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Most migrating cells extrude their front by the force of actin polymerization. Polymerization requires an initial nucleation step, which is mediated by factors establishing either parallel filaments in the case of filopodia or branched filaments that form the branched lamellipodial network. Branches are considered essential for regular cell motility and are initiated by the Arp2/3 complex, which in turn is activated by nucleation-promoting factors of the WASP and WAVE families. Here we employed rapid amoeboid crawling leukocytes and found that deletion of the WAVE complex eliminated actin branching and thus lamellipodia formation. The cells were left with parallel filaments at the leading edge, which translated, depending on the differentiation status of the cell, into a unipolar pointed cell shape or cells with multiple filopodia. Remarkably, unipolar cells migrated with increased speed and enormous directional persistence, while they were unable to turn towards chemotactic gradients. Cells with multiple filopodia retained chemotactic activity but their migration was progressively impaired with increasing geometrical complexity of the extracellular environment. These findings establish that diversified leading edge protrusions serve as explorative structures while they slow down actual locomotion.

In this paper, a method is presented that may be used for the analysis of axially loaded piles in isotropic or crossanisotropic soils. The method employs the finitelayer theory for the analysis of the soil, and because the method requires very little computer storage, it is particularly suited to microcomputers. The performance and versatility of the method is studied by comparing the finitelayer solutions with those obtained from other continuum approaches. For all of the cases studied which involved isotropic, layered, or nonhomogencous soils, the finitelayer results are found to be in very close agreement with the results of the other methods. The method is shown to be capable of producing accurate results, and some original solutions are obtained to the problem of a pile bearing onto a thin layer of soil that is cither softer or stiffcr than the rest of the soil. The effect of the crossanisotropic soil model used in the pile analysis is demonstrated with parametric solutions and also with a field case study of a pile embedded in overconsolidated London clay.

Acute suppression of Arp2/3 complex activity in lamellipodia demonstrates its essential role in actin network treadmilling and filament organization and geometry. Arp2/3 complex activity also defines the recruitment of crucial independent factors, including capping protein and cofilin, and is essential for lamellipodia-based keratocyte migration.

Dynamic actin rearrangements are initiated and maintained by actin filament nucleators, including the Arp2/3-complex. This protein assembly is activated in vitro by distinct nucleation-promoting factors such as Wiskott-Aldrich syndrome protein/Scar family proteins or cortactin, but the relative in vivo functions of each of them remain controversial. Here, we report the conditional genetic disruption of murine cortactin, implicated previously in dynamic actin reorganizations driving lamellipodium protrusion and endocytosis. Unexpectedly, cortactin-deficient cells showed little changes in overall cell morphology and growth. Ultrastructural analyses and live-cell imaging studies revealed unimpaired lamellipodial architecture, Rac-induced protrusion, and actin network turnover, although actin assembly rates in the lamellipodium were modestly increased. In contrast, platelet-derived growth factor-induced actin reorganization and Rac activation were impaired in cortactin null cells. In addition, cortactin deficiency caused reduction of Cdc42 activity and defects in random and directed cell migration. Reduced migration of cortactin null cells could be restored, at least in part, by active Rac and Cdc42 variants. Finally, cortactin removal did not affect the efficiency of receptor-mediated endocytosis. Together, we conclude that cortactin is fully dispensable for Arp2/3-complex activation during lamellipodia protrusion or clathrin pit endocytosis. Furthermore, we propose that cortactin promotes cell migration indirectly, through contributing to activation of selected Rho-GTPases.

Dynamic actin rearrangements are initiated and maintained by actin filament nucleators, including the Arp2/3-complex. This protein assembly is activated in vitro by distinct nucleation-promoting factors such as Wiskott-Aldrich syndrome protein/Scar family proteins or cortactin, but the relative in vivo functions of each of them remain controversial. Here, we report the conditional genetic disruption of murine cortactin, implicated previously in dynamic actin reorganizations driving lamellipodium protrusion and endocytosis. Unexpectedly, cortactin-deficient cells showed little changes in overall cell morphology and growth. Ultrastructural analyses and live-cell imaging studies revealed unimpaired lamellipodial architecture, Rac-induced protrusion, and actin network turnover, although actin assembly rates in the lamellipodium were modestly increased. In contrast, platelet-derived growth factor-induced actin reorganization and Rac activation were impaired in cortactin null cells. In addition, cortactin deficiency caused reduction of Cdc42 activity and defects in random and directed cell migration. Reduced migration of cortactin null cells could be restored, at least in part, by active Rac and Cdc42 variants. Finally, cortactin removal did not affect the efficiency of receptor-mediated endocytosis. Together, we conclude that cortactin is fully dispensable for Arp2/3-complex activation during lamellipodia protrusion or clathrin pit endocytosis. Furthermore, we propose that cortactin promotes cell migration indirectly, through contributing to activation of selected Rho-GTPases.

The pushing structures of cells include laminar sheets, termed lamellipodia, made up of a meshwork of actin filaments that grow at the front and depolymerise at the rear, in a treadmilling mode. We here develop a mathematical model to describe the turnover and the mechanical properties of this network.Our basic modeling assumptions are that the lamellipodium is idealized as a two-dimensional structure, and that the actin network consists of two families of possibly bent, but locally parallel filaments. Instead of dealing with individual polymers, the filaments are assumed to be continuously distributed.The model has the potential to include the effects of (de)polymerization, of the mechanical effects of cross-linking, bundling, and motor proteins, of cell-substrate adhesion, as well as of the leading edge of the membrane.In the first version presented here, the total amount of F-actin is prescribed by assuming a constant polymerisation speed at the leading edge and a fixed total number and length distribution of filaments. We assume that cross-links at filament crossing points as well as integrin linkages with the matrix break and reform in response to incremental changes in network organisation. In this first treatment, the model successfully simulates the persistence of the treadmilling network in radially spread cells.

Cell migration is initiated by plasma membrane protrusions, in the form of lamellipodia and filopodia. The latter rod-like projections may exert sensory functions and are found in organisms as distant in evolution as mammals and amoeba such as Dictyostelium discoideum. In mammals, lamellipodia protrusion downstream of the small GTPase Rac1 requires a multimeric protein assembly, the WAVE-complex, which activates Arp2/3-mediated actin filament nucleation and actin network assembly. A current model of filopodia formation postulates that these structures arise from a dendritic network of lamellipodial actin filaments by selective elongation and bundling. Here, we have analyzed filopodia formation in mammalian cells abrogated in expression of essential components of the lamellipodial actin polymerization machinery. Cells depleted of the WAVE-complex component Nck-associated protein 1 (Nap1), and, in consequence, of lamellipodia, exhibited normal filopodia protrusion. Likewise, the Arp2/3-complex, which is essential for lamellipodia protrusion, is dispensable for filopodia formation. Moreover, genetic disruption of nap1or the WAVE-orthologue suppressor of cAMP receptor (scar) in Dictyosteliumwas also ineffective in preventing filopodia protrusion. These data suggest that the molecular mechanism of filopodia formation is conserved throughout evolution from Dictyosteliumto mammals and show that lamellipodia and filopodia formation are functionally separable.

The UKAEA Prototype Fast Reactor at Dounreay had a liquid sodium-cooled core. Following its shutdown in 1994, the liquid metal is being removed from the reactor and other vessels by means of specialized equipment and reacted with an aqueous solution of sodium hydroxide in a special vessel. The reaction products are neutralized with hydrochloric acid to produce a saline solution.The reactor sodium delivery and processing equipment is all of novel design. As sodium has been withdrawn from the vessel, it has been necessary to switch off the primary sodium pumps (used to heat the sodium), and the reactor is now kept at temperature by a purpose-designed electric heater and a NaK loop heater.A primary sodium extract pump has currently removed [approximately]450 tonnes of primary sodium from the reactor. As the level falls special equipment will be used to punch a hole in the primary circuit pipe work and to drill the strongback to allow trapped sodium to drain for extraction.

Functionally different subsets of actin filament arrays contribute to cellular organization and motility. We report the identification of a novel subset of loose actin filament arrays through regulated association with the widely expressed protein SWAP-70. These loose actin filament arrays were commonly located behind protruding lamellipodia and membrane ruffles. Visualization of these loose actin filament arrays was dependent on lamellipodial protrusion and the binding of the SWAP-70 PH-domain to a 3'-phosphoinositide. SWAP-70 with a functional pleckstrin homology-domain lacking the C-terminal 60 residues was targeted to the area of the loose actin filament arrays, but it did not associate with actin filaments. The C-terminal 60 residues were sufficient for actin filament association, but they provided no specificity for the subset of loose actin filament arrays. These results identify SWAP-70 as a phosphoinositide 3-kinase signaling-dependent marker for a distinct, hitherto unrecognized, array of actin filaments. Overexpression of SWAP-70 altered the actin organization and lamellipodial morphology. These alterations were dependent on a proper subcellular targeting of SWAP-70. We propose that SWAP-70 regulates the actin cytoskeleton as an effector or adaptor protein in response to agonist stimulated phosphatidylinositol (3,4)-bisphosphate production and cell protrusion.

Functionally different subsets of actin filament arrays contribute to cellular organization and motility. We report the identification of a novel subset of loose actin filament arrays through regulated association with the widely expressed protein SWAP-70. These loose actin filament arrays were commonly located behind protruding lamellipodia and membrane ruffles. Visualization of these loose actin filament arrays was dependent on lamellipodial protrusion and the binding of the SWAP-70 PH-domain to a 3′-phosphoinositide. SWAP-70 with a functional pleckstrin homology-domain lacking the C-terminal 60 residues was targeted to the area of the loose actin filament arrays, but it did not associate with actin filaments. The C-terminal 60 residues were sufficient for actin filament association, but they provided no specificity for the subset of loose actin filament arrays. These results identify SWAP-70 as a phosphoinositide 3-kinase signaling-dependent marker for a distinct, hitherto unrecognized, array of actin filaments. Overexpression of SWAP-70 altered the actin organization and lamellipodial morphology. These alterations were dependent on a proper subcellular targeting of SWAP-70. We propose that SWAP-70 regulates the actincytoskeletonasaneffectororadaptorproteininresponsetoagoniststimulatedphosphatidylinositol (3,4)-bisphosphate production and cell protrusion.

Cell motility is driven by the sum of asymmetric traction forces exerted on the substrate through adhesion foci that interface with the actin cytoskeleton. Establishment of this asymmetry involves microtubules, which exert a destabilising effect on adhesion foci via targeting events. Here, we demonstrate the existence of a mechano-sensing mechanism that signals microtubule polymerisation and guidance of the microtubules towards adhesion sites under increased stress. Stress was applied either by manipulating the body of cells moving on glass with a microneedle or by stretching a flexible substrate that cells were migrating on. We propose a model for this mechano-sensing phenomenon whereby microtubule polymerisation is stimulated and guided through the interaction of a microtubule tip complex with actin filaments under tension.

Cell motility is driven by the sum of asymmetric traction forces exerted on the substrate through adhesion foci that interface with the actin cytoskeleton. Establishment of this asymmetry involves microtubules, which exert a destabilising effect on adhesion foci via targeting events. Here, we demonstrate the existence of a mechano-sensing mechanism that signals microtubule polymerisation and guidance of the microtubules towards adhesion sites under increased stress. Stress was applied either by manipulating the body of cells moving on glass with a microneedle or by stretching a flexible substrate that cells were migrating on. We propose a model for this mechano-sensing phenomenon whereby microtubule polymerisation is stimulated and guided through the interaction of a microtubule tip complex with actin filaments under tension.

An understanding of the mechanisms driving cell motility requires clarification of the structural organisation of actin filament arrays in the regions of cell protrusion termed lamellipodia. Currently, there is a lack of consensus on lamellipodia organisation stemming from the application of alternative procedures for ultrastructural visualisation of cytoskeleton networks. In this study, we show that cryo-electron microscopy of extracted cytoskeletons embedded in a thin layer of vitreous ice can reveal the organisation of cytoskeletal elements at high resolution. Since this method involves no dehydration, drying and contrasting steps that can potentially introduce subtle distortions of filament order and interactions, its application opens the way to resolving the controversial details of lamellipodia architecture.

Neurofibromatosis type 1 (NF1) is one of the most common neurogenetic diseases affecting adults and children. Neurofibromas are one of the most common of the protean manifestations of NF1. Plexiform neurofibromas, which will frequently cause cosmetic abnormalities, pain, and neurologic deficits, are composed of “neoplastic” Schwann cells accompanied by other participating cellular and noncellular components. There is increasing evidence that loss of NF1expression in neoplastic Schwann cells is associated with elevated levels of activated RAS, supporting the notion that the NF1gene product, neurofibromin, acts as a growth regulator by inhibiting ras growth-promoting activity. In addition, there is increasing evidence that other cooperating events, which may be under cytokine modulation, are important for neurofibroma development and growth. Treatment of plexiform neurofibromas has been empiric, with surgery being the primary option for those with progressive lesions causing a major degree of morbidity. The efficacy of alternative treatment approaches, including the use of antihistamines, maturation agents, and antiangiogenic drugs, has been questionable. More recently, biologic-based therapeutic approaches, using drugs that target the molecular genetic underpinnings of plexiform neurofibromas or cytokines believed important in tumor growth, have been initiated. Evaluation of such trials is hindered by the unpredictable natural history of plexiform neurofibromas and difficulties in determining objective response in tumors that are notoriously large and irregular in shape. Innovative neuroimaging techniques and the incorporation of quality-of-life scales may be helpful in evaluation of therapeutic interventions. The ability to design more rational therapies for NF1-associated neurofibromas is heavily predicated on an improved understanding of the molecular and cellular biology of the cells involved in neurofibroma formation and growth.

An understanding of the mechanisms driving cell motility requires clarification of the structural organisation of actin filament arrays in the regions of cell protrusion termed lamellipodia. Currently, there is a lack of consensus on lamellipodia organisation stemming from the application of alternative procedures for ultrastructural visualisation of cytoskeleton networks. In this study, we show that cryo-electron microscopy of extracted cytoskeletons embedded in a thin layer of vitreous ice can reveal the organisation of cytoskeletal elements at high resolution. Since this method involves no dehydration, drying and contrasting steps that can potentially introduce subtle distortions of filament order and interactions, its application opens the way to resolving the controversial details of lamellipodia architecture.

Actin polymerization is accompanied by the formation of protein complexes that link extracellular signals to sites of actin assembly such as membrane ruffles and focal adhesions. One candidate recently implicated in these processes is the LIM domain protein zyxin, which can bind both Ena/vasodilator-stimulated phosphoprotein (VASP) proteins and the actin filament cross-linking protein α-actinin. To characterize the localization and dynamics of zyxin in detail, we generated both monoclonal antibodies and a green fluorescent protein (GFP)-fusion construct. The antibodies colocalized with ectopically expressed GFP-VASP at focal adhesions and along stress fibers, but failed to label lamellipodial and filopodial tips, which also recruit Ena/VASP proteins. Likewise, neither microinjected, fluorescently labeled zyxin antibodies nor ectopically expressed GFP-zyxin were recruited to these latter sites in live cells, whereas both probes incorporated into focal adhesions and stress fibers. Comparing the dynamics of zyxin with that of the focal adhesion protein vinculin revealed that both proteins incorporated simultaneously into newly formed adhesions. However, during spontaneous or induced focal adhesion disassembly, zyxin delocalization preceded that of either vinculin or paxillin. Together, these data identify zyxin as an early target for signals leading to adhesion disassembly, but exclude its role in recruiting Ena/VASP proteins to the tips of lamellipodia and filopodia.

Actin polymerization is accompanied by the formation of protein complexes that link extracellular signals to sites of actin assembly such as membrane ruffles and focal adhesions. One candidate recently implicated in these processes is the LIM domain protein zyxin, which can bind both Ena/vasodilator-stimulated phosphoprotein (VASP) proteins and the actin filament cross-linking protein alpha-actinin. To characterize the localization and dynamics of zyxin in detail, we generated both monoclonal antibodies and a green fluorescent protein (GFP)-fusion construct. The antibodies colocalized with ectopically expressed GFP-VASP at focal adhesions and along stress fibers, but failed to label lamellipodial and filopodial tips, which also recruit Ena/VASP proteins. Likewise, neither microinjected, fluorescently labeled zyxin antibodies nor ectopically expressed GFP-zyxin were recruited to these latter sites in live cells, whereas both probes incorporated into focal adhesions and stress fibers. Comparing the dynamics of zyxin with that of the focal adhesion protein vinculin revealed that both proteins incorporated simultaneously into newly formed adhesions. However, during spontaneous or induced focal adhesion disassembly, zyxin delocalization preceded that of either vinculin or paxillin. Together, these data identify zyxin as an early target for signals leading to adhesion disassembly, but exclude its role in recruiting Ena/VASP proteins to the tips of lamellipodia and filopodia.

ThetamAgene ofAspergillus nidulansencodes a 739-amino acid protein with similarity to Uga35p/Dal81p/DurLp ofSaccharomyces cerevisiae. It has been proposed that TamA functions as a co-activator of AreA, the major nitrogen regulatory protein inA. nidulans. Because AreA functions as a transcriptional activator under nitrogen-limiting conditions, we investigated whether TamA was also present in the nucleus. We found that a GFP-TamA fusion protein was predominantly localised to the nucleus in the presence and absence of ammonium, and that AreA was not required for this distribution. As the predicted DNA-binding domain of TamA is not essential for function, we have used a number of approaches to further define functionally important regions. We have cloned thetamAgene ofA. oryzaeand compared its functional and sequence characteristics with those ofA. nidulans tamAandS. cerevisiae UGA35/DAL81/DURL. TheAspergillushomologues are highly conserved and functionally interchangeable, whereas theS. cerevisiaegene does not complement atamAmutant when expressed inA. nidulans. Uga35p/Dal81p/DurLp was also found to be unable to recruit AreA. The sequence changes in a number oftamAmutant alleles were determined, and altered versions of TamA were tested fortamAcomplementation and interaction with AreA. Changes in most regions of TamA appeared to destroy its function, suggesting that the overall conformation of the protein may be critical for its activity.

Abstract: We present a case of trans-sphenoidal encephalomeningocele in association with a posterior cranial fossa malformation which fulfils the criteria for the Dandy-Walker complex [1]. Congenital cardiovascular defects were also present. An abnormality of neural crest development may be responsible for the combined occurrence of these anomalies.

This paper presents a variation of Biot's consolidation theory for analysing problems involving unsaturated soils, and implemented using the finite element method. The numerical method is applied to a few geotechnical problems as examples and the results obtained are compared to some published data. The illustrative examples show how the numerical method can be used to analyse seepage and consolidation problems associated with unsaturated soils and demonstrate the flexibility and applicability of the presented method. Copyright © 2000 John Wiley & Sons, Ltd.

Observations of internal waves were made at the Malin shelf edge during SESAME (Shelf Edge Studies Acoustic Measurement Experiment), a part of the NERC LOIS-SES experiment, in August-September 1996. These measurements provide a high resolution dataset demonstrating internal wave generation and propagation. This note presents observations of the evolution of an internal bore. The process is shown clearly in a sequence of thermistor chain tows across the shelf break covering a complete tidal cycle, as the double-sided bore transforms into a group of undulations and eventually into more distinct solitary waveforms. Current structures associated with the bore and waves were also observed by ship-mounted ADCP. Analysis of the waveforms in terms of the linear modes and empirical orthogonal functions (EOFs) indicate the dominance of the first mode, which is typical of a shallow water seasonal thermocline environment. Determination of the phase speed of the waves from the consecutive ship surveys enabled the Doppler shift in the towed data to be removed, allowing analysis of the real length scales of the waves. The bore evolution has been modelled using a first order non-linear KdV model for the first mode, initialised with the waveform in the first survey. Comparison of the model and the observations show close agreement in the amplitudes, length scales, phase speeds and separations of the leading internal waves as they evolve. Finally, analysis of the observed internal wave shapes indicates that, within the uncertainties of measurement, the wave-lengths lie between those predicted by first and second order soliton theory.

In this paper, we introduce a new approach for altering the properties of bridged polysilsesquioxane xerogels using postprocessing modification of the polymeric network. The bridging organic group contains latent functionalities that can be liberated thermally, photochemically, or chemically after the gel has been processed to a xerogel. These modifications can produce changes in density, solubility, porosity, and or chemical properties of the material. Since every monomer possesses two latent functional groups, the technique allows for the introduction of high levels of functionality in hybrid organic−inorganic materials. Dialkylene carbonate-bridged polysilsesquioxane gels were prepared by the sol−gel polymerization of bis(triethoxysilylpropyl) carbonate (1) and bis(triethoxysilylisobutyl) carbonate (2). Thermal treatment of the resulting nonporous xerogels and aerogels at 300−350 °C resulted in quantitative decarboxylation of the dialkylene carbonate bridging groups to give new hydroxyalkyl and olefinic substituted polysilsesquioxane monolithic xerogels and aerogels that cannot be directly prepared through direct sol−gel polymerization of organotrialkoxysilanes.

The actin cytoskeleton is a dynamic filamentous network whose formation and remodeling underlies the fundamental processes of cell motility and shape determination. To serve these roles, different compartments of the actin cytoskeleton engage in forming specific coupling sites between neighbouring cells and with the underlying matrix, which themselves serve signal transducing functions. In this review, we focus on methods used to visualise the actin cytoskeleton and its dynamics, embracing the use of proteins tagged with conventional fluorophores and green fluorescent protein. Included also is a comparison of cooled CCD technology, confocal and 2‐photon fluorescence microscopy of living and fixed cells, as well as a critique of current procedures for electron microscopy. Microsc. Res. Tech. 47:3–17, 1999. © 1999 Wiley‐Liss, Inc.

The actin cytoskeleton is a dynamic filamentous network whose formation and remodeling underlies the fundamental processes of cell motility and shape determination. To serve these roles, different compartments of the actin cytoskeleton engage in forming specific coupling sites between neighbouring cells and with the underlying matrix, which themselves serve signal transducing functions. In this review, we focus on methods used to visualise the actin cytoskeleton and its dynamics, embracing the use of proteins tagged with conventional fluorophores and green fluorescent protein. Included also is a comparison of cooled CCD technology, confocal and 2-photon fluorescence microscopy of living and fixed cells, as well as a critique of current procedures for electron microscopy. Microsc. Res. Tech. 47:3–17, 1999. © 1999 Wiley-Liss, Inc.

We reviewed 34 patients with 65 zone 2 flexor tendon injuries in 39 digits whose outcome had been prospectively studied in an earlier investigation at a mean of 10.6 years after repair. Grip strength was assessed using a Jamar dynamometer. Outcome of grip strength in relation to normal data indicated an excellent or good outcome in 94% of patients, fair in 3% and poor in 3%. Using the grading system recommended by the American Society for Surgery of the Hand, the active range of motion was graded as excellent or good in 75% of digits, fair in 15% and poor in 10%. These results compare favourably with those of the original study with 5/39 of digits showing an improvement from good to excellent. Sixteen of the 34 of patients continued to suffer from cold sensitivity.Copyright 1999 The British Society for Surgery of the Hand

We reviewed 34 patients with 65 zone 2 flexor tendon injuries in 39 digits whose outcome had been prospectively studied in an earlier investigation at a mean of 10.6 years after repair. Grip strength was assessed using a Jamar dynamometer. Outcome of grip strength in relation to normal data indicated an excellent or good outcome in 94% of patients, fair in 3% and poor in 3%. Using the grading system recommended by the American Society for Surgery of the Hand, the active range of motion was graded as excellent or good in 75% of digits, fair in 15% and poor in 10%. These results compare favourably with those of the original study with 5/39 of digits showing an improvement from good to excellent. Sixteen of the 34 of patients continued to suffer from cold sensitivity.

Metavinculin is a higher mol. wt variant of vinculin expressed only in muscle tissue. Using amino acid sequencing methods on the intact molecules and their proteolytic subfragments, together with a polyclonal antibody specific only for metavinculin from porcine stomach, we have been able to identify and sequence the difference peptide in the porcine metavinculin molecule. By alignment with the complete sequence of chick fibroblast vinculin (communicated by G.J. Price, P. Jones, M.D. Davison, R. Bendori, S. Griffiths, B. Patel, B. Geiger and D.R. Critchley, prior to publication) the exact location of the insert could be determined. In porcine metavinculin, this insert lies between the 90‐kd protease‐resistant amino‐terminal core and the carboxy terminus of the molecule. It contains 68 amino acids and is flanked by KWSSK sequences, one of which is present in vinculin. The identity of the mapped vinculin and metavinculin sequences outside this difference peptide is consistent with the two proteins arising via alternative splicing at the mRNA level. The lack of reactivity of the porcine metavinculin antibody with metavinculin from chicken as well as the finding of different proteolytic cleavage sites in avian metavinculin indicate a species‐specific amino acid sequence in the difference piece of the metavinculin molecule.

Summary: The ultrastructural localization of sympathetic axons was investigated in normal rat sciatic nerves and experimental sciatic nerve neuromas. The best ultrastructural localization of noradrenaline in the dense-cored vesicles of sympathetic axons was accomplished following pretreatment of rats with nialamide and 5-hydroxy dopamine, followed by fixation according to the modified chromaffin technique of Tranzer and Richards (1976). After such preparation, sympathetic axons containing 5-hydroxy dopamine-labelled dense-cored vesicles could be identified in normal sciatic nerve. Large accumulations of labelled dense-cored vesicles were also found in acute neuromas, up to 1 week after nerve section. Much smaller numbers of densecored vesicles could be identified in chronic neuromas from 2 to 3 weeks following nerve section. Sympathetic axons could also be identified following electron probe X-ray microanalysis of the tissue sections, using chromium detection as the marker for the noradrenaline-containing dense-cored vesicles. Unusual configurations of Schwann cell subunits, which enclosed myelinated fibres and sympathetic axon sprouts within the same basal lamina, were identified in the acute neuromas, 3–7 days after nerve section. Such configurations may be of relevance to the pathophysiological interaction which develops between sympathetic efferent and sensory fibres in peripheral nerve neuromas.

Summary In response to concanavalin A (Con A), neuroblastoma cells undergo marked morphological changes which involve the retraction of neurites and the induction of broad and extensive lamellar regions around the cell periphery. From the use of FITC-Con A it was shown that the membrane formed on the induced lamellar regions lacked receptors to Con A from the onset of lamella formation. These receptors were confined to the cell body; they initially showed a uniform distribution and were subsequently collected into patches and finally into aggregates or caps. When the aggregates occurred on the cell periphery their position coincided with areas free of lamellae.

Measurement of summit metabolism (the maximum rate of heat production) in lambs aged 1 or 4h revealed considerable between animal variation. Summit metabolism per unit body weight decreased as body weight increased whereas summit metabolism per unit body surface area was independent of body weight. Severe pre-partum hypoxia was apparently associated with a low summit metabolism at 1 or 4h of age which made such lambs very susceptible to hypothermia. This deficiency in heat production capacity did not appear to be a permanent featuresince most lambs so affected recovered full thermoregulatory ability by 12h of age. Feeding of colostrum conferred an immediate 18% increase in summit metabolism. The significance of these findings to the prevention of hypothermia in the newborn lamb is discussed.

Lactuca serriola L., which was found to enter skotodormancy at a relatively low temperature, was used to try and ascertain differences in thermo- and skotodormancy.

By conventional indices, the standards of living and education in New Zealand are among the highest in the world. Guidance and counselling services have been developing since the 1920s, and a sound research literature is now emerging. The main divisions of non-medical personnel are clinical psychologists, vocational counsellors, social workers, educational psychologists, guidance counsellors and visiting teachers. In schools, guidance networks have been developed to formalize functions and relationships for effective delivery of services. For social workers, training is provided mainly within government departments, but there are also university programmes for them and for all personnel except visiting teachers. Current needs include more programme evaluation and further development of professional supervision, but a sound indigenous basis in research, training, and service delivery has been laid.

Differential settlements often occur between bridge abutments and the approach embankments either because the soil underlying the embankment consolidates or because the pavement and embankment materials are compressible and the bridge deck is essentially rigid. This causes a “bump” to form at the approach abutment, and so many bridges are provided with approach slabs, the purpose of which is to span across any difference in level caused by differential settlement between the embankment and the bridge. Repeated traffic loadings can, however, produce a new bump at the end of the slab. To overcome this problem, approach slabs can be constructed at an angle to the horizontal, sloping down beneath the pavement. The varying thickness of base course above the slab produces a graded deformation in the pavement and results in a smoother riding surface. Model pavements and approach slabs have been tested in a laboratory‐scale test track, and these tests have shown the effectiveness of sloping approach slabs in reducing the problem of a bump forming at the end of the slab.

The usual assumption, namely that the underlying biochemical reactions in an organism tend to a unique steady-state, is shown to be not always correct. There are certain pathway mechanisms (e.g. positive feedback) which allow the system to exists in two alternative stable steady states. This bistability implies that environmental perturbations can “switch” the system from either state to the other. Such a switch takes place at the metabolic level and hence a single genotype can display two different, alternative, phenotypes without involving any changes in gene expression. The infective transmission of Scrapie-type diseases is explained here by such a mechanism involving protein-only changes.