Antifreeze agents in the body fluid of winter active insects and spiders

Summary Insects and spiders which are active at subzero temperatures on snow in winter are found to be protected against internal freezing by antifreeze agents present in their body fluid. The body fluid has a melting point of about –1 °C, but the antifreeze agents prevent growth of ice crystals at temperatures down to –6 to –7 °C.Acknowledgment. We would like to thank Anne Lohrman for her kind help in collecting insects and spiders.     

Coupling of fructose-1,6-P2 to aminated agarose by Schiff base reduction. Affinity chromatography of yeast aldolase.

Fructose-1,6-P2 was immobilized by sodium borohydride reduction of the Schiff base formed with aminated agarose (AH-Sepharose 4B). The coupling occurs with high yield (25 mumoles immobilized fructose-1,6-P2 per ml packed gel) at neutral pH and room temperature. Schiff base reduction thus provides a convenient and mild coupling prodecure for sugar phosphates preserving their labile phospho ester bonds. As exemplified by a new isolation procedure for fructose-1,6-P2 aldolase from yeast, sugar phosphates insolubilized in this manner may be used for affinity chromatography of the corresponding enzymes, provided that contaminating unspecific phosphatases are removed in a preceding fractionation step.     

Isolation of an antifreeze peptide from the Antarctic sponge <Emphasis Type="Italic">Homaxinella balfourensis</Emphasis>

Polar plants and animals survive in subzero waters (-2 degrees C) and many of these marine organisms produce antifreeze proteins (AFPs) to better adapt themselves to these conditions. AFPs prevent the growth of ice crystals which disrupt cellular membranes and destroy cells by inhibiting crystallization of water within the organism. The hydrophilic extract of an Antarctic sponge Homaxinella balfourensis exhibited a non-colligative freezing point depression effect on the crystal morphology of water. The extract was purified by repeated reverse phase high-pressure liquid chromatography, then assayed and shown to contain several AFPs. The major peptide was isolated, analyzed using matrix-assisted laser desorption ionization mass spectrometry and the partial structure of the peptide identified through amino acid sequencing. AFPs have potential applications in agriculture, medicine and the food industry.     

Application of the ion-thinning methods to the high-resolution study of human dental enamel crystals]

Ion thinning technique applied to high resolution T.E.M. study of human dental enamel crystallites makes it possible to visualize a great number of dislocations. Dislocations proved not to be due to artifacts induced by the ultramicrotome diamond knife. They are related to the crystal apartite structure.     

La structure de certaines terminaisons nerveuses montre des variations cycliques

Summary All sensory endings of the trigeminal nerve in the dura mater of the rat are formed by a small swelling, from which continues themetaterminal apparatus, fine filaments at the limit of visibility. Periodically the fine granules at the extremity of the filaments swell, whereas the fibrils themselves disappear, leaving a small, round, argentophil mass. Later the filaments reappear, then redeposit their debris, which persists for a time.Thus the metaterminal apparatus manifests cyclic variations, which recall the transitory existence of collaterals observed duringin vivo orin vitro development of nerve fibres.     

An hypothesis for the process of selection at the subcellular level leading to the appearance of new clones]

According to the hypothesis we propose, the stimulation of one lymphocyte by an antigen induces the simultaneous expression of a great diversity of immunoglobulins of different specificities. Each molecular species is associated with the corresponding mRNA within a subcellular structure: the ergastoplasmic cisterna. It has been shown that in some responding lymphocytes at an early stage of the immune response a few such cisternae are loaded with antibodies while most of the cisternae are synthesizing non specific immunoglobulins. The main point of our proposal is that the selective action of antigen bears on these cisternae and that the mRNA corresponding to the immunoglobulins fitting best to the antigen is transcribed to DNA which is then inserted into the genome. This cell and its progeny become thereafter a monospecific clone submitted to regulation as an element of the network.     

Regulation of cross-bridge detachment by Ca ions at high ionic strength in molluscan catch muscle

Changes in the profile of equatorial intensities of X-ray diffraction from an intact, anterior byssal retractor muscle (ABRM) of Mytilus were examined at rest, during contracture brought about by acetylcholine (ACh) and a subsequent rigor-like contraction caused by raising the tonicity of the external solution, and after returning the tonicity to normal. The results suggest that the cross-bridges formed between thick and thin actin filaments during the ACh-contracture were maintained in the hypertonic solution and broken on decreasing the tonicity before the recovery of spacing of the actin filament lattice. A similar rigor-like contraction was induced in glycerinated ABRM by increasing salt concentration during active contraction. The rigor-like force declined rapidly when Ca++ concentration decreased. The results suggest that the detachment of the cross-bridge from the actin filament is regulated by Ca++ at high ionic strength in the ABRM.     

Fold-Hopf bifurcation in a simplified four-neuron BAM (bidirectional associative memory) neural network with two delays

The bidirectional associative memory (BAM) neural network with four neurons and two delays is considered in the present paper. A linear stability analysis for the trivial equilibrium is firstly employed to provide a possible critical point at which a zero and a pair of pure imaginary eigenvalues occur in the corresponding characteristic equation. A fold-Hopf bifurcation is proved to happen at this critical point by the nonlinear analysis. The coupling strength and the delay are considered as bifurcation par...     

Regulation of cross-bridge detachment by Ca ions at high ionic strength in molluscan catch muscle

Summary Changes in the profile of equatorial intensities of X-ray diffraction from an intact, anterior byssal retractor muscle (ABRM) ofMytilus were examined at rest, during contracture brought about by acetylcholine (ACh) and a subsequent rigor-like contraction caused by raising the tonicity of the external solution, and after returning the tonicity to normal. The results suggest that the cross-bridges formed between thick and thin actin filaments during the ACh-contracture were maintained in the hypertonic solution and broken on decreasing the tonicity before the recovery of spacing of the actin filament lattice. A similar rigor-like contraction was induced in glycerinated ABRM by increasing salt concentration during active contraction. The rigor-like force declined rapidly when Ca⁺⁺ concentration decreased. The results suggest that the detachment of the cross-bridge from the actin filament is regulated by Ca⁺⁺ at high ionic strength in the ABRM.     

Chemical properties of alcohols and their protein binding sites

Alcohols affect a wide array of biological processes including protein folding, neurotransmission and immune responses. It is becoming clear that many of these effects are mediated by direct binding to proteins such as neurotransmitter receptors and signaling molecules. This review summarizes the unique chemical properties of alcohols which contribute to their biological effects. It is concluded that alcohols act mainly as hydrogen bond donors whose binding to the polypeptide chain is stabilized by hydrophobic interactions. The electronegativity of the O atom may also play a role in stabilizing contacts with the protein. Properties of alcohol binding sites have been derived from X-ray crystal structures of alcohol-protein complexes and from mutagenesis studies of ion channels and enzymes that bind alcohols. Common amino acid sequences and structural features are shared among the protein segments that are involved in alcohol binding. The alcohol binding site is thought to consist of a hydrogen bond acceptor in a turn or loop region that is often situated at the N-terminal end of an alpha-helix. The methylene chain of the alcohol molecule appears to be accommodated by a hydrophobic groove formed by two or more structural elements, frequently a turn and an alpha-helix. Binding at these sites may alter the local protein structure or displace bound solvent molecules and perturb the function of key proteins.     

Variation in the number and size of myelinated nerve fibers regenerated after repeated localized freezing of the sciatic nerve of rats]

The number and size of myelinated fibres have been determined in the nerve to medial head of the gastrocnemius muscle of Rats in which the sciatic nerve had been frozen locally 1 to 5 times at monthly intervals. The contralateral nerve was used as a control. When the measurements were made one month after the last freezing, the number of fibres increased progressively until the 3rd freezing, reaching about 220% of the normal value. No higher values were observed after 4 or 5 freezings. The mean diameter of fibres decreased with the number of operations. When the measurements were made 3, 6, 12 or 18 months after the 3rd freezing, the number of fibres decreased by about 30% between the 1st and the 3rd month and then stabilized. The mean diameter of fibres increased progressively. However, at the 18th month, the size of the myelinated fibres had not reached the normal value.     

Cryo-bioorganic chemistry: molecular interactions at low temperature

Freezing of aqueous or organic solutions plays a pivotal role in enhancement of rate and/or yield of biomolecular reactions. The smooth conditions of the frozen state at low temperature can also suppress racemization and side-product formation of the reactions. Molecular interactions in liquid undercooled solutions, on the other hand, offer the possibility to study enzyme activity mechanisms in vitro and a chance for survival of organisms in vivo. This review illustrates the differences between frozen and liquid conditions on several small and large biomolecules, together with the synthetic use of freezing. In relation to the freezing effect on enzyme activity, a peculiar phenomenon is discussed: 'cryo-oscillations' are temporal motions of trypsin activity in frozen solution in the presence of Mn2+ ion. The molecular basis of cold adaptation is also discussed, which points to mechanisms evolved by organisms living at subzero temperatures. The factors involved in the freezing effect are shown; i.e. the role of freeze-concentration and frozen solvent surface is demonstrated and elucidated using several examples.     

聚合物超薄膜结晶相关的形貌形成及链行为

聚合物超薄膜结晶是高分子物理领域的一个新研究课题,近来已引起人们的关注,其为人们在真实时空下研究聚合物结晶及相关链行为提供了可能。聚合物超薄膜结晶与膜厚(D)有很强的相关性,尤其是D<20nm的薄膜在结晶形貌和结晶动力学方面与本体结晶差别很大。已有的结果主要集中在结晶形貌、晶体尺寸、晶体生长速度和习性、晶体的熔融和结晶度等方面,涉及温度、膜厚、基片性质和膜的组成和结构对结晶的影响;然而,有些实验结果和解释彼此不完全一致,甚至有时相互矛盾。本文综述和讨论了近年来在超薄膜结晶方面的研究,重点在于结晶形貌的形成和相关的聚合物链行为。     

Coupling of fructose-1,6-P2 to aminated agarose by Schiff base reduction. Affinity chromatography of yeast aldolase

Summary Fructose-1,6-P₂ was immobilized by sodium borohydride reduction of the Schiff base formed with aminated agarose (AH-Sepharose 4B^®). The coupling occurs with high yield (25 moles immobilized fructose-1,6-P₂ per ml packed gel) at neutral pH and room temperature. Schiff base reduction thus provides a convenient and mild coupling procedure for sugar phosphates preserving their labile phospho ester bonds. As exemplified by a new isolation procedure for fructose-1,6-P₂ aldolase from yeast, sugar phosphates insolubilized in this manner may be used for affinity chromatography of the corresponding enzymes, provided that contaminating unspecific phosphatases are removed in a preceding fractionation step.This work was supported by the Swiss National Science Foundation, grant No. 3.620-0.75. A preliminary account has been presented at the 10th Int. Congress of Biochemistry, Hamburg, 1976, Abstracts. p. 194.Acknowledgment. The excellent technical assistance of Alice Gasser is gratefully acknowledged     

Some observations on bipolar filaments formed by non-muscle myosins.

Adrenal medullary and retinal myosins formed bipolar filaments in vitro. These filaments showed features suggesting flexibility in the rod region of the myosin molecules composing such filaments; in certain cases the myosin heads spread away from the filament backbone, in others the backbone itself was twisted. In addition the bare central backbone showed transverse striations.     

Modifications in the type distribution of muscle fibers induced by repeated local freezings the rat sciatic nerve]

In the Rat, after a localized freezing of the sciatic nerve inducing a complete denervation of the medial head of the gastrocnemius muscle, the reinnervation took place within 16-18 days under our experimental conditions. After only one freezing, a limited and stable "type-grouping" of muscle fibres is observed from 30 to 360 days. After 2 to 5 freezing repeated every three weeks, the muscular changes observed one month after the last freezing are much more pronounced than when only one freezing is performed. These changes consist of a progressive increase in the number of type I and type II C fibres. This transformation is not stable: 3 months after the 3rd freezing, the muscle pattern does not differ from that which is noted after a single freezing.     

The muscle ultrastructure: a structural perspective of the sarcomere

Muscle ultrastructure is characterised by a complex arrangement of many protein-protein interactions. The sarcomere is the basic repeating unit of muscle, formed by two transverse filament systems: the thick and thin filaments. While actin and myosin are the main contractile elements of the sarcomere, other proteins act as scaffolds, control ultrastructure composition, regulate muscle contraction, and transmit tension between sarcomeres and hence to the whole myofibril. Elucidation of the structures of muscle proteins by X-ray crystallography and nuclear magnetic resonance spectroscopy has been essential in understanding muscle contraction, enabling us to relate biological to structural information. These structures reveal how components of the muscle interact, how different factors influence conformational changes within these proteins, and how mutant muscle proteins may interfere with the regulatory fine-tuning of the contractile machinery, hence leading to disease in some cases. Here, structures solved within the sarcomere have been reviewed in order to put the numerous components into context.Received 28 June 2004; received after revision 25 July 2004; accepted 28 July 2004     

Tropomodulins and tropomodulin/tropomyosin interactions

The tropomodulins are a family of proteins that cap the slow-growing (pointed) end of actin filaments and require tropomyosin for optimal function. Tropomodulin is an elongated molecule with a molecular mass of about 40 kDa. The C-terminal half of tropomodulin contains one compact cooperatively melting domain, whereas the N-terminal half has no cooperatively melting structure. The N-terminal half of tropomodulin contains two tropomysin-binding sites and a tropomyosin-dependent actin-binding site, the tropomyosin-independent actin-binding site being located at the C terminus. One tropomodulin molecule binds two tropomyosin molecules, and thus one molecule of tropomodulin is necessary and sufficient for capping at the pointed end. Tropomyosin/tropomodulin interactions are isoform specific. Differences in tropomyosin affinity for the two binding sites in tropomodulin may regulate its correct positioning at the pointed end as well as effectiveness of capping the actin filament. Received 30 July 2007; received after revision 2 October 2007; accepted 10 October 2007     

三圈冻结法在淤泥质土深基坑工程的应用

以往的冻结法在淤泥质土及特殊工程土体中,由于所需的冻结温度低、冻结扩展速度慢、形成冻结壁的强度低等原因,在施工时容易出现片帮、底鼓、突水、涌砂等事故.轻则影响工程进度,重则造成严重的经济损失.针对此类问题,提出了用三圈冻结法优化淤泥质土深基坑开挖的方案,并应用大型数值模拟软件ADINA进行模拟分析.对普通冻结法、连续桩及三圈冻结法等围护结构的位移进行了对比,得出三圈冻结法围护结构不仅最大位移小,而且位移随深度变化平稳.     

Mining electron density for functionally relevant protein polysterism in crystal structures

This review focuses on conceptual and methodological advances in our understanding and characterization of the conformational heterogeneity of proteins. Focusing on X-ray crystallography, we describe how polysterism, the interconversion of pre-existing conformational substates, has traditionally been analyzed by comparing independent crystal structures or multiple chains within a single crystal asymmetric unit. In contrast, recent studies have focused on mining electron density maps to reveal previously ‘hidden’ minor conformational substates. Functional tests of the importance of minor states suggest that evolutionary selection shapes the entire conformational landscape, including uniquely configured conformational substates, the relative distribution of these substates, and the speed at which the protein can interconvert between them. An increased focus on polysterism may shape the way protein structure and function is studied in the coming years.