Proteomic reactors and their applications in biology

Proteomic analysis requires the combination of an extensive suite of technologies including protein processing and separation, micro-flow HPLC, MS and bioinformatics. Although proteomic technologies are still in flux, approaches that bypass gel electrophoresis (gel-free approaches) are dominating the field of proteomics. Along with the development of gel-free proteomics, came the development of devices for the processing of proteomic samples termed proteomic reactors. These microfluidic devices provide rapid, robust and efficient pre-MS sample procession by performing protein sample preparation/concentration, digestion and peptide fractionation. The proteomic reactor has advanced in two major directions: immobilized enzyme reactor and ion exchange-based proteomic reactor. This review summarizes the technical developments and biological applications of the proteomic reactor over the last decade.     

Electroporation: basic principles, practical considerations and applications in molecular biology

There are many approaches available for introducing foreign DNA into eukaryotes and prokaryotes. The most commonly used technique and currently of more practical interest is the electroporation. This review will focus on the electroporation as a promising, highly efficient and effective means of gene transfer. We summarize a detailed assessment of the various parameters and conditions that govern electroporation of a wide range of cell types.     

Confocal microscopy: principles and applications to the field of reproductive biology

Confocal microscopy allows analysis of fluorescent labeled thick specimens without physical sectioning. Optical sections are generated by eliminating out-of-focus fluorescence and displayed as digitalized images. It allows 3-dimensional reconstruction (XYZ) and time-analysis (XYT), thus providing unique chance to link morphology with cell function. Since images are obtained by scanning, excess illumination of the specimen and quick decrease of the fluorescent signal are avoided. Resolution obtained with a Laser Scanning Confocal Microscopy (LSCM) is theoretically better than that of a conventional microscope. The preparation of the specimen may be based on standard techniques, such as immunocytochemistry applied to fixed cells, or on staining of living cells, following the use of different fluorescent probes at the same time (colocalization). In our laboratory, we use the LSCM system Fluoview version 2.1 (Olympus) to study reproductive biology of animals and humans. We work on stainings of oocytes and blastocysts (mouse, bovine, human), and human ovarian tissues. We study mitochondrial distribution, cortical granule migration, calcium oscillations and spindle quality to link culture conditions and oocyte quality. Staining of F-actin is used to check transzonal projections (in zona pellucida) or to detect abnormalities following experimental treatment. Blastocyst quality is analyzed in sequential optical sections for microfilament organization and counting of total cell number (staining with phalloidin (actin) and picogreen (DNA). Trophectoderm and inner cell mass distribution (differential staining), apoptotic cells (TUNEL method) and viable cells (live/dead test) are also evaluated. Confocal imaging can be helpful for rapid determination of follicle density (staining with AM Calcein) and follicle morphology (picogreen) in ovarian cortical biopsies. The current review describes the principles of confocal microscopy and illustrates its applications to the field of reproductive biology by a large collection of pictures.     

Covalent inhibitors of glycosidases and their applications in biochemistry and biology

Glycoside hydrolases are important enzymes in a number of essential biological processes. Irreversible inhibitors of this class of enzyme have attracted interest as probes of both structure and function. In this review we discuss some of the compounds used to covalently modify glycosidases, their use in residue identification, structural and mechanistic investigations, and finally their applications, both in vitro and in vivo, to complex biological systems.     

Novel computational biology methods and their applications to drug discovery

Computational biology methods are now firmly entrenched in the drug discovery process. These methods focus on modeling and simulations of biological systems to complement and direct conventional experimental approaches. Two important branches of computational biology include protein homology modeling and the computational biophysics method of molecular dynamics. Protein modeling methods attempt to accurately predict three-dimensional (3D) structures of uncrystallized proteins for subsequent structure-based drug design applications. Molecular dynamics methods aim to elucidate the molecular motions of the static representations of crystallized protein structures. In this review we highlight recent novel methodologies in the field of homology modeling and molecular dynamics. Selected drug discovery applications using these methods conclude the review.     

Chaos: principles and implications in biology

In this paper we review some of the basic principles of thetheory of dynamical systems. We introduce the reader to thedefinition of chaos and strange attractors and we discuss theirimplications in biology. Received on June 9, 1988; accepted on October 24, 1988     

Toshiaki Osawa: biochemistry of lectins and their applications in immunochemistry and cellular biology

Lectins are proteins that agglutinate cells and exhibit an antibody like, sugar-binding specificity. Professor Toshiaki Osawa has discovered, purified and characterized many plant lectins that display diverse biological activities. Using lectins as biochemical tools, he developed methods to determine the biochemical structures of glycoprotein glycans that react with lectins; separated and characterized glycoproteins and cell populations; analysed the mechanisms by which lectins activate cells; and characterized several cytokines produced by immune cells stimulated by lectins. The studies on lectins, the field he took strong leadership, developed into an essential hub of the biology of multicellular organisms.     

Mathematical models in mammalian cell biology

A report on the Conference on Systems Biology of Mammalian Cells, Dresden, Germany, 22-24 May 2008.     

指示种、伞护种与旗舰种:有关概念及其在保护生物学中的应用

指示种、伞护种和旗舰种等的运用常常可以作为解决保护生物学问题的捷径。指示种被用来评价环境中人为干扰因素的程度,监测其他物种的种群动态,或用于确定生物多样性较高的区域;伞护种则常用来确定需保护生境的类型和面积;而旗舰种被用于引起公众对保护行为的关注。本文试图对上述术语和概念在使用上加以明确区分,并对其应用提出一些指导性原则。认为针对上述不同类型的代理种,使用的目的和选择的标准应有所不同,彼此不能混淆使用。     

Mathematical biology of social behavior

When an individual grows up in a society, he learns certain behavior patterns which are “accepted” by that society. He may in general have a tendency toward behavior patterns other than those which are “accepted” by the society. This tendency toward such unaccepted behavior may be due to a process of cerebration which results in doubt as to the “correctness” of the accepted behavior. Thus, on the one hand, the individual learns to follow the accepted rules almost automatically; on the other hand, he may tend to consciously break those rules. Using a neural circuit, suggested by H. D. Landahl in his theory of learning, a neurobiophysical interpretation of the above situation is outlined. Mathematical expressions are derived which describe the social behavior of an individual as a function of his age, social status, and some neurobiophysical parameters.     

Mathematical biology of automobile driving

Continuing a previous study (Bull. Math. Biophysics, 28, 645–654, 1966), the biophysical mechanism of a corrective turn is investigated for the case where the stimulus for the corrective turn is produced not only by the perception of the nearness of an edge of the lane, but also by the rate of approach of the car towards the edge. In that case it is found that the tracking curve of the car may consist of a series of damped sinusoids and safe driving would be possible at any speed if it were not for the endogenous fluctuation in the driver's central nervous system. If the effect of the rate of approach increases sufficiently rapidly as the distance to the edge of the lane decreases, then a stable undamped oscillating tracking curve is possible. The case is also studied where the driver makes a corrective turn in response to a direct perception of the angle between the direction of the lane and the longitudinal axis of the car.     

Bovine oviductal epithelial cells: their cell culture and applications in studies for reproductive biology

Epithelial cells of the mammalian oviduct play an important role in reproductive and developmental events that occur there. Oviductal epithelial cells from several mammalian species can be isolated and cultured in serum or serum-free medium in vitro and cell culture of bovine oviductal epithelial cells (BOEC) has been described by many investigators. Cultured BOEC show a wide variety of secretory activities and these secretory factors may influence early embryonic development or sperm function. Monolayer cultures of BOEC have been widely used for in vitro co-culture of bovine preimplantation embryos. The use of BOEC co-culture systems has improved embryonic development in nearly all the studies conducted. In addition, interaction of bovine spermatozoa with BOEC, in a similar manner to that observed for spermatozoa in vivo, induced specific changes in sperm capacitation and consequently improved the fertilizing capacity of bovine spermatozoa in vitro. Thus co-culture systems with BOEC may not only offer an excellent model for studying the mechanisms of capacitation and acrosome reaction of bovine spermatozoa but also provide a useful tool for the improvement of embryo development in vitro.     

Practical application of synthetic biology principles

Synthetic biology can be defined as the “repurposing and redesign of biological systems for novel purposes or applications, ” and the field lies at the interface of several biological research areas. This broad definition can be taken to include a variety of investigative endeavors, and successful design of new biological paradigms requires integration of many scientific disciplines including (but not limited to) protein engineering, metabolic engineering, genomics, structural biology, chemical biology, systems biology, and bioinformatics. This review focuses on recent applications of synthetic biology principles in three areas: (i) the construction of artificial biomolecules and biomaterials; (ii) the synthesis of both fine and bulk chemicals (including biofuels); and (iii) the construction of “smart” biological systems that respond to the surrounding environment.