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PubMed1994

The new cell physiology: an outline, presented against its full historical background, beginning from the beginning.

The primary objective of this review is to present a brief outline of the new cell physiology. First introduced in 1962 under the title of the association-induction hypothesis, the theory is now confirmed in its essence, after more than thirty years of world-wide experimental testing. While minor, and even more than minor, improvements and revisions may yet be needed, the weight and scope of the confirmatory evidence have left little doubt that a comprehensive and coherent new theory of cell physiology is established. For the first time in history we have available a realistic cell physiology, awaiting to unite and to guide future biomedical research and teaching. The review also demonstrates in passing how, as well as why, the old cell physiology has fizzled so miserably. Instead of providing the guiding light for all biomedical research it has degenerated into a fairy tale of make-believe. This degradation began with a wrong theory. The situation has grown steadily worse with time by the impact of premature progressive fragmentation on something inherently indivisible; and by the universal adoption of the peer-review system which gives to those with strong vested interests in the preservation of the status quo absolute power in determining who prospers and who is cast out (see Endnote 9). It is thus hardly surprising that up to now, despite superlative chemical engineering know-how available and the lure of great financial awards, not a single drug has been designed from understanding, an understanding that will remain forever beyond reach as long as the guiding theory of the cell physiology is basically wrong. In contrast, in its brief history, and despite its minuscule list of subscribers and their endless debilitating struggle merely to stay alive (see Endnote 9), the new cell physiology has already generated one mankind-enhancing medical technology of major proportions, magnetic resonance imaging or MRI (see Ling, 1984, p. vii; 1992, p. xxv). To this day, most biomedical research scientists, teachers as well as students, still operate under the delusion that the old cell physiology, a little battered perhaps, is still basically sound. By far the greatest majority of them simply do not know enough; and it is hard for them to find a convenient source of relevant information that will help in this regard. A second purpose of the present review is to fill this need. To a few "true believers" of the old cell physiology the preservation of the status quo is of overriding importance.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1997

[Development of a job-exposure matrix in the heavy-metal industry in France].

A job exposure matrix (JEM) was developed by a committee of experts using the DELPHI method, in the French hard metal industry, in order to assess occupational exposures to cobalt along with tungsten carbide resulting from the industrial process. This JEM is part of a nested case-control study, carried out within the historical cohort of workers ever employed in these factories, aimed at assessing lung cancer risk. The committee included 8 experts: hygienists, chemical engineers, occupational physicians and epidemiologists. The JEM was developed in four stages: (i) visit of factories, (ii) definition of lines (job-periods) and columns (exposures) of the JEM, (iii) definition of coding procedures, (iv) coding the cells of the JEM. This last stage used a method derived from the DELPHI method. Throughout the study period 1945-1994. 320 job-periods and 21 agents were defined. A quantitative assignment (level 0 to level 9) along with a frequency code (1 to 3) was attempted for 4 agents, whereas only a qualitative assignment (non exposed/exposed, i.e. 0/l) was done for the other agents. An additional probability code (1 to 3) was assigned to all agents. This procedure led to 46 columns and 14,720 cells in the JEM. When applying the DELPHI method, the consensus of the committee was obtained for 85% of all cells after the first individual assignment of experts, 88% after the second individual assignment and 100% after the third assignment by the experts all together. In order to validate the JEM, these expert assignments will be brought together with the results of exposure measurements that were performed in some workplaces of these factories. The JEM will also be linked with the data base of the case-control study for the exposure assessment of cases and controls.

Case-Control Studies↗
PubMed2000

The commercial production of chemicals using pathway engineering.

Integration of metabolic pathway engineering and fermentation production technologies is necessary for the successful commercial production of chemicals. The 'toolbox' to do pathway engineering is ever expanding to enable mining of biodiversity, to maximize productivity, enhance carbon efficiency, improve product purity, expand product lines, and broaden markets. Functional genomics, proteomics, fluxomics, and physiomics are complementary to pathway engineering, and their successful applications are bound to multiply product turnover per cell, channel carbon efficiently, shrink the size of factories (i.e., reduce steel in the ground), and minimize product development cycle times to bring products to market.

Alcohols↗
PubMed2006

Existing and emerging detection technologies for DNA (Deoxyribonucleic Acid) finger printing, sequencing, bio- and analytical chips: a multidisciplinary development unifying molecular biology, chemical and electronics engineering.

The current status and research trends of detection techniques for DNA-based analysis such as DNA finger printing, sequencing, biochips and allied fields are examined. An overview of main detectors is presented vis-à-vis these DNA operations. The biochip method is explained, the role of micro- and nanoelectronic technologies in biochip realization is highlighted, various optical and electrical detection principles employed in biochips are indicated, and the operational mechanisms of these detection devices are described. Although a diversity of biochips for diagnostic and therapeutic applications has been demonstrated in research laboratories worldwide, only some of these chips have entered the clinical market, and more chips are awaiting commercialization. The necessity of tagging is eliminated in refractive-index change based devices, but the basic flaw of indirect nature of most detection methodologies can only be overcome by generic and/or reagentless DNA sensors such as the conductance-based approach and the DNA-single electron transistor (DNA-SET) structure. Devices of the electrical detection-based category are expected to pave the pathway for the next-generation DNA chips. The review provides a comprehensive coverage of the detection technologies for DNA finger printing, sequencing and related techniques, encompassing a variety of methods from the primitive art to the state-of-the-art scenario as well as promising methods for the future.

Chemical Engineering↗
PubMed2000

Biosynthetic phage display: a novel protein engineering tool combining chemical and genetic diversity.

BACKGROUND: Molecular diversity in nature is developed through a combination of genetic and chemical elements. We have developed a method that permits selective manipulation of both these elements in one protein engineering tool. It combines the ability to introduce non-natural amino acids into a protein using native chemical ligation with exhaustive targeted mutagenesis of the protein via phage-display mutagenesis. RESULTS: A fully functional biosynthetic version of the protease inhibitor eglin c was constructed. The amino-terminal fragment (residues 8-40) was chemically synthesized with a non-natural amino acid at position 25. The remaining carboxy-terminal fragment was expressed as a 30-residue peptide extension of gIIIp or gVIIIp on filamentous phage in a phage-display mutagenesis format. Native chemical ligation was used to couple the two fragments and produced a protein that refolded to its active form. To facilitate the packing of the introduced non-natural amino acid, residues 52 and 54 in the carboxy-terminal fragment were fully randomized by phage-display mutagenesis. Although the majority of the observed solutions for residues 52 and 54 were hydrophobic - complementing the stereochemistry of the introduced non-natural amino acid - a significant number of residues (unexpected because of stereochemical and charge criteria) were observed in these positions. CONCLUSIONS: Peptide synthesis and phage-display mutagenesis can be combined to produce a very powerful protein engineering tool. The physical properties of the environment surrounding the introduced non-natural residue can be selected for by evaluating all possible combinations of amino acid types at a targeted set of sites using phage-display mutagenesis.

Bacteriophage M13↗
PubMed2003

Identification of genotype-selective antitumor agents using synthetic lethal chemical screening in engineered human tumor cells.

We used synthetic lethal high-throughput screening to interrogate 23,550 compounds for their ability to kill engineered tumorigenic cells but not their isogenic normal cell counterparts. We identified known and novel compounds with genotype-selective activity, including doxorubicin, daunorubicin, mitoxantrone, camptothecin, sangivamycin, echinomycin, bouvardin, NSC146109, and a novel compound that we named erastin. These compounds have increased activity in the presence of hTERT, the SV40 large and small T oncoproteins, the human papillomavirus type 16 (HPV) E6 and E7 oncoproteins, and oncogenic HRAS. We found that overexpressing hTERT and either E7 or LT increased expression of topoisomerase 2alpha and that overexpressing RAS(V12) and ST both increased expression of topoisomerase 1 and sensitized cells to a nonapoptotic cell death process initiated by erastin.

Antineoplastic Agents↗
PubMed2006

Engineering of protein thermodynamic, kinetic, and colloidal stability: Chemical Glycosylation with monofunctionally activated glycans.

In this work we establish the relationship between chemical glycosylation and protein thermodynamic, kinetic, and colloidal stability. While there have been reports in the literature that chemical glycosylation modulates protein stability, mechanistic details still remain uncertain. To address this issue, we designed and coupled monofunctional activated glycans (lactose and dextran) to the model protein alpha-chymotrypsin (alpha-CT). This resulted in a series of glycoconjugates with variations in the glycan size and degree of glycosylation. Thermodynamic unfolding, thermal inactivation, and temperature-induced aggregation experiments revealed that chemical glycosylation increased protein thermodynamic (Delta G(25 degrees C)), kinetic (t(1/2)(45 degrees C)), and colloidal stability. These results highlight the potential of chemical glycosylation with monofunctional activated glycans as a technology for increasing the long-term stability of liquid protein formulations for industrial and biotherapeutic applications.

Chemical Engineering↗
PubMedDate not supplied

In situ magnetic resonance measurement of conversion, hydrodynamics and mass transfer during single- and two-phase flow in fixed-bed reactors.

In recent years there has been increasing interest in applying magnetic resonance (MR) techniques in areas of engineering and chemical technology. The science that underpins many of these applications is the physics and chemistry of transport and reaction processes in porous materials. Key to the exploitation of MR methods will be our ability to demonstrate that MR yields information that cannot be obtained using conventional measurement techniques in engineering research. This article describes two case studies that highlight the power of MR to give new insights to chemical engineers. First, we demonstrate the application of MR techniques to explore both mass transfer and chemical conversion in situ within a fixed bed of catalyst, and we then use these data to identify the rate-controlling step of the chemical conversion. Second, we implement a rapid imaging technique to study the stability of the gas-liquid distribution in the low- and high-interaction two-phase flow regimes in a trickle-bed reactor.

Acetic Acid↗