De radio aeternitatis seu honestate e suo fonte scilicet jure naturae, emanante

Variantti B.

Tetramethylpyrazine potentiates arsenic trioxide activity against HL-60 cell lines

The objective of this study was to evaluate the effects of tetramethylpyrazine (TMP) in combination with arsenic trioxide (As2O3) on the proliferation and differentiation of HL-60 cells. The HL-60 cells were treated with 300 µg/mL TMP, 0.5 µM As2O3, and 300 µg/mL TMP combined with 0.5 µM As2O3, respectively. The proliferative inhibition rates were determined with MTT. Differentiation was detected by the nitroblue tetrazolium (NBT) reduction test, Wright’s staining and the distribution of CD11b and CD14. Flow cytometry was used to analyze cell cycle distribution. RT-PCR and Western blot assays were employed to detect the expressions of c-myc, p27, CDK2, and cyclin E1. Combination treatment had synergistic effects on the proliferative inhibition rates. The rates were increased gradually after the combination treatment, much higher than those treated with the corresponding concentration of As2O3 alone. The cells exhibited characteristics of mature granulocytes and a higher NBT-reducing ability, being a 2.6-fold increase in the rate of NBT-positive ratio of HL-60 cells within the As2O3 treatment versus almost a 13-fold increase in the TMP + As2O3 group. Cells treated with both TMP and As2O3 expressed far more CD11b antigens, almost 2-fold compared with the control group. Small doses of TMP potentiate As2O3-induced differentiation of HL-60 cells, possibly by regulating the expression and activity of G0/G1 phase-arresting molecules. Combination treatment of TMP with As2O3 has significant synergistic effects on the proliferative inhibition of HL-60 cells.

Berberine induces apoptosis via ROS generation in PANC-1 and MIA-PaCa2 pancreatic cell lines

Pancreatic cancer is the fourth leading cause of cancer death. Gemcitabine is widely used as a chemotherapeutic agent for the treatment of pancreatic cancer, but the prognosis is still poor. Berberine, an isoquinoline alkaloid extracted from a variety of natural herbs, possesses a variety of pharmacological properties including anticancer effects. In this study, we investigated the anticancer effects of berberine and compared its use with that of gemcitabine in the pancreatic cancer cell lines PANC-1 and MIA-PaCa2. Berberine inhibited cell growth in a dose-dependent manner by inducing cell cycle arrest and apoptosis. After berberine treatment, the G1 phase of PANC-1 cells increased by 10% compared to control cells, and the G1 phase of MIA-PaCa2 cells was increased by 2%. Whereas gemcitabine exerts antiproliferation effects through S-phase arrest, our results showed that berberine inhibited proliferation by inducing G1-phase arrest. Berberine-induced apoptosis of PANC-1 and MIA-PaCa2 cells increased by 7 and 2% compared to control cells, respectively. Notably, berberine had a greater apoptotic effect in PANC-1 cells than gemcitabine. Upon treatment of PANC-1 and MIA-PaCa2 with berberine at a half-maximal inhibitory concentration (IC50), apoptosis was induced by a mechanism that involved the production of reactive oxygen species (ROS) rather than caspase 3/7 activation. Our findings showed that berberine had anti-cancer effects and may be an effective drug for pancreatic cancer chemotherapy.

De radio aeternitatis seu honestate e suo fonte scilicet jure naturae, emanante

Variantti A.

Iron and zinc availability in maize lines

The aim of this study was to characterize the Zn and Fe availability by phytic acid/Zn and phytic acid/Fe molar ratios, in 22 tropical maize inbred lines with different genetic backgrounds. The Zn and Fe levels were determined by atomic absorption spectrophotometry and the P through colorimetry method. Three screening methods for phytic acid (Phy) analysis were tested and one, based on the 2,2'-bipyridine reaction, was select. There was significant variability in the contents of zinc (17.5 to 42 mg.kg-1), iron (12.2 to 36.7 mg.kg-1), phosphorus (230 to 400 mg.100 g-1), phytic acid (484 to 1056 mg.100 g-1), phytic acid P (140 to 293 mg.100 g-1) and available-P (43.5 to 199.5 mg.100 g-1), and in the available-P/total-P ratio (0.14 to 0.50), Phy/Zn (18.0 to 43.5) and Phy/Fe (16.3 to 45.5) molar ratios. Lines 560977, 560978 and 560982 had greater availability of Zn and lines 560975, 560977, 561010 and 5610111 showed better Fe availability. Lines 560975, 560977 and 560978 also showed better available-P/total-P ratio. Thus, the lines 560975, 560977 and 560978 were considered to have the potential for the development of cultivars of maize with high availability of Fe and/or Zn.

Kognitiivinen radio sotilaallisen maanpuolustuksen kontekstissa

Kognitiiviteknologialla on ennustettu olevan disruptiivisia vaikutuksia sodankäyntiin. Kognitiiviteknologian avulla pyritään löytämään myös ratkaisu kasvavista datansiirtovaatimuksista aiheutuvaan radiotaajuisen spektrin ahtauteen. Tämän tutkimuksen keskiössä on kognitiivinen radio. Diplomityössä ennakoidaan niitä muutoksia, joita radion kognitiiviset ominaisuudet tuovat sotilaalliseen toimintaympäristöön. Tutkimuksessa etsitään kognitiivisen radion suorituskykyyn, suorituskyvyn rakentamiseen ja käyttöön liittyviä tekijöitä, jotka ovat merkityksellisiä sotilaallisesta näkökulmasta. Radion kognitiivisuutta käsitellään kongitiivisten ominaisuuksien, kuten adaptiivisuuden tai tilannetietoisuuden, kautta. Painopisteenä on kognitiivisten ominaisuuksien keskinäinen arvottaminen, mahdollisten vaikutusten tarkastelu operatiivisessa viitekehyksessä sekä tarvittavien toimenpiteiden kartoittaminen kognitiivisen tekniikan operatiivisen käytön mahdollistamiseksi. Tutkimuksen pohjalta esitetään kognitiivisten ominaisuuksien tärkeysjärjestys suorituskyvyn ja johtamisjärjestelmien kehittämisen näkökulmista ja alustavia operatiivisia vaatimuksia kognitiiviselle radiolle sekä kuvataan spektrin hallinnan kehittämismahdollisuuksia. Operatiivisia vaikutuksia tutkitaan myös uhkalähtöisesti eli arvioimalla, miten tietty kognitiivinen ominaisuus vaikuttaisi yleisimpiin tiedonsiirtoverkkoihin kohdistuviin uhkiin. Lisäksi työssä arvioidaan kognitiivisen radion käyttöönoton esteitä Puolustusvoimissa. Tutkimuksen päämenetelmä on delfoi, jota on täydennetty asiantuntijahaastatteluin sekä -arvioin. Kognitiiviselle radiolle on useita erilaisia määritelmiä. Kaikkia kriteerejä täyttävää kognitiivista radiota ei ole vielä olemassa, vaikka kognitiivisia ominaisuuksia radioissa on ollut pitkään. Kognitiivinen radio, tai jo pelkästään ohjelmistoradio kehittyneillä kognitiivisilla ominaisuuksilla, tuo huomattavia mahdollisuuksia parantaa langattoman tiedonsiirron toimintakykyä sotilaallisessa käyttöympäristössä. Merkittävin suorituskykylisä sotilaallisessa käyttöympäristössä saadaan seuraavista kognitiivisista ominaisuuksista: 1) dynaaminen spektrin hyväksikäyttö (DSA) 2) yhteyksien adaptiivisuus ja radioresurssien hallinta (SLA ja RRM) sekä 3) älykäs verkonmuodostus (SON ja RBR). Tiedonsiirtojärjestelmien ylläpito ja rakentaminen vaativat aiempaa enemmän suunnittelua ja ammattitaitoa, mutta toisaalta loppukäyttäjän osaamistasovaatimuksia voidaan laskea sitä mukaa, kun lähestytään 0-konfiguraatiojärjestelmiä. Radioiden kognitiivisuuden myötä voidaan myös passiivisiin uhkiin, esimerkiksi elektroniseen tiedusteluun, varautua aiempaa tehokkaammin. Aktiivisiin uhkiin, kuten häirintään, kognitiiviset ominaisuudet vaikuttavat jopa toiminnan fundamentteja muuttavasti. Tiedonsiirtojärjestelmien ohjelmistopohjaisuus nostaa kyberin merkitttävyyttä arvioitaessa eri uhkien kokonaismerkityksiä. Onnistunut kyberoperaatio voi tarkoittaa koko järjestelmän lamautumista tai vakavaa tietovuotoa. Kognitiivisten ominaisuuksien liittäminen osaksi sotilasradiojärjestelmiä on haasteellista. Käyttöönoton esteitä ovat osaamisen puute, lainsäädännön ja regulaation jäykkyys, tekniikan osittainen kypsymättömyys sekä tietoturvan hallinta. Spektri on muutoksessa, mikä tulee näkymään spektrin sirpaloitumisena vuoteen 2020 mennessä. Paine taajuushallinnan muuttamisesta kognitiivitekniikkaa sallivampaan suuntaan nousee erityisesti siviilipuolen kehittyvien mobiililaitteiden datansiirtomäärien kasvaessa. Lainsäädäntö antaa viranomai-sille suuret valtuudet säädellä spektrin käyttöä poikkeusoloissa. Tämä mahdollistaa sotilaallisten järjestelmien suunnittelun ja kehittämisen taajuusriippumattomasti jo normaalioloissa. Haasteena on spektrin hallinnoinnin kehittäminen esimerkiksi ennakoitua nopeammassa tilannekehityksessä. Spektriä tulisi hallita kokonaisturvallisuuden näkökulmasta joustavasti viranomaisten yhteisillä päätöksillä. Kognitiivisen radion tulisi tulevaisuudessa voida käyttää muille järjestelmille osoitettuja taajuusresursseja taajuuksien ollessa käyttämättömänä. Kognitiivista radiota kannattaisi kehittää taajuusvapaasti. Spektri tullee olemaan tulevaisuudessa yhä ahtaampi, ja tähän haasteeseen kognitiivinen radio tarjoaa osaltaan ratkaisun.

Development of packaging cell lines for rescuing BAV2 viral vectors

The construction of adenovirus vectors for cloning and foreign gene expression requires packaging cell lines that can complement missing viral functions caused by sequence deletions and/or replacement with foreign DNA sequences. In this study, packaging cell lines were designed to provide in trans the missing bovine adenovirus functions, so that recombinant viruses could be generated. Fetal bovine kidney and lUng cells, acquired at the trimester term from a pregnant cow, were tranfected with both digested wild type BAV2 genomic DNA and pCMV-EI. The plasmid pCMV-EI was specifically constructed to express El of BAV2 under the control of the cytomegalovirus enhancer/promoter (CMV). Selection for "true" transformants by continuous passaging showed no success in isolating immortalised cells, since the cells underwent crisis resulting in complete cell death. Moreover, selection for G418 resistance, using the same cells, also did not result in the isolation of an immortalised cell line and the same culture-collapse event was observed. The lack of success in establishing an immortalised cell line from fetal tissue prompted us to transfect a pre-established cell line. We began by transfecting MDBK (Mardin-Dardy bovine kidney) cells with pCMV-El-neo, which contain the bacterial selectable marker neo gene. A series of MDBK-derived cell lines, that constitutively express bovine adenoviral (BAV) early region 1 (El), were then isolated. Cells selected for resistance to the drug G418 were isolated collectively for full characterisation to assess their suitability as packaging cell lines. Individual colonies were isolated by limiting dilution and further tested for El expression and efficiency of DNA uptake. Two cell lines, L-23 and L-24, out of 48 generated foci tested positive for £1 expression using Northern Blot analysis. DNA uptake studies, using both lipofectamine and calcium phosphate methods, were performed to compare these cells, their parental MDBK cells, 8 and the unrelated human 293 cells as a benchmark. The results revealed that the new MDBKderived clones were no more efficient than MDBK cells in the transient expression of transfected DNA and that they were inferior to 293 cells, when using lacZ as the reporter gene. In view of the inherently poor transfection efficiency of MDBK cells and their derivatives, a number of other bovine cells were investigated for their potential as packaging cells. The cell line CCL40 was chosen for its high efficiency in DNA uptake and subsequently transfected with the plasmid vector pCMV El-neo. By selection with the drug G418, two cell lines were isolated, ProCell 1 and ProCell 2. These cell lines were tested for El expression, permissivity to BAV2 and DNA uptake efficiency, revealing a DNA uptake efficiency of 37 % , comparable to that of CCL40. Attempts to rescue BAV2 mutants carrying the lacZ gene in place of £1 or £3 were carried out by co-transfecting wild type viral DNA with either the plasmid pdlElE-Z (which contains BAV2 sequences from 0% to 40.4% with the lacZ gene in place of the £1 region from 1.1% to 8.25%) or with the plasmid pdlE3-5-Z (which contains BAV2 sequences from 64.8% to 100% with the lacZ gene in place of the E3 region from 75.8% to 81.4%). These cotransfections did not result in the generation of a viral mutant. The lack of mutant generation was thought to be caused by the relative inefficiency ofDNA uptake. Consequently, cosBAV2, a cosmid vector carrying the BAV2 genome, was modified to carry the neo reporter gene in place of the £3 region from 75.8% to 81.4%. The use of a single cosmid vector earring the whole genome would eliminate the need for homologous recombination in order to generate a viral vector. Unfortunately, the transfection of cosBAV2- neo also did not result in the generation of a viral mutant. This may have been caused by the size of the £3 deletion, where excess sequences that are essential to the virus' survival might have been deleted. As an extension to this study, the spontaneous E3 deletion, accidently discovered in our viral stock, could be used as site of foreign gene insertion.

The establishment of two novel bovine cell lines by transfection with the putative transforming region of bovine adenovirus type 3

Human adenoviruses (Ads), members of the family adenoviridae, are medium-sized DNA viruses which have been used as valuable research tools for the study of RNA processing, oncogenic transformation, and for the development of viral vectors for use in gene delivery and immunization technology. The left 12% of the linear Ad genollle codes for products which are necessary for the efficient replication of the virus, as well as being responsible for the forlllation of tumors in animallllodels. The establishlllent of the 293 cell line, by immortalization of human embryonic kidney cells with th~ E1 region of Ad type S (AdS), has facilitated extensive manipulation of the Ads and the development of recombinant Ad vectors. The study of bovine adenoviruses (BAVs), which cause mild respiratory and gastrointestinal infections in cattle has, on the other hand, been limited primarily to that of infectivity, immunology and clinicallllanifestations. As a result, any potential as gene delivery vehicles has not yet been realized. Continued research into the molecular biolo~gy of BAVs and the development of recolllbinant vectors would benefit from the development of a cell line analogous to that of the 293 cells. In an attelllpt to establish such a cell line, the recombinant plaslllid pKC-neo was constructed, containing the left 0-19.7% of the BAV type 3 (BAV3) genome, and the selectable marker for resistance to the aminoglycoside G418, a neomycin derivative. The plasmid construct was then used to transfect both the Madin-Darby bovine kidney (MDBK) -iicell line and primary bovine lung cells, after which G418-resistant foci were selected for analysis. Two cell lines, E61 (MDBK) and E24 (primary lung), were subsequently selected and analysed for DNA content, revealing the presence of the pKC-neo sequences in their respective genomes. In addition, BAV3 RNA transcripts were detected in the E61 cells. Although the presence of E1 products has yet to be confirmed in both cell lines, the E24 cells exhibit a phenotype characteristic of partial transformation by E1. The apparent immortalization of the primary lung cells will permit exploitation of their ability to take up exogenous DNA at high efficiency.

A Survey of hMLH1 mRNA Splicing Profiles in Human Cell Lines: Comparing Primary Cultured Fibroblasts Before and After Oxidative Stress and Transiently versus Stably Demethylated Colon Cancer Derived Cell Lines

Most human genes undergo alternative splicing and loss of splicing fidelity is associated with disease. Epigenetic silencing of hMLH 1 via promoter cytosine methylation is causally linked to a subset of sporadic non-polyposis colon cancer and is reversible by 5-aza-2' -deoxycytidine treatment. Here I investigated changes in hMLHI mRNA splicing profiles in normal fibroblasts and colon cancer-derived human cell lines. I established the types and frequencies of hMLHI mRNA transcripts generated under baseline conditions, after hydrogen peroxide induced oxidative stress, and in acutely 5-aza-2' -deoxycytidine-treated and stably derepressed cancer cell lines. I found that hMLHI is extensively spliced under all conditions including baseline (50% splice variants), the splice variant distribution changes in response to oxidative stress, and certain splice variants are sensitive to 5- aza-2' -deoxycytidine treatment: Splice variant diversity and frequency of exon 17 skipping correlates with the level of hMLHI promoter methylation suggesting a link between promoter methylation and mRNA splicing.

Canada Steamship Lines Ltd. Niagara to the Sea (1923)

Sailing schedule and trip guide for Canada Steamship Lines Limited for the year 1923

Canada Steamship Lines Limited Niagara to the Sea 1917

Sailing schedule and trip guide for Canada Steamship Lines Limited for the year 1917.

Canada Steamship Lines Limited Richelieu and Ontario Division Niagara to the Sea July 1916

Sailing schedule and trip guide for Canada Steamship Lines Limited for July 1916.

Canada Steamship Lines Limited Richelieu (1915)

Sailing schedule and trip guide for Canada Steamship Lines Limited for July 1915.