Isolation and partial characterization of a complementary protein from the mycoparasite, Piptocephalis virginiana, which specifically binds to two glycoproteins b and c of the host cell surface

Presence of surface glycoprotein in Piptocephalis virginiana that recognizes the host glycoproteins band c, reported earlier from our laboratory, was detected by immunofluorescence microscopy. Germinated spores of P. virginiana treated with Mortierella pusilla cell wall protein extract, primary antibodies prepared against glycoproteins band c and FITC-goat anti-rabbit IgG conjugate showed fluorescence. This indicated that on the surfaces of the biotrophic mycoparasite P. virginiana , there might be a complementary molecule which recognizes the glycoproteins band c from M. pusilla. Immunobinding analysis identified a glycoprotein of Mr 100 kDa from the mycoparasite which binds with the host glycoproteins band c, separately as well as collectively. Purification of this glycoprotein was achieved by (i) 60% ammonium sulfate precipitation, (ii) followed by heat treatment, and (iii) Sephadex G-IOO gel filtration. The glycoprotein was isolated by preparative polyacrylamide gel electrophoresis by cutting and elution. The purity of the protein ·was ascertained by SDS-PAGE and Western blot analysis. Positive reaction to periodic acid-Schiff reagent revealed the glycoprotein nature of this 100 kDa protein. Mannose was identified as a major sugar component of this glycoprotein by using a BoehringerMannheim Glycan Differentiation Kit. Electrophoretically purified glycoprotein was used to raIse polyclonal antibody in rabbit. The specificity of the antibody was determined by dot-immunobinding test and western-blot analysis. Immunofluorescence mIcroscopy revealed surface localization of the protein on the germ tube of Piptocephalis virginiana. Fluorescence was also observed at the surfaceJ of the germinated spores and hyphae of the host, M. pusilla after treatment with complementary protein from P. virginiana, primary antibody prepared against the complementary protein and FITC-goat anti-rabbit IgG conjugate.

Partial characterization of the cloned dihydrofolate reductase gene of Saccharomyces cerevisiae

The cloned dihydrofolate reductase gene of Saccharomyces cerevisiae (DFR 1) is expressed in Escherichia coli. Bacterial strain JF1754 transformed with plasmids containing DFR 1 is at least 5X more resistant to inhibition by the folate antagonist trimethoprim. Expression of yeast DFR 1 in E. coli suggests it is likely that the gene lacks intervening sequences. The 1.8 kbp DNA fragment encoding yeast dhfr activity probably has its own promotor, as the gene is expressed in both orientations in E. coli. Expression of the yeast dhfr gene cloned into M13 viral vectors allowed positive selection of DFR 1 - M13 bacterial transfectants in medium supplemented with trimethoprim. A series of nested deletions generated by nuclease Bal 31 digestion and by restriction endonuclease cleavage of plasmids containing DFR 1 physically mapped the gene to a 930 bp region between the Pst 1 and Sal 1 cut sites. This is consistent with the 21,000 molecular weight attributed to yeast dhfr in previous reports. From preliminary DNA sequence analysis of the dhfr DNA fragment the 3' terminus of DFR 1 was assigned to a position 27 nucleotides from the Eco Rl cut site on the Bam Hi - Eco Rl DNA segment. Several putative yeast transcription termination consensus sequences were identified 3' to the opal stop codon. DFR 1 is expressed in yeast and it confers resistance to the antifolate methotrexate when the gene is present in 2 - 10 copies per cell. Plasmid-dependent resistance to methotrexate is also observed in a rad 6 background although the effect is somewhat less than that conferred to wild-type or rad 18 cells. Integration of DFR 1 into the yeast genome showed an intermediate sensitivity to folate antagonists. This may suggest a gene dosage effect. No change in petite induction in these yeast strains was observed in transformed cells containing yeast dhfr plasmids. The sensitivity of rad 6 , rad 18 and wild-type cell populations to trimethoprim were unaffected by the presence of DFR 1 in transformants. Moreover, trimethoprim did not induce petites in any strain tested, which normally results if dhfr is inhibited by other antifolates such as methotrexate. This may suggest that the dhfr enzyme is not the only possible target of trimethoprim in yeast. rad 6 mutants showed a very low level of spontaneous petite formation. Methotrexate failed to induce respiratory deficient mutants in this strain which suggested that rad 6 might be an obligate grande. However, ethidium bromide induced petites to a level approximately 50% of that exhibited by wild-type and rad 18 strains.

The isolation, partial purification and preliminary characterization of a growth factor from chick embryo brains

Chicl( brain growth factor (CBGF) is a mitogen isolated from embryonic chick brains thought to have a potential role as a trophic factor involved in nerve dependent amphibian limb regeneration. In addition, CBGF stimulates 3H-thymidine incorporation in chick embryo brain astrocytes in vitro. In this study, cultured chick embryo brain non-neuronal cells were employed in a bioassay to monitor CBGF activity throughout various stages of its pllrification. Cell culture and assay conditions were optimized. Nonneuronal cells grew best on collagen-coated culture dishes in complete medium, were most responsive to a growth stimulus [10% fetal bovine serum (FBS)] at the second and third subcultures, and were healthiest when rendered "quiescent" in medium supplemented with 1% FBS. The most effective bioassay conditions consisted of a minimum 14.5 hour "quiescence" time (24 hours was used), a 6 hour "prestimulation" time, and a 24 hour 3H-thymidine labeling time. Four-day subconfluent primary non-neuronal cells consisted of 6.63% GFAP positive cells; as a result cultures were thought to be mainly composed of astroblasts. CBGF was purified from 18-day chick embryo brains by ultrafiltration through Amicon PM-30 and YM-2 membranes, size exclusion chromatography through a Biogel P6 column, and analytical reverse-phase high-performance liquid chromatography (rp-HPLC). The greatest activity resided in rp-HPLC fraction #7 (10 ng/ml) which was as effective as 10% FBS at stimulating 3H-thymidine incorporation in chick embryo brain nonneuronal cells. Although other researchers report the isolation of a mitogenic fraction consisting of 5'-GMP from the embryonic chick brain, UV absorbance spectra, rp-HPLC elution profiles, and fast atom bombardment (FAB) mass spectra indicated that CBGF is neither 5'-GMP nor 51-AMP. 2 Moreover, commercially available 5t-GMP was inhibitory to 3H-thymidine incorporation in the chick non-neuronal cells, while Sf-AMP had no effect. Upon treatment with pronase, the biological activity of fraction P6-3 increased; this increase was nearly 30% greater than what would be expected from a simple additive effect of any mitogenic activity of pronase alone together with P6-3 alone. This may suggest the presence of an inhibitor protein. The bioactive component may be a protein protected by a nucleoside/nucleotide or simply a nucleoside/nucleotide acting alone. While the FAB mass spectrum of rp-HPLC fraction #7 did not reveal molecular weight or sequence information, the ion of highest molecular weight was observed at m/z 1610; this is consistent with previous estimations of CBGF's size. 3

Partial piece of post office registration to J.M. Ball of Toronto, Ont.

Partial piece of post office registration to J.M. Ball of Toronto, Ont. Half of ticket is missing. Text is affected, Jul. 5, 1875.

Envelope (partial) to Colonel Nelles, 4th Lincoln Militia, Grimsby from M.A. Hale of Quebec

Envelope (partial) to Colonel Nelles, 4th Lincoln Militia, Grimsby from M.A. Hale of Quebec. The envelope is stained and torn, but text is not affected, Feb. 7, 1839.

Does friendship give us non-derivative partial reasons ?

Une des façons d’approcher la question de l’existence de raisons partiales non-dérivatives d’une quelconque sorte consiste à expliquer ce que sont les raisons partiales et ensuite à chercher à savoir s’il y a des raisons de cette sorte. Si de telles raisons existent, alors il est au moins possible qu’il y ait des raisons partiales d’amitié. C’est cette approche que j’adopterai ici, et elle produit des résultats intéressants. Le premier a trait à la structure des raisons partiales. C’est au moins une condition nécessaire pour qu’une raison soit partiale qu’elle aie une composante relationnelle explicite. Cette composante, techniquement parlant, est un relatum dans la relation d’être une raison qui elle-même est une relation entre la personne à qui la raison s’applique et la personne concernée par l’action pour laquelle il y a une raison. La deuxième conclusion de ce texte est que cette composante relationnelle est aussi requise dans de nombreuses sortes de raisons admises comme impartiales. Afin d’éviter de banaliser la distinction entre raisons partiales et impartiales nous devons appliquer une condition suffisante additionnelle. Finalement, bien qu’il pourrait s’avérer possible de distinguer les raisons impartiales ayant une composante relationnelle des raisons partiales, cette approche suggère que la question de savoir si l’éthique est partiale ou impartiale devra se régler au niveau de l’éthique normative, ou à tout le moins, qu’elle ne pourra se régler au niveau du discours sur la nature des raisons d’agir.

Essays in partial identification and applications to treatment effects and policy evaluation.

Dans cette thèse, je me suis interessé à l’identification partielle des effets de traitements dans différents modèles de choix discrets avec traitements endogènes. Les modèles d’effets de traitement ont pour but de mesurer l’impact de certaines interventions sur certaines variables d’intérêt. Le type de traitement et la variable d’intérêt peuvent être défini de manière générale afin de pouvoir être appliqué à plusieurs différents contextes. Il y a plusieurs exemples de traitement en économie du travail, de la santé, de l’éducation, ou en organisation industrielle telle que les programmes de formation à l’emploi, les techniques médicales, l’investissement en recherche et développement, ou l’appartenance à un syndicat. La décision d’être traité ou pas n’est généralement pas aléatoire mais est basée sur des choix et des préférences individuelles. Dans un tel contexte, mesurer l’effet du traitement devient problématique car il faut tenir compte du biais de sélection. Plusieurs versions paramétriques de ces modèles ont été largement étudiées dans la littérature, cependant dans les modèles à variation discrète, la paramétrisation est une source importante d’identification. Dans un tel contexte, il est donc difficile de savoir si les résultats empiriques obtenus sont guidés par les données ou par la paramétrisation imposée au modèle. Etant donné, que les formes paramétriques proposées pour ces types de modèles n’ont généralement pas de fondement économique, je propose dans cette thèse de regarder la version nonparamétrique de ces modèles. Ceci permettra donc de proposer des politiques économiques plus robustes. La principale difficulté dans l’identification nonparamétrique de fonctions structurelles, est le fait que la structure suggérée ne permet pas d’identifier un unique processus générateur des données et ceci peut être du soit à la présence d’équilibres multiples ou soit à des contraintes sur les observables. Dans de telles situations, les méthodes d’identifications traditionnelles deviennent inapplicable d’où le récent développement de la littérature sur l’identification dans les modèles incomplets. Cette littérature porte une attention particuliere à l’identification de l’ensemble des fonctions structurelles d’intérêt qui sont compatibles avec la vraie distribution des données, cet ensemble est appelé : l’ensemble identifié. Par conséquent, dans le premier chapitre de la thèse, je caractérise l’ensemble identifié pour les effets de traitements dans le modèle triangulaire binaire. Dans le second chapitre, je considère le modèle de Roy discret. Je caractérise l’ensemble identifié pour les effets de traitements dans un modèle de choix de secteur lorsque la variable d’intérêt est discrète. Les hypothèses de sélection du secteur comprennent le choix de sélection simple, étendu et généralisé de Roy. Dans le dernier chapitre, je considère un modèle à variable dépendante binaire avec plusieurs dimensions d’hétérogéneité, tels que les jeux d’entrées ou de participation. je caractérise l’ensemble identifié pour les fonctions de profits des firmes dans un jeux avec deux firmes et à information complète. Dans tout les chapitres, l’ensemble identifié des fonctions d’intérêt sont écrites sous formes de bornes et assez simple pour être estimées à partir des méthodes d’inférence existantes.

Invariant discretizations of partial differential equations

Un algorithme permettant de discrétiser les équations aux dérivées partielles (EDP) tout en préservant leurs symétries de Lie est élaboré. Ceci est rendu possible grâce à l'utilisation de dérivées partielles discrètes se transformant comme les dérivées partielles continues sous l'action de groupes de Lie locaux. Dans les applications, beaucoup d'EDP sont invariantes sous l'action de transformations ponctuelles de Lie de dimension infinie qui font partie de ce que l'on désigne comme des pseudo-groupes de Lie. Afin d'étendre la méthode de discrétisation préservant les symétries à ces équations, une discrétisation des pseudo-groupes est proposée. Cette discrétisation a pour effet de transformer les symétries ponctuelles en symétries généralisées dans l'espace discret. Des schémas invariants sont ensuite créés pour un certain nombre d'EDP. Dans tous les cas, des tests numériques montrent que les schémas invariants approximent mieux leur équivalent continu que les différences finies standard.

Zeolite-encapsulated Co(II), Ni(II) and Cu(II) complexes as catalysts for partial oxidation of benzyl alcohol and ethylbenzene

Co(II), Ni(II) and Cu(II) complexes of dimethylglyoxime and N,N-ethylenebis(7-methylsalicylideneamine) have been synthesized in situ in Y zeolite by the reaction of ion-exchanged metal ions with the flexible ligand molecules that had diffused into the cavities. The hybrid materials obtained have been characterized by elemental analysis, SEM, XRD, surface area, pore volume, magnetic moment, FTIR, UV-Vis and EPR techniques. Analysis of data indicates the formation of complexes in the pores without affecting the zeolite framework structure, the absence of any extraneous species and the geometry of encapsulated complexes. The catalytic activities for hydrogen peroxide decomposition and oxidation of benzyl alcohol and ethylbenzene of zeolite complexes are reported. Zeolite Cu(II) complexes were found to be more active than the corresponding Co(II) and Ni(II) complexes for oxidation reactions. The catalytic properties of the complexes are influenced by their geometry and by the steric environment of the active sites. Zeolite complexes are stable enough to be reused and are suitable to be utilized as partial oxidation catalysts.

Production, purification and partial characterization of a novel protease from marine Engyodontium album BTMFS10 under solid state fermentation

Engyodontium album isolated from marine sediment produced protease, which was active at pH 11. Process parameters influencing the production of alkaline protease by marine E. album was optimized. Particle size of <425 mm, 60% initial moisture content and incubation at 25 8C for 120 h were optimal for protease production under solid state fermentation (SSF) using wheat bran. The organism has two optimal pH (5 and 10) for maximal enzyme production. Sucrose as carbon source, ammonium hydrogen carbonate as additional inorganic nitrogen source and amino acid leucine enhanced enzyme production during SSF. The protease was purified and partially characterized. A 16-fold purified enzyme was obtained after ammonium sulphate precipitation and ion-exchange chromatography. Molecular weight of the purified enzyme protein was recorded approximately 38 kDa by SDS-PAGE. The enzyme showed maximum activity at pH 11 and 60 8C. Activity at high temperature and high alkaline pH suggests suitability of the enzyme for its application in detergent industry

Lipase from marine Aspergillus awamori BTMFW032: Production, partial purification and application in oil effluent treatment

Marine fungus BTMFW032, isolated from seawater and identified as Aspergillus awamori, was observed to produce an extracellular lipase, which could reduce 92% fat and oil content in the effluent laden with oil. In this study, medium for lipase production under submerged fermentation was optimized statistically employing response surface method toward maximal enzyme production. Medium with soyabean meal- 0.77% (w/v); (NH4)2SO4-0.1 M; KH2PO4-0.05 M; rice bran oil-2% (v/v); CaCl2-0.05 M; PEG 6000-0.05% (w/v); NaCl-1% (w/v); inoculum-1% (v/v); pH 3.0; incubation temperature 35 8C and incubation period-five days were identified as optimal conditions for maximal lipase production. The time course experiment under optimized condition, after statistical modeling, indicated that enzyme production commenced after 36 hours of incubation and reached a maximum after 96 hours (495.0 U/ml), whereas maximal specific activity of enzyme was recorded at 108 hours (1164.63 U/mg protein). After optimization an overall 4.6- fold increase in lipase production was achieved. Partial purification by (NH4)2SO4 precipitation and ion exchange chromatography resulted in 33.7% final yield. The lipase was noted to have a molecular mass of 90 kDa and optimal activity at pH 7 and 40 8C. Results indicated the scope for potential application of this marine fungal lipase in bioremediation.

Characterizations of some continuous distributions using partial moments

Inthis paper,we define partial moments for a univariate continuous random variable. A recurrence relationship for the Pearson curve using the partial moments is established. The interrelationship between the partial moments and other reliability measures such as failure rate, mean residual life function are proved. We also prove some characterization theorems using the partial moments in the context of length biased models and equilibrium distributions

The role of lower partial moments in stochastic modeling

Lower partial moments plays an important role in the analysis of risks and in income/poverty studies. In the present paper, we further investigate its importance in stochastic modeling and prove some characterization theorems arising out of it. We also identify its relationships with other important applied models such as weighted and equilibrium models. Finally, some applications of lower partial moments in poverty studies are also examined

Quantile based reliability aspects of partial moments

Partial moments are extensively used in literature for modeling and analysis of lifetime data. In this paper, we study properties of partial moments using quantile functions. The quantile based measure determines the underlying distribution uniquely. We then characterize certain lifetime quantile function models. The proposed measure provides alternate definitions for ageing criteria. Finally, we explore the utility of the measure to compare the characteristics of two lifetime distributions