Transient production of recombinant proteins by chinese hamster ovary cells using polyethyleneimine/DNA complexes in combination with microtubule disrupting anti-mitotic agents

We have developed a simple and robust transient expression system utilizing the 25 kDa branched cationic polymer polyethylenimine (PEI) as a vehicle to deliver plasmid DNA into suspension-adapted Chinese hamster ovary cells synchronized in G2/M phase of the cell cycle by anti-mitotic microtubule disrupting agents. The PEI-mediated transfection process was optimized with respect to PEI nitrogen to DNA phosphate molar ratio and the plasmid DNA mass to cell ratio using a reporter construct encoding firefly luciferase. Optimal production of luciferase was observed at a PEI N to DNA P ratio of 10:1 and 5 mug DNA 10(6) cells(-1). To manipulate transgene expression at mitosis, we arrested cells in G2/M phase of the cell cycle using the microtubule depolymerizing agent nocodazole. Using secreted human alkaline phosphatase (SEAP) and enhanced green fluorescent protein (eGFP) as reporters we showed that continued inclusion of nocodazole in cell culture medium significantly increased both transfection efficiency and reporter protein production. In the presence of nocodazole, greater than 90% of cells were eGFP positive 24 h post-transfection and qSEAP was increased almost fivefold, doubling total SEAP production. Under optimal conditions for PEI-mediated transfection, transient production of a recombinant chimeric IgG(4) encoded on a single vector was enhanced twofold by nocodazole, a final yield of approximately 5 mug mL(-1) achieved at an initial viable cell density of 1 x 10(6) cells mL(-1). The glycosylation of the recombinant antibody at Asn(297) was not significantly affected by nocodazole during transient production by this method. (C) 2004 Wiley Periodicals, Inc.

Douglas Shire Water Quality Monitoring Strategy: Final Report: a Report to Douglas Shire Council and the Department of the Environment and Heritage

This report describes results and conclusions from the monitoring component of the Douglas Shire Council (DSC) water quality project. The components of this project that this report addresses are: • Site selection and installation of in-stream and off-paddock automatic water quality monitoring equipment in the Douglas Shire. • Design of appropriate sampling strategies for automatic stations. • Estimation of loads of suspended sediment, total nitrogen and total phosphorus in rivers and also estimation of the changes in nutrient loads from sugar cane under different fertilizer application rates. • Development of a community-based water quality sampling program to complement the automatic sampling efforts. • Design of an optimised, long-term water quality monitoring strategy.

A Classification of Homogeneous Operators in the Cowen-Douglas Class

A complete list of homogeneous operators in the Cowen-Douglas class B-n(D) is given. This classification is obtained from an explicit realization of all the homogeneous Hermitian holomorphic vector bundles on the unit disc under the action of the universal covering group of the bi-holomorphic automorphism group of the unit disc.

A classification of homogeneous operators in the Cowen-Douglas class

An explicit construction of all the homogeneous holomorphic Hermitian vector bundles over the unit disc D is given. It is shown that every such vector bundle is a direct sum of irreducible ones. Among these irreducible homogeneous holomorphic Hermitian vector bundles over D, the ones corresponding to operators in the Cowen-Douglas class B-n(D) are identified. The classification of homogeneous operators in B-n(D) is completed using an explicit realization of these operators. We also show how the homogeneous operators in B-n(D) split into similarity classes. (C) 2011 Elsevier Inc. All rights reserved.

Infinitely divisible metrics and curvature inequalities for operators in the Cowen-Douglas class

The curvature (T)(w) of a contraction T in the Cowen-Douglas class B-1() is bounded above by the curvature (S*)(w) of the backward shift operator. However, in general, an operator satisfying the curvature inequality need not be contractive. In this paper, we characterize a slightly smaller class of contractions using a stronger form of the curvature inequality. Along the way, we find conditions on the metric of the holomorphic Hermitian vector bundle E-T corresponding to the operator T in the Cowen-Douglas class B-1() which ensures negative definiteness of the curvature function. We obtain a generalization for commuting tuples of operators in the class B-1() for a bounded domain in C-m.

Flag structure for operators in the Cowen-Douglas class

The explicit description of homogeneous operators and localization of a Hilbert module naturally leads to the definition of a class of Cowen-Douglas operators possessing a flag structure. These operators are irreducible. We show that the flag structure is rigid in the sense that the unitary equivalence class of the operator and the flag structure determine each other. We obtain a complete set of unitary invariants which are somewhat more tractable than those of an arbitrary operator in the Cowen-Douglas class. (C) 2014 Academie des sciences. Published by Elsevier Masson SAS. All rights reserved.

Homogeneous Hermitian holomorphic vector bundles and the Cowen-Douglas class over bounded symmetric domains

It is known that all the vector bundles of the title can be obtained by holomorphic induction from representations of a certain parabolic group on finite-dimensional inner product spaces. The representations, and the induced bundles, have composition series with irreducible factors. We write down an equivariant constant coefficient differential operator that intertwines the bundle with the direct sum of its irreducible factors. As an application, we show that in the case of the closed unit ball in C-n all homogeneous n-tuples of Cowen-Douglas operators are similar to direct sums of certain basic n-tuples. (c) 2015 Academie des sciences. Published by Elsevier Masson SAS. All rights reserved.

The identification of British adult specimens of Sigara lateralis (Leach), Sigara concinna (Fieber), Callicorixa praeusta (Fieber) and Callicorixa wollastoni (Douglas & Scott) (Hemiptera Heteroptera: Corixidae)

The aim of this communication is to briefly review nomenclature in the genus Callicorixa, describe the variation in the dark markings on the posterior legs of all four species, describe alternative diagnostic features, and provide a key to identification based on these alternative features. Attention is also drawn to a small error in FBA Scientific Publication 50 (Adults of the British aquatic Hemiptera Heteroptera: a key with ecological notes).

Report on the 1995 stock assessment of the Douglas catchment

The River Douglas has a long industrial heritage beginning in the early 18th century with its use by boats carrying goods between Wigan and Tarleton. The River and its tributaries have also historically been, and to a certain extent are still, subject to polluting inputs from the urban, agricultural and industrialised areas located within its catchment. During the early stages in the production of the River Douglas Catchment Management Plan, it became apparent that very little data existed on the populations of coarse and salmonid fish species within the River Douglas system. The data that did exist was largely anecdotal, consisting of catch reports from anglers or water bailiffs, or of dead and distressed fish following pollution incidents. This study was initiated to assess the status of coarse and salmonid fish species within the River Douglas system and so address the lack of knowledge. Eighty two sites were surveyed by electric fishing, including 14 sites using an electric fishing punt and up to four anodes. The data was analysed according to a new National Fisheries Classification Scheme. This classified the sites by the fish stock s present and compared the results with a database containing information from sites around the country that have similar habitat types. A stocking experiment was also undertaken in the River Lostock using chub reared at the Leyland Hatchery. These were marked with an identifiable blue spot in the spring of 1995 and then released into three, previously surveyed, locations in the river. These sites were then resurveyed during the summer stock assessment. This report also Site Reports with details on monitored sites, habitat features and fishery classification.

Holocene fossil pollen records of Douglas fir in northwestern California: reconstruction of past climate

Douglas fir is one of the most important trees in northwestern California. Regional pollen records suggest that it has become prominent only in the late Holocene. The primary cause for this change is probably southward stabilization of the mean airstream.

Arthur Douglas : missionary on Lake Nyasa, the story of his life

http://www.archive.org/details/arthurdouglasmi00douguoft