotope dilution enables the separation of inter- and intramolecular contributions to this function and allows one to extract the segmental mean-squared displacements in the millisecond time range, which is hardly accessible by other experimental methods. The proposed technique in combination with time-temperature superposition yields information about segmental translation in polyethylene-alt-propylene over 6 decades in time from 10-6 s up to 1 s. The time dependence of the mean-squared displacement obtained in this time range clearly shows three regimes of power law with exponents, which are in good agreement with the tube-reptation model predictions for the Rouse model, incoherent reptation and coherent reptation regimes. The results at short times coincide with the fast-field cycling relaxometry and neutron spin echo data, yet, significantly extending the probed time range. Furthermore, the obtained data are verified as well by the use of the dipolar-correlation effect on the Hahn echo, which was developed before by the co-authors. At the same time, the amplitude ratio of the intermolecular part of the proton dynamic dipole-dipole correlation function over the intramolecular part obtained from the experimental data is not in agreement with the predictions of the tube-reptation model for the regimes of incoherent and coherent reptation.

  14. Perception and demand for artificial reef by relevant local groups in Altınoluk (Turkey.

    Directory of Open Access Journals (Sweden)

    Sezgin Tunca

    2015-12-01

    Full Text Available Bu çalışmada, Türkiye Ulusal Yapay Resif Master Planı’nın pilot bölgesi olan Altınoluk’da yerleştirilmesi planlanan yapay resiflere olan talebin belirlenmesi amaçlanmaktadır. Çalışma kapsamında, Altınoluk’daki ticari balıkçılar, rekreasyonel balıkçılar ve yöre sakinleri ile yüz yüze görüşmeler gerçekleştirilmiştir. Veri toplama çalışmaları, yapay resif yerleştirmesi yapılmadan önce, Nisan, 2011’de tamamlanmıştır. Çalışma hedef gruplarının yapay resif algısı özel olarak tasarlanan 13 ifade Likert ölçeği yardımıyla analiz edilmiştir. Elde edilen sonuçlara göre, yapay resifler ile ilgili olumlu algı gözlenmiştir. Öte yandan, yapay resif yerleştirilmesi ile birlikte hem rekreasyonel balıkçılık (%158 artış hem de ticari balıkçılık (31% artış günlerinde artış tespit edilmiştir. Sonuç olarak, yapay resiflerle ilişkili grupların daha iyi anlaşılması ve yönetim süreçlerine dahil edilmesi yapay resiflerin etkinliği açısından önem taşımaktadır

  15. Effects of a subdermal levonorgestrel contraceptive implant (Norplant) on serum cholesterol, triglycerides, ALT and AST in Iranian women.

    Science.gov (United States)

    Taheri, Mohsen; Rahimi, Majid; Naderi, Mohammad; Ghavami, Saied; Mokhtari, Mojgan; Rashidi, Homaira; Hashemi, Mohammad

    2006-01-01

    To evaluate the effects of Norplant (36 mg of levonorgestrel, six capsules) on serum cholesterol, triglycerides, alanine transaminase (ALT) and aspartate transminase (AST), we enrolled 465 healthy women from Zahedan, Iran, into a longitudinal study. Blood samples were collected after an overnight fast before implant insertion and after 3, 6, 9 and 12 months of use. Total cholesterol and triglyceride levels did not significantly change during Norplant use. Although there were statistically significant increases in ALT and AST levels during Norplant use, the values were within the reference range.

  16. The effect of N-acetyl-l-cysteine and ascorbic acid on visible-light-irradiated camphorquinone/N,N-dimethyl-p-toluidine-induced oxidative stress in two immortalized cell lines.

    Science.gov (United States)

    Pagoria, D; Geurtsen, W

    2005-11-01

    Recent studies have revealed that visible-light (VL)-irradiated camphorquinone (CQ), in the presence of a tertiary amine (e.g., N,N-dimethyl-p-toluidine, DMT), generates initiating radicals that may indiscriminately react with molecular oxygen forming reactive oxygen species (ROS). In this study, the ability of the antioxidants N-acetyl-l-cysteine (NAC) and ascorbic acid (AA) to reduce intracellular oxidative stress induced by VL-irradiated CQ/DMT or VL-irradiated hydrogen peroxide (H(2)O(2)) was assessed in an immortalized Murine cementoblast cell line (OCCM.30) and an immortalized Murine fibroblast cell line, 3T3-Swiss albino (3T3). Intracellular oxidative stress was measured with the membrane permeable dye, 2',7'-dichlorodihydrofluorescein diacetate (H(2)DCF-DA). VL-irradiated CQ/DMT and VL-irradiated H(2)O(2) each produced significantly (p<0.001) elevated intracellular oxidative levels in both cell types compared to intracellular ROS levels in VL-irradiated untreated cells. OCCM.30 cementoblasts were found to be almost twice as sensitive to VL-irradiated CQ/DMT and VL-irradiated H(2)O(2) treatment compared to 3T3 fibroblasts. Furthermore, 10mm NAC and 10mm AA each eliminated oxidative stress induced by VL-irradiated CQ/DMT and VL-irradiated H(2)O(2) in both cell types. Our results suggest that NAC and AA may effectively reduce or eliminate oxidative stress in cells exposed to VL-irradiated CQ/DMT following polymerization.

  17. Can widely used cell type markers predict the suitability of immortalized or primary mammary epithelial cell models?

    Directory of Open Access Journals (Sweden)

    Edgar Corneille Ontsouka

    Full Text Available BACKGROUND: Mammary cell cultures are convenient tools for in vitro studies of mammary gland biology. However, the heterogeneity of mammary cell types, e.g., glandular milk secretory epithelial or myoepithelial cells, often complicates the interpretation of cell-based data. The present study was undertaken to determine the relevance of bovine primary mammary epithelial cells isolated from American Holstein (bMEC US or Swiss Holstein-Friesian (bMEC CH cows, and of primary bovine mammary alveolar epithelial cells stably transfected with simian virus-40 (SV-40 large T-antigen (MAC-T for in vitro analyses. This was evaluated by testing their expression pattern of cytokeratin (CK 7, 18, 19, vimentin, and α-smooth muscle actin (α-SMA. RESULTS: The expression of the listed markers was assessed using real-time quantitative PCR, flow cytometry and immunofluorescence microscopy. Characteristic markers of the mesenchymal (vimentin, myoepithelial (α-SMA and glandular secretory cells (CKs showed differential expression among the studied cell cultures, partly depending on the analytical method used. The relative mRNA expression of vimentin, CK7 and CK19, respectively, was lower (P < 0.05 in immortalized than in primary mammary cell cultures. The stain index (based on flow cytometry of CK7 and CK19 protein was lower (P < 0.05 in MAC-T than in bMECs, while the expression of α-SMA and CK18 showed an inverse pattern. Immunofluorescence microscopy analysis mostly confirmed the mRNA data, while partly disagreed with flow cytometry data (e.g., vimentin level in MAC-T. The differential expression of CK7 and CK19 allowed discriminating between immortal and primary mammary cultures. CONCLUSIONS: The expression of the selected widely used cell type markers in primary and immortalized MEC cells did not allow a clear preference between these two cell models for in vitro analyses studying aspects of milk composition. All tested cell models exhibited to a variable

  18. Energy Level Tuning of Poly(phenylene-alt-dithienobenzothiadiazole)s for Low Photon Energy Loss Solar Cells

    NARCIS (Netherlands)

    Heuvel, R.; van Franeker, J. J.; Janssen, R. A. J.

    2017-01-01

    Six poly(phenylene-alt-dithienobenzothiadiazole)-based polymers have been synthesized for application in polymer–fullerene solar cells. Hydrogen, fluorine, or nitrile substitution on benzo­thiadiazole and alkoxy or ester substitution on the phenylene moiety are investigated to reduce the energy loss

  19. Kas erivajadustega lapsed saavad õigel ajal abi? / Ene Mägi, Urve Raudsepp-Alt, Ale Sprenk, Peeter Aas

    Index Scriptorium Estoniae

    2009-01-01

    Küsimusele vastavad: Tallinna Ülikooli Kasvatusteaduste Instituudi eri- ja sotsiaalpedagoogika osakonna juhataja Ene Mägi, Tallinna Haridusameti üldhariduse osakonna peaspetsialist Urve Raudsepp-Alt, Krabi põhikooli direktor Ale Sprenk, Põlva Maavalitsuse haridus-, kultuuri- ja sotsiaalosakonna juhataja Peeter Aas

  20. Liver enzymes: interaction analysis of smoking with alcohol consumption or BMI, comparing AST and ALT to γ-GT.

    Directory of Open Access Journals (Sweden)

    Lutz P Breitling

    Full Text Available BACKGROUND: A detrimental interaction between smoking and alcohol consumption with respect serum γ-glutamyltransferase (γ-GT has recently been described. The underlying mechanisms remain unknown. The present work aimed to provide further insights by examining similar interactions pertaining to aspartate and alanine transaminase (AST, ALT, routine liver markers less prone to enzyme induction. METHODOLOGY/PRINCIPAL FINDINGS: The present cross-sectional analysis was based on records from routine occupational health examinations of 15,281 male employees predominantly of the construction industry, conducted from 1986 to 1992 in Southern Germany. Associations of smoking intensity with log-transformed activities of γ-GT, AST, and ALT were examined in regression models adjusted for potential confounders and including an interaction of smoking with alcohol consumption or body mass index (BMI. Statistically significant interactions of smoking were observed with both alcohol consumption (AST and ALT, each with P<0.0001 and BMI (AST only, P<0.0001. The interactions all were in the same directions as for γ-GT, i.e. synergistic with alcohol and opposite with BMI. CONCLUSION: The patterns of interaction between smoking and alcohol consumption or BMI with respect to AST and ALT resembled those observed for γ-GT. This renders enzyme induction a less probable mechanism for these associations, whereas it might implicate exacerbated hepatocellular vulnerability and injury.

  1. A combination of raised serum AST:ALT ratio and erythrocyte mean cell volume level detects excessive alcohol consumption.

    Science.gov (United States)

    Kawachi, I; Robinson, G M; Stace, N H

    1990-04-11

    The usefulness of the serum aspartate aminotransferase (AST): serum alanine aminotransferase (ALT) ratio as a guide to the presence of alcoholism was evaluated in four groups of patients. In alcoholics with elevated transaminases the mean AST:ALT ratio was found to be 1.50 (95% confidence interval (CI): 1.49-1.51), in hepatitis B infection 0.51 (95% CI: 0.50-0.52), in liver cancer 1.25 (95% CI: 1.20-1.29), and in nonmalignant obstructive jaundice 0.59 (95% CI: 0.57-0.61). In alcoholics with normal transaminases the AST:ALT ratio was 1.64 (95% CI: 1.61-1.67). The combination of an AST:ALT ratio of greater than 1.00 with an erythrocyte mean cell volume (MCV) above 90.0 fL resulted in a sensitivity of 97.3% and a specificity of 88.9% for detecting alcoholism in these four groups of patients.

  2. Liver enzymes: interaction analysis of smoking with alcohol consumption or BMI, comparing AST and ALT to γ-GT.

    Science.gov (United States)

    Breitling, Lutz P; Arndt, Volker; Drath, Christoph; Brenner, Hermann

    2011-01-01

    A detrimental interaction between smoking and alcohol consumption with respect serum γ-glutamyltransferase (γ-GT) has recently been described. The underlying mechanisms remain unknown. The present work aimed to provide further insights by examining similar interactions pertaining to aspartate and alanine transaminase (AST, ALT), routine liver markers less prone to enzyme induction. The present cross-sectional analysis was based on records from routine occupational health examinations of 15,281 male employees predominantly of the construction industry, conducted from 1986 to 1992 in Southern Germany. Associations of smoking intensity with log-transformed activities of γ-GT, AST, and ALT were examined in regression models adjusted for potential confounders and including an interaction of smoking with alcohol consumption or body mass index (BMI). Statistically significant interactions of smoking were observed with both alcohol consumption (AST and ALT, each with P<0.0001) and BMI (AST only, P<0.0001). The interactions all were in the same directions as for γ-GT, i.e. synergistic with alcohol and opposite with BMI. The patterns of interaction between smoking and alcohol consumption or BMI with respect to AST and ALT resembled those observed for γ-GT. This renders enzyme induction a less probable mechanism for these associations, whereas it might implicate exacerbated hepatocellular vulnerability and injury.

  3. ACTIVITIES OF ALT, AST, ALP AND LDH ENZYMES IN CATTLE BLOOD PLASMA DEPENDING ON REPRODUCTIVE CYCLE AND SEASONAL CHANGES

    Directory of Open Access Journals (Sweden)

    Nejra Hadžimusić

    2013-03-01

    Full Text Available The study was aimed at determining levels of alanin aminotransferase (ALT, aspartate aminotransferase (AST, alkaline phosphatase (ALP and lactate dehydrogenase (LDH in the plasma of 229 dairy Holstein-Friesian breed cows in different stages of the re¬production cycle and regarding the seasonal changes (diet conditions. Cows were divided into the groups: lactation (Group A, postpartal period within 15 days from calving (Group B and dry period (Group C. Seasonal variations affected AST activity within Groups A and B, while Group C showed no seasonal influence. Differences between Groups A and B were determined during winter, as well as among Groups A and C during the same season. The highest activity of ALT was measured in cows from Group A during the winter season. ALP activity showed no statistical difference among groups, though statistical significance was noted within seasonal changes. The activity of LDH showed no statistical significance within Group C with regard to the seasonal changes. Statistically significant difference was found between cows of Group A and C during the winter season. Differences were considered statistically significant when p<0,05.Research results showed a significant influence of milk production and dry period on the activities of measured enzymes in the blood plasma so we can conclude that there is a need for constant monitoring of the described parameters during the production period.Key words: dairy cows, ALT, AST, ALT, LDH

  4. Naised käsu korras firmade juhatusse! Jah või ei? / Kairi Alt, Aveli Kippari, Karl Koort... [jt.

    Index Scriptorium Estoniae

    2011-01-01

    Küsimusele vastavad Columbus IT Partner Eesti AS personalijuht Kairi Alt, Stele ja Riveli omanik Aveli Kippari, AS Panaviatic'i turundusjuht Karl Koort, OÜ Tarkvara Tehnoloogia Arenduskeskuse tegevjuht Indrek Vainu, AS PricewaterhouseCoopers auditiosakonna direktor Eva Jansen, Kalev Chocolate Factory tootmisjuht Hardo Reinike

  5. Immortalization of CD4(+) and CD8(+) T lymphocytes by human T-cell leukemia virus type 1 Tax mutants expressed in a functional molecular clone.

    Science.gov (United States)

    Robek, M D; Ratner, L

    1999-06-01

    The human T-cell leukemia virus type 1 (HTLV-1) transcriptional trans-activator Tax has been demonstrated to have transforming activity in multiple cell culture and transgenic-mouse models. In addition to activating transcription from the viral long terminal repeat (LTR) through the cyclic AMP response element binding protein/activating transcription factor (CREB/ATF) family of transcription factors, Tax activates the expression of multiple cellular promoters through the NF-kappaB pathway of transcriptional activation. The Tax mutants M22 and M47 have previously been demonstrated to selectively abrogate the ability of Tax to activate transcription through the NF-kappaB or CREB/ATF pathway, respectively. These mutations were introduced in the tax gene of the ACH functional molecular clone of HTLV-1, and virus produced from the mutant ACH clones was examined for the ability to replicate and immortalize primary human lymphocytes. While virus derived from the clone containing the M47 mutation retained the ability to immortalize T lymphocytes, the M22 mutant lost the ability to immortalize infected cells. These results indicate that activation of the CREB/ATF pathway by Tax is dispensable for the immortalization of T cells by HTLV-1, whereas activation of the NF-kappaB pathway may be critical.

  6. Perspectives on Immortality and Eternal Life in the Book of Wisdom and the Gospel of John: A Conceptual Analysis Based on Metaphorical Structuring

    NARCIS (Netherlands)

    Valentin, R.I.

    2016-01-01

    The similarities between the Fourth Gospel and wisdom literature have been addressed by many scholars. This study is also intended to continue along this line by looking at the particular link between the Wisdom of Solomon and the Gospel of John with a specific focus on the concepts of immortality

  7. Immortalization of T-Cells Is Accompanied by Gradual Changes in CpG Methylation Resulting in a Profile Resembling a Subset of T-Cell Leukemias

    Directory of Open Access Journals (Sweden)

    Sofie Degerman

    2014-07-01

    Full Text Available We have previously described gene expression changes during spontaneous immortalization of T-cells, thereby identifying cellular processes important for cell growth crisis escape and unlimited proliferation. Here, we analyze the same model to investigate the role of genome-wide methylation in the immortalization process at different time points pre-crisis and post-crisis using high-resolution arrays. We show that over time in culture there is an overall accumulation of methylation alterations, with preferential increased methylation close to transcription start sites (TSSs, islands, and shore regions. Methylation and gene expression alterations did not correlate for the majority of genes, but for the fraction that correlated, gain of methylation close to TSS was associated with decreased gene expression. Interestingly, the pattern of CpG site methylation observed in immortal T-cell cultures was similar to clinical T-cell acute lymphoblastic leukemia (T-ALL samples classified as CpG island methylator phenotype positive. These sites were highly overrepresented by polycomb target genes and involved in developmental, cell adhesion, and cell signaling processes. The presence of non-random methylation events in in vitro immortalized T-cell cultures and diagnostic T-ALL samples indicates altered methylation of CpG sites with a possible role in malignant hematopoiesis.

  8. Maintenance of vascular endothelial cell-specific properties after immortalization with an amphotrophic replication-deficient retrovirus containing human papilloma virus 16 E6/E7 DNA

    NARCIS (Netherlands)

    Fontijn, R.; Hop, C.; Brinkman, H. J.; Slater, R.; Westerveld, A.; van Mourik, J. A.; Pannekoek, H.

    1995-01-01

    Primary human vascular endothelial cells were immortalized by the integration of a single DNA copy of an amphotrophic, replication-deficient retrovirus containing the E6/E7 genes of human papilloma virus. To date, the resulting cell lines, designated EC-RF7 and EC-RF24, have been cultured for more

  9. The influence of magnesium on the activity of some enzymes (AST, ALT, ALP) and lead content in some tissues.

    Science.gov (United States)

    Todorovic, Tatjana; Vujanovic, Dragana

    2002-12-01

    Many authors in different studies have reported the antagonism between Mg and Pb. Our previous results suggested that oral Mg treatment have better effect on investigation biochemical parameters (protoporphyrins, aminolevulinic acid--ALA and d-aminolevulinic dehydratase ALA-D) used in evaluating Pb intoxication, then CaNa2EDTA, chelation agents, currently used in therapy of Pb intoxication. The toxic effect of Pb induced considerably modifies the activity of many other enzymes. In this work we have examined the influence of Mg (as alternative therapy of Pb poisoning) on enzymes activity--biochemical markers for general health conditions--aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) in condition of lead intoxication. Many studies showed disturbances of activity ALT, AST and ALP. The aim of this study was to confirm positive effects of Mg intake in condition of such intoxication at the level on activity of investigated enzymes. The experiment was performed on 45 male Wister rats, divided in three groups. I--control group; II--group treated daily for 30 days with 100 mg Pb, per kg body weight and next 60 without Pb treatment (spontaneous detoxication); III group--the same treatment as II group for the first 30 days, but next 60 days rats were treated orally with 40 mg Mg/kg body weight. Activity of AST and ALT was significant increased in condition of Pb poisoning, but ALP activity was significant reduced. Influence of excessive oral Mg treatment was positive: decrease of AST activity and ALT activity, which was probably in correlation with significant elimination of Pb from liver and increase of ALT enzyme activity at the normal level.

  10. The endless writing exercise. Multiplied characters and wishes of immortality in the short story production of Carlos Liscano

    Directory of Open Access Journals (Sweden)

    Giuseppe Gatti Riccardi

    2017-12-01

    La historia interminable maintains a thematic continuity with Liscano’s entire narrative production with the leitmotiv of how to match life with writing and does so in two ways. First it creates a series of characters who in turn multiply infinitely, which can be read as reflecting a human need to create a potentially infinite literary corpus as means of attaining “immortality through writing”. Second, the short story offers a reflection on the human obsession with absolute knowledge and the desire to know everything and to narrate everything and indeed seems to present a critical re-elaboration of this obsession; in the story, the identity of the writer dissipates after his death, precisely because nobody can cover all his work. Writing, in this case, becomes merely a “store of experiences” that no one can use.

  11. Man should not let death attain the dominion of his thoughts: An Essay on Subjectivity, Self-Preservation and Immortality

    Directory of Open Access Journals (Sweden)

    Kasper Lysemose

    2010-12-01

    Full Text Available Mortality seems to have no place in the theories on subjectivity in Kant and Husserl. It is suggested that this entente cordiale is an expression of the shared principle at the heart of their philosophy, i.e. the principle of selfpreservation. Self-preservation is a principle that in a certain sense excludes mortality. It is argued that the primary sense of this exclusion is not theoretical but practical. Kant and Husserl are both endorsing the imperative that man should not let death attain the dominion of his thoughts (cf. Mann 1976, 600. The positive correlate to this is to be found in the demand that man should think of himself as if he was immortal. With special regard to Husserl the predicament that accompanies his relentless attempt to fulfill this demand is described as a sluice through which anthropology threatens to flow into the phenomenological enterprise.

  12. A telomerase immortalized human proximal tubule cell line with a truncation mutation (Q4004X in polycystin-1.

    Directory of Open Access Journals (Sweden)

    Brittney-Shea Herbert

    Full Text Available Autosomal dominant polycystic kidney disease (ADPKD is associated with a variety of cellular phenotypes in renal epithelial cells. Cystic epithelia are secretory as opposed to absorptive, have higher proliferation rates in cell culture and have some characteristics of epithelial to mesenchymal transitions. In this communication we describe a telomerase immortalized cell line that expresses proximal tubule markers and is derived from renal cysts of an ADPKD kidney. These cells have a single detectable truncating mutation (Q4004X in polycystin-1. These cells make normal appearing but shorter cilia and fail to assemble polycystin-1 in the cilia, and less uncleaved polycystin-1 in membrane fractions. This cell line has been maintained in continuous passage for over 35 passages without going into senescence. Nephron segment specific markers suggest a proximal tubule origin for these cells and the cell line will be useful to study mechanistic details of cyst formation in proximal tubule cells.

  13. Artichoke compound cynarin differentially affects the survival, growth and stress response of normal, immortalized and cancerous human cells

    DEFF Research Database (Denmark)

    Gezer, Ceren; Yücecan, Sevinç; Rattan, Suresh Inder Singh

    2015-01-01

    of CYN on the proliferative potential, survival, morphology, and stress response (SR) markers haemoxygenase-1 (HO-1) and heat shock protein-70 (HSP70) in normal human skin fibroblasts (FSF-1), telomerase-immortalized mesenchymal stem cells (hTERT-MSC) and cervical cancer cells, HeLa. Effects of CYN....... Furthermore, CYN induced oxidative SR marker HO-1 in both fibroblasts and stem cells in a biphasic manner, but a slight induction of HSP70 was observed only in the stem cells. Thus, CYN may be useful as a protection against the growth and survival of potentially cancerous cells and may promote longevity...... of normal cells by induction of SR proteins. Further advanced researches related with CYN and artichoke are recommended....

  14. [Low ALT activity amongst patients hospitalized in internal medicine wards is a widespread phenomenon associated with low vitamin B6 levels in their blood].

    Science.gov (United States)

    Ramati, Erez; Israel, Ariel; Tal Kessler; Petz-Sinuani, Naama; Sela, Ben-Ami; Idan Goren; Grinfeld, Atar; Lavi, Bruno; Segal, Gad

    2015-02-01

    Monitoring the activity of ALT (SGPT) in the blood is part of the routine, clinical-laboratory follow-up in hospitalized patients. In most cases, activity levels which are above the normal range are considered pathology, mostly related to lysis of hepatocytes, as in cases of hepatitis. Little has been investigated and published in regard to cases in which the ALT activity level is lower than normal. Since normal ALT activity is regarded essential for normal metabolism and homeostasis, we decided to evaluate the extent to which low ALT levels are found in healthy and hospitalized patient populations and to characterize its circumstances and etiology. Furthermore, we measured the blood concentration of vitamin B6 (being the source for the ALT co-factor, Pyridoxal-5-Phosphate) in a random sample of patients with lower than normal ALT activity level. The results of the current study showed a high prevalence, exceeding a third of hospitalized patients in internal medicine departments have low levels of ALT in the serum, and that a linear correlation (p = 0.0004, r = 0.47) exists between lower than normal ALT activity and low concentrations of vitamin B6 in the serum. The authors attribute these findings to a high prevalence of frailty amongst hospitalized patients. We aim to conduct further investigations intended to better characterize quantifiable parameters of frailty amongst our patient population.

  15. Association between ALT level and the rate of cardio/cerebrovascular events in HIV-positive individuals: the D: A: D study

    NARCIS (Netherlands)

    Sabin, Caroline A.; Ryom, Lene; Kovari, Helen; Kirk, Ole; de Wit, Stephane; Law, Matthew; Reiss, Peter; Dabis, Francois; Pradier, Christian; El-Sadr, Wafaa; Monforte, Antonella d'Arminio; Kamara, David; Phillips, Andrew N.; Lundgren, Jens D.

    2013-01-01

    An inverse association between serum alanine aminotransferase (ALT) levels and the risk of myocardial infarction (MI) has been reported in the general population. We investigated associations between ALT levels and the risk of various cardiovascular and cerebrovascular outcomes in a large cohort

  16. Self-healing pH-sensitive poly[(methyl vinyl ether)-alt-(maleic acid)]-based supramolecular hydrogels formed by inclusion complexation between cyclodextrin and adamantane.

    Science.gov (United States)

    Ma, Xiaoe; Zhou, Naizhen; Zhang, Tianzhu; Hu, Wanjun; Gu, Ning

    2017-04-01

    Self-healing materials are of interest for drug delivery, cell and gene therapy, tissue engineering, and other biomedical applications. In this work, on the base of biocompatible polymer poly(methyl vinyl ether-alt-maleic acid) (P(MVE-alt-MA)), host polymer β-cyclodextrin-grafted P(MVE-alt-MA) (P(MVE-alt-MA)-g-β-CD) and guest polymer adamantane-grafted P(MVE-alt-MA) (P(MVE-alt-MA)-g-Ad) were first prepared. Then through taking advantage of the traditional host-guest interaction of β-cyclodextrin and adamantane, a novel self-healing pH-sensitive physical P(MVE-alt-MA)-g-β-CD/P(MVE-alt-MA)-g-Ad supramolecular hydrogels were obtained after simply mixing the aqueous solution of host polymer and guest polymer. This kind of supramolecular hydrogels not only possess pH-sensitivity, but also possess the ability to repair themselves after being damaged. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. TNF-α-induced IL-6 and MMP-9 expression in immortalized ameloblastoma cell line established by hTERT.

    Science.gov (United States)

    Ohta, K; Naruse, T; Ishida, Y; Shigeishi, H; Nakagawa, T; Fukui, A; Nishi, H; Sasaki, K; Ogawa, I; Takechi, M

    2017-03-01

    Ameloblastoma (AM) shows locally invasive behaviour. However, biological investigations regarding regulation of gene expression associated with AM pathological features are difficult to perform, because AM cells can be passaged for a few generations due to senescence. We report a newly established immortalized AM cell line, AMB cells, by transfection with human telomerase reverse transcriptase (hTERT). Furthermore, we examined whether TNF-α modulates bone resorption-related genes, IL-6 and MMP-9 in cooperation with TGF-β or IFN-γ. Following transfection of an hTERT expression vector into AM cells using a non-viral method, the effects of cytokines on the expressions of IL-6 and MMP-9 mRNA were examined using real-time PCR. TNF-α-induced NF-κB activity was examined by western blotting and transcription factor assays. AMB cells continued to grow for more than 100 population doublings. Stimulation with TNF-α increased IL-6 and MMP-9 mRNA expressions, as well as NF-κB activation. Furthermore, TGF-β and IFN-γ dramatically increased TNF-α-mediated expressions of MMP-9 and IL-6 mRNA, respectively, while those responses were suppressed by NF-κB inhibitor. We established an immortalized AM cell line by hTERT transfection. TNF-α-mediated regulation of MMP-9 and IL-6 via NF-κB may play an important role in the pathological behaviour of AMs, such as bone resorption. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  18. Cerebrospinal fluid oxytocin correlated with peripheral ALT and AST in Chinese female subjects.

    Science.gov (United States)

    Yu, Yang; Kang, Yimin; Qiu, Guoshi; Lin, Hong; Xu, Jinzhong; Wang, Xiaofang; Xie, Longteng; Jiang, Yongsheng; Jia, Baofu; Wang, Pengxiang; Wang, Geng; Li, Qiujun; Yang, Xiaoyu; Zuo, Wei; Li, Cunbao

    2015-12-01

    Oxytocin (OT) is primarily synthesized in the paraventricular nucleus of the hypothalamus and supraoptic nucleus of the hypothalamus in the central nervous system and exhibits a wide spectrum of central and peripheral activities. OT is involved in lipid metabolism and glucose homeostasis and plays a protective role against liver damage. In this study, we investigated whether CSF OT levels correlates with peripheral glucose, lipid profiles, and/or liver enzymes in Chinese subjects. Sixty-nine subjects (n=36 males; n=33 females) who were recruited from Beijing Jishuitan Hospital participated in the study. Their levels of CSF OT and peripheral parameters were assayed by radioimmunoassay and continuous monitoring assay, respectively. There was no significant difference in CSF OT levels between males (53.09±6.88 nmol/mL) and females (52.34±6.87 nmol/mL), and no correlation found between CSF OT levels and peripheral glucose and lipid profiles. Significant negative correlation was observed between CSF OT levels and peripheral ALT and AST concentration in females but not in males. Our results support the physiological role of neuropeptides acting on brain sites to regulate liver enzymes, and shed new light on the brain-liver interaction.

  19. Cleaning of metal-contaminated graphite tiles of the TEXTOR ALT-II pump limiter blades

    Energy Technology Data Exchange (ETDEWEB)

    Dippel, K.H.; Kohlhaas, W.; Stickelmann, C. (Kernforschungsanlage Juelich GmbH (Germany, F.R.). Inst. fuer Plasmaphysik); Wallura, E. (Kernforschungsanlage Juelich GmbH (Germany, F.R.). Inst. fuer Reaktorwerkstoffe)

    Graphite is the preferred material for plasma facing components in today's fusion experiments and it is a prime candidate material also for coming fusion devices. At least in the existing tokamaks it cannot be excluded that the graphite surface will become contaminated, e.g. with metals, which leads to undesired effects on the plasma. Instead of an expensive exchange for new graphite parts, an appropriate cleaning method should be applied. This paper describes comparative tests of different mechanical cleaning methods applied to metal contaminated graphite tiles of the ALT-II toroidal belt pump limiter in the TEXTOR tokamak. Scanning electron microscope techniques have been used to characterize the cleaning efficiency. Best results have been obtained by sandblasting the graphite surface with boron carbide grains, which proves to be a very effective, inexpensive and fast cleaning method. After reassembly of the tiles, in-situ tests by exposure to tokamak discharges showed undetectably low ({le}10{sup 5}) metal concentrations in the TEXTOR plasma. (orig.).

  20. Flexible Polyimide Aerogel Cross-linked by Poly(maleic Anhydride-alt-alkylene)

    Science.gov (United States)

    Guo, Haiquan; Meador, Mary Ann B.; Wilkewitz, Brittany Marie

    2014-01-01

    Aerogels are potential materials for aerospace applications due to their lower thermal conductivity, lighter weight, and low dielectric constant. However, silica aerogels are restricted due to their inherent fragility, hygroscopic nature, and poor mechanical properties, especially in extreme aerospace environments. In order to fit the needs of aerospace applications, developing new thermal insulation materials that are flexible, and moisture resistant is needed. To this end, we fabricated a series of polyimide aerogels crosslinked with different poly(maleic anhydride-alt-alkylene)s as seen in Scheme 1. The polyimide oligomers were made with 3,3,4,4-biphenyltetracarboxylic dianhydride (BPDA), and different diamines or diamine combinations. The resulting aerogels have low density (0.06 gcm3 to 0.16 gcm3) and high surface area (240-440 m2g). The effect of the different backbone structures on density, shrinkage, porosity, surface area, mechanical properties, moisture resistance and thermal properties will be discussed. These novel polyalkylene-imide aerogels may be potential candidates for applications such as space suit insulation for planetary surface missions, insulation for inflatable structures for habitats, inflatable aerodynamic decelerators for entry, descent and landing (EDL) operations, and cryotank insulation for advance space propulsion systems. Scheme 1. Network of polyimide aerogels crosslinked with deifferent poly(maleic anhydride).

  1. Plasma influence on throat conductance of the TEXTOR pump limiter ALT-I

    International Nuclear Information System (INIS)

    Hardtke, A.; Finken, K.H.; Reiter, D.; Dippel, K.H.; Goebel, D.M.; McGrath, R.T.; Sagara, A.

    1989-01-01

    On the TEXTOR pump limiter ALT-I conductance measurements for the backstreaming of gas from the pump limiter vessel through the pump limiter entrance have been performed. In these experiments neutral gas has been injected into the pump limiter plenum during a short pulse. The influence of the instreaming plasma results in a reduction of the conductance of the outstreaming gas. For helium the conductance is reduced to about 40% of the molecular conductance when a plasma flux of 0.8 A/cm 2 (T e =T i =11 eV) is streaming into the pump limiter throat. The reduction of the conductance for backstreaming hydrogen and deuterium under the same plasma conditions is smaller; about 70% of the molecular conductance is obtained. This reduction can be explained by an increased recycling of ions which have been produced in the throat back to the neutralizer plate. The experimental results can be reproduced by Monte Carlo neutral transport code calculations if the recycling coefficient is about 0.85 for hydrogen and deuterium and about 0.95 for helium ions. Processes causing these high recycling coefficients are discussed and their influence is estimated. (orig.)

  2. Change in ALT levels after administration of HMG-CoA reductase inhibitors to subjects with pretreatment levels three times the upper normal limit in clinical practice.

    Science.gov (United States)

    Kim, Hyunah; Lee, Hyeseon; Kim, Tong Min; Yang, So Jung; Baik, Seo Yeon; Lee, Seung-Hwan; Cho, Jae-Hyoung; Lee, Hyunyong; Yim, Hyeon Woo; Choi, In Young; Yoon, Kun-Ho; Kim, Hun-Sung

    2018-02-21

    Prescription of statins to patients with chronic liver disease whose alanine transaminase (ALT) is over three times the upper normal limit (UNL) is not recommended. In this study, we attempted to evaluate patients with baseline ALT levels > 3 ×  UNL who were prescribed statins without ethical problems, using electronic medical records. We enrolled subjects with ALT levels > 3 ×  UNL. The patients were divided into three groups consisting of those who had been taking agents affecting liver function (HEPA) and continued to do so after the statin prescription (HepCon), those who had not previously taken HEPA and began doing so after statin prescription (HepNew), and those who had never taken HEPA (HepNo). All ALT levels were determined within 3 months of statins administration, and changes were monitored. From January 2009 to December 2012, 61 patients with baseline ALT levels > 3 ×  UNL were prescribed statins for the first time. During the 3-month ALT monitoring, levels of the HepCon, HepNew, and HepNo groups decreased by 45 ± 8%, 64 ± 10%, and 42 ± 8%, respectively; however, intergroup differences were not significant (P = .386). All the subjects who were administered statins showed improvement or maintained their ALT levels, except for two subjects, which showed deterioration. However, the ALT levels of the two subjects subsequently remained stable. It is not clear whether it is safe to prescribe statins to patients with ALT > 3 times the UNL. Our study showed that prescription of statins in combination with HEPA did not cause deleterious effects, suggesting that ALT levels > 3 times the UNL do not have harmful effects. © 2018 John Wiley & Sons Ltd.

  3. Simple synthesis of P(Cbz-alt-TBT) and PCDTBT by combining direct arylation with suzuki polycondensation of heteroaryl chlorides.

    Science.gov (United States)

    Lombeck, Florian; Matsidik, Rukiya; Komber, Hartmut; Sommer, Michael

    2015-01-01

    Direct arylation (DA) of 2-chlorothiophene and 2-chloro-3-hexylthiophene with 4,7-dibromo-2,1,3-benzothiadiazole is used to synthesize 4,7-bis(5-chloro-2-thienyl)-2,1,3-benzothiadiazole (TBTCl2) and 4,7-bis(5-chloro-4-hexyl-2-thienyl)-2,1,3-benzothiadiazole (DH-TBTCl2) in one step. Suitable conditions of the Suzuki polycondensations (SPC) of TBTCl2 and DH-TBTCl2 with the carbazole comonomer CbzPBE2 are established, furnishing PCDTBT and P(Cbz-alt-TBT) with high molecular weight and yield. Compared with control samples made from the corresponding dibromides, high-temperature NMR and UV-vis spectroscopy indicate similar properties for PCDTBT but an increased content of Cbz-Cbz homocouplings for P(Cbz-alt-TBT). © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. The immunomodulatory, antitumor and antimetastatic responses of melanoma-bearing normal and alcoholic mice to sunitinib and ALT-803: a combinatorial treatment approach.

    Science.gov (United States)

    Gaither, Kari A; Little, Alexander A; McBride, Alisha A; Garcia, Savanna R; Brar, Kiranjot K; Zhu, Zhaohui; Platt, Amity; Zhang, Faya; Meadows, Gary G; Zhang, Hui

    2016-09-01

    ALT-803, a novel IL-15/IL-15 receptor alpha complex, and the tyrosine kinase inhibitor, sunitinib, were examined for their single and combined effects on the growth of subcutaneous B16BL6 melanoma and on lymph node and lung metastasis. The study was conducted in immunocompetent C57BL/6 mice drinking water (Water mice) and in mice that chronically consumed alcohol (Alcohol mice), which are deficient in CD8(+) T cells. Sunitinib inhibited melanoma growth and was more effective in Alcohol mice. ALT-803 did not alter tumor growth or survival in Water or Alcohol mice. Combined ALT-803 and sunitinib inhibited melanoma growth and increased survival, and these effects were greater than sunitinib alone in Water mice. ALT-803 and alcohol independently suppressed lymph node and lung metastasis, whereas sunitinib alone or in combination with ALT-803 increased lymph node and lung metastasis in Water and Alcohol mice. Initially, ALT-803 increased IFN-γ-producing CD8(+)CD44(hi) memory T cells and CD8(+)CD44(hi)CD62L(lo) effector memory T cells and sunitinib decreased immunosuppressive MDSC and T regulatory cells (Treg). However, the impact of these treatments diminished with time. Subcutaneous tumors from Water mice showed increased numbers of CD8(+) T cells, CD8(+)CD44(hi) T cells, NK cells, and MDSC cells and decreased Treg cells after ALT-803 treatment.

  5. CASUÍSTICA DE ALTE NUM HOSPITAL TERCIÁRIO – O QUE MUDOU NOS ÚLTIMOS 5 ANOS?

    Directory of Open Access Journals (Sweden)

    Daniel Meireles

    2016-07-01

    Conclusões: O ALTE cursa frequentemente com evolução favorável. Apesar de não haver recomendação ou consensos na literatura para a investigação etiológica, esta mantém-se uma prática comum, sem aparente modificação na abordagem nos últimos anos, em comparação com estudo prévio realizado em 2009.

  6. Clinico-biochemical factors to early predict biliary etiology of acute pancreatitis: age, female gender, and ALT.

    Science.gov (United States)

    Zarnescu, N O; Costea, R; Zarnescu Vasiliu, E C; Neagu, S

    2015-01-01

    Background/ Aims: Despite the existence of an easy tool to diagnose biliary tract disease as an etiology for acute pancreatitis (AP), the sensitivity of abdominal ultrasound is around 80%, which can be even lower in certain conditions. We have retrospectively reviewed data of 146 patients admitted for acute pancreatitis between 1999 and 2013. Bivariate analysis for clinical and biochemical variables was performed with respect to etiology of AP (biliary versus non-biliary). Multivariate analysis was performed by using binary logistic regression. There were 87 males (59.6%) and 59 females (40.4%), with a median age of 51. The etiology of acute pancreatitis was biliary in 71 patients (48.6%). Bivariate analysis found the following as significant association (p=0.001) with biliary pancreatitis: older age, female gender, and elevated AST, ALT. A binary logistic regression analysis identified as predictor factors for biliary etiology of acute pancreatitis: age OR = 1.031 (95% CI 1.004 - 1.059, p = 0.024), sex (female) OR = 2.34 (95% CI 1.022 - 5.359, p = 0.044) and ALT OR = 1.004 (95% CI 1.001 - 1.007, p =0.004). The two clinical scores included the three variables (A.S.ALT scores) in categorical format were generated and then checked with the ROC curves (areas under curve are 0.768 and 0.778). Age, female gender, and elevated ALT can help identifying cases with biliary etiology of acute pancreatitis.

  7. Unambiguous Tracking Method Based on Combined Correlation Functions for sine/cosine-BOC CBOC and AltBOC Modulated Signals

    OpenAIRE

    J.W. Ren; G.T. Yang; W.M. Jia; M.L. Yao

    2014-01-01

    Unambiguous tracking for Binary Offset Carrier (BOC) modulated signals is an important requirement of modern Global Navigation Satellite System (GNSS) receivers. An unambiguous tracking method based on combined correlation functions for even/odd order sine/cosine-BOC, Composite BOC(CBOC) and Alternate BOC(AltBOC) modulated signals is proposed. Firstly, a unitary mathematical formulation for all kinds of BOC modulations is introduced. Then an unambiguous tracking method is proposed based on th...

  8. The IL-15-Based ALT-803 Complex Enhances FcγRIIIa-Triggered NK Cell Responses and In Vivo Clearance of B Cell Lymphomas.

    Science.gov (United States)

    Rosario, Maximillian; Liu, Bai; Kong, Lin; Collins, Lynne I; Schneider, Stephanie E; Chen, Xiaoyue; Han, Kaiping; Jeng, Emily K; Rhode, Peter R; Leong, Jeffrey W; Schappe, Timothy; Jewell, Brea A; Keppel, Catherine R; Shah, Keval; Hess, Brian; Romee, Rizwan; Piwnica-Worms, David R; Cashen, Amanda F; Bartlett, Nancy L; Wong, Hing C; Fehniger, Todd A

    2016-02-01

    Anti-CD20 monoclonal antibodies (mAb) are an important immunotherapy for B-cell lymphoma, and provide evidence that the immune system may be harnessed as an effective lymphoma treatment approach. ALT-803 is a superagonist IL-15 mutant and IL-15Rα-Fc fusion complex that activates the IL-15 receptor constitutively expressed on natural killer (NK) cells. We hypothesized that ALT-803 would enhance anti-CD20 mAb-directed NK-cell responses and antibody-dependent cellular cytotoxicity (ADCC). We tested this hypothesis by adding ALT-803 immunostimulation to anti-CD20 mAb triggering of NK cells in vitro and in vivo. Cell lines and primary human lymphoma cells were utilized as targets for primary human NK cells. Two complementary in vivo mouse models were used, which included human NK-cell xenografts in NOD/SCID-γc (-/-) mice. We demonstrate that short-term ALT-803 stimulation significantly increased degranulation, IFNγ production, and ADCC by human NK cells against B-cell lymphoma cell lines or primary follicular lymphoma cells. ALT-803 augmented cytotoxicity and the expression of granzyme B and perforin, providing one potential mechanism for this enhanced functionality. Moreover, in two distinct in vivo B-cell lymphoma models, the addition of ALT-803 to anti-CD20 mAb therapy resulted in significantly reduced tumor cell burden and increased survival. Long-term ALT-803 stimulation of human NK cells induced proliferation and NK-cell subset changes with preserved ADCC. ALT-803 represents a novel immunostimulatory drug that enhances NK-cell antilymphoma responses in vitro and in vivo, thereby supporting the clinical investigation of ALT-803 plus anti-CD20 mAbs in patients with indolent B-cell lymphoma. ©2015 American Association for Cancer Research.

  9. AST/ALT ratio is not an index of liver fibrosis in chronic hepatitis C when aminotransferase activities are determinate according to the international recommendations.

    Science.gov (United States)

    Guéchot, Jérôme; Boisson, Renée Claude; Zarski, Jean-Pierre; Sturm, Nathalie; Calès, Paul; Lasnier, Elisabeth

    2013-11-01

    The aspartate aminotransferase activity (AST)/alanine aminotransferase activity (ALT) ratio is used as liver fibrosis index whereas the reported data are conflicting. In chronic hepatitis C (CHC), reported diagnostic accuracies range from none to good for significant fibrosis and to excellent for cirrhosis. Assuming that AST/ALT increases are mainly due to vitamin B6 defects since pyridoxal phosphate (PLP), active form of B6, acts as coenzyme in transamination reactions, we evaluated the diagnostic accuracy of the AST/ALT ratio using standardized methods for AST and ALT activities, with PLP addition as recommended, in a prospective multicenter cohort of CHC patients. ALT and AST activities were measured using the recommended IFCC methods with addition of pyridoxal 5'-phosphate. We evaluated the AST/ALT ratio for the diagnosis of liver fibrosis or cirrhosis in a cohort of CHC patients included in a multicenter prospective study. A liver biopsy was performed in each patient and reviewed by two independent pathologists in order to determine the fibrosis stage according to Metavir classification which was the reference standard. AST/ALT ratio significantly increased with histological stage of liver fibrosis and there was a significant correlation between Metavir fibrosis stage and AST/ALT ratio (r=0.129, PALT ratio does not discriminate significant fibrosis (F≥2) (AUROC=0.531) and had only very poor diagnostic accuracies for severe fibrosis (F≥3) (AUROC=0.584) or cirrhosis (F4) (AUROC=0.626). AST/ALT ratio is not a good and discriminative index of liver fibrosis in CHC when aminotransferase activities are determinate according to the international recommendations. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

  10. ZnO nanoparticles augment ALT, AST, ALP and LDH expressions in C2C12 cells.

    Science.gov (United States)

    Pandurangan, Muthuraman; Kim, Doo Hwan

    2015-11-01

    The present study aimed to investigate the effect of ZnO nanoparticles on alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) enzyme expressions in C2C12 cells. ZnO nanoparticles are widely used in the several cosmetic lotions and other biomedical products. Several studies report on ZnO nanoparticle mediated cytotoxicity. However, there are no reports on the effect of ZnO nanoparticles on ALT, AST, ALP and LDH enzyme expressions in C2C12 cells. A cytotoxicity assay was carried out to determine the effect of ZnO nanoparticles (1-5 mg/ml) on C2C12 cell viability at 48 and 72 h. ZnO nanoparticles increased ALT, AST, ALP and LDH enzyme mRNA expression and their activities in C2C12 cells. In conclusion, the present study showed that ZnO nanoparticles increased these enzyme activities and its mRNA expression in C2C12 cells in a dose-dependent manner.

  11. Differences in levels of albumin, ALT, AST, γ-GT and creatinine in frail, moderately healthy and healthy elderly individuals.

    Science.gov (United States)

    Edvardsson, Maria; Sund-Levander, Märtha; Milberg, Anna; Wressle, Ewa; Marcusson, Jan; Grodzinsky, Ewa

    2018-02-23

    Reference intervals are widely used as decision tools, providing the physician with information about whether the analyte values indicate ongoing disease process. Reference intervals are generally based on individuals without diagnosed diseases or use of medication, which often excludes elderly. The aim of the study was to assess levels of albumin, alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatinine and γ-glutamyl transferase (γ-GT) in frail, moderately healthy and healthy elderly indivuduals. Blood samples were collected from individuals >80 years old, nursing home residents, in the Elderly in Linköping Screening Assessment and Nordic Reference Interval Project, a total of 569 individuals. They were divided into three cohorts: frail, moderately healthy and healthy, depending on cognitive and physical function. Albumin, ALT, AST, creatinine and γ-GT were analyzed using routine methods. Linear regression predicted factors for 34% of the variance in albumin were activities of daily living (ADL), gender, stroke and cancer. ADLs, gender and weight explained 15% of changes in ALT. For AST levels, ADLs, cancer and analgesics explained 5% of changes. Kidney disease, gender, Mini Mental State Examination (MMSE) and chronic obstructive pulmonary disease explained 25% of the variation in creatinine levels and MMSE explained three per cent of γ-GT variation. Because a group of people are at the same age, they should not be assessed the same way. To interpret results of laboratory tests in elderly is a complex task, where reference intervals are one part, but far from the only one, to take into consideration.

  12. Preparation of a Sepia Melanin and Poly(ethylene-alt-maleic Anhydride) Hybrid Material as an Adsorbent for Water Purification.

    Science.gov (United States)

    Panzarasa, Guido; Osypova, Alina; Consolati, Giovanni; Quasso, Fiorenza; Soliveri, Guido; Ribera, Javier; Schwarze, Francis W M R

    2018-01-23

    Meeting the increasing demand of clean water requires the development of novel efficient adsorbent materials for the removal of organic pollutants. In this context the use of natural, renewable sources is of special relevance and sepia melanin, thanks to its ability to bind a variety of organic and inorganic species, has already attracted interest for water purification. Here we describe the synthesis of a material obtained by the combination of sepia melanin and poly(ethylene- alt -maleic anhydride) (P(E- alt -MA)). Compared to sepia melanin, the resulting hybrid displays a high and fast adsorption efficiency towards methylene blue (a common industrial dye) for a wide pH range (from pH 2 to 12) and under high ionic strength conditions. It is easily recovered after use and can be reused up to three times. Given the wide availability of sepia melanin and P(E- alt -MA), the synthesis of our hybrid is simple and affordable, making it suitable for industrial water purification purposes.

  13. Preparation of a Sepia Melanin and Poly(ethylene-alt-maleic Anhydride Hybrid Material as an Adsorbent for Water Purification

    Directory of Open Access Journals (Sweden)

    Guido Panzarasa

    2018-01-01

    Full Text Available Meeting the increasing demand of clean water requires the development of novel efficient adsorbent materials for the removal of organic pollutants. In this context the use of natural, renewable sources is of special relevance and sepia melanin, thanks to its ability to bind a variety of organic and inorganic species, has already attracted interest for water purification. Here we describe the synthesis of a material obtained by the combination of sepia melanin and poly(ethylene-alt-maleic anhydride (P(E-alt-MA. Compared to sepia melanin, the resulting hybrid displays a high and fast adsorption efficiency towards methylene blue (a common industrial dye for a wide pH range (from pH 2 to 12 and under high ionic strength conditions. It is easily recovered after use and can be reused up to three times. Given the wide availability of sepia melanin and P(E-alt-MA, the synthesis of our hybrid is simple and affordable, making it suitable for industrial water purification purposes.

  14. ALT flares during nucleotide analogue therapy are associated with HBsAg loss in genotype A HBeAg-positive chronic hepatitis B.

    Science.gov (United States)

    Wong, Darren; Littlejohn, Margaret; Edwards, Rosalind; Jackson, Kathy; Revill, Peter; Gaggar, Anuj; Kitrinos, Kathryn; Subramanian, Mani; Marcellin, Patrick; Buti-Ferret, Maria; Janssen, Harry; Gane, Ed; Locarnini, Stephen; Thompson, Alexander

    2018-02-10

    Alanine aminotransferase (ALT) flares during NA therapy are uncommon but occur. Evaluation of ALT flares during nucleos(t)ide analogue (NA) therapy is important as new immunomodulatory therapies for hepatitis B virus (HBV) are developed. We evaluated the association between ALT flares and HBsAg loss during long-term therapy for genotype A CHB. This analysis included genotype A subjects from a phase III study of tenofovir vs adefovir in HBeAg-positive HBV. ALT flare was defined as (i) a rise in ALT >2x ULN from normal ALT levels; or (ii) a rise in ALT >2x baseline ALT level. HBsAg response at week 384 was recorded as one of HBsAg loss vs HBsAg decline (≥1 log 10  IU/mL decline) vs non-response. The primary analysis evaluated the association between ALT flare and HBsAg response. 54 subjects were included. 23/54 (43%) subjects experienced an on-treatment ALT flare. 45% achieved an HBsAg reduction ≥1 log 10  IU/mL, and of these 67% achieved HBsAg loss at a median of 102 weeks [IQR: 64-156]. Flare was associated with HBsAg decline vs non-response (67% vs 23%, P = .002), and were more common in subjects who achieved HBsAg loss vs non-response (56% vs 23%), P = .049). There was a median delay of 56 weeks [IQR: 40-80] between a flare and HBsAg loss. In genotype A subjects undergoing long-term NA therapy, ALT flares predict for HBsAg response. The delay between ALT flare and HBsAg loss has implications for clinical trial design for early phase development of immunomodulatory strategies aiming for HBsAg loss. © 2018 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  15. Viral DNA-Dependent Induction of Innate Immune Response to Hepatitis B Virus in Immortalized Mouse Hepatocytes.

    Science.gov (United States)

    Cui, Xiuji; Clark, Daniel N; Liu, Kuancheng; Xu, Xiao-Dong; Guo, Ju-Tao; Hu, Jianming

    2016-01-01

    Hepatitis B virus (HBV) infects hundreds of millions of people worldwide and causes acute and chronic hepatitis, cirrhosis, and hepatocellular carcinoma. HBV is an enveloped virus with a relaxed circular (RC) DNA genome. In the nuclei of infected human hepatocytes, conversion of RC DNA from the incoming virion or cytoplasmic mature nucleocapsid (NC) to the covalently closed circular (CCC) DNA, which serves as the template for producing all viral transcripts, is essential to establish and sustain viral replication. A prerequisite for CCC DNA formation is the uncoating (disassembly) of NCs to expose their RC DNA content for conversion to CCC DNA. We report here that in an immortalized mouse hepatocyte cell line, AML12HBV10, in which RC DNA exposure is enhanced, the exposed viral DNA could trigger an innate immune response that was able to modulate viral gene expression and replication. When viral gene expression and replication were low, the innate response initially stimulated these processes but subsequently acted to shut off viral gene expression and replication after they reached peak levels. Inhibition of viral DNA synthesis or cellular DNA sensing and innate immune signaling diminished the innate response. These results indicate that HBV DNA, when exposed in the host cell cytoplasm, can function to trigger an innate immune response that, in turn, modulates viral gene expression and replication. Chronic infection by hepatitis B virus (HBV) afflicts hundreds of millions worldwide and is sustained by the episomal covalently closed circular (CCC) DNA in the nuclei of infected hepatocytes. Release of viral genomic DNA from cytoplasmic nucleocapsids (NCs) (NC disassembly or uncoating) is a prerequisite for its conversion to CCC DNA, which can also potentially expose the viral DNA to host DNA sensors and trigger an innate immune response. We have found that in an immortalized mouse hepatocyte cell line in which efficient CCC DNA formation was associated with enhanced

  16. Stromal cell-associated hematopoiesis: immortalization and characterization of a primate bone marrow-derived stromal cell line.

    Science.gov (United States)

    Paul, S R; Yang, Y C; Donahue, R E; Goldring, S; Williams, D A

    1991-04-15

    An elucidation of the interaction between the bone marrow microenvironment and hematopoietic stem cells is critical to the understanding of the molecular basis of stem cell self renewal and differentiation. This interaction is dependent, at least in part, on direct cell to cell contact or cellular adhesion to extracellular matrix proteins. Long-term bone marrow cultures (LTMC) provide an appropriate microenvironment for maintenance of primitive hematopoietic stem cells and a means of analyzing this stem cell-stromal cell interaction in vitro. Although LTMC have been successfully generated from murine and human bone marrow, only limited success has been reported in a primate system. In addition, few permanent stromal cell lines are available from nonmurine bone marrow. Because the primate has become a useful model for large animal bone marrow transplant studies and, more specifically, retroviral-mediated gene transfer analysis, we have generated immortalized bone marrow stromal cell lines from primate bone marrow using gene transfer of the Simian virus large T (SV40 LT) antigen. At least one stromal cell line has demonstrated the capacity to maintain early hematopoietic cells in long-term cultures for up to 4 weeks as measured by in vitro progenitor assays. Studies were undertaken to characterize the products of extracellular matrix biosynthesis and growth factor synthesis of this cell line, designated PU-34. In contrast to most murine bone marrow-derived stromal cell lines capable of supporting hematopoiesis in vitro that have been examined, the extracellular matrix produced by this primate cell line includes collagen types I, laminin. Growth factor production analyzed through RNA blot analysis, bone marrow cell culture data, and factor-dependent cell line proliferation assays includes interleukin-6 (IL-6), IL-7, granulocyte-macrophage colony-stimulating factor (GM-CSF), G-CSF, M-CSF, leukemia inhibitory factor, and a novel cytokine designated IL-11. This

  17. The combined effects of irradiation and herpes simplex virus type 1 infection on an immortal gingival cell line

    Science.gov (United States)

    2014-01-01

    Background Oral mucosa is frequently exposed to Herpes simplex virus type 1 (HSV-1) infection and irradiation due to dental radiography. During radiotherapy for oral cancer, the surrounding clinically normal tissues are also irradiated. This prompted us to study the effects of HSV-1 infection and irradiation on viability and apoptosis of oral epithelial cells. Methods Immortal gingival keratinocyte (HMK) cells were infected with HSV-1 at a low multiplicity of infection (MOI) and irradiated with 2 Gy 24 hours post infection. The cells were then harvested at 24, 72 and 144 hours post irradiation for viability assays and qRT-PCR analyses for the apoptosis-related genes caspases 3, 8, and 9, bcl-2, NFκB1, and viral gene VP16. Mann–Whitney U-test was used for statistical calculations. Results Irradiation improved the cell viability at 144 hours post irradiation (P = 0.05), which was further improved by HSV-1 infection at MOI of 0.00001 (P = 0.05). Simultaneously, the combined effects of infection at MOI of 0.0001 and irradiation resulted in upregulation in NFκB1 (P = 0.05). The combined effects of irradiation and HSV infection also significantly downregulated the expression of caspases 3, 8, and 9 at 144 hours (P = 0.05) whereas caspase 3 and 8 significantly upregulated in non-irradiated, HSV-infected cells as compared to uninfected controls (P = 0.05). Infection with 0.0001 MOI downregulated bcl-2 in non-irradiated cells but was upregulated by 27% after irradiation when compared to non-irradiated infected cells (P = 0.05). Irradiation had no effect on HSV-1 shedding or HSV gene expression at 144 hours. Conclusions HSV-1 infection may improve the viability of immortal cells after irradiation. The effect might be related to inhibition of apoptosis. PMID:25005804

  18. Characterization of a novel telomerase-immortalized human endometrial stromal cell line, St-T1b

    Directory of Open Access Journals (Sweden)

    Brosens Jan J

    2009-07-01

    Full Text Available Abstract Background Coordinated differentiation of the endometrial compartments in the second half of the menstrual cycle is a prerequisite for the establishment of pregnancy. Endometrial stromal cells (ESC decidualize under the influence of ovarian progesterone to accommodate implantation of the blastocyst and support establishment of the placenta. Studies into the mechanisms of decidualization are often hampered by the lack of primary ESC. Here we describe a novel immortalized human ESC line. Methods Primary ESC were immortalized by the transduction of telomerase. The resultant cell line, termed St-T1b, was characterized for its morphological and biochemical properties by immunocytochemistry, RT-PCR and immunoblotting. Its progestational response was tested using progesterone and medroxyprogesterone acetate with and without 8-Br-cAMP, an established inducer of decidualization in vitro. Results St-T1b were positive for the fibroblast markers vimentin and CD90 and negative for the epithelial marker cytokeratin-7. They acquired a decidual phenotype indistinguishable from primary ESC in response to cAMP stimulation. The decidual response was characterized by transcriptional activation of marker genes, such as PRL, IGFBP1, and FOXO1, and enhanced protein levels of the tumor suppressor p53 and the metastasis suppressor KAI1 (CD82. Progestins alone had no effect on St-T1b cells, but medroxyprogesterone acetate greatly enhanced the cAMP-stimulated expression of IGFBP-1 after 3 and 7 days. Progesterone, albeit more weakly, also augmented the cAMP-induced IGFBP-1 production but only after 7 days of treatment. The cell line remained stable in continuous culture for more than 150 passages. Conclusion St-T1b express the appropriate phenotypic ESC markers and their decidual response closely mimics that of primary cultures. Decidualization is efficiently induced by cAMP analog and enhanced by medroxyprogesterone acetate, and, to a lesser extent, by natural

  19. Cell-based assay for the detection of chemically induced cellular stress by immortalized untransformed transgenic hepatocytes

    Directory of Open Access Journals (Sweden)

    Vezzoni Paolo

    2004-03-01

    Full Text Available Abstract Background Primary hepatocytes, one of the most widely used cell types for toxicological studies, have a very limited life span and must be freshly derived from mice or even humans. Attempts to use stable cell lines maintaining the enzymatic pattern of liver cells have been so far unsatisfactory. Stress proteins (heat shock proteins, HSPs have been proposed as general markers of cellular injury and their use for environmental monitoring has been suggested. The aim of this work is to develop a bi-transgenic hepatocyte cell line in order to evaluate the ability of various organic and inorganic chemicals to induce the expression of the HSP70 driven reporter gene. We previously described transgenic mice (Hsp70/hGH secreting high levels of human Growth Hormone (hGH following exposure to toxic compounds in vivo and in vitro in primary cultures derived from different organs. In addition, we also reported another transgenic model (AT/cytoMet allowing the reproducible immortalization of untransformed hepatocytes retaining in vitro complex liver functions. Results The transgenic mouse line Hsp70/hGH was crossed with the AT/cytoMet transgenic strain permitting the reproducible immortalization of untransformed hepatocytes. From double transgenic animals we derived several stable hepatic cell lines (MMH-GH which showed a highly-differentiated phenotype as judged from the retention of epithelial cell polarity and the profile of gene expression, including hepatocyte-enriched transcription factors and detoxifying enzymes. In these cell lines, stresses induced by exposure to inorganic [Sodium Arsenite (NaAsO2 and Cadmium Chloride (CdCl2], and organic [Benzo(aPyrene (BaP, PentaChloroPhenol (PCP, TetraChloroHydroQuinone (TCHQ, 1-Chloro-2,4-DiNitro-Benzene (CDNB] compounds, specifically induced hGH release in the culture medium. Conclusions MMH-GH, an innovative model to evaluate the toxic potential of chemical and physical xenobiotics, provides a simple

  20. Establishment of a luciferase assay-based screening system: Fumitremorgin C selectively inhibits cellular proliferation of immortalized astrocytes expressing an active form of AKT

    International Nuclear Information System (INIS)

    Wang Lei; Sasai, Ken; Akagi, Tsuyoshi; Tanaka, Shinya

    2008-01-01

    The AKT pathway is frequently activated in glioblastoma, and as such, inhibitors of this pathway could prove very useful as anti-glioblastoma therapies. Here we established immortalized astrocytes expressing Renilla luciferase as well as those expressing both an active form of AKT and firefly luciferase. Since both luciferase activities represent the numbers of corresponding cell lines, novel inhibitors of the AKT pathway can be identified by treating co-cultures containing the two types of luciferase-expressing cells with individual compounds. Indeed, such a screening system succeeded in identifying fumitremorgin C as an efficient inhibitor of the AKT pathway, which was further confirmed by the ability of fumitremorgin C to selectively inhibit the growth of immortalized astrocytes expressing an active form of AKT. The present study proposes a broadly applicable approach for identifying therapeutic agents that target the pathways and/or molecules responsible for cancer development

  1. Immortalization of human melanocytes does not alter the de novo properties of nitric oxide to induce cell detachment from extracellular matrix components via cGMP.

    Science.gov (United States)

    Ivanova, Krassimira; Lambers, Britta; van den Wijngaard, Rene; Le Poole, I Caroline; Grigorieva, Olga; Gerzer, Rupert; Das, Pranab K

    2008-01-01

    Nitric oxide (NO) is an important mediator in many (patho)physiological processes including inflammation and skin cancer. A key transducer in NO signaling is the soluble guanylyl cyclase (sGC) that catalyzes the formation of guanosine 3',5'-cyclic monophosphate (cGMP). The basic mechanism of NO-cGMP signaling in melanocytic cells is, however, not well elucidated. A setback for such studies is the limited availability of patient-derived melanocytes. Here, we report that immortalized human normal and vitiliginous cell lines generated via cell transfection with human papilloma virus 16 genes E6 and E7 express NO synthase and guanylyl cyclase isoforms and the multidrug resistance-associated proteins 4 and 5 as selective cGMP exporters. Donors of NO (e.g., the NONOate (Z)-1-[N-(3-ammoniopropyl)-N-(n-propyl)amino]diazen-1-ium-1,2-diolate (PAPA-NO) and reactive nitrogen oxygen species (RNOS) like 3-morpholino-sydnonimine (SIN-1) as a donor of peroxynitrite as well as YC-1 as a NO-independent sGC stimulator increased intracellular cGMP levels in immortalized melanocytes (up to eightfold over controls), indicating the expression of functional sGC in these cells. PAPA-NO and SIN-1 also reduced the attachment of immortalized melanocytes to extracellular matrix (ECM) components like fibronectin which was dependent on cellular melanin content and cGMP. Such effects on melanoma cells were positively related to metastatic potential and were cGMP independent. Intriguingly, nonpigmented metastatic melanoma cells were more sensitive to exogenous sources of RNOS than of NO. Thus, immortalized melanocytes can be used as a tool for further research on differences in cell signaling between the different melanocytic lineages in particular towards impairment of cell-ECM adhesion by NO or RNOS, which may be important in metastasis and vitiligo pathogenesis.

  2. Large-scale analysis of protein expression changes in human keratinocytes immortalized by human papilloma virus type 16 E6 and E7 oncogenes

    Directory of Open Access Journals (Sweden)

    Arnouk Hilal

    2009-08-01

    Full Text Available Abstract Background Infection with high-risk type human papilloma viruses (HPVs is associated with cervical carcinomas and with a subset of head and neck squamous cell carcinomas. Viral E6 and E7 oncogenes cooperate to achieve cell immortalization by a mechanism that is not yet fully understood. Here, human keratinocytes were immortalized by long-term expression of HPV type 16 E6 or E7 oncoproteins, or both. Proteomic profiling was used to compare expression levels for 741 discrete protein features. Results Six replicate measurements were performed for each group using two-dimensional difference gel electrophoresis (2D-DIGE. The median within-group coefficient of variation was 19–21%. Significance of between-group differences was tested based on Significance Analysis of Microarray and fold change. Expression of 170 (23% of the protein features changed significantly in immortalized cells compared to primary keratinocytes. Most of these changes were qualitatively similar in cells immortalized by E6, E7, or E6/7 expression, indicating convergence on a common phenotype, but fifteen proteins (~2% were outliers in this regulatory pattern. Ten demonstrated opposite regulation in E6- and E7-expressing cells, including the cell cycle regulator p16INK4a; the carbohydrate binding protein Galectin-7; two differentially migrating forms of the intermediate filament protein Cytokeratin-7; HSPA1A (Hsp70-1; and five unidentified proteins. Five others had a pattern of expression that suggested cooperativity between the co-expressed oncoproteins. Two of these were identified as forms of the small heat shock protein HSPB1 (Hsp27. Conclusion This large-scale analysis provides a framework for understanding the cooperation between E6 and E7 oncoproteins in HPV-driven carcinogenesis.

  3. A kidney injury molecule‐1 (Kim‐1) gene reporter in a mouse artificial chromosome: the responsiveness to cisplatin toxicity in immortalized mouse kidney S3 cells

    Science.gov (United States)

    Kokura, Kenji; Kuromi, Yasushi; Endo, Takeshi; Anzai, Naohiko; Kazuki, Yasuhiro; Oshimura, Mitsuo

    2016-01-01

    Abstract Background Kidney injury molecule‐1 (Kim‐1) has been validated as a urinary biomarker for acute and chronic renal damage. The expression of Kim‐1 mRNA is also activated by acute kidney injury induced by cisplatin in rodents and humans. To date, the measurement of Kim‐1 expression has not fully allowed the detection of in vitro cisplatin nephrotoxicity in immortalized culture cells, such as human kidney‐2 cells and immortalized proximal tubular epithelial cells. Methods We measured the augmentation of Kim‐1 mRNA expression after the addition of cisplatin using immortalized S3 cells established from the kidneys of transgenic mice harboring temperature‐sensitive large T antigen from Simian virus 40. Results A mouse Kim‐1 gene luciferase reporter in conjunction with an Hprt gene reporter detected cisplatin‐induced nephrotoxicity in S3 cells. These two reporter genes were contained in a mouse artificial chromosome, and two luciferases that emitted different wavelengths were used to monitor the respective gene expression. However, the Kim‐1 reporter gene failed to respond to cisplatin in A9 fibroblast cells that contained the same reporter mouse artificial chromosome, suggesting cell type‐specificity for activation of the reporter. Conclusions We report the feasibility of measuring in vitro cisplatin nephrotoxicity using a Kim‐1 reporter gene in S3 cells. PMID:27591740

  4. A kidney injury molecule-1 (Kim-1) gene reporter in a mouse artificial chromosome: the responsiveness to cisplatin toxicity in immortalized mouse kidney S3 cells.

    Science.gov (United States)

    Kokura, Kenji; Kuromi, Yasushi; Endo, Takeshi; Anzai, Naohiko; Kazuki, Yasuhiro; Oshimura, Mitsuo; Ohbayashi, Tetsuya

    2016-10-01

    Kidney injury molecule-1 (Kim-1) has been validated as a urinary biomarker for acute and chronic renal damage. The expression of Kim-1 mRNA is also activated by acute kidney injury induced by cisplatin in rodents and humans. To date, the measurement of Kim-1 expression has not fully allowed the detection of in vitro cisplatin nephrotoxicity in immortalized culture cells, such as human kidney-2 cells and immortalized proximal tubular epithelial cells. We measured the augmentation of Kim-1 mRNA expression after the addition of cisplatin using immortalized S3 cells established from the kidneys of transgenic mice harboring temperature-sensitive large T antigen from Simian virus 40. A mouse Kim-1 gene luciferase reporter in conjunction with an Hprt gene reporter detected cisplatin-induced nephrotoxicity in S3 cells. These two reporter genes were contained in a mouse artificial chromosome, and two luciferases that emitted different wavelengths were used to monitor the respective gene expression. However, the Kim-1 reporter gene failed to respond to cisplatin in A9 fibroblast cells that contained the same reporter mouse artificial chromosome, suggesting cell type-specificity for activation of the reporter. We report the feasibility of measuring in vitro cisplatin nephrotoxicity using a Kim-1 reporter gene in S3 cells. © 2016 The Authors. The Journal of Gene Medicine Published by John Wiley & Sons, Ltd.

  5. Generation of human monoclonal antibodies against HIV-1 proteins; electrofusion and Epstein-Barr virus transformation for peripheral blood lymphocyte immortalization.

    Science.gov (United States)

    Buchacher, A; Predl, R; Strutzenberger, K; Steinfellner, W; Trkola, A; Purtscher, M; Gruber, G; Tauer, C; Steindl, F; Jungbauer, A

    1994-04-01

    Electrofusion and EBV transformation were studied by immortalizing human PBLs from blood of HIV-1-positive volunteers. A panel of 33 cell lines producing human monoclonal antibodies (Hu-MAbs) against HIV-1 was established by cell fusion or EBV transformation. For the first fusion experiments the source of B lymphocytes was peripheral blood of HIV-1-infected donors in CDC stages II or III with CD4 cell counts higher than 500/mm3. Later on, from these patients only, those with high anti-HIV titers were chosen as blood donors. By that means the yield of stable specific hybridomas was increased twofold. In our experiments electrofusion turned out to be a more efficient immortalization method than EBV transformation, due to a high and constant immortalization rate. The hybridomas were stable after intensive subcloning and could be cultivated over a period of 8 months without loss in monoclonal antibody production. Immunoglobulin class, subtype, reactivity against HIV-1 proteins, Western blot patterns, immunofluorescence, and epitopes were characterized. The subtype of all antibodies was IgG1 or IgG3. The light chain was predominantly kappa. All antibodies showed reactivity against HIV-1 envelope or core protein. All hybridomas were stable and suited for mass production. Several Hu-MAbs are becoming an important tool in the field of diagnosis, research, and immunotherapy.

  6. Radiation induced, raft mediated grafting of styrene onto poly(ethylene-alt-tetrafluoroethylene) (ETFE)

    International Nuclear Information System (INIS)

    Celik, G.; Barsbay, M.; Gueven, O.

    2011-01-01

    Complete text of publication follows. The development of cost-effective proton exchange membranes to replace the state-of-the-art and expensive perfluorinated membranes such as Nafion, Flemion, and Aciplex is one of the main challenges for commercialization of polymer electrolyte fuel cell technology. Recently, partially fluorinated poly(ethlyene-alt-tetrafluoroethylene) (ETFE) has been identified as a promising alternative base polymer film to prepare low-cost polymer electrolyte membranes because of its advantageous characters like superior mechanical properties and high resistance to radiation-induced damage. The radiation-induced grafting technique, based on the utilization of a polymer material such as ETFE in combination with further chemical modification steps (sulfonation) allows the functionalization of the base material and the introduction of the desired property (proton conductivity) for preparing a fuel cell membrane. However this simple conventional method suffers from one simple flaw: The molecular weight and the polydispersity of the grafted chains cannot be controlled. Predetermined molecular weights and low dispersities as well as homogeneous composition and desired architecture can be achieved by grafting of monomer onto base polymer under living/controlled free radical polymerization (CRP) conditions. Among the CRP methods, Reversible Addition Fragmentation-Chain Transfer (RAFT) is of particular interest as a very wide range of functional monomers can be polymerized in a controlled manner under non-demanding reaction conditions (e.g., tolerance to oxygen and low temperatures). The present study deals with the RAFT mediated radiation-induced (0.032 kGyh -1 , 60 Co) grafting of styrene on ETFE films followed by the sulfonation of the polystyrene grafts. The effect of monomer concentration, absorbed dose and RAFT agent concentration on the grafting were investigated. The synthesized films were characterized by ATR-FTIR, XPS, DSC and TGA methods

  7. Specific regulation of point-mutated K-ras-immortalized cell proliferation by a photodynamic antisense strategy.

    Science.gov (United States)

    Higuchi, Maiko; Yamayoshi, Asako; Kato, Kiyoko; Kobori, Akio; Wake, Norio; Murakami, Akira

    2010-02-01

    It has been reported that point mutations in genes are responsible for various cancers, and the selective regulation of gene expression is an important factor in developing new types of anticancer drugs. To develop effective drugs for the regulation of point-mutated genes, we focused on photoreactive antisense oligonucleotides. Previously, we reported that photoreactive oligonucleotides containing 2'-O-psoralenylmethoxyethyl adenosine (2'-Ps-eom) showed drastic photoreactivity in a strictly sequence-specific manner. Here, we demonstrated the specific gene regulatory effects of 2'-Ps-eom on [(12)Val]K-ras mutant (GGT --> GTT). Photo-cross-linking between target mRNAs and 2'-Ps-eom was sequence-specific, and the effect was UVA irradiation-dependent. Furthermore, 2'-Ps-eom was able to inhibit K-ras-immortalized cell proliferation (K12V) but not Vco cells that have the wild-type K-ras gene. These results suggest that the 2'-Ps-eom will be a powerful nucleic acid drug to inhibit the expression of disease-causing point mutation genes, and has great therapeutic potential in the treatment of cancer.

  8. Comparative QTL analysis of maize seed artificial aging between an immortalized F2 population and its corresponding RILs

    Directory of Open Access Journals (Sweden)

    Bin Wang

    2016-02-01

    Full Text Available Seed aging decreases the quality and vigor of crop seeds, thereby causing substantial agricultural and economic losses in crops. To identify genetic differences in seed aging between homozygotes and heterozygotes in maize, the seeds of a set of recombinant inbred lines (RILs and an immortalized F2 (IF2 population were subjected to artificial aging treatments for 0, 2, 3, and 4 days under 45 ºC and 85% relative humidity and seed vigor was then evaluated in a field experiment. Seed vigor of all entries tested decreased sharply with longer aging treatment and seed vigor decreased more slowly in heterozygotes than in homozygotes. Forty-nine QTL were detected for four measured seed vigor traits in the RIL (28 QTL and IF2 (21 QTL populations. Only one QTL, qGP5, was detected in both populations, indicating that the genes involved in anti-aging mechanisms differed between inbred lines and hybrids. Several QTL were identified to be responsible for multiple seed vigor traits simultaneously in the RIL and IF2 populations under artificial aging conditions. These QTL may include major genes for seed vigor or seed aging. QTL qVI4b and qGE3a detected in the RIL population coincided with genes ZmLOX1 and ZmPLD1 in the same respective chromosomal regions. These QTL would be useful for screening for anti-aging genes in maize breeding.

  9. Renal carcinoma cell-derived exosomes induce human immortalized line of Jurkat T lymphocyte apoptosis in vitro.

    Science.gov (United States)

    Yang, Lin; Wu, Xiaohou; Wang, Dan; Luo, Chunli; Chen, Lixue

    2013-01-01

    Tumor-derived exosomes usually contain some molecules that can help immune evasion by tumors. This study is aimed at investigating the potential effect of exosomes from human kidney adenocarcinoma cells on a human immortalized line of Jurkat T lymphocytes in vitro. Exosomes were purified from human kidney adenocarcinoma ACHN cells by sequential centrifugations and ultrafiltrations, and characterized by transmission electron microscopy. The effects of exosomes on the proliferation, cytokine production and apoptosis of Jurkat T cells were determined using flow cytometry and enzyme-linked immunosorbent assay. The relative levels of pro- and anti-apoptotic molecules were determined by Western blotting. Exosomes were purified from ACHN cells and exhibited typical characteristics. Treatment with exosomes inhibited Jurkat T cell proliferation and induced Jurkat T cell apoptosis in a dose- and time-dependent manner. Treatment with exosomes reduced spontaneous interleukin-2 (IL-2), interferon-γ, IL-6 and IL-10 production by Jurkat T cells. Treatment with exosomes increased the relative levels of cleaved caspase-3, -8 and -9 as well as Bax, but reduced the levels of Bcl-2 in Jurkat T cells. The purified exosomes contained Fas ligand, and treatment with soluble Fas abrogated exosome-mediated Jurkat T cell apoptosis. Our data indicate that exosomes from kidney adenocarcinoma cells contain Fas ligand and trigger Jurkat T cell apoptosis, contributing to the immune evasion of tumors. Copyright © 2013 S. Karger AG, Basel.

  10. Ski protein levels increase during in vitro progression of HPV16-immortalized human keratinocytes and in cervical cancer.

    Science.gov (United States)

    Chen, Yi; Pirisi, Lucia; Creek, Kim E

    2013-09-01

    We compared the levels of the Ski oncoprotein, an inhibitor of transforming growth factor-beta (TGF-β) signaling, in normal human keratinocytes (HKc), HPV16 immortalized HKc (HKc/HPV16), and differentiation resistant HKc/HPV16 (HKc/DR) in the absence and presence of TGF-β. Steady-state Ski protein levels increased in HKc/HPV16 and even further in HKc/DR, compared to HKc. TGF-β treatment of HKc, HKc/HPV16, and HKc/DR dramatically decreased Ski. TGF-β-induced Ski degradation was delayed in HKc/DR. Ski and phospho-Ski protein levels are cell cycle dependent with maximal Ski expression and localization to centrosomes and mitotic spindles during G2/M. ShRNA knock down of Ski in HKc/DR inhibited cell proliferation. More intense nuclear and cytoplasmic Ski staining and altered Ski localization were found in cervical cancer samples compared to adjacent normal tissue in a cervical cancer tissue array. Overall, these studies demonstrate altered Ski protein levels, degradation and localization in HPV16-transformed human keratinocytes and in cervical cancer. Copyright © 2013 Elsevier Inc. All rights reserved.

  11. Unique inhibition of bile salt-induced apoptosis by lecithins and cytoprotective bile salts in immortalized mouse cholangiocytes.

    Science.gov (United States)

    Komichi, Daisuke; Tazuma, Susumu; Nishioka, Tomoji; Hyogo, Hideyuki; Une, Mizuho; Chayama, Kazuaki

    2003-12-01

    Bile duct epithelium is physiologically exposed to high concentrations of bile salts, suggesting the presence of a cytoprotective mechanism(s). The aim of this study was to clarify whether bile salts cause bile duct cell damage and to elucidate the mechanism(s) providing protection against such an action of bile salts. Immortalized mouse cholangiocytes were incubated with taurocholate, taurochenodeoxycholate, glycochenodeoxycholate (GCDC), taurodeoxycholate, and tauroursodeoxycholate (TUDC), followed by flow-cytometric analysis and caspase activity assay to evaluate the induction of apoptosis. GCDC time-dependently induced caspase 3 (3.4-fold)- and caspase 9 (1.4-fold)-mediated apoptosis of cholangiocytes, but this was inhibited by lecithins and TUDC. Further, expression of cholangiocyte bile salt transporters (apical sodium-dependent bile salt transporter [Asbt] and multidrug resistance protein 3 [Mrp3]) was examined by RT-PCR and western blotting, and cholangiocyte bile salt uptake was determined using radiolabeled bile salts. Expression of cholangiocyte Asbt and Mrp3 was increased by bile salts, whereas lecithins interestingly reduced bile salt uptake to inhibit cholangiocyte apoptosis. In conclusion, bile salts themselves cause cholangiocyte apoptosis when absorbed by and retained inside the cell, but this is inhibited by washing out cytotoxic bile salts according to Mrp3, a rescue exporting molecule. Biliary lecithin is seemingly another cytoprotective player against cytotoxic bile salts, reducing their uptake, and this is associated with a reduced expression of Mrp3.

  12. Medium Calcium Concentration Determines Keratin Intermediate Filament Density and Distribution in Immortalized Cultured Thymic Epithelial Cells (TECs)

    Science.gov (United States)

    Sands, Sandra S.; Meek, William D.; Hayashi, Jun; Ketchum, Robert J.

    2005-08-01

    Isolation and culture of thymic epithelial cells (TECs) using conventional primary tissue culture techniques under conditions employing supplemented low calcium medium yielded an immortalized cell line derived from the LDA rat (Lewis [Rt1l] cross DA [Rt1a]) that could be manipulated in vitro. Thymi were harvested from 4 5-day-old neonates, enzymically digested using collagenase (1 mg/ml, 37°C, 1 h) and cultured in low calcium WAJC404A medium containing cholera toxin (20 ng/ml), dexamethasone (10 nM), epidermal growth factor (10 ng/ml), insulin (10 [mu]g/ml), transferrin (10 [mu]g/ml), 2% calf serum, 2.5% Dulbecco's Modified Eagle's Medium (DMEM), and 1% antibiotic/antimycotic. TECs cultured in low calcium displayed round to spindle-shaped morphology, distinct intercellular spaces (even at confluence), and dense reticular-like keratin patterns. In high calcium (0.188 mM), TECs formed cobblestone-like confluent monolayers that were resistant to trypsinization (0.05%) and displayed keratin intermediate filaments concentrated at desmosomal junctions between contiguous cells. Changes in cultured TEC morphology were quantified by an analysis of desmosome/membrane relationships in high and low calcium media. Desmosomes were significantly increased in the high calcium medium. These studies may have value when considering the growth conditions of cultured primary cell lines like TECs.

  13. Efficient transfer of HTLV-1 tax gene in various primary and immortalized cells using a flap lentiviral vector.

    Science.gov (United States)

    Royer-Leveau, Christelle; Mordelet, Elodie; Delebecque, Frédéric; Gessain, Antoine; Charneau, Pierre; Ozden, Simona

    2002-08-01

    Human T cell leukemia virus type 1 (HTLV-1) causes two major diseases: adult T-cell leukemia-lymphoma and tropical spastic paraparesis/HTLV-1 associated myelopathy (TSP/HAM). In order to understand the involvement of Tax protein in HTLV-1 pathogenesis, we constructed a HIV-1 based lentiviral vector containing the central DNA flap sequence and either the green fluorescent protein (GFP) or the HTLV-1 tax genes. Using these vectors, GFP and tax genes were introduced in several primary and immortalized cells of endothelial, lymphoid, astrocytic or macrophagic origin. As assessed by GFP expression, up to 100% efficiency of transduction was obtained for all cell types tested. Tax expression was detected by Western blot and immuno-fluorescence in the transduced cells. After transduction, the Tax transcriptional activity was confirmed by the transactivation of HTLV-1 LTR-lacZ or HTLV-1 LTR-GFP reporter genes. Increased CD25 and HLA DR expression was observed in human peripheral blood lymphocytes transduced with the Tax vector. These results indicate that both pathways of Tax transactivation, CREB (viral LTR) and NF-kappa B (CD25 and HLA DR), are functional after transduction by TRIP Tax vector. Therefore, this vector provides a useful tool for investigating the role of the Tax viral protein in the pathogenesis of diseases linked to HTLV-1 infection.

  14. Induction of chromosomal aberrations in human primary fibroblasts and immortalized cancer cells exposed to extremely-low-frequency electromagnetic fields

    International Nuclear Information System (INIS)

    Seyyedi, S. S.; Mozdarani, H.; Rezaei Tavirani, M.; Heydari, S.

    2010-01-01

    Rapidly increasing possibilities of exposure to environmental extremely low-frequency electromagnetic fields have become a topic of worldwide investigation. Epidemiological and laboratory studies suggest that exposure to extremely low-frequency electromagnetic fields may increase cancer risk therefore assessment of chromosomal damage in various cell lines might be of predictive value for future risk estimation. Materials and Methods: Primary cultures of fibroblasts from human skin biopsy were exposed to continuous extremely low-frequency electromagnetic fields (3, 50 and 60 Hz, sinusoidal, 3h, and 4 m T). Also immortalized cell lines, SW480, MCF-7 and 1321N1 were exposed to continuous extremely low-frequency electromagnetic fields (50 Hz, sinusoidal, 3 h, 4 m T). Metaphase plates Were prepared according to standard methods and stained in 5% Giemsa solution. Chromosomal aberrations of both chromosome and chromatid types were scored to evaluate the effects of extremely low-frequency electromagnetic fields on primary or established cell lines. Results: Results indicate that by increasing the frequency of extremely low-frequency electromagnetic fields, chromosomal aberrations were increased up to 7-fold above background levels in primary human fibroblast cells. In addition, continuous exposure to a 50 Hz electromagnetic field led to a significant increase in chromosomal aberrations in SW480, MCF-7 and 1321N1 cell lines compared to sham control. Conclusion: Results obtained indicate that extremely low-frequency electromagnetic fields has the potential for induction of chromosomal aberrations in all cell types.

  15. A Styrene-alt-Maleic Acid Copolymer Is an Effective Inhibitor of R5 and X4 Human Immunodeficiency Virus Type 1 Infection

    Directory of Open Access Journals (Sweden)

    Vanessa Pirrone

    2010-01-01

    Full Text Available An alternating copolymer of styrene and maleic acid (alt-PSMA differs from other polyanionic antiviral agents in that the negative charges of alt-PSMA are provided by carboxylic acid groups instead of sulfate or sulfonate moieties. We hypothesized that alt-PSMA would have activity against human immunodeficiency virus type 1 (HIV-1 comparable to other polyanions, such as the related compound, poly(sodium 4-styrene sulfonate (PSS. In assays using cell lines and primary immune cells, alt-PSMA was characterized by low cytotoxicity and effective inhibition of infection by HIV-1 BaL and IIIB as well as clinical isolates of subtypes A, B, and C. In mechanism of action assays, in which each compound was added to cells and subsequently removed prior to HIV-1 infection (“washout” assay, alt-PSMA caused no enhancement of infection, while PSS washout increased infection 70% above control levels. These studies demonstrate that alt-PSMA is an effective HIV-1 inhibitor with properties that warrant further investigation.

  16. Immunomodulation of ALT-2 and TLR may collude in antigen specific T cell hyporesponsiveness: proposed mechanism for elevated IL-10 levels in Balb/C mice.

    Science.gov (United States)

    Aparnaa, Ramanathan; Kaliraj, Perumal

    2014-03-01

    ALT-2, a novel antigen belonging to the chromadorea ALT-2 family of the filarial nematode is proved to clear filarial parasites in Jirds. In order to increase the protection efficacy by stimulating the cell mediated immunity, MPLA a detoxified derivative of LPS known to induce the cellular response, was used in this study as an adjuvant on mice models. ALT-2+MPLA formulation elicited a high titer of total IgG antibody, with profoundly increased levels of IgG2b. Reduced splenocyte proliferation was observed in immunized group when compared to control groups which could be attributed to many in vivo factors. The levels of IFN-γ were high in unstimulated MPLA group compared to ALT-2 stimulated MPLA group, suggesting that the ALT-2 antigen suppressed the IFN-γ levels. A high level of IL-10 was induced by the ALT-2+MPLA formulation, which inhibited the production of Th2 cytokines (IL-4, IL-5) and also reduced the Th1 cytokine (IFN-γ, IL-2) levels which are not in vogue with the classical MPLA adjuvant formulation. We propose a mechanism for this immunomodulation which involves a diminished expression of TLR-4, by which the filarial parasites have evolved to evade host immune mechanism.

  17. The Preoperative AST/ALT (De Ritis) Ratio Represents a Poor Prognostic Factor in a Cohort of Patients with Nonmetastatic Renal Cell Carcinoma.

    Science.gov (United States)

    Bezan, Angelika; Mrsic, Edvin; Krieger, Daniel; Stojakovic, Tatjana; Pummer, Karl; Zigeuner, Richard; Hutterer, Georg C; Pichler, Martin

    2015-07-01

    Aminotransaminases, which are strongly involved in cellular metabolism and cancer cell turnover, represent easily measureable, potential blood based biomarkers. We evaluated the prognostic value of the preoperatively assessed AST/ALT (De Ritis) ratio on clinically meaningful end points in a large European cohort of patients with nonmetastatic renal cell carcinoma. We retrospectively evaluated clinicopathological data on 698 patients with nonmetastatic renal cell carcinoma operated on between 2005 and 2013 at a single tertiary academic center. The potential prognostic value of the AST/ALT ratio was analyzed using the Kaplan-Meier method, and univariate and multivariate Cox proportional regression models. The impact of the ratio on the predictive accuracy of the Leibovich prognosis score was determined by the Harrell c-index. An increased (1.26 or greater) preoperative AST/ALT ratio was statistically significantly associated with several well established prognostic factors, including pathological T stage, as well as with histological tumor necrosis (p ALT ratio was an independent prognostic factor for metastasis-free survival (HR 1.61, 95% CI 1.25-2.07, p ALT was added. In our study cohort with nonmetastatic renal cell carcinoma the preoperatively assessed AST/ALT ratio represented an independent prognostic factor. This ratio might further improve the predictive accuracy of well established prognosis scores. Copyright © 2015 American Urological Association Education and Research, Inc. Published by Elsevier Inc. All rights reserved.

  18. Comparative Cytotoxicity of Glycyrrhiza glabra Roots from Different Geographical Origins Against Immortal Human Keratinocyte (HaCaT), Lung Adenocarcinoma (A549) and Liver Carcinoma (HepG2) Cells.

    Science.gov (United States)

    Basar, Norazah; Oridupa, Olayinka Ayotunde; Ritchie, Kenneth J; Nahar, Lutfun; Osman, Nashwa Mostafa M; Stafford, Angela; Kushiev, Habibjon; Kan, Asuman; Sarker, Satyajit D

    2015-06-01

    Glycyrrhiza glabra L. (Fabaceae), commonly known as 'liquorice', is a well-known medicinal plant. Roots of this plant have long been used as a sweetening and flavouring agent in food and pharmaceutical products, and also as a traditional remedy for cough, upper and lower respiratory ailments, kidney stones, hepatitis C, skin disorder, cardiovascular diseases, diabetes, gastrointestinal ulcers and stomach ache. Previous pharmacological and clinical studies have revealed its antitussive, antiinflammatory, antiviral, antimicrobial, antioxidant, immunomodulatory, hepatoprotective and cardioprotective properties. While glycyrrhizin, a sweet-tasting triterpene saponin, is the principal bioactive compound, several bioactive flavonoids and isoflavonoids are also present in the roots of this plant. In the present study, the cytotoxicity of the methanol extracts of nine samples of the roots of G. glabra, collected from various geographical origins, was assessed against immortal human keratinocyte (HaCaT), lung adenocarcinoma (A549) and liver carcinoma (HepG2) cell lines using the in vitro 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazoliumbromide cell toxicity/viability assay. Considerable variations in levels of cytotoxicity were observed among various samples of G. glabra. Copyright © 2015 John Wiley & Sons, Ltd.

  19. Effects of Ashwagandha (Withania somnifera Root Extract On Some Serum Liver Marker Enzymes (AST, ALT In Gentamicin Intoxicated Rats

    Directory of Open Access Journals (Sweden)

    Nayma Sultana

    2012-06-01

    Full Text Available Background: Liver is an essential metabolic organ. It can be damaged due to prolonged use and higher doses of drugs, exposure to some chemicals, toxins, or infectious agents. Herbal plants as ashwagandha (Withania somnifera may have free radical scavenging activity thereby can be used for the prevention and treatment of liver damage.Objective: To observe the effect of ashwagandha (Withania somnifera root extract on gentamicin induced changes of some liver marker enzymes e,g serum aspartate amino transferase (AST and alanine amino transferase (ALT in Wistar albino rats.Methods: This experimental study was carried out in the Department of Physiology, Sir Salimullah Medical College (SSMC, Dhaka from 1st July 2010 to 30th June 2011. A total number of 35 Wistar albino rats, aged 90 to 120 days, weighing between 150 to 200 grams were selected for the study. After acclimatization for 14 days, they were divided into control group (Group A and experimental group (Group B. Control group was again subdivided into group A1 (baseline control, consisted of 10 rats and group A2 (gentamicin treated control group, consisted of 10 rats. Again, experimental group (Group B-ashwagandha pretreated and gentamicin treated group consisted of 15 rats. All groups of animals received basal diet for 22 consecutive days. In addition to this, group A2 also received gentamicin subcutaneously (100mg /kg body weight/day for the last eight (15th to 22nd day consecutive days. Again, group B received ashwagandha root extract (500mg/kg body weight/day, orally for 22 consecutive days and gentamicin subcutaneously (100mg/kg body weight /day for last eight (15th to 22nd day days. All the animals were sacrificed on 23rd day. Then blood and liver samples were collected. For assessment of liver function, serum AST, ALT and bilirubin levels were estimated. All these tests were done by standard Laboratory technique. The statistical analysis was done by one way ANOVA and Bonferroni test as

  20. Essai préliminaire de corrélations entre paramètres altérologiques ...

    African Journals Online (AJOL)

    débouche sur la détermination de sept paramètres altérologiques qui permettent des caractérisations ponctuelles, des études évolutives et/ou prospectives de géologie appliquée au Génie civil et même à la recherche minière. Pour une mise en œuvre optimale, les résultats de l'analyse chimique doivent faire apparaître.

  1. Enabling a robust scalable manufacturing process for therapeutic exosomes through oncogenic immortalization of human ESC-derived MSCs

    Directory of Open Access Journals (Sweden)

    Choo Andre

    2011-04-01

    Full Text Available Abstract Background Exosomes or secreted bi-lipid vesicles from human ESC-derived mesenchymal stem cells (hESC-MSCs have been shown to reduce myocardial ischemia/reperfusion injury in animal models. However, as hESC-MSCs are not infinitely expansible, large scale production of these exosomes would require replenishment of hESC-MSC through derivation from hESCs and incur recurring costs for testing and validation of each new batch. Our aim was therefore to investigate if MYC immortalization of hESC-MSC would circumvent this constraint without compromising the production of therapeutically efficacious exosomes. Methods The hESC-MSCs were transfected by lentivirus carrying a MYC gene. The transformed cells were analyzed for MYC transgene integration, transcript and protein levels, and surface markers, rate of cell cycling, telomerase activity, karyotype, genome-wide gene expression and differentiation potential. The exosomes were isolated by HPLC fractionation and tested in a mouse model of myocardial ischemia/reperfusion injury, and infarct sizes were further assessed by using Evans' blue dye injection and TTC staining. Results MYC-transformed MSCs largely resembled the parental hESC-MSCs with major differences being reduced plastic adherence, faster growth, failure to senesce, increased MYC protein expression, and loss of in vitro adipogenic potential that technically rendered the transformed cells as non-MSCs. Unexpectedly, exosomes from MYC-transformed MSCs were able to reduce relative infarct size in a mouse model of myocardial ischemia/reperfusion injury indicating that the capacity for producing therapeutic exosomes was preserved. Conclusion Our results demonstrated that MYC transformation is a practical strategy in ensuring an infinite supply of cells for the production of exosomes in the milligram range as either therapeutic agents or delivery vehicles. In addition, the increased proliferative rate by MYC transformation reduces the time

  2. Human papillomavirus-16 E7 interacts with Siva-1 and modulates apoptosis in HaCaT human immortalized keratinocytes.

    Science.gov (United States)

    Severino, Anna; Abbruzzese, Claudia; Manente, Lucrezia; Valderas, Alvaro Avivar; Mattarocci, Stefano; Federico, Antonio; Starace, Giuseppe; Chersi, Alberto; Mileo, Anna Maria; Paggi, Marco G

    2007-07-01

    The viral factor E7 plays a key role in the well-established association between "high-risk" Human Papillomavirus (HPV) infection and the development of epithelial malignant tumors, as uterine cervix and ano-genital cancer. To delve into the molecular mechanisms of HPV-mediated cell transformation, we searched for novel potential cellular targets of the HPV-16 E7 oncoprotein, by means of the yeast two-hybrid technique, identifying a protein-protein interaction between HPV-16 E7 and the pro-apoptotic cellular factor Siva-1. Using co-precipitation assays and the "PepSets" technique, we confirmed this physical interaction and mapped accurately, for both proteins, the amino acid residues involved. Additionally, we found that HPV-16 E7 competed in vitro with the binding of the Bcl-X(L) anti-apoptotic factor to Siva-1, an interaction that has a major inference in UV radiation-induced apoptosis. In HaCaT immortalized human keratinocytes, forced HPV-16 E7 expression by retroviral infection caused Siva-1 transcript up-regulation, detected by cDNA macroarray hybridization and real-time quantitative PCR, paralleled by an increased amount of protein. Confirming the anti-apoptotic role of HPV-16 E7 in the HaCaT cellular model, evaluated by nuclear morphology, we also found that Siva-1 expression produced a significant increase of the apoptotic rate in UV radiation-exposed HaCaT cells, and that this effect resulted explicitly counteracted by HPV-16 E7. Being apoptosis a key physiological process for the elimination of irreversibly injured cells, the anti-apoptotic role of HPV-16 E7, performed at least by its interference with Siva-1, can be considered an additional mechanism for the survival of damaged, potentially transforming, cell clones.

  3. Hepatocyte nuclear factor 4A improves hepatic differentiation of immortalized adult human hepatocytes and improves liver function and survival.

    Science.gov (United States)

    Hang, Hua-Lian; Liu, Xin-Yu; Wang, Hai-Tian; Xu, Ning; Bian, Jian-Min; Zhang, Jian-Jun; Xia, Lei; Xia, Qiang

    2017-11-15

    Immortalized human hepatocytes (IHH) could provide an unlimited supply of hepatocytes, but insufficient differentiation and phenotypic instability restrict their clinical application. This study aimed to determine the role of hepatocyte nuclear factor 4A (HNF4A) in hepatic differentiation of IHH, and whether encapsulation of IHH overexpressing HNF4A could improve liver function and survival in rats with acute liver failure (ALF). Primary human hepatocytes were transduced with lentivirus-mediated catalytic subunit of human telomerase reverse transcriptase (hTERT) to establish IHH. Cells were analyzed for telomerase activity, proliferative capacity, hepatocyte markers, and tumorigenicity (c-myc) expression. Hepatocyte markers, hepatocellular functions, and morphology were studied in the HNF4A-overexpressing IHH. Hepatocyte markers and karyotype analysis were completed in the primary hepatocytes using shRNA knockdown of HNF4A. Nuclear translocation of β-catenin was assessed. Rat models of ALF were treated with encapsulated IHH or HNF4A-overexpressing IHH. A HNF4A-positive IHH line was established, which was non-tumorigenic and conserved properties of primary hepatocytes. HNF4A overexpression significantly enhanced mRNA levels of genes related to hepatic differentiation in IHH. Urea levels were increased by the overexpression of HNF4A, as measured 24h after ammonium chloride addition, similar to that of primary hepatocytes. Chromosomal abnormalities were observed in primary hepatocytes transfected with HNF4A shRNA. HNF4α overexpression could significantly promote β-catenin activation. Transplantation of HNF4A overexpressing IHH resulted in better liver function and survival of rats with ALF compared with IHH. HNF4A improved hepatic differentiation of IHH. Transplantation of HNF4A-overexpressing IHH could improve the liver function and survival in a rat model of ALF. Copyright © 2017 Elsevier Inc. All rights reserved.

  4. Inhibition of chronic prostate inflammation by hyaluronic acid through an immortalized human prostate stromal cell line model.

    Science.gov (United States)

    Liu, Ming-Che; Chen, Wei-Hong; Chiou, Chi-Sheng; Lo, Wen-Cheng; Dubey, Navneet Kumar; Chen, Yu-Chin; Lai, Wen-Fu T; Yeh, Shauh-Der; Chiang, Han-Sun; Deng, Win-Ping

    2017-01-01

    Benign prostatic hyperplasia (BPH) is the most common urologic disease among elderly men. A well-established in vitro cell model is required to determine the therapeutic mechanism of BPH inflammation. In this study, we attempted to establish an immortalized human prostate stromal cell line by transfecting with HPV-16 E6/E7 and designated as ihPSC. No significant difference was found in fibroblast-like morphology between primary hPSC and ihPSC. The ihPSC possessed a significantly higher cell proliferation rate than primary hPSC. The prostate-specific markers and proteins including cytoskeleton (α-SMA and vimentin) and smooth muscle (calponin), especially the androgen receptor (AR) were also examined in ihPSC, almost identical to the primary hPSC. To create an in vitro model featuring chronic prostatic inflammation, ihPSC was stimulated with IFN-γ+IL-17 and then treated with the high molecular weight hyaluronic acid hylan G-F 20 as an alternative strategy for inhibiting BPH inflammation. Hylan G-F 20 could dose-dependently diminish the inflammation-induced proliferation in ihPSC. The enhanced expressions of inflammatory molecules including IL-1β, IL-6, IL-8, cyclooxygenase 2 (COX2), inducible nitrogen oxide synthase (iNOS), and Toll-like receptor 4 (TLR4) were all abolished by hylan G-F 20. For inflammatory signaling, hylan G-F 20 can also diminish the IFN-γ+IL-17-increased expression of iNOS and p65 in ihPSC. These findings suggest that ihPSC could provide a mechanism-based platform for investigating prostate inflammation. The hylan G-F 20 showed strong anti-inflammatory effects by decreasing inflammatory cytokines and signalings in the ihPSC, indicating its therapeutic potentials in BPH treatment in the future.

  5. Inhibition of chronic prostate inflammation by hyaluronic acid through an immortalized human prostate stromal cell line model

    Science.gov (United States)

    Liu, Ming-Che; Chen, Wei-Hong; Chiou, Chi-Sheng; Lo, Wen-Cheng; Dubey, Navneet Kumar; Chen, Yu-Chin; Lai, Wen-Fu T.; Yeh, Shauh-Der; Chiang, Han-Sun; Deng, Win-Ping

    2017-01-01

    Benign prostatic hyperplasia (BPH) is the most common urologic disease among elderly men. A well-established in vitro cell model is required to determine the therapeutic mechanism of BPH inflammation. In this study, we attempted to establish an immortalized human prostate stromal cell line by transfecting with HPV-16 E6/E7 and designated as ihPSC. No significant difference was found in fibroblast-like morphology between primary hPSC and ihPSC. The ihPSC possessed a significantly higher cell proliferation rate than primary hPSC. The prostate-specific markers and proteins including cytoskeleton (α-SMA and vimentin) and smooth muscle (calponin), especially the androgen receptor (AR) were also examined in ihPSC, almost identical to the primary hPSC. To create an in vitro model featuring chronic prostatic inflammation, ihPSC was stimulated with IFN-γ+IL-17 and then treated with the high molecular weight hyaluronic acid hylan G-F 20 as an alternative strategy for inhibiting BPH inflammation. Hylan G-F 20 could dose-dependently diminish the inflammation-induced proliferation in ihPSC. The enhanced expressions of inflammatory molecules including IL-1β, IL-6, IL-8, cyclooxygenase 2 (COX2), inducible nitrogen oxide synthase (iNOS), and Toll-like receptor 4 (TLR4) were all abolished by hylan G-F 20. For inflammatory signaling, hylan G-F 20 can also diminish the IFN-γ+IL-17-increased expression of iNOS and p65 in ihPSC. These findings suggest that ihPSC could provide a mechanism-based platform for investigating prostate inflammation. The hylan G-F 20 showed strong anti-inflammatory effects by decreasing inflammatory cytokines and signalings in the ihPSC, indicating its therapeutic potentials in BPH treatment in the future. PMID:28558037

  6. Self-reported rate of eating is associated with higher circulating ALT activity in middle-aged apparently healthy Japanese men.

    Science.gov (United States)

    Mochizuki, Kazuki; Miyauchi, Rie; Hariya, Natsuyo; Misaki, Yasumi; Kasezawa, Nobuhiko; Tohyama, Kazushige; Goda, Toshinao

    2013-04-01

    Elevated circulating activities of alanine aminotransferase (ALT), a marker for liver injury, and the lifestyle of a higher rate of eating in healthy and preclinical subjects are associated with increased risk of obesity and diabetes. In this study, we examined the associations between self-reported rate of eating and circulating ALT activity in middle-aged apparently healthy Japanese men. We conducted a cross-sectional study of 3,929 apparently healthy men aged 40-59 years (mean ± SD age, 49.2 ± 5.8 years; BMI, 23.5 ± 2.8 kg/m²) who participated in health checkups in Japan. We analyzed their clinical serum parameters and lifestyle factors, including self-reported rate of eating. Associations between self-reported rate of eating and liver injury markers [ALT, γ-glutamyl transpeptidase (GTP), and aspartate aminotransferase (AST)], other clinical parameters or lifestyle factors were determined using analysis of variance followed by Tukey's test. Multivariate logistic regression analyses (MLRA) were performed with ALT activity as the dependent variable and independent variables that included self-reported rate of eating. MLRA showed that ALT activity showed trends for higher self-reported rate of eating after adjustment for age, energy intake, and smoking status. The association between ALT activity and self-reported rate of eating disappeared after adjustment for BMI. The results of this study show that ALT activity is positively associated with self-reported rate of eating in middle-aged apparently healthy Japanese men.

  7. Revised cutoff values of ALT and HBV DNA level can better differentiate HBeAg (-) chronic inactive HBV patients from active carriers.

    Science.gov (United States)

    Ijaz, Bushra; Ahmad, Waqar; Javed, Fouzia T; Gull, Sana; Hassan, Sajida

    2011-02-27

    ELISA is still used as primary test for diagnosis HBV disease. However, ELISA-positive patients were marked as HBV inactive after confirmation with PCR and vice versa. Our aim was to assess the performance of new cut-off value of ALT, HBV DNA load and significance of AST as screening tool for HBeAg (-) chronic active or inactive patients in Pakistani population. In a cross-sectional, cohort study, 567 HBeAg (-) patients followed for one year were selected. Patients with persistent elevated ALT than normal and HBV DNA ≥ 100,000 copies/mL were taken as active chronic. Diagnostic values for ALT, AST and HBV DNA load in HBV HBeAg (-) chronic active and inactive patients compared using receiver operation characteristic (ROC) curves. Of 567 HBeAg (-) patients, 228 were classified as chronic inactive and 339 as active. HBV infection was dominant in male. Serum ALT, AST and HBV DNA levels showed significant and high AUROC to differentiate chronic HBeAg (-) inactive patients from active. AUROC for Serum ALT, AST and HBV DNA were observed 0.997, 0.969 and 1.000, respectively. For revised cut off value for ALT (30 IU/L for male and 19 IU/L for female) and HBV DNA load ≥100,000 copies/mL, a PPV of 97%, NPV of 94%, a sensitivity of 98%, and a specificity of 92% was observed to discriminate active carriers from inactive carriers. We also observed 93.5% specificity, 83.1% sensitivity, 82% PPV and 89.5% NPV for AST ≤20 IU/L to differentiate inactive carriers from active ones in our study group. Revised cut off value of ALT and NIH derived HBV DNA value can better discriminate between HBeAg (-) chronic active and inactive patients.

  8. Relation of ALT and AST levels to the histopathological changes in liver biopsies of patients with chronic hepatitis C genotype 4.

    Science.gov (United States)

    Khattab, Hany; Fouad, Ahmed; Hamza, Maya; Mohey, Mohammad A; El-Akel, Wafaa; Ghoneim, Hossam; Abul-Fotouh, Amr; Esmat, Gamal

    2015-06-01

    Worldwide, Egypt has a high prevalence of adult hepatitis C virus (HCV) infection. Serum alanine aminotransferase (ALT) activity is most commonly measured to assess hepatic disease. The revision of the definition of the normal limits for the ALT level is advisable. The aim of this work was to compare the histopathological changes in the liver tissue biopsies of HCV-infected patients, clinically presenting with ALT levels below normal, based on the conventional, previously used upper limit of normal (ULN) of ALT (40U/L for men and 30U/L for women) with the proposed new ULN (30U/L for men, and 19U/L for women). This is a retrospective cross-sectional study. A total of 668 cases of chronic hepatitis C genotype 4 were included. Patients were classified according to grades of histological activity and fibrosis stages (by the Metavir scoring system). They were also classified into normal and high groups according to the old and new cutoffs of both aspartate transaminase (AST) and ALT levels. The results of our study showed that the serum AST level in our study showed a better correlation with the histopathological changes in liver biopsy rather than ALT, especially when using the old cutoff of the ULN for AST. The serum ALT level in our study (both the old and the new cutoffs) did not show a significant correlation with the histopathological status in the liver biopsies of our patients. This study concluded that the old cutoff of the ULN AST is a better predictor of fibrosis. Copyright © 2015 Arab Journal of Gastroenterology. Published by Elsevier B.V. All rights reserved.

  9. Rangelia vitalii: changes in the enzymes ALT, CK and AST during the acute phase of experimental infection in dogs.

    Science.gov (United States)

    Costa, Márcio Machado; França, Raqueli Teresinha; Da Silva, Aleksandro Schafer; Paim, Carlos Breno; Paim, Francine; do Amaral, Carlos Henrique; Dornelles, Guilherme Lopes; da Cunha, João Paulo Monteiro Carvalho Mori; Soares, João Fabio; Labruna, Marcelo Bahia; Mazzanti, Cinthia Melazzo Andrade; Monteiro, Silvia Gonzalez; Lopes, Sonia Terezinha Dos Anjos

    2012-01-01

    Rangelia vitalii is a protozoon that causes diseases in dogs, and anemia is the most common laboratory finding. However, few studies on the biochemical changes in dogs infected with this protozoon exist. Thus, this study aimed to investigate the biochemical changes in dogs experimentally infected with R. vitalii, during the acute phase of the infection. For this study, 12 female dogs (aged 6-12 months and weighing between 4 and 7 kg) were used, divided in two groups. Group A was composed of healthy dogs (n = 5); and group B consisted of infected animals (n = 7). Blood samples were collected on days 0, 10, 20 and 30 after infection, using tubes without anticoagulant to obtain serum and analyze the biochemical parameters. An increase in alanine aminotransferase (ALT) on day 20 (P < 0.05) was observed. Also, increased creatine kinase (CK) and aspartate aminotransferase (AST) levels were observed throughout the experimental period (P < 0.05). No changes in the serum gamma-glutamyltransferase, urea and creatinine levels were observed. Thus, is possible to conclude that experimental infection with R. vitalii in dogs causes changes to the biochemical profile, with increased ALT, AST and CK enzyme levels.

  10. #whitegenocide, the Alt-right and Conspiracy Theory: How Secrecy and Suspicion Contributed to the Mainstreaming of Hate

    Directory of Open Access Journals (Sweden)

    Andrew F. Wilson,

    2018-02-01

    Full Text Available This article considers the relationship between “hashtag activism” as it is currently being used by the alt-right and the tendency to draw on conspiracy theory that Richard Hofstadter identified as being prevalent among what he termed “pseudo-conservatives” half a century earlier. Both the alt-right and Hofstadter’s “pseudo-conservatives” can be characterised by a pronounced populist nationalism that understands its aims as protecting a particular way of life whilst drawing on an aggrieved sense of injustice at being conspired against by an unseen enemy. That this “enemy” is typically foreign in actuality or in spirit confirms the cultural dimension on which their politics is played out. It is argued here that this paranoid populist nationalism has been figuratively drawn upon in the rhetoric of Donald Trump and that this apparent openness to the “pseudo-conservative” discourse on nationalism has provided a bridging effect via which far right elements are seeking to normalize extremist viewpoints

  11. Frederick W. Alt received the 2015 Szent-Györgi Prize for Progress in Cancer Research.

    Science.gov (United States)

    Scully, Peter; Zhao, Jie; Ba, Sujuan

    2016-02-03

    The Szent-Györgyi Prize for Progress in Cancer Research is a prestigious scientific award established by the National Foundation for Cancer Research (NFCR)--a leading cancer research charitable organization in the United States that is committed to supporting scientific research and public education relating to the prevention, early diagnosis, better treatments, and ultimately, a cure for cancer. Each year, the Szent-Györgyi Prize honors an outstanding researcher, nominated by colleagues or peers, who has contributed outstanding, significant research to the fight against cancer, and whose accomplishments have helped improve treatment options for cancer patients. The Prize also promotes public awareness of the importance of basic cancer research and encourages the sustained investment needed to accelerate the translation of these research discoveries into new cancer treatments. This report highlights the pioneering work led by the 2015 Prize winner, Dr. Frederick Alt. Dr. Alt's work in the area of cancer genetics over four decades has helped to shape the very roots of modern cancer research. His work continues to profoundly impact the approaches that doctors around the globe use to diagnose and treat cancer. In particular, his seminal discoveries of gene amplification and his pioneering work on molecular mechanisms of DNA damage repair have helped to usher in the era of genetically targeted therapy and personalized medicine.

  12. ELEVATED ALANINE AMINOTRANSFERASE (ALT IN BLOOD DONORS: AN ASSESSMENT OF THE MAIN ASSOCIATED CONDITIONS AND ITS RELATIONSHIP TO THE DEVELOPMENT OF HEPATITIS C

    Directory of Open Access Journals (Sweden)

    Fernando Lopes GONÇALES Jr.

    1998-07-01

    Full Text Available The determination of aminotranferases levels is very useful in the diagnosis of hepatopathies. In recent years, an elevated serum ALT level in blood donors has been associated with an increased risk of post-transfusion hepatitis (PTH. The purpose of the study was to research the factors associated with elevated ALT levels in a cohort of voluntary blood donors and to evaluate the relationship between increased ALT levels and the development of hepatitis C (HCV infection. 166 volunteer blood donors with elevated ALT at the time of their first donation were studied. All of the donors were questioned about previous hepatopathies, exposure to hepatitis, exposure to chemicals, use of medication or drugs, sexual behaviour, contact with blood or secretions and their intake of alcohol. Every three months, the serum levels of AST, ALT, alkaline phosphatase, gamma glutamyl transpeptidase, cholesterol, triglyceride and glycemia are assessed over a two year follow-up. The serum thyroid hormone levels as well as the presence of auto-antibodies were also measured. Abdominal ultrasound was performed in all patients with persistently elevated ALT or AST levels. A needle biopsy of liver was performed in 9 donors without definite diagnostic after medical investigation. The presence of anti-HCV antibodies in 116 donors were assayed again the first clinical evaluation. At the end of follow-up period (2 years later 71 donors were tested again for the presence of anti-HCV antibodies. None of donors resulted positive for hepatitis B or hepatitis C markers during the follow-up. Of the 116 donors, 101 (87% had persistently elevated ALT serum levels during the follow-up. Obesity and alcoholism were the principal conditions related to elevated ALT serum levels in 91/101 (90.1% donors. Hypertriglyceridemia, hypercholesterolemia, hypothyroidism and diabetes mellitus also were associated with increased ALT levels. Only 1/101 (0.9% had mild chronic active non A-G viral

  13. Association between ALT level and the rate of cardio/cerebrovascular events in HIV-positive individuals: the D: A: D study.

    Science.gov (United States)

    Sabin, Caroline A; Ryom, Lene; Kovari, Helen; Kirk, Ole; de Wit, Stephane; Law, Matthew; Reiss, Peter; Dabis, Francois; Pradier, Christian; El-Sadr, Wafaa; Monforte, Antonella d'Arminio; Kamara, David; Phillips, Andrew N; Lundgren, Jens D

    2013-08-01

    An inverse association between serum alanine aminotransferase (ALT) levels and the risk of myocardial infarction (MI) has been reported in the general population. We investigated associations between ALT levels and the risk of various cardiovascular and cerebrovascular outcomes in a large cohort study of HIV-positive individuals. Using Poisson regression, we investigated associations between the latest ALT level and MI, coronary heart disease (CHD), and stroke, after adjusting for known confounders and cumulative/recent exposure to antiretroviral drugs. Analyses were also performed for the end points of all-cause/liver-related mortality and new-onset diabetes mellitus. By February 2011, participants had experienced 541 MIs, 804 CHD, and 258 stroke events. The MI rate decreased from 3.1/1000 person-years among those with ALT ≤18 U/L to 2.1/1000 person-years among those with ALT >60 U/L. After adjustment for confounders, each 2-fold increment in ALT was associated with a 19% drop in the MI rate {relative rate, 0.81 [95% confidence interval (CI): 0.74 to 0.89], P = 0.0001}. A weaker inverse association was seen for CHD with no indication of a linear association between ALT levels and stroke (P = 0.72). Adjusted relative rates were 0.88 (95% CI: 0.81 to 0.97) and 0.70 (95% CI: 0.54 to 0.92) in those who were hepatitis C virus negative and hepatitis C virus positive, respectively, and 0.72 (95% CI: 0.58 to 0.89) and 0.84 (0.77 to 0.93) in injection drug users and non-injection drug users, respectively. Liver-related mortality and diabetes both demonstrated a positive association with ALT levels, whereas all-cause mortality showed a U-shaped relationship. Higher ALT levels are associated with lower MI risk in HIV-positive individuals, but with higher risks of liver-related mortality and diabetes mellitus.

  14. Immortalization and Characterization of Porcine Macrophages That Had Been Transduced with Lentiviral Vectors Encoding the SV40 Large T Antigen and Porcine Telomerase Reverse Transcriptase

    Directory of Open Access Journals (Sweden)

    Takato Takenouchi

    2017-08-01

    Full Text Available The domestic pig is an important agricultural animal, and thus, infectious diseases that affect pigs can cause severe economic losses in the global swine industry. Various porcine pathogens target macrophages, which are classical innate immune cells. Although macrophages basically protect the host from pathogens, they also seem to contribute to infectious processes. Therefore, cultured macrophages can be used to develop in vitro models for studying not only genes associated with porcine innate immunity but also the infectious processes of porcine pathogens. However, the availability of porcine macrophage cell lines is limited. In this study, we describe a novel immortalized porcine kidney-derived macrophage (IPKM cell line, which was generated by transferring the SV40 large T antigen (SV40LT and porcine telomerase reverse transcriptase (pTERT genes into primary porcine kidney-derived macrophages using lentiviral vectors. The IPKM displayed a typical macrophage morphology and was routinely passaged (doubling time: about 4 days. These cells were immunostained for macrophage markers. In addition, they exhibited substantial phagocytosis of polystyrene microbeads and released inflammatory cytokines upon lipopolysaccharide (LPS stimulation. Furthermore, the maturation and secretion of interleukin-1β were observed after nigericin-induced inflammasome activation in LPS-primed IPKM. These findings suggest that IPKM exhibit the typical inflammatory characteristics of macrophages. By transferring the SV40LT and pTERT genes using lentiviral vectors, we also successfully immortalized macrophages derived from the peripheral blood of a low-density lipoprotein receptor-deficient pig. These results suggest that the co-expression of SV40LT and pTERT is an effective way of immortalizing porcine macrophages.

  15. Immortalized keratinocytes derived from patients with epidermolytic ichthyosis reproduce the disease phenotype: a useful in vitro model for testing new treatments.

    Science.gov (United States)

    Chamcheu, J C; Pihl-Lundin, I; Mouyobo, C E; Gester, T; Virtanen, M; Moustakas, A; Navsaria, H; Vahlquist, A; Törmä, H

    2011-02-01

    Epidermolytic ichthyosis (EI) is a skin fragility disorder caused by mutations in genes encoding suprabasal keratins 1 and 10. While the aetiology of EI is known, model systems are needed for pathophysiological studies and development of novel therapies. To generate immortalized keratinocyte lines from patients with EI for studies of EI cell pathology and the effects of chemical chaperones as putative therapies. We derived keratinocytes from three patients with EI and one healthy control and established immortalized keratinocytes using human papillomavirus 16-E6/E7. Growth and differentiation characteristics, ability to regenerate organotypic epidermis, keratin expression, formation of cytoskeletal aggregates, and responses to heat shock and chemical chaperones were assessed. The cell lines EH11 (K1_p.Val176_Lys197del), EH21 (K10_p.156Arg>Gly), EH31 (K10_p.Leu161_Asp162del) and NKc21 (wild-type) currently exceed 160 population doublings and differentiate when exposed to calcium. At resting state, keratin aggregates were detected in 9% of calcium-differentiated EH31 cells, but not in any other cell line. Heat stress further increased this proportion to 30% and also induced aggregates in 3% of EH11 cultures. Treatment with trimethylamine N-oxide and 4-phenylbutyrate (4-PBA) reduced the fraction of aggregate-containing cells and affected the mRNA expression of keratins 1 and 10 while 4-PBA also modified heat shock protein 70 (HSP70) expression. Furthermore, in situ proximity ligation assay suggested a colocalization between HSP70 and keratins 1 and 10. Reconstituted epidermis from EI cells cornified but EH21 and EH31 cells produced suprabasal cytolysis, closely resembling the in vivo phenotype. These immortalized cell lines represent a useful model for studying EI biology and novel therapies. © 2011 The Authors. BJD © 2011 British Association of Dermatologists.

  16. Interaction of CtBP with adenovirus E1A suppresses immortalization of primary epithelial cells and enhances virus replication during productive infection

    Energy Technology Data Exchange (ETDEWEB)

    Subramanian, T.; Zhao, Ling-jun; Chinnadurai, G., E-mail: chinnag@slu.edu

    2013-09-01

    Adenovirus E1A induces cell proliferation, oncogenic transformation and promotes viral replication through interaction with p300/CBP, TRRAP/p400 multi-protein complex and the retinoblastoma (pRb) family proteins through distinct domains in the E1A N-terminal region. The C-terminal region of E1A suppresses E1A/Ras co-transformation and interacts with FOXK1/K2, DYRK1A/1B/HAN11 and CtBP1/2 (CtBP) protein complexes. To specifically dissect the role of CtBP interaction with E1A, we engineered a mutation (DL→AS) within the CtBP-binding motif, PLDLS, and investigated the effect of the mutation on immortalization and Ras cooperative transformation of primary cells and viral replication. Our results suggest that CtBP–E1A interaction suppresses immortalization and Ras co-operative transformation of primary rodent epithelial cells without significantly influencing the tumorigenic activities of transformed cells in immunodeficient and immunocompetent animals. During productive infection, CtBP–E1A interaction enhances viral replication in human cells. Between the two CtBP family proteins, CtBP2 appears to restrict viral replication more than CtBP1 in human cells. - Highlights: • Adenovirus E1A C-terminal region suppresses E1A/Ras co-transformation. • This E1A region binds with FOXK, DYRK1/HAN11 and CtBP cellular protein complexes. • We found that E1A–CtBP interaction suppresses immortalization and transformation. • The interaction enhances viral replication in human cells.

  17. Laboratory tests in patients treated with isotretinoin: occurrence of liver and muscle abnormalities and failure of AST and ALT to predict liver abnormality.

    Science.gov (United States)

    Webster, Guy F; Webster, Timothy G; Grimes, Lorraine R

    2017-05-15

    Current laboratory monitoring may not be optimal. A retrospective chart review was performed on thelaboratory results of 246 patients who were treated with isotretinoin for acne over a 9-year period. Tests obtained were CBC, lipid panel, AST, ALT, CK, GGT,and C-reactive protein. Thirty-five patients had an elevated AST and 35 of these had an elevated CK; 32 had an elevated ALT and 11 of these had an elevated CK. Thirteen patients had an elevated GGT; in 5 this was the only abnormality, whereas 8 had a GGT elevation accompanied by an elevated AST or ALT. Two had an elevated GGT and an elevated CK with normal AST and ALT. Fifty-two patients had a single episode of elevated CK, of which 22 were female. However, 57 had multiple CK elevations and only one was female. Thirty-five patients had CK elevations <2 times normal; 38 had levels between 2 and 3 times normal, 18 had levels between 3 and 4 times normal, and 18 had levels greater than 4 times normal. We suggest that ALT and AST are not useful for monitoring isotretinoin therapy and that GGT and CK may be of greater value in managing patients.

  18. Post-operative delayed elevation of ALT correlates with early death in patients with HBV-related hepatocellular carcinoma and Post-hepatectomy Liver Failure.

    Science.gov (United States)

    Yu, Liang-He; Yu, Wen-Long; Zhao, Teng; Wu, Meng-Chao; Fu, Xiao-Hui; Zhang, Yong-Jie

    2018-01-16

    Post-hepatectomy Liver Failure (PHLF) remains the primary cause of perioperative death. The kinetics of transaminase levels are usually measured as markers of hepatocellular injury following partial hepatectomy, but their correlation with PHLF and post-operative mortality is unclear. The aim of study was to compare the post-operative transaminase kinetics with short term survival in those patients that developed PHLF. A retrospective review of patients with HBV-related HCC and who developed PHLF was performed. Logistic regression analysis was conducted to analyze risk factors for postoperative delayed elevation of ALT (PDE-ALT) PHLF and lethal PHLF. Of the 69 patients who developed PHLF 36 (52%) died. In those patients who died the mean ± SD ALT and AST rose from day (POD) 1-3 and continued to fluctuate with highly abnormal levels beyond day 3 with a mean ± SD peak ALT level beyond POD 3 of 1851 ± 1644 U/L (p ALT indicates an increased risk of death in HBV-related HCC patients with PHLF. Copyright © 2017 International Hepato-Pancreato-Biliary Association Inc. Published by Elsevier Ltd. All rights reserved.

  19. The impact of intraoperative vascular occlusion during liver surgery on postoperative peak ALT levels: A systematic review and meta-analysis.

    Science.gov (United States)

    Guo, Tao; Xiao, Yusha; Liu, Zhisu; Liu, Quanyan

    2016-03-01

    Intraoperative vascular occlusion techniques during liver surgeries have been performed and refined for decades. However, the impact of these techniques on postoperative peak ALT levels remains uncertain. Thus, we performed a literature review and meta-analysis to determine the impact of intraoperative vascular occlusion during liver surgery on postoperative peak ALT levels. A systematic literature search of the PubMed database was conducted to discover relevant controlled clinical trials. Studies that reported postoperative peak ALT values for both an observation group and a control group were included. The Q statistic and the I(2) index statistic were used to assess heterogeneity. Publication bias was evaluated using Egger's test and Orwin's fail-safe N test. Of the 281 retrieved articles, 10 articles fulfilled the inclusion criteria. These 10 articles involved 12 randomized controlled trials with a total of 1443 records. The pooled estimation results indicated that intraoperative vascular occlusion significantly elevated postoperative peak ALT levels (test for SMD: Z = 4.09, P ALT values than non-occlusion procedures for liver surgeries. Copyright © 2016 IJS Publishing Group Limited. Published by Elsevier Ltd. All rights reserved.

  20. Alt er tilladt, men ikke alt gavner

    DEFF Research Database (Denmark)

    Andersen, Kirsten Margrethe

    2016-01-01

    Det fyger med forslag om staten skal møde de religiøse påbud med forbud - mod halalslagtet kød, tørklæder og kønsopdelt svømning. Faktisk var de første kristne optaget af problemer, vi igen står midt i. Paulus havde en løsning. Vi stadig kan lære af....

  1. A importância do índice AST/ALT no diagnóstico da esteatohepatite não-alcoólica The importance of AST/ALT rate in nonalcoholic steatohepatitis diagnosis

    OpenAIRE

    Idilio ZAMIN Jr.; Angelo Alves de MATTOS; Christiano PERIN; Gabriel Zatti RAMOS

    2002-01-01

    Racional - A esteatohepatite não-alcoólica e a doença hepática alcoólica apresentam similaridade histológica e em algumas situações, o diagnóstico diferencial das mesmas pode ser difícil, pois alguns pacientes não revelam o consumo abusivo de álcool. Objetivo- Avaliar a utilidade da determinação do índice AST/ALT no diagnóstico diferencial da esteatohepatite não-alcoólica e da doença hepática alcoólica. Pacientes e Métodos -Foram estudados 29 pacientes obesos com esteatohepatite não-alcoólica...

  2. Bioethics in popular science: evaluating the media impact of The Immortal Life of Henrietta Lacks on the biobank debate.

    Science.gov (United States)

    Nisbet, Matthew C; Fahy, Declan

    2013-02-28

    The global expansion of biobanks has led to a range of bioethical concerns related to consent, privacy, control, ownership, and disclosure. As an opportunity to engage broader audiences on these concerns, bioethicists have welcomed the commercial success of Rebecca Skloot's 2010 bestselling book The Immortal Life of Henrietta Lacks. To assess the impact of the book on discussion within the media and popular culture more generally, we systematically analyzed the ethics-related themes emphasized in reviews and articles about the book, and in interviews and profiles of Skloot. We conducted a content analysis of a population of relevant English-language articles and transcripts (n = 125) produced by news organizations and publications in the U.S., Canada, Great Britain/Ireland, and Australia/New Zealand. We scored each article for the emphasis and appearance of 9 ethics-related themes. These were informed consent, welfare of the vulnerable, compensation, scientific progress, control/access, accountability/oversight, privacy, public education, and advocacy. The informed consent theme dominated media discussion, with almost 39.2 percent of articles/transcripts featuring the theme as a major focus and 44.8 percent emphasizing the theme as a minor focus. Other prominent themes and frames of reference focused on the welfare of the vulnerable (18.4 percent major emphasis; 36.0 percent minor emphasis), and donor compensation (19.2 percent major; 52.8 percent minor). Ethical themes that comprised a second tier of prominence included those of scientific progress, control/access, and accountability/oversight. The least prominent themes were privacy, public education, and advocacy. The book has been praised as an opportunity to elevate media discussion of bioethics, but such claims should be re-considered. The relatively narrow focus on informed consent in the media discussion generated by Skloot's book may limit the ability of ethicists and advocates to elevate attention to

  3. Bioethics in popular science: evaluating the media impact of The Immortal Llife of Henrietta Lacks on the biobank debate

    Science.gov (United States)

    2013-01-01

    Background The global expansion of biobanks has led to a range of bioethical concerns related to consent, privacy, control, ownership, and disclosure. As an opportunity to engage broader audiences on these concerns, bioethicists have welcomed the commercial success of Rebecca Skloot’s 2010 bestselling book The Immortal Life of Henrietta Lacks. To assess the impact of the book on discussion within the media and popular culture more generally, we systematically analyzed the ethics-related themes emphasized in reviews and articles about the book, and in interviews and profiles of Skloot. Methods We conducted a content analysis of a population of relevant English-language articles and transcripts (n = 125) produced by news organizations and publications in the U.S., Canada, Great Britain/Ireland, and Australia/New Zealand. We scored each article for the emphasis and appearance of 9 ethics-related themes. These were informed consent, welfare of the vulnerable, compensation, scientific progress, control/access, accountability/oversight, privacy, public education, and advocacy. Results The informed consent theme dominated media discussion, with almost 39.2 percent of articles/transcripts featuring the theme as a major focus and 44.8 percent emphasizing the theme as a minor focus. Other prominent themes and frames of reference focused on the welfare of the vulnerable (18.4 percent major emphasis; 36.0 percent minor emphasis), and donor compensation (19.2 percent major; 52.8 percent minor). Ethical themes that comprised a second tier of prominence included those of scientific progress, control/access, and accountability/oversight. The least prominent themes were privacy, public education, and advocacy. Conclusions The book has been praised as an opportunity to elevate media discussion of bioethics, but such claims should be re-considered. The relatively narrow focus on informed consent in the media discussion generated by Skloot’s book may limit the ability of

  4. Soma, food of the immortals according to the Bower Manuscript (Kashmir, 6th century A.D.).

    Science.gov (United States)

    Leonti, Marco; Casu, Laura

    2014-08-08

    Food is medicine and vice versa. In Hindu and Ayurvedic medicine, and among human cultures of the Indian subcontinent in general, the perception of the food-medicine continuum is especially well established. The preparation of the exhilarating, gold-coloured Soma, Amrita or Ambrosia, the elixir and food of the 'immortals'-the Hindu pantheon-by the ancient Indo-Aryans, is described in the Rigveda in poetic hymns. Different theories regarding the botanical identity of Soma circulate, but no pharmacologically and historically convincing theory exists to date. We intend to contribute to the botanical, chemical and pharmacological characterisation of Soma through an analysis of two historical Amrita recipes recorded in the Bower Manuscript. The recipes are referred therein as panaceas (clarified butter) and also as a medicine to treat nervous diseases (oil), while no exhilarating properties are mentioned. Notwithstanding this, we hypothesise, that these recipes are related to the ca. 1800 years older Rigvedic Soma. We suppose that the psychoactive Soma ingredient(s) are among the components, possibly in smaller proportions, of the Amrita recipes preserved in the Bower Manuscript. The Bower Manuscript is a medical treatise recorded in the 6th century A.D. in Sanskrit on birch bark leaves, probably by Buddhist monks, and unearthed towards the end of the 19th century in Chinese Turkestan. We analysed two Amrita recipes from the Bower Manuscript, which was translated by Rudolf Hoernle into English during the early 20th century. A database search with the updated Latin binomials of the herbal ingredients was used to gather quantitative phytochemical and pharmacological information. Together, both Amrita recipes contain around 100 herbal ingredients. Psychoactive alkaloid containing species still important in Ayurvedic, Chinese and Thai medicine and mentioned in the recipe for 'Amrita-Prâsa clarified butter' and 'Amrita Oil' are: Tinospora cordifolia (Amrita, Guduchi), three

  5. Expression of a TGF-{beta} regulated cyclin-dependent kinase inhibitor in normal and immortalized airway epithelial cells

    Energy Technology Data Exchange (ETDEWEB)

    Tierney, L.A.; Bloomfield, C.; Johnson, N.F. [and others

    1995-12-01

    Tumors arising from epithelial cells, including lung cancers are frequently resistant to factors that regulate growth and differentiation in normal in normal cells. Once such factor is transforming growth factor-{Beta} (TGF-{Beta}). Escape from the growth-inhibitory effects of TGF-{Beta} is thought to be a key step in the transformation of airway epithelial cells. most lung cancer cell lines require serum for growth. In contrast, normal human bronchial epithelial (NHBE) cells are exquisitely sensitive to growth-inhibitory and differentiating effects of TGF-{Beta}. The recent identification of a novel cyclin-dependent kinase inhibitor, p15{sup INK4B}, which is regulated by TGF-{Beta}, suggests a mechanism by which TGF-{Beta} mediates growth arrest in NHBE cells. The purpose of this study was two-fold: (1) to determine if p15{sup INK4B} is induced by TGF-{Beta} in NHBE cells or immortalized bronchial epithelial (R.1) cells and if that induction corresponds to a G1/S cell-cycle arrest; (2) to determine the temporal relationship between p15{sup INK4B} induction, cell-cycle arrest, and the phosphorylation state of the pRB because it is thought that p15{sup INK4B} acts indirectly by preventing phosphorylation of the RB gene product. In this study, expression of p15{sup INK4B} was examined in NHBE cells and R.1 cells at different time intervals following TGF-{Beta} treatment. The expression of this kinase inhibitor and its relationship to the cell and the pRb phosphorylation state were examined in cells that were both sensitive (NHBE) and resistant (R.1) to the effects of TGF-{Beta}. These results suggest that the cyclin-dependent kinase inhibitor, p15{sup INK4B}, is involved in airway epithelial cell differentiation and that loss or reduction of expression plays a role in the resistance of transformed or neoplastic cells to the growth-inhibitory effects of TGF-{Beta}.

  6. Mathematics revealed

    CERN Document Server

    Berman, Elizabeth

    1979-01-01

    Mathematics Revealed focuses on the principles, processes, operations, and exercises in mathematics.The book first offers information on whole numbers, fractions, and decimals and percents. Discussions focus on measuring length, percent, decimals, numbers as products, addition and subtraction of fractions, mixed numbers and ratios, division of fractions, addition, subtraction, multiplication, and division. The text then examines positive and negative numbers and powers and computation. Topics include division and averages, multiplication, ratios, and measurements, scientific notation and estim

  7. Elevated levels of AST, ALT, and CPK · no family history of liver disease · Dx?

    Science.gov (United States)

    Patel, Mitesh B; Maddur, Haripriya

    2017-07-01

    A 26-year-old healthy male veteran with bipolar disorder and post-traumatic stress disorder was referred for a gastroenterology consultation after a routine laboratory evaluation revealed elevated levels of aspartate aminotransferase, 1040 IU/L (normal range, 10-40 IU/L), and alanine aminotransferase, 334 IU/L (normal range, 7-56 IU/L). He had been taking divalproex and ziprasidone for the previous 2 years, during which time liver test results had been normal.

  8. Aging, fertility, and immortality.

    Science.gov (United States)

    Rauser, Casandra L; Mueller, Laurence D; Rose, Michael R

    2003-01-01

    Evolutionary theory suggests that fecundity rates will plateau late in life in the same fashion as mortality rates. We demonstrate that late-life plateaus arise for fecundity in Drosophila melanogaster. The result qualitatively fits the evolutionary theory of late life based on the force of natural selection. But there are a number of alternative interpretations. Fecundity plateaus could be secondary consequences of mortality-rate plateaus. Female fecundity plateaus might arise from diminished male sexual function. Another alternative hypothesis is analogous to male sexual inadequacy: nutritional shortfalls. These may arise later in life because of a decline in female feeding or digestion. If some females have a life-long tendency to lay eggs at a faster rate, but die earlier, then aging for fecundity could arise from the progressive loss of the fast-layers, with the late-life plateau simply the laying patterns of individual females who were slow-layers throughout adult life. If this type of model is generally applicable to late life, then we should find that the females who survive to lay at a slow but steady rate in late life have a similar laying pattern in mid-life.

  9. Space shuttle engineering and operations support. ALT separation reference trajectories for tailcone on orbiter forward and aft CG configurations. Mission planning, mission analysis and software formulation

    Science.gov (United States)

    Glenn, G. M.

    1977-01-01

    A preflight analysis of the ALT separation reference trajectories for the tailcone on, forward, and aft cg orbiter configurations is documented. The ALT separation reference trajectories encompass the time from physical separation of the orbiter from the carrier to orbiter attainment of the maximum ALT interface airspeed. The trajectories include post separation roll maneuvers by both vehicles and are generated using the final preflight data base. The trajectories so generated satisfy all known separation design criteria and violate no known constraints. The requirement for this analysis is given along with the specifications, assumptions, and analytical approach used to generate the separation trajectories. The results of the analytical approach are evaluated, and conclusions and recommendations are summarized.

  10. Improvement of ALT decay kinetics by all-oral HCV treatment: Role of NS5A inhibitors and differences with IFN-based regimens.

    Science.gov (United States)

    Cento, Valeria; Nguyen, Thi Huyen Tram; Di Carlo, Domenico; Biliotti, Elisa; Gianserra, Laura; Lenci, Ilaria; Di Paolo, Daniele; Calvaruso, Vincenza; Teti, Elisabetta; Cerrone, Maddalena; Romagnoli, Dante; Melis, Michela; Danieli, Elena; Menzaghi, Barbara; Polilli, Ennio; Siciliano, Massimo; Nicolini, Laura Ambra; Di Biagio, Antonio; Magni, Carlo Federico; Bolis, Matteo; Antonucci, Francesco Paolo; Di Maio, Velia Chiara; Alfieri, Roberta; Sarmati, Loredana; Casalino, Paolo; Bernardini, Sergio; Micheli, Valeria; Rizzardini, Giuliano; Parruti, Giustino; Quirino, Tiziana; Puoti, Massimo; Babudieri, Sergio; D'Arminio Monforte, Antonella; Andreoni, Massimo; Craxì, Antonio; Angelico, Mario; Pasquazzi, Caterina; Taliani, Gloria; Guedj, Jeremie; Perno, Carlo Federico; Ceccherini-Silberstein, Francesca

    2017-01-01

    Intracellular HCV-RNA reduction is a proposed mechanism of action of direct-acting antivirals (DAAs), alternative to hepatocytes elimination by pegylated-interferon plus ribavirin (PR). We modeled ALT and HCV-RNA kinetics in cirrhotic patients treated with currently-used all-DAA combinations to evaluate their mode of action and cytotoxicity compared with telaprevir (TVR)+PR. Mathematical modeling of ALT and HCV-RNA kinetics was performed in 111 HCV-1 cirrhotic patients, 81 treated with all-DAA regimens and 30 with TVR+PR. Kinetic-models and Cox-analysis were used to assess determinants of ALT-decay and normalization. HCV-RNA kinetics was biphasic, reflecting a mean effectiveness in blocking viral production >99.8%. The first-phase of viral-decline was faster in patients receiving NS5A-inhibitors compared to TVR+PR or sofosbuvir+simeprevir (pALT-normalization, noted λ, was slower in patients receiving TVR+PR or sofosbuvir+simeprevir compared to NS5A-inhibitors (0.17 vs 0.27 d-1, respectively, pALT normalization rate and, for a given level of viral reduction, ALT-normalization was more profound in patients receiving DAA, and NS5A in particular, than TVR+PR. Our data support a process of HCV-clearance by all-DAA regimens potentiated by NS5A-inhibitor, and less relying upon hepatocyte death than IFN-containing regimens. This may underline a process of "cell-cure" by DAAs, leading to a fast improvement of liver homeostasis.

  11. Therapeutic administration of IL-15 superagonist complex ALT-803 leads to long-term survival and durable antitumor immune response in a murine glioblastoma model.

    Science.gov (United States)

    Mathios, Dimitrios; Park, Chul-Kee; Marcus, Warren D; Alter, Sarah; Rhode, Peter R; Jeng, Emily K; Wong, Hing C; Pardoll, Drew M; Lim, Michael

    2016-01-01

    Glioblastoma is the most aggressive primary central nervous system malignancy with a poor prognosis in patients. Despite the need for better treatments against glioblastoma, very little progress has been made in discovering new therapies that exhibit superior survival benefit than the standard of care. Immunotherapy has been shown to be a promising treatment modality that could help improve clinical outcomes of glioblastoma patients by assisting the immune system to overcome the immunosuppressive tumor environment. Interleukin-15 (IL-15), a cytokine shown to activate several effector components of the immune system, may serve as an excellent immunotherapeutic candidate for the treatment of glioblastoma. Thus, we evaluated the efficacy of an IL-15 superagonist complex (IL-15N72D:IL-15RαSu-Fc; also known as ALT-803) in a murine GL261-luc glioblastoma model. We show that ALT-803, as a single treatment as well as in combination with anti-PD-1 antibody or stereotactic radiosurgery, exhibits a robust antitumor immune response resulting in a prolonged survival including complete remission in tumor bearing mice. In addition, ALT-803 treatment results in long-term immune memory against glioblastoma tumor rechallenge. Flow cytometric analysis of tumor infiltrating immune cells shows that ALT-803 leads to increased percentage of CD8+-cell infiltration, but not the NK cells, and IFN-γ production into the tumor microenvironment. Cell depletion studies, in accordance with the flow cytometric results, show that the ALT-803 therapeutic effect is dependent on CD4+ and CD8+ cells. These results provide a rationale for evaluating the therapeutic activity of ALT-803 against glioblastoma in the clinical setting. © 2015 UICC.

  12. Intravesical ALT-803 and BCG treatment reduces tumor burden in a carcinogen induced bladder cancer rat model; a role for cytokine production and NK cell expansion.

    Science.gov (United States)

    Gomes-Giacoia, Evan; Miyake, Makito; Goodison, Steve; Sriharan, Aravindhan; Zhang, Ge; You, Lijing; Egan, Jack O; Rhode, Peter R; Parker, Alexander S; Chai, Karl X; Wong, Hing C; Rosser, Charles J

    2014-01-01

    Intravesical Bacillus Calmette-Guérin (BCG) has been shown to induce a specific immunologic response (i.e., activation of IL-2 and effector T-cells), while preclinical studies using ALT-803 (mutated IL-15 analogue combined with IL-15Rα-Fc fusion) have shown promising results by prolonging the agent's half-life and stimulating CD8+ T-cells. Based on these results, we hypothesized that the intravesical administration of ALT-803 along with BCG will generate an immunologic response leading to significant bladder tumor burden reduction. Using a well-established carcinogen induced rat non-muscle invasive bladder cancer (NMIBC) model, we studied the effects of intravesical ALT-803 with and without BCG. Rat tissues were evaluated to document treatment response. Intravesical ALT-803 was safe and well tolerated alone and in combination with BCG. As a single treatment agent, ALT-803 reduced tumor burden by 35% compared to control whereas BCG alone only reduced tumor burden by 15%. However, the combination of ALT-803 plus BCG reduced tumor burden by 46% compared to control. Immune monitoring suggested that the antitumor response was linked to the production and secretion of IL-1α, IL-1β and RANTES, which in turn, induced the proliferation and activation of NK cells. Lastly, tumoral responses of the combinational treatment were associated with 76% reduction in angiogenesis, which is significantly higher than when assessed with either agent alone. The enhanced therapeutic index seen with this duplet provides justification for the development of this regimen for future clinical trials.

  13. Chitosan: poly( N-vinylpyrrolidone- alt-itaconic anhydride) nanocapsules—a promising alternative for the lung cancer treatment

    Science.gov (United States)

    Raţă, Delia Mihaela; Chailan, Jean-François; Peptu, Cătălina Anişoara; Costuleanu, Marcel; Popa, Marcel

    2015-07-01

    This study reports the preparation of novel polymeric nanocapsules based on a natural polymer, chitosan and a synthetic one, poly( N-vinylpyrrolidone- alt-itaconic anhydride) [(poly(NVPAI)] using an interfacial condensation technique. The infrared spectroscopy studies confirmed the crosslinking through the presence of amide bonds, formed between the two polymers chains. The diameter of nanocapsules was found in the range of 126-214 nm and it was determined by dynamic light scattering method. Morphological characterization demonstrated their nano size, the spherical shape of the nanocapsules and the formation of hollow particles. The nanocapsules presented good swelling capacity in aqueous solutions. 5-Fluorouracil (5-FU) loading and release capacity was studied, the processes being controlled by the drug diffusion through the polymeric membrane. The obtained results were encouraging, showing that 5-FU-loaded nanocapsules had 70 % higher apoptotic effect on A549 tumour cells than the drug in free state or mixed with the nanocapsules.

  14. Chitosan: poly(N-vinylpyrrolidone-alt-itaconic anhydride) nanocapsules—a promising alternative for the lung cancer treatment

    International Nuclear Information System (INIS)

    Raţă, Delia Mihaela; Chailan, Jean-François; Peptu, Cătălina Anişoara; Costuleanu, Marcel; Popa, Marcel

    2015-01-01

    This study reports the preparation of novel polymeric nanocapsules based on a natural polymer, chitosan and a synthetic one, poly(N-vinylpyrrolidone-alt-itaconic anhydride) [(poly(NVPAI)] using an interfacial condensation technique. The infrared spectroscopy studies confirmed the crosslinking through the presence of amide bonds, formed between the two polymers chains. The diameter of nanocapsules was found in the range of 126–214 nm and it was determined by dynamic light scattering method. Morphological characterization demonstrated their nano size, the spherical shape of the nanocapsules and the formation of hollow particles. The nanocapsules presented good swelling capacity in aqueous solutions. 5-Fluorouracil (5-FU) loading and release capacity was studied, the processes being controlled by the drug diffusion through the polymeric membrane. The obtained results were encouraging, showing that 5-FU-loaded nanocapsules had 70 % higher apoptotic effect on A549 tumour cells than the drug in free state or mixed with the nanocapsules

  15. Chitosan: poly(N-vinylpyrrolidone-alt-itaconic anhydride) nanocapsules—a promising alternative for the lung cancer treatment

    Energy Technology Data Exchange (ETDEWEB)

    Raţă, Delia Mihaela, E-mail: iureadeliamihaela@yahoo.com [„Apollonia” University of Iasi, Faculty of Medical Dentistry, „Academician Ioan Haulică” Research Institute (Romania); Chailan, Jean-François, E-mail: chailan@univ-tln.fr [University of Sud Toulon-Var, « Matériaux-Polymères-Interfaces-Environnement Marin (MAPIEM) Laboratory (France); Peptu, Cătălina Anişoara, E-mail: catipeptu@yahoo.co.uk [“Gheorghe Asachi” Technical University of Iasi, Department of Natural and Synthetic Polymers, Faculty of Chemical Engineering and Environmental Protection (Romania); Costuleanu, Marcel, E-mail: mcostuleanu@yahoo.com [University of Medicine and Pharmacy “Grigore T. Popa”- Iaşi, Department of General Pathology, Faculty of Dental Medicine (Romania); Popa, Marcel, E-mail: marpopa2001@yahoo.fr [“Gheorghe Asachi” Technical University of Iasi, Department of Natural and Synthetic Polymers, Faculty of Chemical Engineering and Environmental Protection (Romania)

    2015-07-15

    This study reports the preparation of novel polymeric nanocapsules based on a natural polymer, chitosan and a synthetic one, poly(N-vinylpyrrolidone-alt-itaconic anhydride) [(poly(NVPAI)] using an interfacial condensation technique. The infrared spectroscopy studies confirmed the crosslinking through the presence of amide bonds, formed between the two polymers chains. The diameter of nanocapsules was found in the range of 126–214 nm and it was determined by dynamic light scattering method. Morphological characterization demonstrated their nano size, the spherical shape of the nanocapsules and the formation of hollow particles. The nanocapsules presented good swelling capacity in aqueous solutions. 5-Fluorouracil (5-FU) loading and release capacity was studied, the processes being controlled by the drug diffusion through the polymeric membrane. The obtained results were encouraging, showing that 5-FU-loaded nanocapsules had 70 % higher apoptotic effect on A549 tumour cells than the drug in free state or mixed with the nanocapsules.

  16. Preserved GLP-1 and exaggerated GIP secretion in type 2 diabetes and relationships with triglycerides and ALT

    DEFF Research Database (Denmark)

    Alssema, Marjan; Rijkelijkhuizen, Josina M; Holst, Jens Juul

    2013-01-01

    -based study. METHODS: A total of 163 persons with normal glucose metabolism (NGM), 20 with intermediate hyperglycaemia and 20 with type 2 diabetes aged 40-65 years participated. Participants received a mixed meal and oral glucose load on separate occasions. Glucagon-like peptide 1 (GLP-1), glucose......AUC per hour) was inversely related to fasting triglycerides. GIP (tAUC per hour) was positively related to fasting and postprandial triglycerides. Higher fasting GIP levels were related to higher fasting and postprandial triglyceride levels and ALT. CONCLUSION: This study confirms that in type 2 diabetes......, GLP-1 secretion is generally preserved and that GIP secretion is exaggerated. The mechanism underlying the divergent associations of GLP-1 and GIP metabolism with fat metabolism and liver fat accumulation warrants further study....

  17. Liver stiffness measurement and biochemical markers in Senegalese chronic hepatitis B patients with normal ALT and high viral load.

    Directory of Open Access Journals (Sweden)

    Papa Saliou Mbaye

    Full Text Available BACKGROUND AND AIMS: Despite the high prevalence of chronic hepatitis B (CHB in Africa, few studies have been performed among African patients. We sought to evaluate liver stiffness measurement by FibroScan® (LSM and two biochemical scores (FibroTest®, Fibrometer® to diagnose liver fibrosis in Senegalese CHB patients with HBV plasma DNA load ≥3.2 log(10 IU/mL and normal alanine aminotransferase (ALT values. METHODS: LSM and liver fibrosis biochemical markers were performed on 225 consecutive HBV infected Senegalese patients with high viral load. Patients with an LSM range between 7 and 13 kPa underwent liver biopsy (LB. Two experienced liver pathologists performed histological grading using Metavir and Ishak scoring. RESULTS: 225 patients were evaluated (84% male and LB was performed in 69 patients, showing F2 and F3 fibrosis in 17% and 10% respectively. In these patients with a 7-13 kPa range of LSM, accuracy for diagnosis of significant fibrosis according to LB was unsatisfactory for all non-invasive markers with AUROCs below 0.70. For patients with LSM values below 7 kPa, FibroTest® (FT, and Fibrometer® (FM using the cut-offs recommended by the test promoters suggested a fibrosis in 18% of cases for FT (8% severe fibrosis and 8% for FM. For patients with LSM values greater than 13 kPa, FT, FM suggested a possible fibrosis in 73% and 70%, respectively. CONCLUSION: In highly replicative HBV-infected African patients with normal ALT and LSM value below 13 kPa, FibroScan®, FibroTest® or Fibrometer® were unsuitable to predict the histological liver status of fibrosis.

  18. [Aspartate aminotransferase (AST) more than alanine aminotransferase (ALT) levels predict the progression of liver fibrosis in chronic HCV infection].

    Science.gov (United States)

    Stránský, J; Ryzlová, M; Striteský, J; Horák, J

    2002-10-01

    The development and severity of liver fibrosis in patients with chronic HCV infection can be evaluated best according to the staging of fibrosis in blind liver biopsy. So far there is however no biochemical indicator suggesting advanced fibrosis or progression of fibrosis in chronic HCV infection. In 1997 - 1999 60 adult out-patients (32 women) with chronic HCV infection were examined by blind liver biopsy. The grading of hepatitis was scored according to Knodell and staging of fibrosis according to Desmet. All patients were anti-HCV positive, assessed by the ELISA-3 method and 48/60 had positive HCV RNA in serum. The main risk factor of HCV infection was blood transfusion (67%). Of 27 examined patients 20 (74%) had serotype HCV 1. Staging of fibrosis: histologically confirmed fibrosis was not recorded in 11 patients (18.3%), mild and medium fibrosis was recorded in 25 (42%), severe fibrosis in 14 (23%) and cirrhosis in 10 (17%). With confirmed fibrosis correlated more closely AST serum activity (p < 0.002) than ALT activity (p < 0.03). Steatosis of the liver was found in 25 (42%) patients. The mean age of patients with steatosis was significantly higher than that of patients without steatosis (p < 0.0008). Steatosis was more frequent in patients with fibrosis (p < 0.04), in particularin the age group above 60 years. The development of fibrosis in patients with chronic HCV infection is suggested by permanently elevated activity of both transaminases whereby AST has a higher predictive value than ALT activity. A total of 40% histologically tested patients had the highest staging of fibrosis (3 - 4). Steatosis is in chronic HCV infection a very frequent finding (42%), in particular in patients above 60 years and those with serious fibrosis. The finding of fibrosis should stimulate the initiation of antiviral treatment which can lead to regression of fibrosis and improvement of the histological finding.

  19. AST to ALT Ratio is elevated in disseminated histoplasmosis as compared to localized pulmonary disease and other endemic mycoses.

    Science.gov (United States)

    Spec, Andrej; Barrios, Christopher R; Ahmad, Usama; Proia, Laurie A

    2017-07-01

    Severe pulmonary or disseminated histoplasmosis often necessitates presumptive antifungal treatment while awaiting definitive diagnosis. Histoplasma antigen assays have improved sensitivity but results may lag up to 7 days. In order to increase diagnostic certainty, "soft clues" may be looked for in laboratory and radiologic data, such as elevated alkaline phosphatase or ferritin levels and findings of mediastinal adenopathy or hepatosplenomegaly. To determine if elevated aspartate aminotransferase (AST) to alanine aminotransferase (ALT) ratio is specific to histoplasmosis or a non-specific marker for disseminated fungal infection or sepsis in general, we retrospectively examined records of all patients diagnosed with an endemic fungal infection (EFI) at Rush University Medical Center from January of 1997 to October of 2012, and a cohort of septic patients with elevated liver enzymes. We identified 90 cases of EFIs during the study period that met all inclusion criteria (Histoplasma 21, Blastomyces 56, Coccidioides 12, Paracoccidioides 1). We also evaluated 10 control patients with bacterial sepsis. The mean ratio of AST to ALT in patients with disseminated histoplasmosis was 2.69 (95% CI:1.22, 4.16) while for other EFIs, the mean ratio ranged from 0.38 to 1.14 with disseminated coccidioidomycosis and blastomycosis respectively (P histoplasmosis in the appropriate host, and to possibly distinguish cross reactivity of the Histoplasma antigen assay with other EFIs. © The Author 2016. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  20. Altépetl: evidencia arqueológica de una organización político territorial en la Tlaxcala prehispánica

    OpenAIRE

    García Sánchez, Magdalena Amalia

    2007-01-01

    Los estudiosos de la historia antigua de México, entre los que hay historiadores y etnohistoriadores, han dado cuenta del concepto náhuatl altépetl, que tiene connotaciones territoriales (de a: agua y tepetl: cerro) pero también sociales (poblacionales, simbólicas) en Mesoamérica. Sin embargo, no existe un puente que una los datos históricos con los geográficos y arqueológicos, por lo que hasta ahora altépetl no ha tenido evidencia material que lo confirme. Es objetivo de este trabajo present...

  1. Altépetl: evidencia arqueológica de una organización político territorial en la Tlaxcala prehispánica

    OpenAIRE

    García Sánchez, Magdalena Amalia

    2006-01-01

    Los estudiosos de la historia antigua de México, entre los que hay historiadores y etnohistoriadores, han dado cuenta del concepto náhuatl altépetl, que tiene connotaciones territoriales (de a: agua y tepetl: cerro) pero también sociales (poblacionales, simbólicas) en Mesoamérica. Sin embargo, no existe un puente que una los datos históricos con los geográficos y arqueológicos, por lo que hasta ahora altépetl no ha tenido evidencia material que lo confirme. Es objetivo de este trabajo present...

  2. Orbiter altitude at ALT interface based on ALSES (Approach and Landing Shuttle Engineering Simulation) analyses. Mission planning, mission analysis and software formulation

    Science.gov (United States)

    Colwell, C. L.

    1976-01-01

    The details of the orbiter altitude attainable at the approach and landing tests (ALT) interface determined by the approach and landing shuttle engineering simulation (manned) were documented. The analysis culminated in the verification of the trends observed in a similar analysis performed previously on the space vehicle dynamics simulation (unmanned). Altitude variations attributable to pilot steering variability ranged between 492 ft higher to 383 ft lower. The requirement for this parametric analysis is first elaborated. The specifications, assumptions, and analytical approach used to determine the orbiter altitude at the ALT interface are then presented, followed by the results of the analytical approach and the conclusions and recommendations.

  3. The ratio of aspartate aminotransferase to alanine aminotransferase (AST/ALT): the correlation of value with underlying severity of alcoholic liver disease.

    Science.gov (United States)

    Gurung, R B; Purbe, B; Gyawali, P; Risal, P

    2013-01-01

    Alcoholic liver disease is one of the most frequently diagnosed liver problems in the hospitalized patients in most tertiary care hospitals all over the world .The diagnosis of alcoholic liver disease is most of the time clinical. The AST/ALT ratio is a useful and reliable biochemical marker of liver injury due to alcohol. Whether the value of AST/ALT ratio correlates with clinical severity has not been studied. To study values of AST/ALT ratio in correlation with clinical severity of illness due to alcoholic liver disease using Child-Pugh's grading. This is a retrospective study. Inpatient records of all the patients admitted with diagnosis of alcoholic liver disease from July 2009 to 2011 June were analyzed. Data from 174 patients with the diagnosis of alcoholic liver disease-alcoholic hepatitis or alcoholic cirrhosis were retrieved; out of 174 patients, 138 were eligible for the study. The AST/ALT ratio and Child's grading of all the patients were calculated from the documented biochemical and clinical parameters on admission. Demographic profiles of all the patients were also recorded and analyzed. The data was analyzed using software SPPSS 16 version. A total of 138 patients diagnosed as alcoholic liver disease since July 2009 to June 2011 were analyzed. The male-female ratio was found to be 5.34: 1.The mean age of the patients at diagnosis was found to be 47.58 ± 12.83 years. Among 138 patients, Mongolians were found to have the highest prevalence of alcoholic liver disease (38.8%), followed by Newars ( 33.6%), Brahmin and Chhetri (19.1%) and Dalit (7.2%). With respect to AST/ALT ratio and Child's grading of ALD, the mean AST/ALT ratio was found to be 3.03 ± 2.24 in those patients who had Chlild's grade C; likewise the mean AST/ALT ratio was 2.28 ± 1.14, and 1.68 ± 0.83 in patients with Child B and Child A respectively. The higher value of AST/ALT ratio is indicative of more severe liver damage due to alcohol.

  4. Immortalized prairie vole-derived fibroblasts (VMF-K4DTs) can be transformed into pluripotent stem cells and provide a useful tool with which to determine optimal reprogramming conditions.

    Science.gov (United States)

    Katayama, Masafumi; Hirayama, Takashi; Kiyono, Tohru; Onuma, Manabu; Tani, Tetsuya; Takeda, Satoru; Nishimori, Katsuhiko; Fukuda, Tomokazu

    2017-06-21

    The cellular conditions required to establish induced pluripotent stem cells (iPSCs), such as the number of reprogramming factors and/or promoter selection, differ among species. The establishment of iPSCs derived from cells of previously unstudied species therefore requires the extensive optimization of programming conditions, including promoter selection and the optimal number of reprogramming factors, through a trial-and-error approach. While the four Yamanaka factors Oct3/4, Sox2, Klf4, and c-Myc are sufficient for iPSC establishment in mice, we reported previously that six reprogramming factors were necessary for the creation of iPSCs from primary prairie vole-derived cells. Further to this study, we now show detailed data describing the optimization protocol we developed in order to obtain iPSCs from immortalized prairie vole-derived fibroblasts. Immortalized cells can be very useful tools in the optimization of cellular reprogramming conditions, as cellular senescence is known to dramatically decrease the efficiency of iPSC establishment. The immortalized prairie vole cells used in this optimization were designated K4DT cells as they contained mutant forms of CDK4, cyclin D, and telomerase reverse transcriptase (TERT). We show that iPSCs derived from these immortalized cells exhibit the transcriptional silencing of exogenous reprogramming factors while maintaining pluripotent cell morphology. There were no observed differences between the iPSCs derived from primary and immortalized prairie vole fibroblasts. Our data suggest that cells that are immortalized with mutant CDK4, cyclin D, and TERT provide a useful tool for the determination of the optimal conditions for iPSC establishment.

  5. Cytotoxicity and Genotoxicity of Panel of Single- and Multiwalled Carbon Nanotubes: In Vitro Effects on Normal Syrian Hamster Embryo and Immortalized V79 Hamster Lung Cells

    Directory of Open Access Journals (Sweden)

    C. Darne

    2014-01-01

    Full Text Available Carbon nanotubes (CNTs belong to a specific class of nanomaterials with unique properties. Because of their anticipated use in a wide range of industrial applications, their toxicity is of increasing concern. In order to determine whether specific physicochemical characteristics of CNTs are responsible for their toxicological effects, we investigated the cytotoxic and genotoxic effects of eight CNTs representative of each of the commonly encountered classes: single- SW-, double- DW-, and multiwalled (MW CNTs, purified and raw. In addition, because most previous studies of CNT toxicity were conducted on immortalized cell lines, we decided to compare results obtained from V79 cells, an established cell line, with results from SHE (Syrian hamster embryo cells, an easy-to-handle normal cell model. After 24 hours of treatment, MWCNTs were generally found to be more cytotoxic than SW- or DWCNTs. MWCNTs also provoked more genotoxic effects. No correlation could be found between CNT genotoxicity and metal impurities, length, surface area, or induction of cellular oxidative stress, but genotoxicity was seen to increase with CNT width. The toxicity observed for some CNTs leads us to suggest that they might also act by interfering with the cell cycle, but no significant differences were observed between normal and immortalized cells.

  6. Oncoprotein E2A-Pbx1 immortalizes a myeloid progenitor in primary marrow cultures without abrogating its factor-dependence.

    Science.gov (United States)

    Kamps, M P; Wright, D D

    1994-11-01

    E2A-PBX1 is a chimeric homeobox oncogene formed by the t(1;19) translocation of human pre-B cell acute lymphoblastic leukemia (ALL). In a previous study, we found that retroviral expression of E2A-Pbx1 in the marrow of reconstituted mice induced the formation of acute myeloid leukemia (AML) in vivo. Here, we report that E2A-Pbx1 can also immortalize myeloid progenitors in vitro, and that the outgrowth of immortalized myeloblasts is evident only in the presence of the myeloid lymphokine, granulocyte-macrophage colony stimulating factor (GM-CSF). When cultured in the presence of GM-CSF, responsive myeloblasts from normal marrow exhibit concurrent proliferation and differentiation, and undergo terminal differentiation into non-mitotic neutrophils and macrophages within 4 weeks. Infection of identical cultures with a retrovirus encoding E2A-Pbx1 produces a rapid outgrowth of myeloid progenitors that express high levels of E2A-Pbx1 protein. A small fraction of myeloblasts in each population exhibited limited differentiation to neutrophils, and all populations of myeloblasts retained a strict dependence on GM-CSF for both survival and proliferation. This data suggests that the function of E2A-Pbx1 in leukemias is to strongly retard differentiation without affecting growth-factor dependence.

  7. Toxicity evaluation of chlorinated organic compounds using immortalized rat hepatocytes; Fushika rat kansaibo wo mochiita yuki enso kagobutsu no dokusei hyoka no kokoromi

    Energy Technology Data Exchange (ETDEWEB)

    Sone, H.; Nakajima, M.; Yonemoto, J. [National Institute for Environmental Studies, Tsukuba (Japan)

    1997-11-10

    Chlorinated organic compounds has high priority for toxicity screening among environmental hazardous chemicals. In the present study, we used immortalized rat hepatocytes as a liver model in vitro to evaluate the toxicity of nine chlorinated organic compounds. Toxicity of nine chlorinated organic compounds were evaluated to cellular viability of immortalized rat hapatocytes. The potency of the toxicity based on 50% inhibitory concentration (IC50) value was in the following order: triclocalban>triclosan>3,4-dichloroaniline>2,5-diclorophenol> 2,5-dichloroanisole>p-dichlorobenzene> p-chloroaniline>o-dichlorobenzene=tris (2-chloroethyl) phosphate. The rank order of cytotoxic potency of nine chemicals was compared with toxicity information using animals. The rank order of cytotoxic potency did not relative to the order referenced mean lethal dose (LD50) as an index of acute toxicity of rats or mice. However, the rank order of cytotoxic potency relatively correlated non-observed adverse effect level (NOAEL) under the exposure duration adjusted for chronic toxicity in vivo. These data suggests that the origin of testing cell had better to make match target organ of toxic chemicals for extrapolation from data of bioassay in vitro to in vivo. 16 refs., 2 figs., 3 tabs.

  8. Feedback on the Inaugural Meeting of the Cost 347 Accelerated Loading Test (ALT) Action in Europe: June 27th to 29th 2001

    CSIR Research Space (South Africa)

    De Beer, Morris

    2001-11-01

    Full Text Available Describes in summary the attendance of the author at an invited meeting of the COST 347 ALT (Accelerated Loading Testing) Committee in Cologne, Germany, Europe. The meeting was held at Bast, near Cologne, from 27th June 2001 to 29th June 2001...

  9. Predictors of treatment efficacy and ALT non-normalization with sofosbuvir/ribavirin therapy for patients with hepatitis C virus genotype 2.

    Science.gov (United States)

    Watanabe, Takao; Tokumoto, Yoshio; Joko, Kouji; Michitaka, Kojiro; Horiike, Norio; Tanaka, Yoshinori; Tada, Fujimasa; Kisaka, Yoshiyasu; Nakanishi, Seiji; Nonaka, Takashi; Yamauchi, Kazuhiko; Hirooka, Masashi; Abe, Masanori; Hiasa, Yoichi

    2017-09-01

    The tolerability and efficacy of sofosbuvir and ribavirin in patients infected with hepatitis C virus (HCV) genotype 2 were investigated under actual clinical conditions. A total of 208 patients with chronic HCV genotype 2 infection were treated with sofosbuvir 400 mg and ribavirin (weight-based dosing) for 12 weeks. Treatment discontinuation and sustained virological response 12 (SVR12) were evaluated. Moreover, factors associated with SVR12, hemoglobin decreasing to less than 10 g/dL during treatment, and alanine aminotransferase (ALT) non-normalization after treatment were evaluated. In all patients, SVR12 responses were 96.1% (200/208). About 6 of 8 patients (3.8%) who did not achieve SVR12 were re-treatment patients, and eight patients who did not achieve SVR all had liver cirrhosis. Multivariate analysis also identified body mass index (OR = 0.79; P ALT non-normalization after treatment. Cirrhotic patients may have a relatively high rate of treatment failure. In patients whose M2BpGi levels are elevated, their ALT tended to not normalize after treatment completion. These patients who did not achieve normalization of ALT after sofosbuvir plus RBV treatment need more careful observation for emergence of hepatocellular carcinoma even after achievement of SVR. © 2017 Wiley Periodicals, Inc.

  10. Combined introduction of Bmi-1 and hTERT immortalizes human adipose tissue-derived stromal cells with low risk of transformation

    Energy Technology Data Exchange (ETDEWEB)

    Tatrai, Peter, E-mail: peter.tatrai@biomembrane.hu [Institute of Enzymology, Research Center for Natural Sciences, Hungarian Academy of Sciences, Karolina ut 29, H-1113 Budapest (Hungary); Department of Biochemistry and Molecular Biology, Medical and Health Science Center, University of Debrecen, Egyetem ter 1, H-4032 Debrecen (Hungary); Szepesi, Aron, E-mail: aron.szepesi@biomembrane.hu [Creative Cell Ltd., Puskas Tivadar utca 13, H-1119 Budapest (Hungary); Matula, Zsolt, E-mail: matula.zsolt@gmail.com [Creative Cell Ltd., Puskas Tivadar utca 13, H-1119 Budapest (Hungary); Szigeti, Anna, E-mail: anna.szigeti@biomembrane.hu [Creative Cell Ltd., Puskas Tivadar utca 13, H-1119 Budapest (Hungary); Buchan, Gyoengyi, E-mail: buchan@med.unideb.hu [Department of Biochemistry and Molecular Biology, Medical and Health Science Center, University of Debrecen, Egyetem ter 1, H-4032 Debrecen (Hungary); Madi, Andras, E-mail: madi@med.unideb.hu [Department of Biochemistry and Molecular Biology, Medical and Health Science Center, University of Debrecen, Egyetem ter 1, H-4032 Debrecen (Hungary); Stem Cell, Apoptosis and Genomics Research Group of the Hungarian Academy of Sciences, University of Debrecen, Egyetem ter 1, H-4032 Debrecen (Hungary); Uher, Ferenc, E-mail: uher@biomembrane.hu [Stem Cell Laboratory, Hungarian National Blood Transfusion Service, Dioszegi ut 64, H-1113 Budapest (Hungary); and others

    2012-05-25

    Highlights: Black-Right-Pointing-Pointer We immortalized human adipose stromal cells (ASCs) with hTERT, Bmi-1, and SV40T. Black-Right-Pointing-Pointer hTERT-only ASCs are prone to transformation, while Bmi-only ASCs become senescent. Black-Right-Pointing-Pointer SV40T introduced along with hTERT abrogates proliferation control and multipotency. Black-Right-Pointing-Pointer hTERT combined with Bmi-1 yields stable phenotype up to 140 population doublings. -- Abstract: Adipose tissue-derived stromal cells (ASCs) are increasingly being studied for their usefulness in regenerative medicine. However, limited life span and donor-dependent variation of primary cells such as ASCs present major hurdles to controlled and reproducible experiments. We therefore aimed to establish immortalized ASC cell lines that provide steady supply of homogeneous cells for in vitro work while retain essential features of primary cells. To this end, combinations of human telomerase reverse transcriptase (hTERT), murine Bmi-1, and SV40 large T antigen (SV40T) were introduced by lentiviral transduction into ASCs. The resulting cell lines ASC{sup hTERT}, ASC{sup Bmi-1}, ASC{sup Bmi-1+hTERT} and ASC{sup SV40T+hTERT} were tested for transgene expression, telomerase activity, surface immunomarkers, proliferation, osteogenic and adipogenic differentiation, karyotype, tumorigenicity, and cellular senescence. All cell lines have maintained expression of characteristic surface immunomarkers, and none was tumorigenic. However, ASC{sup Bmi-1} had limited replicative potential, while the rapidly proliferating ASC{sup SV40T+hTERT} acquired chromosomal aberrations, departed from MSC phenotype, and lost differentiation capacity. ASC{sup hTERT} and ASC{sup hTERT+Bmi-1}, on the other hand, preserved all essential MSC features and did not senesce after 100 population doublings. Notably, a subpopulation of ASC{sup hTERT} also acquired aberrant karyotype and showed signs of transformation after long-term culture

  11. Alternagin-C (ALT-C), a disintegrin-like protein from Rhinocerophis alternatus snake venom promotes positive inotropism and chronotropism in fish heart.

    Science.gov (United States)

    Monteiro, D A; Kalinin, A L; Selistre-de-Araujo, H S; Vasconcelos, E S; Rantin, F T

    2016-02-01

    Alternagin-C (ALT-C) is a disintegrin-like protein purified from the venom of the snake, Rhinocerophis alternatus. Recent studies showed that ALT-C is able to induce vascular endothelial growth factor (VEGF) expression, endothelial cell proliferation and migration, angiogenesis and to increase myoblast viability. This peptide, therefore, can play a crucial role in tissue regeneration mechanisms. The aim of this study was to evaluate the effects of a single dose of alternagin-C (0.5 mg kg(-1), via intra-arterial) on in vitro cardiac function of the freshwater fish traíra, Hoplias malabaricus, after 7 days. ALT-C treatment increased the cardiac performance promoting: 1) significant increases in the contraction force and in the rates of contraction and relaxation with concomitant decreases in the values of time to the peak tension and time to half- and 90% relaxation; 2) improvement in the cardiac pumping capacity and maximal electrical stimulation frequency, shifting the optimum frequency curve upward and to the right; 3) increases in myocardial VEGF levels and expression of key Ca(2+)-cycling proteins such as SERCA (sarcoplasmic reticulum Ca(2+)-ATPase), PLB (phospholamban), and NCX (Na(+)/Ca(2+) exchanger); 4) abolishment of the typical negative force-frequency relationship of fish myocardium. In conclusion, this study indicates that ALT-C improves cardiac function, by increasing Ca(2+) handling efficiency leading to a positive inotropism and chronotropism. The results suggest that ALT-C may lead to better cardiac output regulation indicating its potential application in therapies for cardiac contractile dysfunction. Copyright © 2015 Elsevier Ltd. All rights reserved.

  12. Predictive value of serum ALT and T-cell receptor beta variable chain for HBeAg seroconversion in chronic hepatitis B patients during tenofovir treatment.

    Science.gov (United States)

    Yang, Jiezuan; Yan, Dong; Guo, Renyong; Chen, Jiajia; Li, Yongtao; Fan, Jun; Fu, Xuyan; Yao, Xinsheng; Diao, Hongyan; Li, Lanjuan

    2017-03-01

    Effective antiviral therapy plays a key role in slowing the progression of chronic hepatitis B (CHB). Identification of serum indices, including hepatitis B e antigen (HBeAg) expression and seroconversion, will facilitate evaluation of the efficacy of antiviral therapy in HBeAg-positive CHB patients. The biochemical, serological, virological parameters, and the frequency of circulating CD4CD25 regulatory T cell (Treg) in 32 patients were measured at baseline and every 12 weeks during 96 weeks of tenofovir disoproxil fumarate (TDF) treatment. The relationship between the hepatitis B virus (HBV) deoxyribonucleic acid (DNA) and Treg and alanine aminotransferase (ALT) levels was analyzed, respectively. The molecular profiles of T-cell receptor beta variable chain (TRBV) were determined using gene melting spectral pattern. For the seroconverted 12 patients, ALT declined to normal levels by week 24 and remained at this level in subsequent treatment; moreover, the predictive cutoff value of ALT for HBeAg seroconversion (SC) was 41.5 U/L at week 24. The positive correlation between HBV DNA and Treg and ALT was significant in SC patients, but not in non-SC patients. Six TRBV families (BV3, BV11, BV12, BV14, BV20, and BV24) were predominantly expressed in SC patients at baseline. The decline of ALT could be used to predict HBeAg seroconversion for CHB patients during TDF treatment. In addition, the profile of Tregs and TRBVs may be associated with HBeAg seroconversion and could also be a potential indicator for predicting HBeAg SC and treatment outcome for CHB patients.

  13. Cooperative therapeutic anti-tumor effect of IL-15 agonist ALT-803 and co-targeting soluble NKG2D ligand sMIC.

    Science.gov (United States)

    Basher, Fahmin; Jeng, Emily K; Wong, Hing; Wu, Jennifer

    2016-01-05

    Shedding of the human NKG2D ligand MIC (MHC class I-chain-related molecule) from tumor cell surfaces correlates with progression of many epithelial cancers. Shedding-derived soluble MIC (sMIC) enables tumor immune escape through multiple immune suppressive mechanisms, such as disturbing natural killer (NK) cell homeostatic maintenance, impairing NKG2D expression on NK cells and effector T cells, and facilitating the expansion of arginase I+ myeloid suppressor cells. Our recent study has demonstrated that sMIC is an effective cancer therapeutic target. Whether targeting tumor-derived sMIC would enhance current active immunotherapy is not known. Here, we determined the in vivo therapeutic effect of an antibody co-targeting sMIC with the immunostimulatory IL-15 superagonist complex, ALT-803, using genetically engineered transplantable syngeneic sMIC+ tumor models. We demonstrate that combined therapy of a nonblocking antibody neutralizing sMIC and ALT-803 improved the survival of animals bearing sMIC+ tumors in comparison to monotherapy. We further demonstrate that the enhanced therapeutic effect with combined therapy is through concurrent augmentation of NK and CD8 T cell anti-tumor responses. In particular, expression of activation-induced surface molecules and increased functional potential by cytokine secretion are improved greatly by the administration of combined therapy. Depletion of NK cells abolished the cooperative therapeutic effect. Our findings suggest that administration of the sMIC-neutralizing antibody can enhance the anti-tumor effects of ALT-803. With ALT-803 currently in clinical trials to treat progressive solid tumors, the majority of which are sMIC+, our findings provide a rationale for co-targeting sMIC to enhance the therapeutic efficacy of ALT-803 or other IL-15 agonists.

  14. Immortalization of Human Fetal Hepatocyte by Ectopic Expression of Human Telomerase Reverse Transcriptase, Human Papilloma Virus (E7) and Simian Virus 40 Large T (SV40 T) Antigen Towards Bioartificial Liver Support.

    Science.gov (United States)

    Giri, Shibashish; Bader, Augustinus

    2014-09-01

    Generation of genetically stable and non-tumoric immortalization cell line from primary cells would be enormously useful for research and therapeutic purposes, but progress towards this goal has so far been limited. It is now universal acceptance that immortalization of human fetal hepatocytes based on recent advances of telomerase biology and oncogene, lead to unlimited population doubling could be the possible source for bioartificial liver device. Immortalization of human fetal hepatocytes cell line by ectopic expression of human telomerase reverse transcriptase (hTERT), human papilloma virus gene (E7) and simian virus 40 large T (SV40 T) antigens is main goal of present study. We used an inducible system containing human telomerase and E7, both of which are cloned into responder constructs controlled by doxycycline transactivator. We characterized the immortalized human fetal hepatocyte cells by analysis of green fluorescent cells (GFP) positive cells using flow cytometry (FACs) cell sorting and morphology, proliferative rate and antigen expression by immunohistochemical analysis. In addition to we analysized lactate formation, glucose consumption, albumin secretion and urea production of immortalized human fetal hepatocyte cells. After 25 attempts for transfection of adult primary hepatocytes by human telomerase and E7 to immortalize them, none of the transfection systems resulted in the production of a stable, proliferating cell line. Although the transfection efficiency was more than 70% on the first day, the vast majority of the transfected hepatocytes lost their signal within the first 5-7 days. The remaining transfected hepatocytes persisted for 2-4 weeks and divided one or two times without forming a clone. After 10 attempts of transfection human fetal hepatocytes using the same transfection system, we obtained one stable human fetal hepatocytes cell line which was able albumin secretion urea production and glucose consumption. We established a

  15. Transduction of an immortalized olfactory ensheathing glia cell line with the green fluorescent protein (GFP) gene: Evaluation of its neuroregenerative capacity as a proof of concept.

    Science.gov (United States)

    Plaza, N; Simón, D; Sierra, J; Moreno-Flores, M T

    2016-01-26

    Olfactory ensheathing glia (OEG) cells are known to foster axonal regeneration of central nervous system (CNS) neurons. Several lines of reversibly immortalized human OEG (ihOEG) have been previously established that enabled to develop models for their validation in vitro and in vivo. In this work, a constitutively GFP-expressing ihOEG cell line was obtained, and named Ts14-GFP. Ts14-GFP neuroregenerative ability was similar to that found for the parental line Ts14 and it can be assayed using in vivo transplantation experimental paradigms, after spinal cord or optic nerve damage. Additionally, we have engineered a low-regenerative ihOEG line, hTL2, using lentiviral transduction of the large T antigen from SV40 virus, denominated from now on Ts12. Ts12 can be used as a low regeneration control in these experiments. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  16. Herpes simplex virus-1 infection or Simian virus 40-mediated immortalization of corneal cells causes permanent translocation of NLRP3 to the nuclei

    Directory of Open Access Journals (Sweden)

    Shu-Long Wang

    2015-01-01

    Full Text Available AIM: To investigate into the potential involvement of pyrin containing 3 gene (NLRP3, a member of the nucleotide-binding oligomerization domain-like receptors with cytosolic pattern recognition, in the host defense of corneas against viruses. METHODS: The herpes viral keratitis model was utilized in BALB/c mice with inoculation of herpes simplex virus-1 (HSV-1. Corneal tissues removed during therapy of patients with viral keratitis as well as a Simian vacuolating virus 40 (SV40-immortalized human corneal epithelial cell line were also examined. Immunohistochemistry was used to detect NLRP3 in these subjects, focusing on their distribution in tissue or cells. Western blot was used to measure the level of NLRP3 and another two related molecules in NLPR3 inflammasome, namely caspase-1 and IL-1β. RESULTS: The NLRP3 activation induced by HSV-1 infection in corneas was accompanied with redistribution of NLRP3 from the cytoplasm to the nucleus in both murine and human corneal epithelial cells. Furthermore, in the SV40-immortalized human corneal epithelial cells, NLRP3 was exclusively located in the nucleus, and treatment of the cells with high concentration of extracellular potassium (known as an inhibitor of NLRP3 activation effectively drove NLRP3 back to the cytoplasm as reflected by both immunohistochemistry and Western blot. CONCLUSION: It is proposed that herpes virus infection activates and causes redistribution of NLRP3 to nuclei. Whether this NLRP3 translocation occurs with other viral infections and in other cell types merit further study.

  17. Immortal time bias in pharmacoepidemiological studies on cancer patient survival: empirical illustration for beta-blocker use in four cancers with different prognosis.

    Science.gov (United States)

    Weberpals, Janick; Jansen, Lina; van Herk-Sukel, Myrthe P P; Kuiper, Josephina G; Aarts, Mieke J; Vissers, Pauline A J; Brenner, Hermann

    2017-11-01

    Immortal time bias (ITB) is still seen frequently in medical literature. However, not much is known about this bias in the field of cancer (pharmaco-)epidemiology. In context of a hypothetical beneficial beta-blocker use among cancer patients, we aimed to demonstrate the magnitude of ITB among 9876 prostate, colorectal, lung and pancreatic cancer patients diagnosed between 1998 and 2011, which were selected from a database linkage of the Netherlands Cancer Registry and the PHARMO Database Network. Hazard ratios (HR) and 95% confidence intervals from three ITB scenarios, defining exposure at a defined point after diagnosis (model 1), at any point after diagnosis (model 2) and as multiple exposures after diagnosis (model 3), were calculated to investigate the association between beta-blockers and cancer prognosis using Cox proportional hazards regression. Results were compared to unbiased estimates derived from the Mantel-Byar model. Ignoring ITB led to substantial smaller HRs for beta-blocker use proposing a significant protective association in all cancer types [e.g. HR 0.18 (0.07-0.43) for pancreatic cancer in model 1], whereas estimates derived from the Mantel-Byar model were mainly suggesting no association [e.g. HR 1.10 (0.84-1.44)]. The magnitude of bias was consistently larger among cancer types with worse prognosis [overall median HR differences between all scenarios in model 1 and Mantel-Byar model of 0.56 (prostate), 0.72 (colorectal), 0.77 (lung) and 0.85 (pancreas)]. In conclusion, ITB led to spurious beneficial associations of beta-blocker use among cancer patients. The magnitude of ITB depends on the duration of excluded immortal time and the prognosis of each cancer.

  18. Differential regulation of iron chelator-induced IL-8 synthesis via MAP kinase and NF-κB in immortalized and malignant oral keratinocytes

    Directory of Open Access Journals (Sweden)

    Lee Suk-Keun

    2007-09-01

    Full Text Available Abstract Background Interleukin-8 (IL-8 is a cytokine that plays an important role in tumor progression in a variety of cancer types; however, its regulation is not well understood in oral cancer cells. In the present study, we examined the expression and mechanism of IL-8 in which it is involved by treating immortalized (IHOK and malignant human oral keratinocytes (HN12 cells with deferoxamine (DFO. Methods IL-8 production was measured by an enzyme-linked immunoabsorbent assay and reverse transcriptase-polymerase chain reaction (RT-PCR analysis. Electrophoretic mobility shift assays was used to determine NF-κB binding activity. Phosphorylation and degradation of the I-κB were analyized by Western blot. Results IHOK cells incubated with DFO showed increased expression of IL-8 mRNA, as well as higher release of the IL-8 protein. The up-regulation of DFO-induced IL-8 expression was higher in IHOK cells than in HN12 cells and was concentration-dependent. DFO acted additively with IL-1β to strongly up-regulate IL-8 in IHOK cells but not in HN12 cells. Accordingly, selective p38 and ERK1/2 inhibitors for both kinases abolished DFO-induced IL-8 expression in both IHOK and HN12 cells. Furthermore, DFO induced the degradation and phosphorylation of IκB, and activation of NF-κB. The IL-8 inducing effects of DFO were mediated by a nitric oxide donor (S-nitrosoglutathione, and by pyrrolidine dithiocarbamate, an inhibitor of NF-κB, as well as by wortmannin, which inhibits the phosphatidylinositol 3-kinase-dependent activation of NAD(PH oxidase. Conclusion This results demonstrate that DFO-induced IL-8 acts via multiple signaling pathways in immortalized and malignant oral keratinocytes, and that the control of IL-8 may be an important target for immunotheraphy against human oral premalignant lesions.

  19. Energy Level Tuning of Poly(phenylene-alt-dithienobenzothiadiazole)s for Low Photon Energy Loss Solar Cells.

    Science.gov (United States)

    Heuvel, Ruurd; van Franeker, Jacobus J; Janssen, René A J

    2017-03-01

    Six poly(phenylene- alt -dithienobenzothiadiazole)-based polymers have been synthesized for application in polymer-fullerene solar cells. Hydrogen, fluorine, or nitrile substitution on benzo-thiadiazole and alkoxy or ester substitution on the phenylene moiety are investigated to reduce the energy loss per converted photon. Power conversion efficiencies (PCEs) up to 6.6% have been obtained. The best performance is found for the polymer-fullerene combination with distinct phase separation and crystalline domains. This improves the maximum external quantum efficiency for charge formation and collection to 66%. The resulting higher photocurrent compensates for the relatively large energy loss per photon ( E loss = 0.97 eV) in achieving a high PCE. By contrast, the poly-mer that provides a reduced energy loss ( E loss = 0.49 eV) gives a lower photocurrent and a reduced PCE of 1.8% because the external quantum efficiency of 17% is limited by a suboptimal morphology and a reduced driving force for charge transfer.

  20. Separator Membrane from Crosslinked Poly(Vinyl Alcohol and Poly(Methyl Vinyl Ether-alt-Maleic Anhydride

    Directory of Open Access Journals (Sweden)

    Charu Vashisth Rohatgi

    2015-03-01

    Full Text Available In this work, we report separator membranes from crosslinking of two polymers, such as poly vinyl alcohol (PVA with an ionic polymer poly(methyl vinyl ether-alt-maleic anhydride (PMVE-MA. Such interpolymer-networked systems were extensively used for biomedical and desalination applications but they were not examined for their potential use as membranes or separators for batteries. Therefore, the chemical interactions between these two polymers and the influence of such crosslinking on physicochemical properties of the membrane are systematically investigated through rheology and by critical gel point study. The hydrogen bonding and the chemical interaction between PMVE-MA and PVA resulted in highly cross-linked membranes. Effect of the molecular weight of PVA on the membrane properties was also examined. The developed membranes were extensively characterized by studying their physicochemical properties (water uptake, swelling ratio, and conductivity, thermal and electrochemical properties using differential scanning calorimetry (DSC, dynamic mechanical analysis (DMA, thermo-gravimetric analysis (TGA and electrochemical impedance spectroscopy (EIS. The DSC study shows the presence of a single Tg in the membranes indicating compatibility of the two polymers in flexible and transparent films. The membranes show good stability and ion conductivity suitable for separator applications.

  1. Separator Membrane from Crosslinked Poly(Vinyl Alcohol) and Poly(Methyl Vinyl Ether-alt-Maleic Anhydride)

    Science.gov (United States)

    Rohatgi, Charu Vashisth; Dutta, Naba K.; Choudhury, Namita Roy

    2015-01-01

    In this work, we report separator membranes from crosslinking of two polymers, such as poly vinyl alcohol (PVA) with an ionic polymer poly(methyl vinyl ether-alt-maleic anhydride) (PMVE-MA). Such interpolymer-networked systems were extensively used for biomedical and desalination applications but they were not examined for their potential use as membranes or separators for batteries. Therefore, the chemical interactions between these two polymers and the influence of such crosslinking on physicochemical properties of the membrane are systematically investigated through rheology and by critical gel point study. The hydrogen bonding and the chemical interaction between PMVE-MA and PVA resulted in highly cross-linked membranes. Effect of the molecular weight of PVA on the membrane properties was also examined. The developed membranes were extensively characterized by studying their physicochemical properties (water uptake, swelling ratio, and conductivity), thermal and electrochemical properties using differential scanning calorimetry (DSC), dynamic mechanical analysis (DMA), thermo-gravimetric analysis (TGA) and electrochemical impedance spectroscopy (EIS). The DSC study shows the presence of a single Tg in the membranes indicating compatibility of the two polymers in flexible and transparent films. The membranes show good stability and ion conductivity suitable for separator applications. PMID:28347019

  2. Altıntop Suyundaki Acılığın Naringinaz Enzimi ile Giderilmesi

    Directory of Open Access Journals (Sweden)

    Zuhal Çeviker

    2015-02-01

    Full Text Available Bu çalışmada MarshSeedless ve Rio Red çeşiti altıntoplardan elde edilen meyve sularındaki acılığın giderilmesi araştırılmıştır. Acılık giderme işlemi farklı naringinaz konsantrasyonlarında (0.25, 0.50, 0.75 ve 1.00 g/L ve farklı sıcaklıklarda (25, 35 ve 40oC gerçekleştirilmiştir. Naringin giderimi, naringinaz miktarı ve sıcaklıktaki artışa paralel olarak artmıştır. Her iki çeşitte de en yüksek naringin giderimi 1 g/L naringinaz konsantrasyonunda 40oC’de ve 6 saatlik inkübasyon sonunda elde edilmiştir. Duyusal değerlendirmede panelistler kontrol örneklerini enzimle muamele edilmiş örneklerden ayırt edebilmişlerdir.

  3. Ethnobotany of the Alt Empordà region (Catalonia, Iberian Peninsula): plants used in human traditional medicine.

    Science.gov (United States)

    Parada, Montse; Carrió, Esperança; Bonet, Maria Angels; Vallès, Joan

    2009-07-30

    This paper provides significant ethnobotanical information on pharmaceutical plant uses from a tourist and industrialised region, where some degree of acculturation exists, so that there is urgency in recording such data. To collect, analyze and evaluate the ethnobotanical knowledge about medicinal plants in a north-eastern Iberian region (Alt Empordà, 1358 km(2), 129,160 inhabitants). We performed 101 semi-structured interviews with 178 informants (mean age 69; 71% women, 29% men), identified the plant taxa reported and analyzed the results, comparing them with those from other territories. The informants reported data on 518 species. Of these, 335, belonging to 80 botanical families, were claimed as medicinal. This work is focused on human medicinal plant uses, which represent 98% of the pharmaceutical uses (3581 out of 3643 use reports). Around 800 medicinal uses, concerning 200 species, have not, or have very rarely been cited as medicinal; of these, 32 uses of 30 species have been reported by three or more independent informants. The folk knowledge about medicinal plant use is still alive in the studied region, and a number of scarcely reported plant uses has been detected, some of them with promising phytotherapeutical applications.

  4. Performance of an Optimized Paper-Based Test for Rapid Visual Measurement of Alanine Aminotransferase (ALT in Fingerstick and Venipuncture Samples.

    Directory of Open Access Journals (Sweden)

    Sidhartha Jain

    Full Text Available A paper-based, multiplexed, microfluidic assay has been developed to visually measure alanine aminotransferase (ALT in a fingerstick sample, generating rapid, semi-quantitative results. Prior studies indicated a need for improved accuracy; the device was subsequently optimized using an FDA-approved automated platform (Abaxis Piccolo Xpress as a comparator. Here, we evaluated the performance of the optimized paper test for measurement of ALT in fingerstick blood and serum, as compared to Abaxis and Roche/Hitachi platforms. To evaluate feasibility of remote results interpretation, we also compared reading cell phone camera images of completed tests to reading the device in real time.96 ambulatory patients with varied baseline ALT concentration underwent fingerstick testing using the paper device; cell phone images of completed devices were taken and texted to a blinded off-site reader. Venipuncture serum was obtained from 93/96 participants for routine clinical testing (Roche/Hitachi; subsequently, 88/93 serum samples were captured and applied to paper and Abaxis platforms. Paper test and reference standard results were compared by Bland-Altman analysis.For serum, there was excellent agreement between paper test and Abaxis results, with negligible bias (+4.5 U/L. Abaxis results were systematically 8.6% lower than Roche/Hitachi results. ALT values in fingerstick samples tested on paper were systematically lower than values in paired serum tested on paper (bias -23.6 U/L or Abaxis (bias -18.4 U/L; a correction factor was developed for the paper device to match fingerstick blood to serum. Visual reads of cell phone images closely matched reads made in real time (bias +5.5 U/L.The paper ALT test is highly accurate for serum testing, matching the reference method against which it was optimized better than the reference methods matched each other. A systematic difference exists between ALT values in fingerstick and paired serum samples, and can be

  5. A importância do índice AST/ALT no diagnóstico da esteatohepatite não-alcoólica

    Directory of Open Access Journals (Sweden)

    ZAMIN Jr. Idilio

    2002-01-01

    Full Text Available Racional - A esteatohepatite não-alcoólica e a doença hepática alcoólica apresentam similaridade histológica e em algumas situações, o diagnóstico diferencial das mesmas pode ser difícil, pois alguns pacientes não revelam o consumo abusivo de álcool. Objetivo- Avaliar a utilidade da determinação do índice AST/ALT no diagnóstico diferencial da esteatohepatite não-alcoólica e da doença hepática alcoólica. Pacientes e Métodos -Foram estudados 29 pacientes obesos com esteatohepatite não-alcoólica, cujo o diagnóstico foi realizado após exclusão de outras causas de doença hepática e que na biopsia apresentasse, no mínimo, esteatose macrovesicular associada a infiltrado inflamatório lobular e injúria hepatocelular. Como grupo controle, foram estudados 28 pacientes com doença hepática alcoólica. Resultados - Nos pacientes com esteatohepatite não-alcoólica, a média de AST foi de 52,3 ± 21,2 U/L e a de ALT de 90,1 ± 37,9 U/L, sendo o índice AST/ALT menor que 1 em todos os casos. No grupo controle, a média de AST foi de 140 ± 82,5 U/L e a de ALT foi de 50,6 ± 40,3 U/L. O índice AST/ALT foi superior a 1 em todos os pacientes e a 2 em 24 (85,7%, o que foi estatisticamente significativo quando comparado aos pacientes com esteatohepatite não-alcoólica. Conclusão- O índice AST/ALT parece ser útil no diagnóstico diferencial das hepatopatias, sendo que valores inferiores a 1 sugerem fortemente a hipótese de esteatohepatite não-alcoólica.

  6. Performance of an Optimized Paper-Based Test for Rapid Visual Measurement of Alanine Aminotransferase (ALT) in Fingerstick and Venipuncture Samples.

    Science.gov (United States)

    Jain, Sidhartha; Rajasingham, Radha; Noubary, Farzad; Coonahan, Erin; Schoeplein, Ryan; Baden, Rachel; Curry, Michael; Afdhal, Nezam; Kumar, Shailendra; Pollock, Nira R

    2015-01-01

    A paper-based, multiplexed, microfluidic assay has been developed to visually measure alanine aminotransferase (ALT) in a fingerstick sample, generating rapid, semi-quantitative results. Prior studies indicated a need for improved accuracy; the device was subsequently optimized using an FDA-approved automated platform (Abaxis Piccolo Xpress) as a comparator. Here, we evaluated the performance of the optimized paper test for measurement of ALT in fingerstick blood and serum, as compared to Abaxis and Roche/Hitachi platforms. To evaluate feasibility of remote results interpretation, we also compared reading cell phone camera images of completed tests to reading the device in real time. 96 ambulatory patients with varied baseline ALT concentration underwent fingerstick testing using the paper device; cell phone images of completed devices were taken and texted to a blinded off-site reader. Venipuncture serum was obtained from 93/96 participants for routine clinical testing (Roche/Hitachi); subsequently, 88/93 serum samples were captured and applied to paper and Abaxis platforms. Paper test and reference standard results were compared by Bland-Altman analysis. For serum, there was excellent agreement between paper test and Abaxis results, with negligible bias (+4.5 U/L). Abaxis results were systematically 8.6% lower than Roche/Hitachi results. ALT values in fingerstick samples tested on paper were systematically lower than values in paired serum tested on paper (bias -23.6 U/L) or Abaxis (bias -18.4 U/L); a correction factor was developed for the paper device to match fingerstick blood to serum. Visual reads of cell phone images closely matched reads made in real time (bias +5.5 U/L). The paper ALT test is highly accurate for serum testing, matching the reference method against which it was optimized better than the reference methods matched each other. A systematic difference exists between ALT values in fingerstick and paired serum samples, and can be addressed by

  7. Astrofisica delle alte energie

    CERN Document Server

    Vietri, Mario

    2006-01-01

    L'insegnamento dell'astrofisica moderna richiede conoscenze di base di svariate discipline, oltre ad alcuni modelli fondamentali. Lo scopo di questo libro è di presentare sia le une che gli altri in maniera possibilmente esaustiva, e di essere dunque utilizzabile sia come prima introduzione alle problematiche astrofisiche, sia come testo di riferimento ulteriore per giovani studiosi.

  8. Riget - alt er tilladt

    DEFF Research Database (Denmark)

    Agger, Gunhild

    2017-01-01

    Riget indtager en nøglestilling, når overgangen mellem faser i Triers produktion skal karakteriseres. Riget danner bro mellem den billedfascination og de radikale stileksperimenter, der præger Triers tidlige produktion, og den genre- og plotbevidsthed, der er typisk for film som Breaking the Waves...

  9. Er alt design?

    DEFF Research Database (Denmark)

    Michelsen, Anders Ib; Engholm, Ida

    2016-01-01

    Design er et ord med mange betydninger. Hvad enten design er det, der kommer ud af en shoppingtur, om det indgår i et dannelsesbegreb, hvor den kultiverede og ”gode stol” har sin plads, er en uddannelse med forskning underneden eller en strategisk indsats for velfærdssamfundets forvandling til et...

  10. Alt koster 10 kr

    DEFF Research Database (Denmark)

    Ejersbo, Lisser Rye

    2014-01-01

    Jeg havde engang en elev, som fortalte, at når han gik i byen for at købe ind, så var det nemt at beregne, om han havde nok penge med i byen. Han sagde bare, at alle varerne kun kostede 10 kr. Det kan lyde som lidt af en tilsnigelse, så klassen og jeg besluttede at undersøge om hans metode virkede....

  11. Familien betyder alt

    DEFF Research Database (Denmark)

    Danneskiold-Samsøe, Sofie; Mørck, Yvonne; Sørensen, Bo Wagner

    Denne bog er et resultat af forskningsprojektet ”Vold mod kvinder i etniske minoritetsfamilier – en kvalitativ undersøgelse med fokus på fremadrettede indsatser” udbudt af TrygFonden og Danner. Målet har været at få viden om volden og de aktuelle støttetilbud, idet omkring halvdelen af de kvinder...

  12. Effect of single and three months treatment with Ukrain on aminotransferases (ALT and AST) and on the serum protein level in rodents.

    Science.gov (United States)

    Jagiełło-Wójtowicz, E; Kleinrok, Z; Surmaczyńska, B; Baran, E; Feldo, M; Nowicky, J W

    1992-01-01

    The influence of Ukrain on the activity of aminotransferases (ALT and AST) and on the serum total protein content was estimated in mice and rats of both sexes receiving single or repeated doses of the drug. It was found that one hour after intraperitoneal (i.p.) administration of Ukrain no characteristic changes were recorded in the activity of the investigated enzymes, or in the serum protein content of animals of either sex. Similar effects were observed after three months treatment with Ukrain in rats of either sex. Only in mice receiving Ukrain for three months was a rise in ALT and AST activity found. No particular changes were observed in the total serum protein level, except for a small decreases in the sera of male mice.

  13. The prognostic significance of preoperatively assessed AST/ALT (De Ritis) ratio on survival in patients underwent radical cystectomy.

    Science.gov (United States)

    Gorgel, Sacit Nuri; Kose, Osman; Koc, Esra Meltem; Ates, Erhan; Akin, Yigit; Yilmaz, Yuksel

    2017-09-01

    We aimed to evaluate prognostic significance of preoperatively assessed aspartate aminotransaminase (AST)/alanine aminotransferase (ALT) (De Ritis) ratio on survival in bladder cancer (BC) patients underwent radical cystectomy (RC). We, respectively, analysed clinical and pathological data of 153 patients who underwent RC for BC between February 2006 and December 2016 at a tertiary level hospital. The potential prognostic value of De Ritis ratio was assessed by using ROC curve analysis. The effect of the De Ritis ratio was analysed by the Kaplan-Meier method and Cox regression hazard models for patients' disease-specific survival (DSS) and overall survival (OAS). We had 149 BC patients, in total. Mean age was 61.65 ± 9.13 years. One hundred and thirty-nine (93.3%) of the patients were men. According to ROC analysis, optimal threshold of De Ritis ratio for DSS was 1.30. In Kaplan-Meier analyses, the high De Ritis ratio group showed worse progression in DSS and OAS (all parameters, p < 0.001). On Cox regression models of clinical and pathological parameters to predict DSS, De Ritis ratio (HR 5.79, 95% CI 2.25-15.13), pathological T stage (HR 15.89, 95% CI 3.92-64.33, in all p < 0.001); and to predict OAS, De Ritis ratio (HR 2.61, 95% CI 1.49-4.56; p < 0.001), pathological T stage (HR 5.42, 95% CI 2.63-11.64; p < 0.001) and age (HR 1.05, 95% CI 1.02-1.08; p = 0.001) were determined as independent prognostic factors. Preoperative elevated De Ritis ratio could be an independent prognostic factor in BC patients underwent RC. Our results should be confirmed by large and properly designed prospective, randomized trials.

  14. Altın Fiyatlarının Yapay Sinir Ağları ile Tahmini ve Bir Uygulama

    Directory of Open Access Journals (Sweden)

    Rıdvan YÜKSEL

    2016-03-01

    Full Text Available Bu çalışmada altın fiyatlarını yapay sinir ağları ile öngörmek amacıyla, altın fiyatlarını etkileyebileceği düşünülen değişkenler olan Gümüş fiyatları, Brent Petrol fiyatları, ABD doları/ EUR paritesi, EuroNext100 endeksi, Amerika Dow Jones Endeksi, 13 Hafta vadeli ABD bonosu faiz oranı ve ABD TÜFE endeksi kullanılarak modeller kurulmuştur. Yapay sinir ağları ile kurulan modellerden elde edilen tahmin sonuçları, gerçek değerler ile R2, RMSE, MAE ve MAPE (% gibi performans kriterleri hesaplanarak karşılaştırılmıştır. Elde edilen bulgular yapay sinir ağlarının altın fiyatlarının tahmininde başarı ile kullanılabileceğini göstermektedir. Yapılan duyarlılık analizinin sonuçları değerlendirildiğinde altın fiyatlarını etkileyen faktörlerin başında gümüş ve petrol fiyatlarının geldiği tespit edilmiştir.

  15. Associations of I148M variant in PNPLA3 gene with plasma ALT levels during 2-year follow-up in normal weight and overweight children: the PANIC Study.

    Science.gov (United States)

    Viitasalo, A; Pihlajamaki, J; Lindi, V; Atalay, M; Kaminska, D; Joro, R; Lakka, T A

    2015-04-01

    PNPLA3 I148M polymorphism (rs738409) has been strongly associated with liver fat content and plasma alanine aminotransferase (ALT) levels in obese adults and children, but little is known about these relationships in normal weight individuals. We studied the associations and interactions of overweight and the PNPLA3 I148M polymorphism with plasma ALT levels during 2-year follow-up in children. Subjects were a population sample of 481 Caucasian children aged 6-8 years examined at baseline and 419 children re-examined after 2-year follow-up. Altogether, 58 (12%) of 481 children at baseline and 71 (17%) of 419 children after 2-year follow-up were overweight. We assessed plasma ALT levels and other cardiometabolic risk factors and genotyped the PNPLA3 I148M polymorphism. Being overweight and carrying PNPLA3 148M allele were associated with increased ALT levels at baseline (P = 0.002; P = 0.033) and after 2-year follow-up (P ALT levels during 2-year follow-up. PNPLA3 148M allele carriers had increased ALT levels at baseline (P = 0.024 for interaction) and after 2-year follow-up (P = 0.002 for interaction) as well as a larger increase in ALT levels during 2-year follow-up (P = 0.002 for interaction) if they were overweight but not if they were normal weight. Further adjustment for clinical puberty, dietary factors, physical activity or sedentary behaviour had little or no effect on these associations. PNPLA3 148M allele carriers had higher plasma ALT levels and larger increase in ALT levels during follow-up than non-carriers only among overweight children. © 2014 The Authors. Pediatric Obesity © 2014 World Obesity.

  16. The effect of AST/ALT (De Ritis) ratio on survival and its relation to tumor histopathological variables in patients with localized renal cell carcinoma.

    Science.gov (United States)

    Canat, Lütfi; Ataly, Hasan Anil; Agalarov, Samir; Alkan, Ilter; Alturende, Fatih

    2017-12-07

    To assess the relationship between De Ritis (aspartate aminotransaminase [AST]/Alanine aminotransaminase [ALT]) ratio and pathological variables and whether it is an independent prognostic factor. We analyzed 298 consecutive patients who underwent radical or partial nephrectomy for non-metastatic renal cell carcinoma (RCC) between 2006 and 2015. The association between De Ritis ratio and pathological variables including tumor size, presence of renal vein invasion, vena cava invasion, renal capsule infiltration, Gerota fascia invasion, renal sinus involvement, renal pelvic invasion, angiolymphatic invasion, adrenal gland involvement, lymph node involvement, tumor necrosis, and Fuhrman's grade was tested. Multivariable Cox analysis was performed to evaluate the impact of this ratio on overall survival and cancer-specific survival. An increased preoperative De Ritis ratio was significantly associated with renal vein invasion, renal capsule infiltration and renal pelvis involvement (p<0.05) in non-metastatic RCC. On multivariate analysis we found that tumor size, Fuhrman grade and lymph node involvement were independent prognostic factors for cancerspecific survival. AST/ALT ratio had no influence on the risk of overall and cancerspecific survival. An increased preoperative AST/ALT ratio had a significant association with renal vein invasion, renal capsule infiltration and renal pelvis involvement in patients with non-metastatic RCC. However, it does not appear to be an independent prognostic marker in non-metastatic RCC. Copyright® by the International Brazilian Journal of Urology.

  17. [Clinical efficacy of AST/ALT ratio and platelet counts as predictors of degree of fibrosis in HBV infected patients without clinically evident liver cirrhosis].

    Science.gov (United States)

    Park, Soo Young; Kang, Kyung Hee; Park, Jee Hyun; Lee, Jong Hyup; Cho, Chang Min; Tak, Won Young; Kweon, Young Oh; Kim, Sung Kook; Choi, Yong Hwan

    2004-04-01

    Hepatic fibrosis is an important prognostic factor in chronic hepatitis B. Liver biopsy is a gold standard diagnostic tool but an invasive procedure, so it cannot be done on all patients. We evaluated the clinical efficacy of AST/ALT ratio and platelet counts as predictors of fibrosis in chronic hepatitis B. We reviewed retrospectively clinical records of 323 patients, who visited Kyungpook National University Hospital for chronic hepatitis B and underwent liver biopsy from September 1998 to May 2002. Correlation with laboratory parameters with hepatic fibrosis stage was identified. Of 323 patients, there were 278 male patients with mean age 27 (9~59). Platelet counts showed a significant correlation (r=-0.343, p=0.000), and AST/ALT ratio showed a weak but significant correlation (r=0.137, p=0.013) with fibrosis stage. Patients with severe fibrosis or cirrhosis (stage 3 and 4) can be identified to have AST/ALT ratio > 1 and platelet counts 1 in combination with platelet counts. However, its sensitivity is too low to replace liver biopsy.

  18. The use of the ALT Flap and Lateral Femoral Cutaneous Nerve for the Reconstruction of Carpal Soft Tissue and Ulnar Nerve Defects: a Case Report.

    Science.gov (United States)

    Karonidis, Athanasios; Bouloumpasis, Serafeim; Apostolou, Konstantinos; Tsoutsos, Dimosthenis

    2015-06-01

    The anterolateral thigh (ALT) flap has become one of the workhorse flaps, with indications including diverse reconstructive problems. The lateral thigh area is also a useful donor site for nerve grafts. The lateral femoral cutaneous (LFC) nerve can be dissected along with the ALT flap for a substantial length, depending on the requirements of the recipient site. The LFC nerve can be used as a vascularized or non-vascularized nerve graft. The technique offers advantages and it can find clinical applications, satisfying the functional and aesthetic reconstructive requirements of a complex defect. We report the case of a patient who presented with traumatic soft tissue defect of the volar aspect of the wrist and ulnar nerve defect as a complication of a fracture of distal radius. An ALT flap was used to reconstruct the soft tissue defect. The ulnar nerve was resected due to necrosis and the gap was repaired with non-vascularized grafts of the anterior branch of the LFC nerve. The soft tissues were resurfaced successfully without complications. Functional recovery was good for the superficial branch of the ulnar nerve, whereas it was variable for the deep branch of the ulnar nerve. The anterolateral thigh area offers significant advantages as donor site in the reconstruction of complex soft tissue defects being a large source of vascularized skin, fat, fascia, muscle and nerve. This availability allows for single donor site dissection, minimizing the operating time and the associated morbidity.

  19. The 1-year and 3-month prognostic utility of the AST/ALT ratio and model for end-stage liver disease score in patients with viral liver cirrhosis.

    Science.gov (United States)

    Giannini, Edoardo; Botta, Federica; Testa, Emanuela; Romagnoli, Paola; Polegato, Simone; Malfatti, Federica; Fumagalli, Alessandra; Chiarbonello, Bruno; Risso, Domenico; Testa, Roberto

    2002-11-01

    The AST/ALT ratio has shown good diagnostic accuracy in patients with chronic viral liver disease. However, its prognostic utility has never been tested. Recently, the Model for End-Stage Liver Disease (MELD) has been proposed as a simple and effective tool to predict survival in patients with liver cirrhosis. The aims of this study were to assess the 3-month and 1-yr prognostic ability of the AST/ALT ratio in a series of patients with virus-related liver cirrhosis, and to evaluate the relationship between the AST/ALT ratio and the MELD score and to compare their prognostic ability. The AST/ALT ratios and MELD scores of 99 patients with liver cirrhosis of viral etiology (73 patients with hepatitis C virus and 26 with hepatitis B virus) who had been followed-up for at least 1 yr were retrospectively calculated and correlated with the patients' 3-month and 1-yr prognosis. Receiver operating characteristic curves were used to determine the AST/ALT ratio and the MELD score cut-offs with the best sensitivity (SS) and specificity (SP) in discriminating between patients who survived and those who died. Univariate survival curves were estimated by the Kaplan-Meier method using the cut-offs identified by means of receiver operating characteristic curves. AST/ALT ratios and MELD scores showed a significant correlation (r(s) = 0.503, p = 0.0001). In all, 8% and 30% of the patients had died after 3 months and 1 yr of follow-up, respectively. AST/ALT ratios and MELD scores were significantly higher among the patients who died during both 3-month and 1-yr follow-up. An AST/ALT ratio cut-off of 1.17 had 87% SS and 52% SP, whereas a MELD cut-off of 9 had 57% SS and 74% SP in discriminating between patients who survived and those who died after I yr. The combined assessment of the AST/ALT ratio and/or MELD score had 90% SS and 78% SP. Survival curves of the patients showed that both parameters clearly discriminated between patients who survived and those who died in the short term

  20. Common reference intervals for aspartate aminotransferase (AST), alanine aminotransferase (ALT) and γ-glutamyl transferase (GGT) in serum: results from an IFCC multicenter study.

    Science.gov (United States)

    Ceriotti, Ferruccio; Henny, Joseph; Queraltó, Josep; Ziyu, Shen; Özarda, Yeşim; Chen, Baorong; Boyd, James C; Panteghini, Mauro

    2010-11-01

    Aspartate aminotransferase (AST), alanine aminotransferase (ALT) and γ-glutamyl transferase (GGT) measurements are important for the assessment of liver damage. The aim of this study was to define the reference intervals (RIs) for these enzymes in adults, paying attention to standardization of the methods used and careful selection of the reference population. AST, ALT and GGT were measured with commercial analytical systems standardized to the IFCC-recommended reference measurement systems. Three centers (two in Italy and one in China) measured their own freshly collected samples; one of these centers also measured frozen samples from the Nordic Countries RI Project and from a Turkish center. RIs were generated using non-parametric techniques from the results of 765 individuals (411 females and 354 males, 18-85 years old) selected on the basis of the results of other laboratory tests and a specific questionnaire. AST results from the four regions (Milan, Beijing, Bursa and Nordic Countries) were statistically different, but these differences were too small to be clinically relevant. Likewise, differences between the upper reference limits for genders was only 1.7 U/L (0.03 μkat/L), allowing a single RI of 11-34 U/L (0.18-0.57 μkat/L) to be defined. Interregional differences were not statistically significant for ALT, but partitioning was required due to significant gender differences. RIs for ALT were 8-41 U/L (0.13-0.68 μkat/L) for females and 9-59 U/L (0.15-0.99 μkat/L) for males, respectively. The upper reference limits for GGT from the Nordic Country population were higher than those from the other three regions and results from this group were excluded from final calculations. The GGT RIs were 6-40 U/L (0.11-0.66 μkat/L) for females and 12-68 U/L (0.20- 1.13 μkat/L) for males, respectively. For AST and ALT, the implementation of common RIs appears to be possible, because no differences between regions were observed. However, a common RI for GGT that is

  1. Fibroscan Compared to FIB-4, APRI, and AST/ALT Ratio for Assessment of Liver Fibrosis in Saudi Patients With Nonalcoholic Fatty Liver Disease.

    Science.gov (United States)

    Fallatah, Hind I; Akbar, Hisham O; Fallatah, Alyaa M

    2016-07-01

    Nonalcoholic fatty liver disease (NAFLD) is being increasingly recognized as a cause of chronic liver disease. It has also been associated with devastating outcomes such as decompensated liver cirrhosis and hepatocellular carcinoma, as well as diabetes and metabolic syndrome. This study was conducted in order to assess liver fibrosis using Fibroscan, and to compare these results to the use of Fibrosis-4 (FIB-4) scores, AST platelet ratio index (APRI scores), and the AST/ALT ratios on NAFLD patients. A cross sectional study was conducted on NAFLD patients who underwent Fibroscan examinations between September 1, 2011 and June 30, 2014. Demographic data was collected, including sex, age, and nationality; serum alanine aminotransferase levels (ALT, 30 - 65 U/L), serum aspartate aminotransferase levels (AST, 15 - 37 U/L), and platelet counts (150 - 400 k/μL) were also determined. The stages of fibrosis (F0 1 - 6, F1 6.1 - 7, F2 7 - 9, F3 9.1 - 10.3, and F4 ≥ 10.4) were defined in kPa. For each patient, the AST/ALT ratio was also measured. The results of APRI and FIB-4 were compared with the Fibroscan fibrosis scores. The results of 122 patients were analyzed, including 65 (53.3%) males with a mean age of 50.2 years (SD: 13.7; range: 18 - 86). The males were significantly younger than the females (48.7 years (SD: 16.03) versus 51.8 years (SD: 10.3 P = 0.05), respectively). The mean stiffness score was 12.02 (SD: 12.7) kPa. Forty-four patients (36%) had advanced fibrosis. The mean platelet and serum ALT levels were normal. There was a significant positive correlation between the Fibroscan results and the AST/ALT ratios, the APRI scores, and the FIB-4 results. Similarly, there was a significant positive correlation between age and fibrosis score, and a significant negative correlation between platelet count and stiffness score. The data showed that more than one-third of the cohort exhibited advanced fibrosis, demonstrating the need for the early diagnosis and

  2. La sémantique du stereotype et la representation topique de l’altérité

    Directory of Open Access Journals (Sweden)

    Afsaneh Pourmazaheri

    2017-12-01

    Full Text Available Ce travail de recherche s’interroge sur la représentation discursive des référents exotiques filtrée par un regard stéréotypé dans les récits de trois voyageurs français en Orient de la première moitié du XIXe siècle. Les énonciateurs-voyageurs, placés dans un contexte linguistique et culturel asymétrique, tentent de saisir et de transcrire leurs observations sur le terrain étranger en fonction de leur intellect, de leur cognition et de leur expérience. Une série d’éléments marqués par l’altérité viennent modifier la représentation que l’auteur-voyageur donne du référent et de l’univers référentiel auquel il est confronté. C’est à cette façon « réductrice » de voir les référents issus d’un univers différent et à l’intervention de la subjectivité de l’auteur que nous nous intéressons dans cette étude. Nous analysons les effets discursifs produits par l’usage des ressources linguistiques ainsi que par les positionnements énonciatifs des auteurs-voyageurs. Cette approche homogénéisante et généralisante de la perception de l’Autre se fait au travers des moules préconstruits et une démarche subjective dans la culture réceptrice que nous tenterons de voir à l’aide des outils d’analyse langagiers et discursifs dans les récits de voyage de Goupil Fesquet (1844, de Rémy Aucher-Eloy (1843 et de J.M. Tancoigne (1819 en Orient, notamment en Perse.

  3. Alternagin-C (ALT-C), a Disintegrin-Like Cys-Rich Protein Isolated from the Venom of the Snake Rhinocerophis alternatus, Stimulates Angiogenesis and Antioxidant Defenses in the Liver of Freshwater Fish, Hoplias malabaricus.

    Science.gov (United States)

    Monteiro, Diana Amaral; Selistre-de-Araújo, Heloisa Sobreiro; Tavares, Driele; Fernandes, Marisa Narciso; Kalinin, Ana Lúcia; Rantin, Francisco Tadeu

    2017-09-28

    Alternagin-C (ALT-C) is a disintegrin-like protein isolated from Rhinocerophis alternatus snake venom, which induces endothelial cell proliferation and angiogenesis. The aim of this study was to evaluate the systemic effects of a single dose of alternagin-C (0.5 mg·kg -1 , via intra-arterial) on oxidative stress biomarkers, histological alterations, vascular endothelial growth factor (VEGF) production, and the degree of vascularization in the liver of the freshwater fish traíra, Hoplias malabaricus , seven days after the initiation of therapy. ALT-C treatment increased VEGF levels and hepatic angiogenesis. ALT-C also enhanced hepatic antioxidant enzymes activities such as superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase, decreasing the basal oxidative damage to lipids and proteins in the fish liver. These results indicate that ALT-C improved hepatic tissue and may play a crucial role in tissue regeneration mechanisms.

  4. [Lange, Thomas. Zwischen Reformation und Untergang Alt-Livlands. Der Rigaer Erzbischof Wilhelm von Brandenburg im Beziehungsgeflecht der livländischen Konföderation und ihrer Nachbarländer] / Madis Maasing

    Index Scriptorium Estoniae

    Maasing, Madis, 1984-

    2014-01-01

    Arvustus: Lange, Thomas. Zwischen Reformation und Untergang Alt-Livlands. Der Rigaer Erzbischof Wilhelm von Brandenburg im Beziehungsgeflecht der livländischen Konföderation und ihrer Nachbarländer

  5. Multifunctional e-spun colloidal nanofiber structures from various dispersed blends of PVA/ODA-MMT with PVP/ODA-MMT, poly(VP-alt-MA) and AgNPs incorporated polymer complexes as electro-active platforms

    OpenAIRE

    U. Bunyatova; Z. M. O. Rzayev; M. Simsek

    2016-01-01

    This work presented a new approach to fabricate polymer nanocomposites films with nanofiber structures from solution blends of poly(vinyl alcohol) + octadecyl amine-montmorillonite (ODA-MMT) (matrix) with poly(N-vinylpyrrolidone) + ODA-MMT (partner-1), poly(N-vinylpyrrolidone-alt-maleic anhydride) ((poly(VP-alt-MA)) + (ODA-MMT) (partner-2) and their silver (Ag)-carrying polymer complexes by electrospinning. Chemical and physical structures, surface morphologies, thermal behaviors, electrical ...

  6. Combined multi-lobed flaps: A series of 39 extensive hand and multi-digit injuries one-staged reconstructions using modified designs of ALT, DPA and chimeric linking flaps.

    Science.gov (United States)

    Tang, Liang; Pafitanis, Georgios; Yang, Peng; Li, Zan; Jia, Ming; Koshima, Isao; Song, Dajiang; Chi, Zhenglin

    2017-07-01

    The use of multiple small flaps linked in a "chain-linked" flap microanastomosed chimeric system is recommended in distal hand and digital defects reconstruction. The aim of this study is to demonstrates our experience utilizing microsurgical fabrication, multi-lobed and linking combined flaps for the reconstruction of hand degloving injuries with complex multidigit soft tissue defects. Microanastomosed chimeric flap systems using ALT and DPA modified designed flaps were combined in five selections to cover extensive soft tissue defects involving the hands and multiple digits of 39 patients (M:F - 36:3) from October 2009 to February 2013. Five different microsurgical combined chimeric flap systems utilised in extensive hand and multidigit injuries; innervated ALT flaps, multilobed DPA flaps, innervated ALT flap with multilobed DPA flap, innervated ALT flap with sensate ALT flap and bilobed ALT flap with multilobed DPA flap. All DPA donor sites were reconstructed using free ALT flap and anterior tibial artery propeller flap. Thirty-nine combined free flap extremities reconstructions on 39 patients (M:F - 36:3) with average age 28.5 (18-45) years sustained traumatic degloving injuries, 24 from road traffic accidents and 15 from industrial devices. Five different designs of combined multi-lobed flaps have be successfully used without any peri-operative complications. Average follow-up of 12 months, all flaps survived without complications. Operated extremities showed favorable functional recovery with restoration of the diminished protective sensation on the flap through reinnervation. All flaps survived uneventfully with coverage matching the texture and color of the recipients. Donor sites healed without complication. The microsurgical fabrication of chimeric ALT flaps and multilobed DPA flaps is a valuable alternative for the reconstruction of hand degloving injury with complex multidigit soft tissue defects. Level IV, therapeutic study. Copyright © 2017 Elsevier

  7. Dienogest, a synthetic progestin, inhibits the proliferation of immortalized human endometrial epithelial cells with suppression of cyclin D1 gene expression.

    Science.gov (United States)

    Shimizu, Yutaka; Takeuchi, Takashi; Mita, Shizuka; Mizuguchi, Kiyoshi; Kiyono, Tohru; Inoue, Masaki; Kyo, Satoru

    2009-10-01

    Dienogest is a specific progesterone receptor agonist with potent oral endometrial activity and is used in the treatment of endometriosis. In this study, we examined the direct effects of dienogest on the proliferation of human endometrial epithelial cells using an immortalized cell line. 5-Bromo-2'-deoxyuridine incorporation into the cells was inhibited by dienogest and by progesterone (P(4)) in dose-dependent fashion at concentrations of 10(-8) mol/l or higher. To identify the target genes of dienogest and P(4), we screened the expression of 84 genes related to cell cycle regulation by real-time polymerase chain reaction after 6 h of treatment at a concentration of 10(-7) mol/l. Results showed that only cyclin D1 expression was significantly down-regulated, although expression of the other genes did not significantly change after dienogest or P(4) treatment compared with the control. In a time-course study during the first 24 h after drug treatment, dienogest and P(4) each produced a lasting decrease in the expression of cyclin D1 mRNA, followed by a decrease in cyclin E1 mRNA but not an increase in the expression of cell cycle inhibitor genes (p21, p27 and p53). These findings suggest that dienogest directly inhibits the proliferation of human endometrial epithelial cells with suppression of cyclin D1 gene expression.

  8. Mapping of QTL for tiller number at different stages of growth in wheat using double haploid and immortalized F2 populations.

    Science.gov (United States)

    Li, Zhuokun; Peng, Tao; Xie, Quangang; Han, Shuxiao; Tian, Jichun

    2010-12-01

    Effective tiller number is one of the most important traits for wheat (Triticum aestivum L.) yield, but the inheritance of tillering is poorly understood. A set of 168 doubled haploid (DH) lines derivatives of a cross between two winter wheat cultivars (Huapei 3 and Yumai 57), and an immortalized F(2) (IF(2)) population generated by randomly permutated intermating of these DHs were investigated, and QTLs of tillering related to the maximum tillering of pre-winter (MTW), maximum tillering in spring (MTS), and effective tillering in harvest (ETH) were mapped. Phenotypic data were collected for the two populations from two different environments. Using inclusive composite interval mapping (ICIM), a total of 9 and 18 significant QTL were detected across environments for tillering in the DH and IF(2) populations, respectively. Four QTLs were common between two populations. A major QTL located on the 5D chromosome with the allele originating from Yumai 57 was detected and increased 1.92 and 3.55 tillers in MTW and MTS, respectively. QTLs (QMts6D, QEth6D) having a neighbouring marker interval at Xswes679.1 and Xcfa2129 on chromosome 6D was detected in MTS and ETH. These results provide a better understanding of the genetic factors for selectively expressing the control of tiller number in different growth stages and facilitate marker-assisted selection strategy in breeding.

  9. An In Vitro System Comprising Immortalized Hypothalamic Neuronal Cells (GT1–7 Cells for Evaluation of the Neuroendocrine Effects of Essential Oils

    Directory of Open Access Journals (Sweden)

    Dai Mizuno

    2015-01-01

    Full Text Available Aromatherapy and plant-based essential oils are widely used as complementary and alternative therapies for symptoms including anxiety. Furthermore, it was reportedly effective for the care of several diseases such as Alzheimer’s disease and depressive illness. To investigate the pharmacological effects of essential oils, we developed an in vitro assay system using immortalized hypothalamic neuronal cells (GT1–7 cells. In this study, we evaluated the effects of essential oils on neuronal death induced by hydrogen peroxide (H2O2, aluminum, zinc, or the antagonist of estrogen receptor (tamoxifen. Among tests of various essential oils, we found that H2O2-induced neuronal death was attenuated by the essential oils of damask rose, eucalyptus, fennel, geranium, ginger, kabosu, mandarin, myrrh, and neroli. Damask rose oil had protective effects against aluminum-induced neurotoxicity, while geranium and rosemary oil showed protective activity against zinc-induced neurotoxicity. In contrast, geranium oil and ginger oil enhanced the neurotoxicity of tamoxifen. Our in vitro assay system could be useful for the neuropharmacological and endocrine pharmacological studies of essential oils.

  10. Retinoids reduce formation of keratin aggregates in heat-stressed immortalized keratinocytes from an epidermolytic ichthyosis patient with a KRT10 mutation*.

    Science.gov (United States)

    Li, Hao; Törmä, Hans

    2013-01-01

    Epidermolytic ichthyosis (EI) is an autosomal dominant epidermal skin fragility disorder caused by mutations in keratin 1 and 10 (K1 and K10) genes. Mutated keratins form characteristic aggregates in vivo and in vitro. Some patients benefit from retinoid therapy, although the mechanism is not fully understood. Our aim was to demonstrate whether retinoids affect the formation of keratin aggregates in immortalized EI cells in vitro. EI keratinocytes were seeded on cover slips, pre-treated or not with retinoids, heat-stressed, and keratin aggregate formation monitored. K10 aggregates were detected in 5% of cells in the resting state, whereas heat stress increased this proportion to 25%. When cells were pre-incubated with all-trans-retinoic acid (ATRA) or retinoic acid receptor (RAR)-α agonists the aggregates decreased in a dose-dependent manner. Furthermore, ATRA decreased the KRT10 transcripts 200-fold as well as diminished the ratio of mutant to wild-type transcripts from 0.41 to 0.35, thus providing a plausible rational for retinoid therapy of EI due to K10 mutations.

  11. Type I Interferon Reaction to Viral Infection in Interferon-Competent, Immortalized Cell Lines from the African Fruit Bat Eidolon helvum

    Science.gov (United States)

    Biesold, Susanne E.; Ritz, Daniel; Gloza-Rausch, Florian; Wollny, Robert; Drexler, Jan Felix; Corman, Victor M.; Kalko, Elisabeth K. V.; Oppong, Samuel; Drosten, Christian; Müller, Marcel A.

    2011-01-01

    Bats harbor several highly pathogenic zoonotic viruses including Rabies, Marburg, and henipaviruses, without overt clinical symptoms in the animals. It has been suspected that bats might have evolved particularly effective mechanisms to suppress viral replication. Here, we investigated interferon (IFN) response, -induction, -secretion and -signaling in epithelial-like cells of the relevant and abundant African fruit bat species, Eidolon helvum (E. helvum). Immortalized cell lines were generated; their potential to induce and react on IFN was confirmed, and biological assays were adapted to application in bat cell cultures, enabling comparison of landmark IFN properties with that of common mammalian cell lines. E. helvum cells were fully capable of reacting to viral and artificial IFN stimuli. E. helvum cells showed highest IFN mRNA induction, highly productive IFN protein secretion, and evidence of efficient IFN stimulated gene induction. In an Alphavirus infection model, O'nyong-nyong virus exhibited strong IFN induction but evaded the IFN response by translational rather than transcriptional shutoff, similar to other Alphavirus infections. These novel IFN-competent cell lines will allow comparative research on zoonotic, bat-borne viruses in order to model mechanisms of viral maintenance and emergence in bat reservoirs. PMID:22140523

  12. Effects of the antimicrobial peptide cathelicidin (LL-37) on immortalized gingival fibroblasts infected with Porphyromonas gingivalis and irradiated with 625-nm LED light.

    Science.gov (United States)

    Kim, JiSun; Kim, SangWoo; Lim, WonBong; Choi, HongRan; Kim, OkJoon

    2015-11-01

    Porphyromonas gingivalis causes chronic inflammatory diseases (periodontal diseases) that destroy the periodontal ligament and alveolar bone. Antimicrobial peptides are crucial components of the host defense response required to maintain cellular homeostasis during microbial invasion. Because light-emitting diode (LED) irradiation influences the host defense response against bacterial infections, we investigated its effect on immortalized gingival fibroblasts (IGFs) infected with P. gingivalis. IGFs were incubated with P. gingivalis following LED irradiation at 425, 525, and 625 nm. The dark 1 group comprised noninfected, nonirradiated IGFs, and the dark 2 group comprised nonirradiated IGFs infected with P. gingivalis. These groups served as controls. Infected cells and controls were assayed for reactive oxygen species (ROS) and were subjected to RT-PCR and Western blotting analyses to determine the levels of expression of antimicrobial peptides. LED irradiation enhanced the bactericidal effects of the antimicrobial peptide LL-37 in cells infected with P. gingivalis. Irradiation at 625 nm decreased inflammatory responses involving the release of prostaglandin E2 induced by ROS in P. gingivalis-infected IGFs. LED irradiation at 625 nm induces an anti-inflammatory response that elicits the production of antimicrobial peptides, providing an efficacious method of treatment for periodontal diseases.

  13. Isolation and characterization of an HIV-1 envelope glycoprotein-specific B-cell from an immortalized human naïve B-cell library.

    Science.gov (United States)

    Sun, Zehua; Lu, Shiqiang; Yang, Zheng; Li, Jingjing; Zhang, Meiyun

    2017-04-01

    With the recent development of single B-cell cloning techniques, an increasing number of human immunodeficiency virus type 1 (HIV-1)-specific broadly neutralizing antibodies have been isolated since 2009. However, knowledge regarding HIV-1-specific B cells in vivo is limited. In this study, an HIV-1-specific B-cell line was established using healthy PBMC donors by the highly efficient Epstein-Barr virus transformation method to generate immortalized human naïve B-cell libraries. The enrichment of HIV-1 envelope-specific B cells was observed after four rounds of cell panning with the HIV-1 envelope glycoprotein. An HIV-1 envelope-specific stable B-cell line (LCL-P4) was generated. Although this cell line acquired a lymphoblastic phenotype, no expression was observed for activation-induced cytidine deaminase, an enzyme responsible for initiating somatic hypermutation and class switch recombination in B cells. This study describes a method that enables fast isolation of HIV-1-specific B cells, and this approach may extend to isolating other B-cell-specific antigens for further experiments.

  14. Quantitative assessment of the relative antineoplastic potential of the n-butanolic leaf extract of Annona muricata Linn. in normal and immortalized human cell lines.

    Science.gov (United States)

    George, V Cijo; Kumar, D R Naveen; Rajkumar, V; Suresh, P K; Kumar, R Ashok

    2012-01-01

    Natural products have been the target for cancer therapy for several years but there is still a dearth of information on potent compounds that may protect normal cells and selectively destroy cancerous cells. The present study was aimed to evaluate the cytotoxic potential of n-butanolic leaf extract of Annona muricata L. on WRL-68 (normal human hepatic cells), MDA-MB-435S (human breast carcinoma cells) and HaCaT (human immortalized keratinocyte cells) lines by XTT assay. Prior to cytotoxicity testing, the extract was subjected to phytochemical screening for detecting the presence of compounds with therapeutic potential. Their relative antioxidant properties were evaluated using the reducing power and DPPH* radical scavenging assay. Since most of the observed chemo-preventive potential invariably correlated with the amount of total phenolics present in the extract, their levels were quantified and identified by HPLC analysis. Correlation studies indicated a strong and significant (Pcancerous cells (IC50 values of 29.2 μg for MDA-MB-435S and 30.1 μg for HaCaT respectively). The study confirms the presence of therapeutically active antineoplastic compounds in the n-butanolic leaf extract of Annona muricata. Isolation of the active metabolites from the extract is in prospect.

  15. Symbiotic Plant Peptides Eliminate Candida albicans Both In Vitro and in an Epithelial Infection Model and Inhibit the Proliferation of Immortalized Human Cells

    Directory of Open Access Journals (Sweden)

    Lilla Ördögh

    2014-01-01

    Full Text Available The increasing number of multidrug-resistant microbes now emerging necessitates the identification of novel antimicrobial agents. Plants produce a great variety of antimicrobial peptides including hundreds of small, nodule-specific cysteine-rich NCR peptides that, in the legume Medicago truncatula, govern the differentiation of endosymbiotic nitrogen fixing bacteria and, in vitro, can display potent antibacterial activities. In this study, the potential candidacidal activity of 19 NCR peptides was investigated. Cationic NCR peptides having an isoelectric point above 9 were efficient in killing Candida albicans, one of the most common fungal pathogens of humans. None of the tested NCR peptides were toxic for immortalized human epithelial cells at concentrations that effectively killed the fungus; however, at higher concentrations, some of them inhibited the division of the cells. Furthermore, the cationic peptides successfully inhibited C. albicans induced human epithelial cell death in an in vitro coculture model. These results highlight the therapeutic potential of cationic NCR peptides in the treatment of candidiasis.

  16. Achievement report on project in fiscal 2000 of public appeal for proposal of international joint research (new industry - No.1). Development of technology to immortalize human cells by co-expression of mortalin and telomerase; 2000 nendo kokusai kyodo kenkyu teian kobo jigyo seika hokokusho (shinki No.1). Mortalin oyobi telomerase no kyohatsugen ni yoru hitosaibo fushika gijutsu no kaihatsu

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2001-03-01

    Researches have been performed on development of a technology to immortalize human cells by co-expression of mortalin and telomerase. The technology was adopted in the public appeal for proposal of international joint research. This paper summarizes the achievements thereof. The research has standardized a method to establish co-expression cell strains of mortalin genes and telomerase genes. Expressing the h TERT genes alone normally was not sufficient to cause immortalization in human fibroblasts. Furthermore, it was found in many cell strains that the co-expression of LTAg and h TERT is not sufficient to immortalize their cells. Expressing SV40stAg simultaneously in these cell strains was proved to have induced the immortalization highly efficiently. Introducing the mot-2 gene in place of virus antigen has resulted in acquisition of human cells regarded immortalized. It was concluded that suppressing initially the action of p53 by using mot-2 initially the immortalization due to the h TERT introduction. (NEDO)

  17. New Fractures, Old Wounds: Africa and the Renewal of South Agency Neue Brüche, alte Wunden: Afrika und die Erneuerung der South agency

    Directory of Open Access Journals (Sweden)

    Carlos Lopes

    2010-01-01

    Full Text Available Africa has recently come to the forefront of world politics as part of the emerging South. Its increased prominence in the global discourse as a “new frontier of development” signals the recognition of its economic potential. Indeed, the continent has registered an average 5 per cent annual GDP growth rate over the past decade. However, there is more to the story than that: The rising profile of the African continent also reveals the growing role of a number of its countries in the emergence of a new South agency. It is argued that South–South cooperation is an opportunity. The discussion of the current situation in Africa understood as a continent in all its diversity including sub-Saharan Africa, but also the Maghreb and Egypt, will therefore be placed into this wider context. The renewal of a South agency witnessed over the past decade is somewhat different from the trilateral alliance of Asia–Africa–Latin America formed in the wake of decolonization. Current mega-trends demonstrate that the global South, driven by a number of regional powers, will play a vital role in shaping the twenty-first century. Understanding the complexities of this renewed agency is vital for addressing old wounds that marked the emergence of a South voice in the not-so-distant past. In jüngster Zeit ist Afrika als Teil des emerging South in den Vordergrund der Weltpolitik gerückt. Seine zunehmende Prominenz im globalen Diskurs als new frontier of development signalisiert die Anerkennung des ökonomischen Potenzials des Kontinents. Tatsächlich konnte Afrika im vergangenen Jahrzehnt eine durchschnittliche jährliche BIP-Wachstumsrate in Höhe von 5 Prozent verzeichnen. Aber es geht um mehr: Die wachsende Wahrnehmung des afrikanischen Kontinents ist Ausdruck der immer wichtigeren Rolle einiger afrikanischer Länder bei der Entstehung einer neuen South agency. Die Süd-Süd-Kooperation wird wieder als Möglichkeit erkannt. Die Diskussion der aktuellen Lage

  18. An inhibitory switch derepressed by pbx, hox, and Meis/Prep1 partners regulates DNA-binding by pbx1 and E2a-pbx1 and is dispensable for myeloid immortalization by E2a-pbx1.

    Science.gov (United States)

    Calvo, K R; Knoepfler, P; McGrath, S; Kamps, M P

    1999-12-23

    The Pbx/Exd family of homeodomain (HD) proteins contribute to the transcriptional and developmental roles of other Hox and Meis/Prep1/Hth HD proteins through heterodimer formation. E2a-Pbx1 is an oncogenic derrivative of Pbx1 produced by the t(1;19) translocation in pediatric pre-B cell acute lymphoblastic leukemia. E2a-Pbx1 heterodimerizes with Hox but not with Meis/Prep1 proteins, produces acute myeloid leukemia in mice, and blocks differentiation of cultured murine myeloid progenitors. Here, we characterize negative and positive regulatory sequences that flank the Pbx1 HD and determine their importance for myeloid immortalization by E2a-Pbx1. A 25 residue predicted alpha helix preceding the Pbx1 HD bound the HD and prevented both its binding to DNA and its ability to heterodimerize with Hox proteins. Addition of 39 residues N-terminal to this inhibitory helix exposed a Pbx dimerization interface that orchestrated cooperative DNA-binding of E2a-Pbx1 and all Pbx proteins as homodimers and heterdimers. Sequences inhibiting DNA-binding and mediating Pbx dimerization coincided with those reported to have nuclear export function. An additional 103 residues N-terminal to the Pbx dimerization interface restored heterodimerization with Hox and Meis1/Prep1 proteins. This negative switch domain - comprised of the inhibitory helix and N-terminal regions required for its partner-mediated derepression - was dispensable for myeloid immortalization by E2a-Pbx1. While stabilizing the heterodimer, the 310 helix C-terminal to the Pbx1 HD was also dispensable for the ability of E2a-Pbx1 to heterodimerize with Hox proteins and immortalize myeloblasts. Retention of myeloid immortalization by E2a-Pbx1 proteins lacking all Pbx1 sequences N- or C-terminal to the HD indicates that Hox proteins, or a yet undefined factor that binds the Pbx1 HD and derepresses DNA-binding by the HD, cooperate with E2a-Pbx1 in myeloid immortalization.

  19. Caspase-3 Expression and ALT, AST, and GGT Activity After 24 Hours of Porcine Liver Cold Storage, Depending on the Type of Transgenesis.

    Science.gov (United States)

    Roman, P; Budziński, G; Suszka-Świtek, A; Caban, A; Oczkowicz, G; Czech, E; Ryszka, F; Wiaderkiewicz, R; Smorąg, Z; Cierpka, L

    2016-06-01

    Because of an insufficient number of human organs for transplantation, xenotransplantation may become an effective alternative. We aimed to analyze if the type of transgenesis has an influence on the hepatic caspase-3 expression, the enzyme that executes apoptosis as well as ALT, AST, and GGT activity after 24 hours of cold storage. The experiment was carried out on the 24 livers of Polish White Landrace pigs carrying human α1,2-fucosyltransferase and/or α-galactosidase (GAL) genes and livers without this genetic modification (control). Livers were perfused, stored for 24 hours in solution, and subsequently re-flushed. Hepatic concentration of the caspase-3 protein and its mRNA expression were measured just after the animal was killed as well as after 30 minutes of perfusion and after 24 hours of cold storage followed by 30 minutes of reperfusion. Caspase-3 mRNA level was detected with the RT-PCR method. Protein concentration (capsase-3 active and inactive) was assessed with the Western blotting technique. Kinetic methods were applied for the analysis of the ALT, AST, and GGT activity. The highest increase of the ALT activity after cold storage was observed in the group with GAL transgenesis, whereas the GGT activity was highest in the unmodified livers. There was no difference in the caspase-3 expression and AST activity after cold storage as compared with the respective initial results (P = .57 and P = .97, respectively). It appears that transgenesis does not aggravate ischemic injury of the liver. Copyright © 2016 Elsevier Inc. All rights reserved.

  20. The effect of endurance training with cinnamon supplementation on plasma concentrations of liver enzymes (ALT, AST in women with type II diabetes

    Directory of Open Access Journals (Sweden)

    Shahla Torabi

    2016-09-01

    Full Text Available Background: Diabetes is associated with many pathological changes and one of the most important consequences of the diabetes is hepatic injury. The present study was performed to investigate the effect of eight weeks endurance training with consumption of cinnamon supplementation on plasma concentrations of liver enzymes, alanine aminotransferase (ALT and aspartate aminotransferase (AST in women with type II diabetes. Methods: In this quasi-experimental study, 36 female volunteers with type II diabetes (age 52.72±2.64 years and body mass index 29.28±2.94 Kg/m2 were participated. The subjects were homogenized regarding their body mass index and then were divided randomly into four groups (each group=9 patients: Training, training-cinnamon, cinnamon, and Control. Endurance training was performed for eight weeks (three sessions per week at the intensity of 60-75% of maximum heart rate for 40-60 minutes. The consumption of cinnamon supplementation was 1.5 gr per day. Plasma concentrations of ALT and AST were measured following 12 hours fasting, 48 hours before and after performing the experiment, by the enzymatic method. Data were analyzed by paired t-test and factorial ANOVA, using SPSS version 21 (Chicago, IL, USA and at the significant level of P0.05. There was no significant difference between groups in pre and posttests. Conclusion: The results confirm that cinnamon supplementation may be effective in improving the plasma levels of ALT but the intensity and duration of an effective exercise training especially with consumption of cinnamon supplementation simultaneously need more study in diabetic patients.

  1. Synthesis and Properties of the Metallo-Supramolecular Polymer Hydrogel Poly[methyl vinyl ether-alt-mono-sodium maleate]∙AgNO3

    KAUST Repository

    Al-Dossary, Mona S.

    2014-05-01

    Gels are a special class of materials which are composed of 3D networks of crosslinked polymer chains that encapsulate liquid/air in the matrix. They can be classified into organogels or hydrogels (organic solvent for organogel and water for hydrogel). For hydrogels that contain metallic elements in the form of ions, the term of metallo-supramolecular polymer hydrogel (MSPHG) is often used. The aim of this project is to develop a kind of new MSPHG and investigate its properties and possible applications. The commercial polymeric anhydride poly(methyl vinyl ether-alt-maleic anhydride) (PVM/MA) is converted by reaction with NaOH to give poly(methyl vinyl ether-alt-monosodium maleate) (PVM/Na-MA). By addition of AgNO3-solution, the formation of the silver(I) supramolecular polymer hydrogel poly[methyl vinyl ether-alt-mono-sodium maleate]∙AgNO3 is obtained. Freeze-dried samples of the hydrogel show a mesoporous network of polycarboxylate ligands that are crosslinked by silver(I) cations. The supercritical CO2 dried silver(I) hydrogel was characterized by FT-IR, SEM-EDAX, TEM, TGA and Physical adsorption (BET) measurements. The intact silver(I) hydrogel was characterized by cryo-SEM. In the intact hydrogel, ion-exchange studies are reported and it is shown that Ag+ ions can be exchanged by copper(II) cations without disintegration of the hydrogel. The silver(I) hydrogel shows effective antibacterial activity and potential application as burn wound dressing.

  2. Acute Effects of on Javdar Supplementation on Asparate Aminotransferase(AST and Alanine Aminotransferase(ALT after Exhaustive Incremental Exercise in Men’s Handball

    Directory of Open Access Journals (Sweden)

    Mojtaba Khansooz

    2017-03-01

    Full Text Available Abstract Background: The aim of this study was to investigate the acute effect of supplementation Jadvar on aspartate aminotransferase (AST and alanine aminotransferase (ALT enzymes after exhaustive incremental exercise in men's handball. Materials and Methods: In this semi-experimental double blinded study 12 handball players with at least 2 years record in league (with average age=21.42, height=186cm, weight=83.25, and body mass index=24.09kg/m2 divided into 2 groups (n=6 accidentally. Both groups performed maximum Bruce protocol until exhausting level. Before (fasting and after performing protocol ,blood samples were taken from middle forearm vein and transferred to lab then supplementation group consumed three 500 mg jadvar capsules and control group consumed three 500 mg maltodextrin for 7 days daily. 24 hours after eating last capsule ,they performed maximum Bruce protocol up to exhausting level and the samples were collected like primary protocol blood samples before (fastingand after performing protocol and were transferred to the laboratory. Results: The results of dependent and independent t-test showed that consuming jadvar supplements for 7days did not have significant effect on aspartate aminotransferase(AST and alanine aminotransferase(ALT(p≥0.05.But alanine aminotransferase in supplementation group (16.83 mg.dl-1 decreased in comparison to placebo group (20.5 mg.dl-1. Also, aspartate aminotransferase was increased from 28 mg.dl-1 to 35.17 mg.dl-1 , but neither decrease nor increase was not meaningful (p≥0.05. Conclusion: It seems that acute consuming of jadvar supplement and one session incremental exercise does not have meaningful effect on AST and ALT in man handball players.

  3. Need for validation of clinical decision aids: use of the AST/ALT ratio in predicting cirrhosis in chronic hepatitis C.

    Science.gov (United States)

    Imperiale, T F; Said, A T; Cummings, O W; Born, L J

    2000-09-01

    A value of > or = 1 for the ratio of aspartate amino-transferase to alanine aminotransferase (the AST/ALT ratio or AAR) has been shown to have a positive predictive value of 100% for the diagnosis of cirrhosis in patients with chronic hepatitis C. If validated on separate cohorts, an AAR > or = 1 might obviate the need for liver biopsy in some patients with hepatitis C. We attempted to validate the AAR by abstracting demographic and clinical data from a database of consecutive patients with hepatitis C who had a liver biopsy between 1993 and 1998. We used definitions, methods of data collection, and analyses comparable to those of the published study. A hepatopathologist blindly reviewed 49 liver biopsies for histological grade and stage. The current cohort of 177 patients and the previous cohort of 139 patients were comparable in mean age (42.3 vs 43.8 yr), percentage of men (63 vs 67), percentage with an AAR > or =1 (20 vs 17), and Child-