The character of a cell is defined by its constituent proteins, which are the result of specific patterns of gene expression. Crucial determinants of gene expression patterns are DNA-binding transcription factors that choose genes for transcriptional activation or repression by recognizing the sequence of DNA bases in their promoter regions. Interaction of these factors with their cognate sequences triggers a chain of events, often involving changes in the structure of chromatin, that leads to the assembly of an active transcription complex (e.g., Cosma et al. 1999). But the types of transcription factors present in a cell are not alone sufficient to define its spectrum of gene activity, as the transcriptional potential of a genome can become restricted in a stable manner during development. The constraints imposed by developmental history probably account for the very low efficiency of cloning animals from the nuclei of differentiated cells (Rideout et al. 2001; Wakayama and Yanagimachi 2001). A “transcription factors only” model would predict that the gene expression pattern of a differentiated nucleus would be completely reversible upon exposure to a new spectrum of factors. Although many aspects of expression can be reprogrammed in this way (Gurdon 1999), some marks of differentiation are evidently so stable that immersion in an alien cytoplasm cannot erase the memory. The genomic sequence of a differentiated cell is thought to be identical in most cases to that of the zygote from which it is descended (mammalian B and T cells being an obvious exception). This means that the marks of developmental history are unlikely to be caused by widespread somatic mutation. Processes less irrevocable than mutation fall under the umbrella term “epigenetic” mechanisms. A current definition of epigenetics is: “The study of mitotically and/or meiotically heritable changes in gene function that cannot be explained by changes in DNA sequence” (Russo et al. 1996). There are two epigenetic systems that affect animal development and fulfill the criterion of heritability: DNA methylation and the Polycomb-trithorax group (Pc-G/trx) protein complexes. (Histone modification has some attributes of an epigenetic process, but the issue of heritability has yet to be resolved.) This review concerns DNA methylation, focusing on the generation, inheritance, and biological significance of genomic methylation patterns in the development of mammals. Data will be discussed favoring the notion that DNA methylation may only affect genes that are already silenced by other mechanisms in the embryo. Embryonic transcription, on the other hand, may cause the exclusion of the DNA methylation machinery. The heritability of methylation states and the secondary nature of the decision to invite or exclude methylation support the idea that DNA methylation is adapted for a specific cellular memory function in development. Indeed, the possibility will be discussed that DNA methylation and Pc-G/trx may represent alternative systems of epigenetic memory that have been interchanged over evolutionary time. Animal DNA methylation has been the subject of several recent reviews (Bird and Wolffe 1999; Bestor 2000; Hsieh 2000; Costello and Plass 2001; Jones and Takai 2001). For recent reviews of plant and fungal DNA methylation, see Finnegan et al. (2000), Martienssen and Colot (2001), and Matzke et al. (2001).

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DNA methylation patterns and epigenetic memory

A review of potentially low-cost sorbents for heavy metals

Protein release from alginate matrices.

Flow Injection Analysis

This critical review summarizes developments in microfluidic platforms that enable the miniaturization, integration, automation and parallelization of (bio-)chemical assays (see S. Haeberle and R. Zengerle, Lab Chip, 2007, 7, 1094–1110, for an earlier review). In contrast to isolated application-specific solutions, a microfluidic platform provides a set of fluidic unit operations, which are designed for easy combination within a well-defined fabrication technology. This allows the easy, fast, and cost-efficient implementation of different application-specific (bio-)chemical processes. In our review we focus on recent developments from the last decade (2000s). We start with a brief introduction into technical advances, major market segments and promising applications. We continue with a detailed characterization of different microfluidic platforms, comprising a short definition, the functional principle, microfluidic unit operations, application examples as well as strengths and limitations of every platform. The microfluidic platforms in focus are lateral flow tests, linear actuated devices, pressure driven laminar flow, microfluidic large scale integration, segmented flow microfluidics, centrifugal microfluidics, electrokinetics, electrowetting, surface acoustic waves, and dedicated systems for massively parallel analysis. This review concludes with the attempt to provide a selection scheme for microfluidic platforms which is based on their characteristics according to key requirements of different applications and market segments. Applied selection criteria comprise portability, costs of instrument and disposability, sample throughput, number of parameters per sample, reagent consumption, precision, diversity of microfluidic unit operations and the flexibility in programming different liquid handling protocols (295 references).

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Microfluidic lab-on-a-chip platforms: requirements, characteristics and applications

Enzymatic substrate analysis is an attractive means of analysis in clinical chemistry because of its sensitivity and specificity. The GeMSAEC Fast Analyzer, in conjunction with a small computer, provides a means of performing routine enzymatic substrate analysis and offers the following advantages: ( a ) selectivity of approaches to enzymatic analysis, i.e., end-point or kinetic; ( b ) essentially parallel analyses of multiple samples, yielding a unique method for performing kinetic fixed-time analysis; ( c ) on-line data reduction, resulting in rapid calculation and output of results and the minimization of data handling errors; and ( d ) a small reagent volume per test (400 µl), which reduces the cost of analysis. The analysis of substrate with enzymatic end-point and kinetic procedures is examined by use of a computer-interfaced Fast Analyzer. Computer programs were written to facilitate this study. Glucose (hexokinase/GPD), urea (urease/GMD), and uric acid (uricase) have been used as examples in evaluating both end-point and kinetic analyses. The advantages and limitations of each type of analysis are presented, with the emphasis being placed on enzymatic substrate analysis and means by which the computer-interfaced Fast Analyzer can facilitate both end-point and kinetic analyses.

Enzymatic Kinetic Rate and End-Point Analyses of Substrate, by Use of a GeMSAEC Fast Analyzer

Immobilized cells : a review of recent literature

Design features and operation of a prototype miniaturized Fast Analyzer are described, and some results obtained with it are presented. The Analyzer occupies only one cubic foot of space. It has a 17-cuvet plastic rotor that rotates through a stationary optical system at speeds up to 5000 rpm. The resulting centrifugal force is utilized to transfer and mix a series of sample(s) and reagent(s) into the cuvets. The ensuing reactions are monitored spectrophotometrically, and the data evaluated in real time by an on-line computer. Samples (1 to 10 µl) and reagents (70 to 110 µl) are loaded into the rotor either discretely or dynamically; various rotor configurations can be used to do this. Many of the standard clinical analyses, including most of the NADH-linked enzymatic analyses, have been adapted for use with this analyzer. Precision obtained ranges from 1 to 4%. This report considers, specifically, analyses of some serum enzymes. Results show that the small analyzer possesses the previously demonstrated advantages of Fast Analyzers and, in addition, has several beneficial features arising from miniaturization.

Development of a miniature fast analyzer.

Reactor systems based on tapered fluidized beds are being developed for aqueous bioprocesses in which adhering microorganisms or immobilized active biological fractions are used. The use of a fluidized bed prevents biomass buildup, accommodates particulates in the feed stream, is compatible with gas sparging, and allows easy removal or addition of the active materials. The tapered reactor tends to stabilize the fluidized bed, thus allowing a much wider range of operating conditions. Preliminary experimental results and an empirical mathematical model of the tapered bed indicate that bed stability is associated with a decreasing velocity and void-fraction profile up the bed and the pressure drop across the bed decreases with increasing flow rates. The tapered fluidized bed bioreactor is being evaluated for use in the enzymatic production of hydrogen, microbiological denitrification, and microbiological degradation of coal conversion aqueous waste streams. The enzyme catalyzed conversion of lactose to glucose and galactose was used in the evaluation of the reactor concept.

Use of a tapered fluidized bed as a Continuous bioreactor

A compact analytical photometer of the rotary cuvette type designed to use small disposable cuvette rotors. A power driven cuvette rotor holder having a generally flat, circular configuration is provided with an integral, upstanding, annular lip for receiving, in an easily insertable and removable fashion, small disposable cuvette rotors. A series of axially extending apertures are disposed in a circular array through the rotor holder in axial alignment with respective cuvettes in the rotor to permit light to be transmitted through the cuvettes as the rotor holder and cuvette rotor rotate between a stationary light source and photodetector. Additional apertures extend through the rotor holder near its periphery to permit light passage through the rotor holder for rotor and cuvette synchronization purposes. Movable rotor and cuvette synchronization detectors are positioned along the periphery of the rotor holder for generating signals which are used to synchronize the photometer output with a computer and to provide rotor speed control. Rapid deceleration of the rotor holder and rotor for sample mixing purposes is accomplished by braking means engaging the rotor holder drive shaft.

Charles D. Scott

Papers

Enzymatic Kinetic Rate and End-Point Analyses of Substrate, by Use of a GeMSAEC Fast Analyzer

Immobilized cells : a review of recent literature

Development of a miniature fast analyzer.

Use of a tapered fluidized bed as a Continuous bioreactor

Compact dynamic multistation photometer utilizing disposable cuvette rotor