“Substrate Inhibition” Is One of the Aspects of Substrate Specificity in Vertebrate and Invertebrate Cholinesterases

An analytical review is performed of the literature data on the hydrolysis rate, affinity of substrate to active center, and constants of the substrate inhibition (K _ss) at hydrolysis of the choline (acetyl-, propyonyl-, butyrylcholine, acetyl--methylcholine) and/or of corresponding thiocholine substrates by 59 cholinesterases from 49 different animals (chordate, insects, molluscs, nematodes). The characteristic peculiarities of enzymes from different groups of animals are revealed. The absence of regular changes of parameters of the cholinesterase substrate specificity in the course of evolutionary development is shown.     

Explosive Expansion of βγ-Crystallin Genes in the Ancestral Vertebrate

In jawed vertebrates, βγ-crystallins are restricted to the eye lens and thus excellent markers of lens evolution. These βγ-crystallins are four Greek key motifs/two domain proteins, whereas the urochordate βγ-crystallin has a single domain. To trace the origin of the vertebrate βγ-crystallin genes, we searched for homologues in the genomes of a jawless vertebrate (lamprey) and of a cephalochordate (lancelet). The lamprey genome contains orthologs of the gnathostome βB1-, βA2- and γN-crystallin genes and a single domain γN-crystallin-like gene. It contains at least two γ-crystallin genes, but lacks the gnathostome γS-crystallin gene. The genome also encodes a non-lenticular protein containing βγ-crystallin motifs, AIM1, also found in gnathostomes but not detectable in the uro- or cephalochordate genome. The four cephalochordate βγ-crystallin genes found encode two-domain proteins. Unlike the vertebrate βγ-crystallins but like the urochordate βγ-crystallin, three of the predicted proteins contain calcium-binding sites. In the cephalochordate βγ-crystallin genes, the introns are located within motif-encoding region, while in the urochordate and in the vertebrate βγ-crystallin genes the introns are between motif- and/or domain encoding regions. Coincident with the evolution of the vertebrate lens an ancestral urochordate type βγ-crystallin gene rapidly expanded and diverged in the ancestral vertebrate before the cyclostomes/gnathostomes split. The β- and γN-crystallin genes were maintained in subsequent evolution, and, given the selection pressure imposed by accurate vision, must be essential for lens function. The γ-crystallin genes show lineage specific expansion and contraction, presumably in adaptation to the demands on vision resulting from (changes in) lifestyle.     

The myth of “minima” and “maxima”, the species of Physalis in the Indian Subcontinent

The status of the names, Physalis minima L. and P. maxima Mill. (Solanaceae), and their alleged presence on the Indian subcontinent are discussed. The issues of nativity and identity of Linnaean Physalis minima are long-debated while the use of the name P. maxima Mill. and its report from India are recent. The available evidence indicates that the name "P. minima L." is misapplied to two different elements, viz., P. angulata L. and P. lagascae Roem. & Schult. The name Physalis minima L. may be rejected as nomen confusum, for which the paper provides the primary information. As on today, it is submerged under the synonymy of P. angulata L. The correct name for the widely known P. minima is P. lagascae. The name "P. maxima Mill." applied to the escape and naturalized weed in the Indian subcontinent and elsewhere is to be substituted by P. pruinosa L., a name misapplied to P. grisea (Waterf.) M. Martínez.     

The myth of “minima” and “maxima”, the species of Physalis in the Indian Subcontinent

The status of the names, Physalis minima L. and P. maxima Mill. (Solanaceae), and their alleged presence on the Indian subcontinent are discussed. The issues of nativity and identity of Linnaean Physalis minima are long-debated while the use of the name P. maxima Mill. and its report from India are recent. The available evidence indicates that the name “P. minima L.” is misapplied to two different elements, viz., P. angulata L. and P. lagascae Roem. & Schult. The name Physalis minima L. may be rejected as nomen confusum, for which the paper provides the primary information. As on today, it is submerged under the synonymy of P. angulata L. The correct name for the widely known P. minima is P. lagascae. The name “P. maxima Mill.” applied to the escape and naturalized weed in the Indian subcontinent and elsewhere is to be substituted by P. pruinosa L., a name misapplied to P. grisea (Waterf.) M. Martínez.     

Cell Adhesion, the Backbone of the Synapse: ��Vertebrate�� and ��Invertebrate�� Perspectives

Synapses are asymmetric intercellular junctions that mediate neuronal communication. The number, type, and connectivity patterns of synapses determine the formation, maintenance, and function of neural circuitries. The complexity and specificity of synaptogenesis relies upon modulation of adhesive properties, which regulate contact initiation, synapse formation, maturation, and functional plasticity. Disruption of adhesion may result in structural and functional imbalance that may lead to neurodevelopmental diseases, such as autism, or neurodegeneration, such as Alzheimer''s disease. Therefore, understanding the roles of different adhesion protein families in synapse formation is crucial for unraveling the biology of neuronal circuit formation, as well as the pathogenesis of some brain disorders. The present review summarizes some of the knowledge that has been acquired in vertebrate and invertebrate genetic model organisms.Synapses are asymmetric, intercellular junctions that are the basic structural units of neuronal transmission. The correct development of synaptic specializations and the establishment of appropriate connectivity patterns are crucial for the assembly of functional neuronal circuits. Improper synapse formation and function may cause neurodevelopmental disorders, such as mental retardation (MsR) and autism spectrum disorders (ASD) (McAllister 2007; Sudhof 2008), and likely play a role in neurodegenerative disorders, such as Alzheimer''s disease (AD) (Haass and Selkoe 2007).At chemical synapses (reviewed in Sudhof 2004; Zhai and Bellen 2004; Waites et al. 2005; McAllister 2007; Jin and Garner 2008), the presynaptic compartment contains synaptic vesicles (SV), organized in functionally distinct subcellular pools. A subset of SVs docks to the presynaptic membrane around protein-dense release sites, named active zones (AZ). Upon the arrival of an action potential at the terminal, the docked and “primed” SVs fuse with the plasma membrane and release neurotransmitter molecules into the synaptic cleft. Depending on the type of synapse (i.e., excitatory vs. inhibitory synapses), neurotransmitters ultimately activate an appropriate set of postsynaptic receptors that are accurately apposed to the AZ.Synapse formation occurs in several steps (Fig. 1) (reviewed in Eaton and Davis 2003; Goda and Davis 2003; Waites et al. 2005; Garner et al. 2006; Gerrow and El-Husseini 2006; McAllister 2007). Spatiotemporal signals guide axons through heterogeneous cellular environments to contact appropriate postsynaptic targets. At their destination, axonal growth cones initiate synaptogenesis through adhesive interactions with target cells. In the mammalian central nervous system (CNS), immature postsynaptic dendritic spines initially protrude as thin, actin-rich filopodia on the surface of dendrites. Similarly, at the Drosophila neuromuscular junction (NMJ), myopodia develop from the muscles (Ritzenthaler et al. 2000). The stabilization of intercellular contacts and their elaboration into mature, functional synapses involves cytoskeletal arrangements and recruitment of pre- and postsynaptic components to contact sites in spines and boutons. Conversely, retraction of contacts results in synaptic elimination. Both stabilization and retraction sculpt a functional neuronal circuitry.Open in a separate windowFigure 1.(A–C) Different stages of synapse formation. (A) Target selection, (B) Synapse assembly, (C) Synapse maturation and stabilization. (D–F) The role of cell adhesion molecules in synapse formation is exemplified by the paradigm of N-cadherin and catenins in regulation of the morphology and strength of dendritic spine heads. (D) At an early stage the dendritic spines are elongated from motile structures “seeking” their synaptic partners. (E) The contacts between the presynaptic and postsynaptic compartments are stabilized by recruitment of additional cell adhesion molecules. Adhesional interactions activate downstream pathways that remodel the cytoskeleton and organize pre- and postsynaptic apparatuses. (F) Cell adhesion complexes, stabilized by increased synaptic activity, promote the expansion of the dendritic spine head and the maturation/ stabilization of the synapse. Retraction and expansion is dependent on synaptic plasticity.In addition to the plastic nature of synapse formation, the vast heterogeneity of synapses (in terms of target selection, morphology, and type of neurotransmitter released) greatly enhances the complexity of synaptogenesis (reviewed in Craig and Boudin 2001; Craig et al. 2006; Gerrow and El-Husseini 2006). The complexity and specificity of synaptogenesis relies upon the modulation of adhesion between the pre- and postsynaptic components (reviewed in Craig et al. 2006; Gerrow and El-Husseini 2006; Piechotta et al. 2006; Dalva et al. 2007; Shapiro et al. 2007; Yamada and Nelson 2007; Gottmann 2008). Cell adhesive interactions enable cell–cell recognition via extracellular domains and also mediate intracellular signaling cascades that affect synapse morphology and organize scaffolding complexes. Thus, cell adhesion molecules (CAMs) coordinate multiple synaptogenic steps.However, in vitro and in vivo studies of vertebrate CAMs are often at odds with each other. Indeed, there are no examples of mutants for synaptic CAMs that exhibit prominent defects in synapse formation. This apparent “resilience” of synapses is probably caused by functional redundancy or compensatory effects among different CAMs (Piechotta et al. 2006). Hence, studies using simpler organisms less riddled by redundancy, such as Caenorhabditis elegans and Drosophila, have aided in our understanding of the role that these molecules play in organizing synapses.In this survey, we discuss the roles of the best characterized CAM families of proteins involved in synaptogenesis. Our focus is to highlight the complex principles that govern the molecular basis of synapse formation and function from a comparative perspective. We will present results from cell culture studies as well as in vivo analyses in vertebrate systems and refer to invertebrate studies, mainly performed in Drosophila and C. elegans, when they have provided important insights into the role of particular CAM protein families. However, we do not discuss secreted factors, for which we refer the reader to numerous excellent reviews (as for example Washbourne et al. 2004; Salinas 2005; Piechotta et al. 2006; Shapiro et al. 2006; Dalva 2007; Yamada and Nelson 2007; Biederer and Stagi 2008; Salinas and Zou 2008).     

The “Lillie Transition”: models of the onset of saltatory conduction in myelinating axons

Almost 90 years ago, Lillie reported that rapid saltatory conduction arose in an iron wire model of nerve impulse propagation when he covered the wire with insulating sections of glass tubing equivalent to myelinated internodes. This led to his suggestion of a similar mechanism explaining rapid conduction in myelinated nerve. In both their evolution and their development, myelinating axons must make a similar transition between continuous and saltatory conduction. Achieving a smooth transition is a potential challenge that we examined in computer models simulating a segmented insulating sheath surrounding an axon having Hodgkin-Huxley squid parameters. With a wide gap under the sheath, conduction was continuous. As the gap was reduced, conduction initially slowed, owing to the increased extra-axonal resistance, then increased (the “rise”) up to several times that of the unmyelinated fiber, as saltatory conduction set in. The conduction velocity slowdown was little affected by the number of myelin layers or modest changes in the size of the “node,” but strongly affected by the size of the “internode” and axon diameter. The steepness of the rise of rapid conduction was greatly affected by the number of myelin layers and axon diameter, variably affected by internode length and little affected by node length. The transition to saltatory conduction occurred at surprisingly wide gaps and the improvement in conduction speed persisted to surprisingly small gaps. The study demonstrates that the specialized paranodal seals between myelin and axon, and indeed even the clustering of sodium channels at the nodes, are not necessary for saltatory conduction.     

Comparative Study of the Absorption,Plasma Levels,and Urinary Excretion of the “New” and the “Old” Lanoxin

A comparative study was performed of the absorption, the plasma level at equilibrium, and the urinary excretion of digoxin using two types of Lanoxin tablets, those produced before and after the 1972 alteration of the tablet manufacture.After a single dose the absorption rate of the new tablets was about twice as great as the old, both in young subjects and in the elderly patients. There were no significant differences in the plasma levels of digoxin for the two tablets 15 hours after the last administration in patients on an equal maintenance dose. The urinary excretion of digoxin increased about 40% when the “old” Lanoxin was replaced by the “new.” In elderly patients a daily dose of 0·125 mg twice daily of the new tablets should be sufficient to reach the therapeutic range. Young people need a higher dosage. If the kidney function is reduced by as much as 50% the dose should be reduced.     

Clarification of the status of Rhipicephalus tricuspis Dönitz, 1906 and Rhipicephalus lunulatus Neumann, 1907 (Ixodoidea,Ixodidae)

The systematic status of Rhipicephalus species whose males have tricuspid adanal plates has been confused for many years. Some authors have regarded Rhipicephalus tricuspis Dönitz, 1906 as the only valid entity with this morphological character and synonymized both Rhipicephalus lunulatus Neumann, 1907 and Rhipicephalus glyphis Dönitz, 1910 with it. Others, however, have always maintained that R. tricuspis and R. lunulatus (syn. R. glyphis) are separate species. Detailed comparative studies, including scanning electron microscopy, of laboratory-reared series as well as numerous field collections of these ticks have now confirmed that the latter view is correct. In addition, a third species, designated here as a Rhipicephalus sp. near tricuspis, has been identified as a member of this group.All stages of R. tricuspis and R. lunulatus are herein redescribed and illustrated by means of scanning electron micrographs. Their life cycles in the laboratory, hosts, distribution and disease relationships are discussed and their differentiation is described with the aid of line drawings. Rhipicephalus tricuspis has been recorded primarily in southern Africa, but also in Zambia and western Zaire, in various types of dry woodland. Its adults are most commonly parasitic on relatively small mammals such as hares, spring hares, jackals and small antelopes. R. lunulatus is much more widespread in the Afrotropical region, most commonly in different types of woodland but also in a variety of other habitats. Its adults parasitize a very wide range of hosts including domestic animals (especially cattle and dogs), the African buffalo, many different antelopes (especially the larger species) and wild pigs. The Rhipicephalus sp. near tricuspis occurs in eastern and parts of central Africa, where its distribution often overlaps with that of R. lunulatus.     

Seasonality and Paleoecology of the Late Cretaceous Multi-Taxa Vertebrate Assemblage of “Lo Hueco” (Central Eastern Spain)

Isotopic studies of multi-taxa terrestrial vertebrate assemblages allow determination of paleoclimatic and paleoecological aspects on account of the different information supplied by each taxon. The late Campanian-early Maastrichtian “Lo Hueco” Fossil-Lagerstätte (central eastern Spain), located at a subtropical paleolatitude of ~31°N, constitutes an ideal setting to carry out this task due to its abundant and diverse vertebrate assemblage. Local δ¹⁸O_PO4 values estimated from δ¹⁸O_PO4 values of theropods, sauropods, crocodyliforms, and turtles are close to δ¹⁸O_H2O values observed at modern subtropical latitudes. Theropod δ¹⁸O_H2O values are lower than those shown by crocodyliforms and turtles, indicating that terrestrial endothermic taxa record δ¹⁸O_H2O values throughout the year, whereas semiaquatic ectothermic taxa δ¹⁸O_H2O values represent local meteoric waters over a shorter time period when conditions are favorable for bioapatite synthesis (warm season). Temperatures calculated by combining theropod, crocodyliform, and turtle δ¹⁸O_H2O values and gar δ¹⁸O_PO4 have enabled us to estimate seasonal variability as the difference between mean annual temperature (MAT, yielded by theropods) and temperature of the warmest months (TWMs, provided by crocodyliforms and turtles). ΔTWMs-MAT value does not point to a significantly different seasonal thermal variability when compared to modern coastal subtropical meteorological stations and Late Cretaceous rudists from eastern Tethys. Bioapatite and bulk organic matter δ¹³C values point to a C₃ environment in the “Lo Hueco” area. The estimated fractionation between sauropod enamel and diet is ~15‰. While waiting for paleoecological information yielded by the ongoing morphological study of the “Lo Hueco” crocodyliforms, δ¹³C and δ¹⁸O_CO3 results point to incorporation of food items with brackish influence, but preferential ingestion of freshwater. “Lo Hueco” turtles showed the lowest δ¹³C and δ¹⁸O_CO3 values of the vertebrate assemblage, likely indicating a diet based on a mixture of aquatic and terrestrial C₃ vegetation and/or invertebrates and ingestion of freshwater.     

Nation-building,masculinity and entrepreneurship: memories of the “liberation of Kosovo” in Switzerland

Since the end of the Kosovo War in 1999 and the “liberation” of the territory from Serbian forces, narratives about the “freedom struggle” have been crafted and defended both in Kosovo and abroad. The transmission of these memories forms part of a broader effort to create a “national” history of Kosovo and constitute an “Albanian imagined community”. This article scrutinizes the memories of the “liberation” produced by Albanian-speaking migrants who were active on behalf of their homeland in Switzerland. It explores the construction of masculinities within narratives collected via oral history interviews. In line with the literature on “nation” and gender in Kosovo, this research acknowledges the presence of two main forms of masculinity: the “heroic fighter” and the “pacifist”. However, it also demonstrates the crystallization of the “entrepreneur”, an alternative type who integrates the transnational “neoliberal” discourses and proposes a more positive image of “Albanian men” in Switzerland.     

“Facilitation” and “Reversal of Response” of Neurones in the Cerebral Cortex

IN the course of experiments in which movements were elicited by direct electrical stimulation of the cerebral cortex, Graham-Brown^1–4 and Sherrington⁵ found that, after each train of stimuli, there were prolonged changes in the excitability of the cortex. They showed that the motor responses of forelimb muscles in primates, even though elicited by identical cortical stimuli, could be extremely variable. Two of the phenomena they described in detail (see also Lilly⁶) were “facilitation” (the increased motor response resulting from a second train of stimuli when it was applied within 20 s after the first train) and “reversal of response” (when repeated, identical stimulation of the same cortical point elicited flexion instead of extension, or vice versa).     

“Tight” junctions in the sheath of normal and regenerating motor nerves of the crayfish,Orconectes virilis

Summary Tight or occluding intercellular junctions occur between adjacent glial processes in normal and regenerating crayfish motor nerve sheaths. Although infrequent, these junctions possess the ridge and groove configuration characteristic of freeze-cleaved occluding junctions. When present, nerve sheath tight junctions consist of a single, or at most a few, parallel intramembrane ridges situated on the EF membrane face of the glial plasma membrane. Consequently, such contacts are rarely recognized in thin sections of plasticembedded nerve sheaths. Crayfish nerve sheath tight junctions are of the fascia occludens type and, therefore, do not impede solute flow across the nerve sheath. Fasciae occludentes of regenerating nerve sheaths occur in close proximity to discoid plaque-like aggregates of particles assumed to represent maculae adhaerentes. This relationship, which was not observed in normal nerve sheaths, suggests a functional association between the two types of junctions, perhaps developmental transformation of one junction type into the other. Although ridges and grooves of tight junctions occur next to crossfractured trans-glial channels, no functional significance is proposed for this relationship. This study is the first report of tight intercellular junctions in crustacean glial nerve sheaths.Supported by the National Research Council of Canada     

Mutagenicity of bi-, tri- and tetra-cyclic aromatic hydrocarbons in the “taped-plate assay” and in the conventional Salmonella mutagenicity assay

Aromatic hydrocarbons in the range of 1–4 nuclear rings were examined for mutagenicity in the so-called “taped-plate assay”. This modification of the Ames assay is particularly equipped for the detection of volatile mutagens. Of the many compounds tested only phenanthrene, pyrene, benzo[c]phenanthrene and benzoacenaphthylene were positive in this assay. The present data underline the exceptional behaviour of fluoranthene by being a rather potent bacterial mutagen with a volatile nature (as found in a previous study).     

Crystallographic studies of the binding of ligands to the dicarboxylate site of Complex II,and the identity of the ligand in the “oxaloacetate-inhibited” state

Mitochondrial Complex II (succinate:ubiquinone oxidoreductase) is purified in a partially inactivated state, which can be activated by removal of tightly bound oxaloacetate (E.B. Kearney, et al., Biochem. Biophys. Res. Commun. 49 1115–1121). We crystallized Complex II in the presence of oxaloacetate or with the endogenous inhibitor bound. The structure showed a ligand essentially identical to the “malate-like intermediate” found in Shewanella Flavocytochrome c crystallized with fumarate (P. Taylor, et al., Nat. Struct. Biol. 6 1108–1112) Crystallization of Complex II in the presence of excess fumarate also gave the malate-like intermediate or a mixture of that and fumarate at the active site. In order to more conveniently monitor the occupation state of the dicarboxylate site, we are developing a library of UV/Vis spectral effects induced by binding different ligands to the site. Treatment with fumarate results in rapid development of the fumarate difference spectrum and then a very slow conversion into a species spectrally similar to the OAA-liganded complex. Complex II is known to be capable of oxidizing malate to the enol form of oxaloacetate (Y.O. Belikova, et al., Biochim. Biophys. Acta 936 1–9). The observations above suggest it may also be capable of interconverting fumarate and malate. It may be useful for understanding the mechanism and regulation of the enzyme to identify the malate-like intermediate and its pathway of formation from oxaloacetate or fumarate.