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.

Compact tunable microfluidic interferometer

We demonstrate a compact tunable filter based on a novel microfluidic single beam Mach-Zehnder interferometer. The optical path difference occurs during propagation across a fluid-air interface ( meniscus), the inherent mobility of which provides tunability. Optical losses are minimized by optimizing the meniscus shape through surface treatment. Optical spectra are compared to a 3D beam propagation method simulations and good agreement is found. Tunability, low insertion loss and strength of the resonance are well reproduced. The device performance displays a resonance depth of - 28 dB and insertion loss maintained at - 4 dB. (C) 2004 Optical Society of America.

Diving behaviour of a reptile (Crocodylus johnstoni) in the wild: Interactions with heart rate and body temperature

The differences in physical properties of air and water pose unique behavioural and physiological demands on semiaquatic animals. The aim of this study was to describe the diving behaviour of the freshwater crocodile Crocodylus johnstoni in the wild and to assess the relationships between diving, body temperature, and heart rate. Time-depth recorders, temperature-sensitive radio transmitters, and heart rate transmitters were deployed on each of six C. johnstoni (4.0-26.5 kg), and data were obtained from five animals. Crocodiles showed the greatest diving activity in the morning (0600-1200 hours) and were least active at night, remaining at the water surface. Surprisingly, activity pattern was asynchronous with thermoregulation, and activity was correlated to light rather than to body temperature. Nonetheless, crocodiles thermoregulated and showed a typical heart rate hysteresis pattern (heart rate during heating greater than heart rate during cooling) in response to heating and cooling. Additionally, dive length decreased with increasing body temperature. Maximum diving length was 119.6 min, but the greatest proportion of diving time was spent on relatively short (

Do socioeconomic gradients in women's health widen over time and with age

A population-based study was conducted to investigate changes over time in women's well-being and health service use by socio-cconomic status and whether these varied by age. Data from 12,328 mid-age women (aged 45-50 years in 1996) and 10,430 older women (aged 70-75 years) from the Australian Longitudinal Study on Women's Health were analysed. The main outcome measures were changes in the eight dimensions of the Short Form General Health Survey (SF-36) adjusted for baseline scores, lifestyle and behavioural factors; health care utilisation at Survey 2; and rate of deaths (older cohort only). Cross-sectional analyses showed clear socioeconomic differentials in well-being for both cohorts. Differential changes in health across tertiles of socioeconomic status (SES) were more evident in the mid-age cohort than in the older cohort. For the mid-aged women in the low SES tertile, declines in physical functioning (adjusted mean change of -2.4, standard error (SE) 1.1) and general health perceptions (-1.5, SE 1.1) were larger than the high SES group (physical functioning -0.8 SE 1.1, general health perceptions -0.8 SE 1.2). In the older cohort, changes in SF-36 scores over time were similar for all SES groups but women in the high SES group had lower death rates than women in the low SES group (relative risk: 0.79, 95% confidence interval 0.64-0.98). Findings suggest that SES differentials in physical health seem to widen during women's mid-adult years but narrow in older age. Nevertheless, SES remains an important predictor of health, health service use and mortality in older Australian women. (C) 2003 Elsevier Ltd. All rights reserved.

The physical and surface chemical characteristics of activated carbons and the adsorption of methylene blue from wastewater

Adsorption of a basic dye, methylene blue, from aqueous solutions onto as-received activated carbons and acid-treated carbons was investigated. The physical and surface chemical properties of the activated carbons were characterized using BET-N-2 adsorption, X-ray photoelectron spectroscopy (XPS), and mass titration. It was found that acid treatment had little effect on carbon textural characteristics but significantly changed the surface chemical properties, resulting in an adverse effect on dye adsorption. The physical properties of activated carbon, such as surface area and pore volume, have little effect on dye adsorption, while the pore size distribution and the surface chemical characteristics play important roles in dye adsorption. The pH value of the solution also influences the adsorption capacity significantly. For methylene blue, a higher pH of solution favors the adsorption capacity. The kinetic adsorption of methylene blue on all carbons follows a pseudo-second-order equation. (c) 2004 Elsevier Inc. All rights reserved.

Validity of accelerometry for measurement of activity in people with brain injury

Purpose: To evaluate the validity of a uniaxial accelerometer (MTI Actigraph) for measuring physical activity in people with acquired brain injury (ABI) using portable indirect calorimetry (Cosmed K4b(2)) as a criterion measure. Methods: Fourteen people with ABI and related gait pattern impairment (age 32 +/- 8 yr) wore an MTI Actigraph that measured activity (counts(.)min-(1)) and a Cosmed K4b(2) that measured oxygen consumption (mL(.)kg(-1.)min(-1)) during four activities: quiet sitting (QS) and comfortable paced (CP), brisk paced (BP), and fast paced (FP) walking. MET levels were predicted from Actigraph counts using a published equation and compared with Cosmed measures. Predicted METs for each of the 56 activity bouts (14 participants X 4 bouts) were classified (light, moderate, vigorous, or very vigorous intensity) and compared with Cosmed-based classifications. Results: Repeated-measures ANOVA indicated that walking condition intensities were significantly different (P < 0.05) and the Actigraph detected the differences. Overall correlation between measured and predicted METs was positive, moderate, and significant (r = 0.74). Mean predicted METs were not significantly different from measured for CP and BP, but for FP walking, predicted METs were significantly less than measured (P < 0.05). The Actigraph correctly classified intensity for 76.8% of all activity bouts and 91.5% of light- and moderate-intensity bouts. Conclusions: Actigraph counts provide a valid index of activity across the intensities investigated in this study. For light to moderate activity, Actigraph-based estimates of METs are acceptable for group-level analysis and are a valid means of classifying activity intensity. The Actigraph significantly underestimated higher intensity activity, although, in practice, this limitation will have minimal impact on activity measurement of most community-dwelling people with ABI.

Multi-types of Skyrmions in SU(N) Quantum Hall System

The skyrmions in SU(N) quantum Hall (QH) system are discussed. By analyzing the gauge field structure and the topological properties of this QH system it is pointed out that in the SU(N) QH system there can exist (N-1) types of skyrmion structures, instead of only one type of skyrmions. In this paper, by means of the Abelian projections according to the (N-1) Cartan subalgebra local bases, we obtain the (N-1) U(1) electromagnetic field tensors in the SU(N) gauge field of the QH system, and then derive (N-1) types of skyrmion structures from these U(1) sub-field tensors. Furthermore, in light of the phi-mapping topological current method, the topological charges and the motion of these skyrmions are also discussed.

A cross-cultural investigation of the use of teaching styles

Teacher beliefs are a major influence on teacher actions. Because context influences beliefs, it was the purpose of this study to explore teachers' beliefs about Mosston's Spectrum of Teaching Styles from an international perspective. Over 1,400 teachers from 7 countries completed a survey related to their self-reported use of and beliefs about various teaching styles. Data suggested a shared core of reproduction teaching style use. The use of and beliefs about the production styles of teaching were more varied. Teachers' use of styles was significantly related to their beliefs about the styles. [PUBLICATION ABSTRACT]

An experimental and finite element poroelastic creep response analysis of an intervertebral hydrogel disc model in axial compression

A hydrogel intervertebral disc (lVD) model consisting of an inner nucleus core and an outer anulus ring was manufactured from 30 and 35% by weight Poly(vinyl alcohol) hydrogel (PVA-H) concentrations and subjected to axial compression in between saturated porous endplates at 200 N for 11 h, 30 min. Repeat experiments (n = 4) on different samples (N = 2) show good reproducibility of fluid loss and axial deformation. An axisymmetric nonlinear poroelastic finite element model with variable permeability was developed using commercial finite element software to compare axial deformation and predicted fluid loss with experimental data. The FE predictions indicate differential fluid loss similar to that of biological IVDs, with the nucleus losing more water than the anulus, and there is overall good agreement between experimental and finite element predicted fluid loss. The stress distribution pattern indicates important similarities with the biological lVD that includes stress transference from the nucleus to the anulus upon sustained loading and renders it suitable as a model that can be used in future studies to better understand the role of fluid and stress in biological IVDs. (C) 2005 Springer Science + Business Media, Inc.

Water and nutrient availabilities do not affect lignotuber growth and sprouting ability of three eucalypt species of south-eastern Queensland

Shoot biomass and lignotuber size of seedlings of three eucalypt species, Eucalyptus acmenoides Schauer, E. siderophloia Benth. and Corymbia variegata [syn. E. maculata (F. Muell.)K. D. Hill and L. A. S. Johnson], were measured for glasshouse-grown seedlings established under two water and nutrient regimes. Seedlings were subjected to shoot removal (clipping) at ages from 9 to 19 weeks, and transferred to the high water treatment for a further 8 weeks to assess shoot emergence from lignotubers. Seedling shoot biomass was greater in both the high than the low nutrient and water treatments, but lignotuber diameter was not affected significantly. C. variegata seedlings had the largest lignotuber diameters, followed by E. siderophloia and E. acmenoides, respectively. Although growth of shoots was influenced by nutrient availability, results suggest that species differences in the growth of lignotubers was less affected. It is suggested that lignotuber growth was strongly influenced by genotype. More than 70% of C. variegata seedlings clipped at 9 weeks sprouted, compared with only 5 and 10% of seedlings of E. siderophloia and E. acmenoides, respectively. All C. variegata seedlings sprouted after being clipped at 19 weeks, but < 80% of E. siderophloia and < 60% of E. acmenoides sprouted when clipped at the same age. It was concluded that seedlings forming part of the regeneration stratum in dry sclerophyll forests need to be protected from damage for at least 4 months (for C. variegata) or at least 6 months (for E. siderophloia and E. acmenoides) if they are to survive by sprouting from lignotubers.

Lessons learned . . . about curriculum: Five years on and half a world away

The knowledge, skills, and attitudes manifested in health and physical education school curricula are an arbitrary selection of that which is known and valued at a particular place and time. Bernstein's (2000) theories of the social construction of knowledge offer a way to better understand the relationship among the production, selection, and reproduction of curricular knowledge. This article overviews contemporary knowledge in the primary field (production) on which curriculum writers in the recontextualizing field might draw. It highlights tensions in the knowledge generated within the primary field and, using a case of the USXs National Standards for Physical Education (NASPE), demonstrates how particular discourses become privileged when translated into curriculum documents in the recontextualizing field

Highly-ordered hybrid organic-inorganic isoporous membranes from polymer modified nanoparticles

atomic force microscopy (AFM); atom transfer radical polymerization (ATRP); block copolymers; self-assembly; silica nanoparticles.

Fusing subunit antigens to interleukin-2 and encapsulating them in liposomes improves their antigenicity but not their protective efficacy

Subunit vaccines commonly lack sufficient immunogenicity to stimulate a comprehensive protective immune response in vivo. We have investigated the potential of specific cytokines (interleukin-2) and particulate delivery systems (liposomes) to enhance antigenicity. Here we report that the IgG1 and IFN-gamma responses to a subunit antigen, consisting of a T and B-cell epitope from Influenza haemagglutinin, can be improved when it is both fused to interelukin-2 and encapsulated in liposomes. However, this vaccine formulation was not able to protect animals against a challenge with live Influenza A/PR/8/34 virus. The addition of more potent immune stimulators may be necessary to improve responses. (c) 2005 Elsevier Ltd. All rights reserved.

The importance of being physical in the middle years

Young people are physical (as are adults) and their bodies are significant in relation to who they are, what and how they learn, and who they can become. Consistent with middle schooling philosophy, but often not reflected in practice, a balanced approach to all aspects of the growth and development of young people is supported. Much research has shown the middle years is an important time assigned to 'identity development' and 'physical development' while it is also a time when many young people become less physically active and less engaged in learning at school. This paper reviews current research about physical activity, physical education and physicality in order to locate the place of the physical in the lives of young people and encourage practices in the middle years that acknowledge this physicality.