The spin label method developed by McConnell 15 years ago is now widely used in studies of the structure and dynamic properties of a variety of the biological systems such as proteins and protein complexes, lipids and membranes, nucleic acids, nucleoproteins, etc.

The ESR spectrum of the nitroxide radcal – the spin label – is very sensitive to its microenvironment and permits easy registration of even subtle alterations in it. If spin labels are attached to different sites of a macromolecule the information can be gained about conformational properties of all these local regions and, as a result, about the dynamic behaviour of the object as a whole.

The anatomy and physiology of cornea has been described in reviews (Maurice, 1969; Davson, 1980). We are primarily concerned with the structure of the stroma of cornea. There are two levels of organization in the stroma: the collagen fibrils and the lamellae. Each lamella contains an array of collagen fibrils which appear as long thin cylinders in electron micrographs. The lamellae are parallel to the cornea surface and are at right angles to the light direction. Adjacent lamellae are approximately at right angles to each other. There are many lamellae (about 200) in the thickness of the stroma so that the different lamellae have on the average all possible orientations.

The rates of reaction and reaction mechanisms in biological systems are the controlling factors in the metabolism and life cycles of all living organisms. Nevertheless, very little is known today about the details of these processes, particularly in more complicated life forms. For example, addition of certain metallic ions to an enzymic matrix can cause gross changes in enzymic activity (Brewer & Weber, 1966); just how these ions specifically enter into enzymic reactions is a subject of intense interest. The nature of the kinetics of bonding in nucleic acids is also the subject of much research (Eigen, 1967).

Nanofluidic structures have over the last two decades emerged as a powerful platform for detailed analysis of DNA on the kilobase pair length scale. When DNA is confined to a nanochannel, the combination of excluded volume and DNA stiffness leads to the DNA being stretched to near its full contour length. Importantly, this stretching takes place at equilibrium, without any chemical modifications to the DNA. As a result, any DNA can be analyzed, such as DNA extracted from cells or circular DNA, and it is straight-forward to study reactions on the ends of linear DNA. In this comprehensive review, we first give a thorough description of the current understanding of the polymer physics of DNA and how that leads to stretching in nanochannels. We then describe how the versatility of nanofabrication can be used to design devices specifically tailored for the problem at hand, either by controlling the degree of confinement or enabling facile exchange of reagents to measure DNA–protein reaction kinetics. The remainder of the review focuses on two important applications of confining DNA in nanochannels. The first is optical DNA mapping, which provides the genomic sequence of intact DNA molecules in excess of 100 kilobase pairs in size, with kilobase pair resolution, through labeling strategies that are suitable for fluorescence microscopy. In this section, we highlight solutions to the technical aspects of genomic mapping, including the use of enzyme-based labeling and affinity-based labeling to produce the genomic maps, rather than recent applications in human genetics. The second is DNA–protein interactions, and several recent examples of such studies on DNA compaction, filamentous protein complexes, and reactions with DNA ends are presented. Taken together, these two applications demonstrate the power of DNA confinement and nanofluidics in genomics, molecular biology, and biophysics.

Amyloid formation of the synaptic brain protein α-synuclein (αS) is related to degeneration of dopaminergic neurons in Parkinson's disease patients. αS is thought to function in vesicle transport and fusion and it binds strongly to negatively charged vesicles in vitro. Here we combined circular dichroism, fluorescence and imaging methods in vitro to characterize the interaction of αS with negatively charged vesicles of DOPS (1,2-dioleoyl-sn-glycero-3-phospho-L-serine, sodium salt) and DOPG (1,2-dioleoyl-sn-glycero-3-phospho-(1′-rac-glycerol), sodium salt) and the consequences of such interactions on αS amyloid formation. We found that lipid head-group chemistry modulates αS interactions and also affects amyloid fiber formation. During the course of the experiments, we made the unexpected discovery that pre-formed αS oligomers, typically present in a small amount in the αS starting material, acted as templates for linear growth of anomalous amyloid fibers in the presence of vesicles. At the same time, the remaining αS monomers were restricted from vesicle-mediated nucleation of amyloid fibers. Although not a dominant process in bulk experiments, this hidden αS aggregation pathway may be of importance in vivo.

Unlike proteins, the RNA backbone has numerous degrees of freedom (eight, if one counts the sugar pucker), making RNA modeling, structure building and prediction a multidimensional problem of exceptionally high complexity. And yet RNA tertiary structures are not infinite in their structural morphology; rather, they are built from a limited set of discrete units. In order to reduce the dimensionality of the RNA backbone in a physically reasonable way, a shorthand notation was created that reduced the RNA backbone torsion angles to two (η and θ, analogous to φ and ψ in proteins). When these torsion angles are calculated for nucleotides in a crystallographic database and plotted against one another, one obtains a plot analogous to a Ramachandran plot (the η/θ plot), with highly populated and unpopulated regions. Nucleotides that occupy proximal positions on the plot have identical structures and are found in the same units of tertiary structure. In this review, we describe the statistical validation of the η/θ formalism and the exploration of features within the η/θ plot. We also describe the application of the η/θ formalism in RNA motif discovery, structural comparison, RNA structure building and tertiary structure prediction. More than a tool, however, the η/θ formalism has provided new insights into RNA structure itself, revealing its fundamental components and the factors underlying RNA architectural form.

Among the unsolved mysteries of modern biology is the nature of a lining of blood vessels called the ‘endothelial surface layer’ or ESL. In venous micro-vessels, it is half a micron in thickness. The ESL is 10 times thicker than the endothelial glycocalyx (eGC) at its base, has been presumed to be comprised mainly of water, yet is rigid enough to exclude red blood cells. How is this possible? Developments in physical chemistry suggest that the venous ESL is actually comprised of nanobubbles of CO2, generated from tissue metabolism, in a foam nucleated in the eGC. For arteries, the ESL is dominated by nanobubbles of O2 and N2 from inspired air. The bubbles of the foam are separated and stabilized by thin layers of serum electrolyte and proteins, and a palisade of charged polymer strands of the eGC. The ESL seems to be a respiratory organ contiguous with the flowing blood, an extension of, and a ‘lung’ in miniature. This interpretation may have far-reaching consequences for physiology.

CryoEM has become the method of choice for determining the structure of large macromolecular complexes in multiple conformations, at resolutions where unambiguous atomic models can be built. Two effects that have limited progress in single-particle cryoEM are (i) beam-induced movement during image acquisition and (ii) protein adsorption and denaturation at the air-water interface during specimen preparation. While beam-induced movement now appears to have been resolved by all-gold specimen support grids with very small holes, surface effects at the air-water interface are a persistent problem. Strategies to overcome these effects include the use of alternative support films and new techniques for specimen deposition. We examine the future potential of recording perfect images of biological samples for routine structure determination at atomic resolution.

Nonlinear dynamics of biopolymers is a new and rapidly developing field of biophysical science. It can be considered as a part of the general dynamics which deals with the internal mobility of biopolymers. Theoreticians define it also as the next (anharmonic or nonlinear) approximation after the first harmonic or linear one.

Biophysicists, who, by definition, apply physical and mathematical principles to their analysis of biological problems for the understanding of the functions of living organisms, are to a great extent concerned with basic characteristics more or less common to all organisms. As pointed out by Griffin (1958) in his book on acoustic orientation, it may well be that ‘this fruitful preoccupation with the universals of protoplasm tends to foster an unduly restricted view of its many intricate potentialities’. The knowledge of the many ways in which animals and plants are adapted for a ‘successful life’ has had a great impact upon the concepts of evolution and natural selection. As a contribution to the proper understanding of these concepts the study of the biophysical principles, by which various geometrically oriented tissues serve different functions, becomes of special interest.

In the study of transmitter mechanisms in the peripheral nervous system and the spinal cord, intracellular recording has been a great advantage. In contrast, studies on central synapses have often used more indirect techniques, often extracellular recording from a cell or a small group of cells. In particular, a large amount of information has been acquired by iontophoretic application of drugs, often with a multibarrel electrode system where the iontophoretic electrodes are coupled to a recording electrode. Because of the difficulties with proper intracellular recording from intact brain preparations, there are relatively few intracellular studies on central transmitter effects. This situation compares unfavourably with the many studies on spinal cord, and, in particular, on peripheral nervous tissue.

A bacterium incorporates amino acids into protein with an error frequency close to one in three thousand (Edelman & Gallant, 1977). Nevertheless, the structural differences between related amino acids are so small that it is difficult to see how they can be distinguished from each other with such accuracy (see, for example, Pauling, 1958).

Indeed, the selection of amino acids during protein synthesis is carried out twice: first by the aminoacyl-tRNA synthetases and then by the codon-programmed ribosome. Each of these substrate selections is in fact a double selection. In the case of the synthetase both a particular amino acid and a corresponding cognate tRNA must be chosen to form the aminoacyl-tRNA. On the ribosome, the aminoacyl-tRNA must be matched with a cognate codon, and then the mRNA must be advanced by exactly one codon length to position the next codon in the appropriate ribosome site so that it too can be translated.

Some species of minerals, when comminuted and aerosolized in the form of respirable-size particulate dusts, are recognized as capable of producing disease in man after their inhalation. Comprehensive listings of such agents are in the current literature (e.g. in Aponte, 1970; Langer & Mackler, 1972; Ehrenreich et al. 1973a, b) and many of these and their effects are well known and receive much attention. Asbestos fibres of all varieties, ampbibole and quartz contaminated talcum powders (see Rohl et al. 1976), fibrous amphiboles and their cleavage fragments which contaminate the ambient environment (e.g., the contamination of Lake Superior with cummingtonite-grunerite, as discussed in Bowes, Langer & Rohl, 1977), have been the subject of recent intensive investigations mostly due to their protential impact to large segments of the general population.

Integrins are large heterodimeric type 1 membrane proteins expressed in all nucleated mammalian cells. Eighteen α-chains and eight β-chains can combine to form 24 different integrins. They are cell adhesion proteins, which bind to a large variety of cellular and extracellular ligands. Integrins are required for cell migration, hemostasis, translocation of cells out from the blood stream and further movement into tissues, but also for the immune response and tissue morphogenesis. Importantly, integrins are not usually active as such, but need activation to become adhesive. Integrins are activated by outside-in activation through integrin ligand binding, or by inside-out activation through intracellular signaling. An important question is how integrin activity is regulated, and this topic has recently drawn much attention. Changes in integrin affinity for ligand binding are due to allosteric structural alterations, but equally important are avidity changes due to integrin clustering in the plane of the plasma membrane. Recent studies have partially solved how integrin cell surface structures change during activation. The integrin cytoplasmic domains are relatively short, but by interacting with a variety of cytoplasmic proteins in a regulated manner, the integrins acquire a number of properties important not only for cell adhesion and movement, but also for cellular signaling. Recent work has shown that specific integrin phosphorylations play pivotal roles in the regulation of integrin activity. Our purpose in this review is to integrate the present knowledge to enable an understanding of how cell adhesion is dynamically regulated.

Genetic engineering has quite rightly an image of science fiction. The time when new species with any wanted combination of genetic properties can be ordered from an animal or plant breeding factory seems far away. The layman's view that the science fiction of today is the reality of tomorrow is certainly an insufficient argument to justify optimism. If this were so, we should by now be able to produce hybrids between members of the animal and plant kingdom as was foreseen by a nineteenth-century equivalent of Fred Hoyle (see Fig. I). Despite the scepsis expressed by the prominent scientist Si.r Macfarlane Burnet in his book Genes, Dreams and Realities (1971), recent advances in molecular genetics have raised new enthusiasm (and uneasiness) which make people speak of genetic engineering as something to aim at as an approach to correct inborn errors of metabolism. This will, however, not be our principal dish if we restrict ourselves to a vegetarian menu. We view genetic engineering of plants not only as a future method to improve species, but also as a fundamental approach to the study of gene expression, especially with respect to cell differentiation. If we consider the term literally, the definition of genetic engineering might be any intentional genetic manipulation to alter species or to make new ones. In this sense genetic engineering was practised long before Mendel presented his laws (1865). In the seventeenth century the bulb growers of the Low Countries produced new varieties of tulips for which prices of a thousand forms each were paid (at the 1636 price index!). We notice here already a strong impact of genetic engineering on society; the so-called crazy tulip trade caused a financial disaster on the Amsterdam stock-exchange comparable to the Wall Street crash in the 1930s.

Water, solutes and membranes have long been the subject of intensive research, and many excellent reviews have brought the major biological problems in focus, have discussed accomplishments, and have outlined unsolved problems. The most interesting results pertain to the role of membranes, whether ‘active’ or ‘passive’, which separate different solutions, in other words, membranes at a water–water inerface Liquid gas interfaces have receied less attention, and I therefore wish to review some biological problem which relate to such systems. I shall discuss a variety of phenomena which may have little in common, except that they all bear onthe transition of water between liquid and gas.

Glucose is one of the substrates that is controlled with the most efficient hormonal mechanisms in higher organisms. The presence of tissues such as the central nervous system which, under normal conditions, depend solely on glucose as substrate, and the sporadic type of food intake with periods of fasting of various lengths in the mammalians necessitate that the distribution of energy-rich substrates among various tissues be continuously adjusted by changes in the secretion of a number of hormones. The efficiency of this system is evidenced by the stability of the blood glucose level in man, in whom after a carbohydrate-rich meal more than 70% of the glucose that has been ingested will be retained in the liver during a single passage of portal blood, resulting in only small changes of the glucose concentration in peripheral blood. Likewise, periods of fasting up to24–36 h are followed by modest to minimal reductions of the blood glucose level, the liver now supplying the circulation with the hexose.

The outer layer of the skin, stratum corneum (SC) is an efficient transport barrier and it tolerates mechanical deformation. At physiological conditions, the majority of SC lipids are solid, while the presence of a small amount of fluid lipids is considered crucial for SC barrier and material properties. Here we use solid-state and diffusion nuclear magnetic resonance to characterize the composition and molecular dynamics of the fluid lipid fraction in SC model lipids, focusing on the role of the essential SC lipid CER EOS, which is a ceramide esterified omega-hydroxy sphingosine linoleate with very long chain. We show that both rigid and mobile structures are present within the same CER EOS molecule, and that the linoleate segments undergo fast isotropic reorientation while exhibiting extraordinarily slow self-diffusion. The characterization of this unusual self-assembly in SC lipids provides deepened insight into the molecular arrangement in the SC extracellular lipid matrix and the role of CER EOS linoleate in the healthy and diseased skin.

In general the characterization of a virus particle depends upon a number of known and independently acquired parameters, all of which require a great deal of time. As a direct method of measurement, the mass of viruses has been largely ignored in virus characterization studies. Where the molecular weight of viruses has been determined by sedimentation and diffusion, sedimentation equilibrium, light scattering, or the electron microscopy-counting procedure, the relationship between this property and mass has been arrived at by employing Avogadro's number. In the present report it will be demonstrated that the disadvantages found in these usual methods of determining molecular weight, such as experimentation, calculation of virus size, and purification of virus preparation, are not present in the quantitative electron microscopy method for determination of mass and molecular weight.

The findings of clinical and experimental studies conducted over the past 25 years provide extensive evidence that in both laboratory animals and man memory can be modified by treatments which affect the central nervous system. Patients with head injuries may suffer from retrograde amnesia, a loss of memory for experiences which occur just prior to the onset of the injury. Findings of laboratory studies using animal subjects indicate that retrograde amnesia can be produced by a wide variety of experimental treatments.