Process development for a recombinant Chinese Hamster Ovary (CHO) cell line utilizing a metal induced and amplified metallothionein expression system

The suspension Chinese Hamster Ovary cell line, 13-10-302, utilizing the metallothionein (MT) expression system producing recombinant human growth hormone (hGH) was studied in a serum-free and cadmium-free medium at different fermentation scales and modes of operation. Initial experiments were carried out to optimize the concentration of metal addition to induce the MT promoter. Subsequently, the cultivation of the 13-10-302 cell line was scaled up from spinner flasks into bioreactors, and the cultivation duration was extended with fed-batch and perfusion strategies utilizing 180 muM zinc to induce the promoter controlling expression of recombinant hGH. It was shown that a fed-batch process could increase the maximum cell numbers twofold, from 3.3 to 6.3 x 10(6) cell/mL, over those obtained in normal batch fermentations, and this coupled with extended fermentation times resulted in a fourfold increase in final hGH titer, from 135 +/- 15 to 670 +/- 70 mg/L at a specific productivity q(hGH) value of 12 pg cell(-1)d(-1). The addition of sodium butyrate increased the specific productivity of hGH in cells to a value of approximately 48 pg cell(-1)d(-1), resulting in a final hGH titer of over a gram per liter during fed-batch runs. A BioSep acoustic cell recycler was used to retain the cells in the bioreactor during perfusion operation. It was necessary to maintain the specific feeding rates (SFR) above a value of 0.2 vvd/(10(6) cell/mL) to maintain the viability and productivity of the 13-10-302 cells; under these conditions the viable cell number increased to over 107 cell/mL and resulted in a volumetric productivity of over 120 mg(hGH) L(-1)d(-1). Process development described in this work demonstrates cultivation at various scales and sustained high levels of productivity under cadmium free condition in a CHO cell line utilizing an inducible metallothionein expression system. (C) 2004 Wiley Periodicals, Inc.

Malformations of Cortical Development in Patients With Mid line Facial Defects and Ocular Hypertelorism

Objectives: We studied the neuroimaging and neurophysiological aspects of 17 patients with midline facial defects with ocular hypertelorism (MFDH). Methods: The investigation protocol included a previous semistructured questionnaire about family history; gestational, neonatal, and postnatal development; and dysmorphologic and neurologic evaluation. Recognized monogenic disorders and individuals with other well-known conditions were excluded. All patients had high resolution magnetic resonance imaging (MRI) with multiplanar reconstruction (MPR) and routine electroencephalograms (EEGs). Results: We detected abnormalities in five patients whose MRIs had been previously reported as normal. MRI showed central nervous system (CNS) structural abnormalities in all patients, which included commissural alterations in 16/17 (94%), malformations of cortical development in 10/17 (58%), disturbances of neural tube closure in 7/17(42%), and posterior fossa anomalies in 6/17 (35%). Some patients had more than one type of malformation occurring at different stages of the embryonary process. EEGs showed epileptiform activity in 4/17 (24%) and background abnormalities in 5/17 (29%) of patients. Conclusion: This study clearly demonstrated the presence of structural and functional neurologic alterations related to MFDH. Therefore, the CNS anomalies cannot be considered incidental findings but an intrinsic part of this condition, which could be related to environmental effects and/or genetic mutations. These findings would provide a basis for future investigations on MFDH and should also be considered when planning rehabilitation.

Development of a real-time RT-PCR assay for plasma membrane calcium ATPase isoform 1 (PMCA1) mRNA levels in a human breast epithelial cell line.

The plasma membrane Ca2+ pump is a key regulator of cytosolic free Ca2+. Recent studies have demonstrated the dynamic expression of the plasma membrane Ca2+ pump in a variety of cell types. Furthermore, alterations in plasma membrane calcium pump activity have now been implicated in human disease. In this study, the development of a technique to quantitatively assess mRNA expression of the human plasma membrane Ca2+ ATPase (PMCA1) isoform of the plasma membrane Ca2+ pump, using a real-time reverse transcriptase-polymerase chain reaction (real-time RT-PCR) assay in a human breast epithelial cell line (MCF-7) is described. The sequences of the PMCA1 primers and probe for real-time RT-PCR are presented. The results also indicate that PMCA1 mRNA can be normalized to both 18S ribosomal RNA (18S rRNA) and human glyceraldehyde-3-phosphate dehydrogenase (hGAPDH) in MCF-7 cells. Real-time RT-PCR will be most useful in assessing PMCA1 mRNA expression in cases where only low amounts of RNA are available and/or when numerous samples must be assessed simultaneously. (C) 2001 Elsevier Science Inc. All rights reserved.

Managing new product development in the Chinese steel industry: An empirical investigation

This study explores several important aspects of the management of new product development (NPD) in the Chinese steel industry. Specifically it explores NPD success factors, the importance of management functions to new product success and measures of new product success from the perspective of the industry's practitioners. Based on a sample of 190 industrial practitioners from 18 Chinese steel companies, the study provides a mixed picture as China makes the transition from a centrally-controlled to market-based economy. On one hand, respondents ranked understanding users' needs as the most important factor influencing the performance of the new products. Further, formulating new product strategy and strengthening market research are perceived as the most important managerial functions in NPD. However, technical performance measures are regarded as more important and are more widely used in industry than market-based or financial measures of success.

Warranty and optimal reliability improvement through product development

For products sold with warranty, the warranty servicing cost can be reduced by improving product reliability through a development process. However, this increases the unit manufacturing cost. Optimal development must achieve a trade-off between these two costs. The outcome of the development process is uncertain and needs to be taken into account in the determination of the optimal development effort. The paper develops a model where this uncertainty is taken into account. (C) 2003 Elsevier Ltd. All rights reserved.

Integrating environmental and economic life cycle analysis in product development: a material selection case study

Purpose Achieving sustainability by rethinking products, services and strategies is an enormous challenge currently laid upon the economic sector, in which materials selection plays a critical role. In this context, the present work describes an environmental and economic life cycle analysis of a structural product, comparing two possible material alternatives. The product chosen is a storage tank, presently manufactured in stainless steel (SST) or in a glass fibre reinforced polymer composite (CST). The overall goal of the study is to identify environmental and economic strong and weak points related to the life cycle of the two material alternatives. The consequential win-win or trade-off situations will be identified via a Life Cycle Assessment/Life Cycle Costing (LCA/LCC) integrated model. Methods The LCA/LCC integrated model used consists in applying the LCA methodology to the product system, incorporating, in parallel, its results into the LCC study, namely those of the Life Cycle Inventory (LCI) and the Life Cycle Impact Assessment (LCIA). Results In both the SST and CST systems the most significant life cycle phase is the raw materials production, in which the most significant environmental burdens correspond to the Fossil fuels and Respiratory inorganics categories. The LCA/LCC integrated analysis shows that the CST has globally a preferable environmental and economic profile, as its impacts are lower than those of the SST in all life cycle stages. Both the internal and external costs are lower, the former resulting mainly from the composite material being significantly less expensive than stainless steel. This therefore represents a full win-win situation. As a consequence, the study clearly indicates that using a thermoset composite material to manufacture storage tanks is environmentally and economically desirable. However, it was also evident that the environmental performance of the CST could be improved by altering its End-of-Life stage. Conclusions The results of the present work provide enlightening insights into the synergies between the environmental and the economic performance of a structural product made with alternative materials. Further, they provide conclusive evidence to support the integration of environmental and economic life cycle analysis in the product development processes of a manufacturing company, or in some cases even in its procurement practices.

Risk management of new product development process

Dissertation submitted to Faculdade de Ciências e Tecnologia of Universidade Nova de Lisboa for the achievement of Integrated Master´s degree in Industrial Management Engineering

Development of a satellite based PM10 concentration product for urban areas

Dissertação para obtenção do Grau de Doutor em Engenharia do Ambiente

Single line for assembly just-in-sequence multiple models

Programa Doutoral em Líderes para as Indústrias Tecnológicas

Propolis: A complex natural product with a plethora of biological activities that can be explored for drug development

The health industry has always used natural products as a rich, promising, and alternative source of drugs that are used in the health system. Propolis, a natural resinous product known for centuries, is a complex product obtained by honey bees from substances collected from parts of different plants, buds, and exudates in different geographic areas. Propolis has been attracting scientific attention since it has many biological and pharmacological properties, which are related to its chemical composition. Several in vitro and in vivo studies have been performed to characterize and understand the diverse bioactivities of propolis and its isolated compounds, as well as to evaluate and validate its potential. Yet, there is a lack of information concerning clinical effectiveness. The goal of this review is to discuss the potential of propolis for the development of new drugs by presenting published data concerning the chemical composition and the biological properties of this natural compound from different geographic origins.

Introducing biomimetic approaches to materials development and product design for engineering students

In this paper, we present a new course entitled “Biomimicry: from life to nanotechnological innovations” at the Mines Nancy Engineering School, Nancy, France, and explain how we developed a specific curriculum covering biomimicry. We discuss strategies that can be followed by teachers to explain selected contents in the multi-disciplinary field of biomimicry and/or bioinspiration to undergraduate students and how practical classroom activities can be conducted as individual or team work. We hope that sharing our experience will help teachers and senior researchers disseminate useful concepts and real examples of biomimetic principles and tools for the development of new materials, new/improved design and fabrication strategies, and innovation methodologies.

In-line rheo-optical microstructural characterization of complex polymer systems

Tese de Doutoramento em Ciência e Engenharia de Polímeros e Compósitos.