The new biochemistry: blending the traditional with the other

Designing and presenting a modern biochemistry course is a challenging task. With an overabundance of topics to cover and a limited amount of class time, instructors must be creative and flexible. Traditionally, biochemical studies have been based on protein structure and function with emphasis on metabolism and bioenergetics. Developing methods in molecular biology are providing a new perspective on biochemistry. To train students for the future, it is imperative that these new topics be blended with the traditional. Model syllabi with this perspective are presented for a typical one-year, (28 weeks, two semester) course for biochemistry majors and the shorter survey course (one half-year, 14 weeks, one semester) for nonmajors, usually health professionals and biological science majors. The major change suggested from the present is to move the coverage of nucleic acid structure and function before metabolism. It is then possible to present metabolism in a more contemporary format with emphasis on regulation and integration.     

临床生物化学与检验教育史话

临床生物化学与检验是在对人体正常、病理状况的生理生化变化过程认识的基础上,通过分析人体标本为疾病的诊断和治疗提供有用的信息。其课程主要内容包括糖类、脂类、核酸、蛋白质、酶、激素、有机小分子、血液气体、电解质、治疗性药物、临床毒物等的检测及其临床应用。临床生物化学与检验课程对临床医学和医学检验专业学生非常重要。本文对临床生物化学与检验课程建设的历史和现状进行了总结。     

Remembering Our Forebears: Albert Jan Kluyver and the Unity of Life

The Dutch microbiologist/biochemist Albert Jan Kluyver (1888–1956) was an early proponent of the idea of biochemical unity, and how that concept might be demonstrated through the careful study of microbial life. The fundamental relatedness of living systems is an obvious correlate of the theory of evolution, and modern attempts to construct phylogenetic schemes support this relatedness through comparison of genomes. The approach of Kluyver and his scientific descendants predated the tools of modern molecular biology by decades. Kluyver himself is poorly recognized today, yet his influence at the time was profound. Through lens of today however, it has been argued that the focus by Kluyver and others to create taxonomic and phylogenetic schemes using morphology and biochemistry distorted and hindered progress of the discipline of microbiology, because of a perception that the older approaches focused too much on a reductionist worldview. This essay argues that in contrast the careful characterization of fundamental microbial metabolism and physiology by Kluyver made many of the advances of the latter part of the twentieth century possible, by offering a framework which in many respects anticipated our current view of phylogeny, and by directly and indirectly training a generation of scientists who became leaders in the explosive growth of biotechnology.     

Comparative biochemistry and physiology in Brazil: a critical appraisal

Brazil stood out as the country with the highest number of submissions to the editorial project dedicated to Latin America by the journal Comparative Biochemistry and Physiology. Therefore, we felt that it was important to critically discuss the state of comparative biochemistry and physiology in this country. Our study is based on data collected from the ISI Web-of-Science. We analyzed publication trends through time, availability of novel approaches and techniques, patterns of collaboration among different geographical regions, patterns of collaboration with researchers abroad, and relative efforts dedicated to the study of biochemical and physiological adaptation of native fauna representing different terrestrial Brazilian biomes. Overall, our data shows that comparative biochemistry and physiology is a lively and productive discipline, but that some biases limit the scope of the field in Brazil. Some important limitations are the very heterogeneous distribution of research nuclei throughout the country and the absence of some important approaches, such as remote sensing and the use of molecular biology techniques in a comparative or evolutionary context. We also noticed that international collaboration far surpasses interregional collaboration, and discuss the possible causes and consequences of this situation. Finally, we found that Brazilian comparative biochemistry and physiology is biome-biased, as the Amazonian fauna has received far more attention than the whole pool of fauna representing other terrestrial biomes. We discuss the possible causes of these biases, and propose some directions that may contribute to invigorate the field in the country.     

Microfluidics Approaches in Modern Developmental Biology

Modern automated microsystems based on microhydrodynamic (microfluidic) technologies— labs on chips—make it possible to solve various basic and applied research problems. In the last 15 years, the development of these approaches in application to the problems of modern quantitative (systems) development biology has been observed. In this field, high-throughput experiments aimed at accumulating ample quantitative data for their subsequent computer analysis are important. In this review, the main directions in the development and application of microfluidics approaches for solving problems of modern developmental biology using the classical model object, Drosophila embryo, as an example is discussed. Microfluidic systems provide an opportunity to perform experiments that can hardly be performed using other approaches. These systems allow automated, rapid, reliable, and proper placing of many live embryos on a substrate for their simultaneous confocal scanning, sorting them, or injecting them with various agents. Such systems make it possible, in particular, to create controlled gradients of microenvironmental parameters along a series of developing embryos or even to introduce discontinuity in parameters within the microenvironment of one embryo, so that the head half is under other conditions compared to the tail half (at continuous scanning). These approaches are used both in basic research of the functions of gene ensembles that control early development, including the problems of resistance of early patterns to disturbances, and in test systems for screening chemical agents on developing embryos. The problems of integration of microfluidic devices in systems for automated performance of experiments simultaneously on many developing embryos under conditions of their continuous scanning using modern fluorescence microscopy instruments will be discussed. The methods and approaches developed for Drosophila are also applicable to other model objects, even mammalian embryos.     

医学留学生生物化学实验教学的实践与探索

随着国际交流与合作的日益频繁,留学生教育已成为中国高等教育的一个重要组成部分,生物化学实验是医学留学生的必修重要课程之一,因此,生物化学实验课的教学质量对学生的全面发展非常重要。通过对留学生生物化学实验教学的实践,根据留学生自身的特点和生物化学实验课程的特点,对我校留学生的生物化学实验教学进行了有益的实践与探索。结果显示留学生生物化学实验课的教学质量取得了显著提高。     

Current developments in optical biochemical sensors

By combining modern fibre optics and opto-electronic instrumentation with chemical and biochemical reagent systems, it has become possible to fabricate optical biosensors. The current state of the art in this development is reviewed in this paper. Many developments describe selective and sensitive methods for sensing bioanalytes and it is likely that such a development will continue to be a very active area of analytical research. However, these biosensing devices can be regarded as successful only if their practicality and reliability can be demonstrated.     

Putting the systems back into systems biology

In recent years the term "systems biology" has become widespread in the biological literature, but most of the papers in which these words appear have surprisingly little to do with older notions of biological systems: they often seem to imply little more than reductionist biology applied on a large scale, with a little attention to interactions between some of the components, but with minimal attention to the kinetic properties of enzymes, which supplied much of the reductionist foundation of biochemistry. A systemic approach to biology ought to put the emphasis on the entire system; insofar as it is concerned with components at all, it is to explain their roles in meeting the needs of the system as a whole. Genuinely systemic thinking allows us to understand how biochemical systems are regulated, and why clumsy attempts to manipulate them for biotechnological purposes may fail. At a more abstract level, it is necessary for understanding the nature of life, because as long as an organism is treated as no more than a collection of components, one cannot ask the right questions, and certainly cannot answer them.     

Foundations of Vectorial Metabolism and Osmochemistry

Chemical transformations, like osmotic translocations, are transport processes when looked at in detail. In chemiosmotic systems, the pathways of specific ligand conduction are spatially orientated through osmoenzymes and porters in which the actions of chemical group, electron and solute transfer occur as vectorial (or higher tensorial order) diffusion processes down gradients of total potential energy that represent real spatially directed fields of force. Thus, it has been possible to describe classical bag-of-enzymes biochemistry as well as membrane biochemistry in terms of transport. But it would not have been possible to explain biological transport in terms of classical transformational biochemistry or chemistry. The recognition of this conceptual asymmetry in favour of transport has seemed to be upsetting to some biochemists and chemists; and they have resisted the shift towards thinking primarily in terms of the vectorial forces and co-linear displacements of ligands in place of their much less informative scalar products that correspond to the conventional scalar energies. Nevertheless, considerable progress has been made in establishing vectorial metabolism and osmochemistry as acceptable biochemical disciplines embracing transport and metabolism, and bioenergetics has been fundamentally transformed as a result.     

Foundations of vectorial metabolism and osmochemistry

Chemical transformations, like osmotic translocations, are transport processes when looked at in detail. In chemiosmotic systems, the pathways of specific ligand conduction are spatially orientated through osmoenzymes and porters in which the actions of chemical group, electron and solute transfer occur as vectorial (or higher tensorial order) diffusion processes down gradients of total potential energy that represent real spatially-directed fields of force. Thus, it has been possible to describe classical bag-of-enzymes biochemistry as well as membrane biochemistry in terms of transport. But it would not have been possible to explain biological transport in terms of classical transformational biochemistry or chemistry. The recognition of this conceptual asymmetry in favour of transport has seemed to be upsetting to some biochemists and chemists; and they have resisted the shift towards thinking primarily in terms of the vectorial forces and co-linear displacements of ligands in place of their much less informative scalar products that correspond to the conventional scalar energies. Nevertheless, considerable progress has been made in establishing vectorial metabolism and osmochemistry as acceptable biochemical disciplines embracing transport and metabolism, and bioenergetics has been fundamentally transformed as a result.     

Visualizing the Behavior of Human Rad51 at the Single-Molecule Level

The repair of double-stranded DNA breaks by homologous recombination is essential for maintaining genome integrity. Much of what we know about this DNA repair pathway in eukaryotes has been gleaned from genetics, in vivo experiments with GFP-tagged proteins and traditional biochemical experiments with purified proteins. However, many questions have remained inaccessible to these experimental approaches. Recent technological advances have made it possible to directly visualize the behaviors of individual DNA and protein molecules in vitro, and it is now becoming feasible to apply these technology-driven approaches to complex biochemical systems, such as those involved in the repair of damaged DNA. This report summarizes the use of total internal reflection fluorescence microscopy to probe fundamental aspects of protein-DNA interactions at the single-molecule level, and specific emphasis is placed on our efforts to develop new methods and techniques for studying DNA repair. Using these new approaches we are investigating the DNA-binding behavior of human Rad51 and we have recently demonstrated that this protein can slide on dsDNA via a one-dimensional random walk mechanism driven solely by thermal fluctuations of the surrounding solvent. Here, we highlight some possible implications of this recent finding, and we also briefly discuss the potential benefits of future single-molecule studies in the study of protein-DNA interactions and DNA repair.     

Reasoning methods in medical consultation systems: artificial intelligence approaches

It has been argued that the problem of medical diagnosis is fundamentally ill-structured, particularly during the early stages when the number of possible explanations for presenting complaints can be immense. This paper discusses the process of clinical hypothesis evocation, contrasts it with the structured decision making approaches used in traditional computer-based diagnostic systems, and briefly surveys the more open-ended reasoning methods that have been used in medical artificial intelligence (AI) programs. The additional complexity introduced when an advice system is designed to suggest management instead of (or in addition to) diagnosis is also emphasized. Example systems are discussed to illustrate the key concepts.     

Enrichment of Bruch's Membrane from Human Donor Eyes

Age-related macular degeneration (AMD) is a leading cause of visual impairment in the developed world. The disease manifests itself by the destruction of the center of the retina, called the macula, resulting in the loss of central vision. Early AMD is characterised by the presence of small, yellowish lesions called soft drusen that can progress onto late AMD such as geographic atrophy (dry AMD) or neovascularisation (wet AMD). Although the clinical changes are well described, and the understanding of genetic influences on conferring AMD risk are getting ever more detailed, one area lacking major progress is an understanding of the biochemical consequences of genetic risk. This is partly due to difficulties in understanding the biochemistry of Bruch’s membrane, a very thin extracellular matrix that acts as a biological filter of material from the blood supply and a scaffold on which the retinal pigment epithelial (RPE) cell monolayer resides. Drusen form within Bruch’s membrane and their presence disrupts nutrient flow to the RPE cells. Only by investigating the protein composition of Bruch’s membrane, and indeed how other proteins interact with it, can researchers hope to unravel the biochemical mechanisms underpinning drusen formation, development of AMD and subsequent vision loss. This paper details methodologies for enriching either whole Bruch’s membrane, or just from the macula region, so that it can be used for downstream biochemical analysis, and provide examples of how this is already changing the understanding of Bruch’s membrane biochemistry.     

Various methodological aspects of biochemistry

Certain methodological problems of present-day biochemistry are considered. Comprehension of the mechanisms of biochemical processes is shown to be possible on the basis of the structural and functional analysis at the molecular level. Ways of development and use of structural and functional approach to studies of supermolecular structures, cells and organisms on the whole are discussed.     

Understanding the evolution of restriction-modification systems: clues from sequence and structure comparisons

Restriction-modification (RM) systems comprise two opposing enzymatic activities: a restriction endonuclease, that targets specific DNA sequences and performs endonucleolytic cleavage, and a modification methyltransferase that renders these sequences resistant to cleavage. Studies on molecular genetics and biochemistry of RM systems have been carried out over the past four decades, laying foundations for modern molecular biology and providing important models for mechanisms of highly specific protein-DNA interactions. Although the number of known, relevant sequences 3D structures of RM proteins is growing steadily, we do not fully understand their functional diversities from an evolutionary perspective and we are not yet able to engineer new sequence specificities based on rational approaches. Recent findings on the evolution of RM systems and on their structures and mechanisms of action have led to a picture in which conserved modules with defined function are shared between different RM proteins and other enzymes involved in nucleic acid biochemistry. On the other hand, it has been realized that some of the modules have been replaced in the evolution by unrelated domains exerting similar function. The aim of this review is to give a survey on the recent progress in the field of structural phylogeny of RM enzymes with special emphasis on studies of sequence-structure-function relationships and emerging potential applications in biotechnology.     

From steroid receptors to cytokines: the thermodynamics of self-associating systems

Since 1987, the Gibbs Conference on Biothermodynamics has maintained a focus on understanding the quantitative aspects of gene regulatory systems. These studies coupled rigorous techniques with exact theory to dissect the linked reactions associated with bacterial and lower eukaryotic gene regulation. However, only in the last ten years has it become possible to apply this approach to clinically relevant, human gene regulatory systems. Here we summarize our work on the thermodynamics of human steroid receptors and their interactions with multi-site promoter sequences, highlighting results not available from more traditional biochemical and structural approaches. Noting that the Gibbs Conference has also served as a vehicle to promote the broader use of thermodynamics in understanding biology, we then discuss collaborative work on the hydrodynamics of a cytokine implicated in tumor suppression, prostate derived factor (PDF).     

PBL与图表相结合的教学法在《基础生物化学》课程教学中的初步实践

针对生物化学课程的特点,以改革当前传统的课堂讲授式教学模式作为突破口,采用基于问题学习（PBL）与图表结合的教学模式在《基础生物化学》中进行教学尝试。初步教学实践表明,通过PBL与图表教学法的互补结合,基本能克服《基础生物化学》教学活动中存在的两类矛盾,使传统的以＂教＂为主的教学模式,转变为以＂学＂与＂教＂相互平衡和促进的教学模式,使学生在掌握所学课程核心内容的同时又能获得学习方法、提高学习兴趣和学习主动性。     

The virtual human: towards a global systems biology of multiscale, distributed biochemical network models

There is an emerging recognition of the importance of modelling large-scale biochemical systems, with the 'digital human' an obviously desirable goal. This will then permit researchers to analyse the behaviour of such systems in silico so as to be able to perform 'what-if?' experiments prior to determining whether they are actually worthwhile or not, and for understanding whether a particular model does in fact describe or predict experimental observations. Existing and developing standards such as SBML are beginning to permit the principled storage and exchange of biochemical network models, while environments for effecting distributed workflows (such as Taverna) will allow us to link together these models and their behaviour. This allows the local experts to work on those parts of cellular or organellar biochemistry on which they have most expertise, while making their results available to the community as a whole. This kind of architecture permits the distributed yet integrated goal of an evolving 'digital human' model to be realized.