Carboxyl terminus structural requirements for glycosyl-phosphatidylinositol anchor addition to cell surface proteins.

Glycosyl phosphatidylinositol (GPI)-anchored proteins contain in their COOH-terminal region a peptide segment that is thought to direct glycolipid addition. This signal has been shown to require a pair of small amino acids positioned 10-12 residues upstream of an hydrophobic C-terminal domain. We analysed the contribution of the region separating the anchor acceptor site and the C-terminal hydrophobic segment by introducing amino acid deletions and substitutions in the spacer element of the GPI-anchored Thy-1 glycoprotein. Deletions of 7 amino acids in this region, as well as the introduction of 2 charged residues, prevented the glycolipid addition to Thy-1, suggesting that the length and the primary sequence of the spacer domain are important determinants in the signal directing GPI anchor transfer onto a newly synthesized polypeptide. Furthermore, we tested these rules by creating a truncated form of the normally transmembranous Herpes simplex virus I glycoprotein D (gDI) and demonstrating that when its C-terminal region displays all the features of a GPI-anchored protein, it is able to direct glycolipid addition onto another cell surface molecule.

Large N phases, gravitational instantons and the nuts and bolts of AdS holography

Recent results in the literature concerning holography indicate that the thermodynamics of quantum gravity (at least with a negative cosmological constant) can be modeled by the large N thermodynamics of quantum field theory. We emphasize that this suggests a completely unitary evolution of processes in quantum gravity, including black hole formation and decay, and even more extreme examples involving topology change. As concrete examples which show that this correspondence holds even when the space-time is only locally asymptotically AdS, we compute the thermodynamical phase structure of the AdS-Taub-NUT and AdS-Taub-bolt spacetimes, and compare them to a (2+1)-dimensional conformal field theory (at large N) compactified on a squashed three-sphere and on the twisted plane.

Trapeziectomy and tendon suspension with or without a mitek anchor fixation in the thumb basal joint osteoarthritis.

Partial trapeziectomy with suspension ligamentoplasty is a commonly performed treatment of thumb osteoarthritis. Nevertheless, the post-operative recovery remains long and critical reason for which different modifications of the surgical technique have been proposed. To compare two suspension ligamentoplasty techniques, one with a mitek anchor and another without, a retrospective study of 55 consecutive operated patients was performed. A detailed clinical analysis of pain, function and a radiologic assessment of the trapeziometacarpal space were performed. Mitek anchor fixation was associated with a shorter convalescence period. However, in spite of an improved radiological maintenance of the scaphometacarpal space, mitek anchor fixation was associated with an impaired postoperative function and residual pain when compared with the conventional suspension ligamentoplasty procedure. Patient's satisfaction was comparable in both groups. In our series stabilization of the suspension ligamentoplasty procedure by the insertion of a mitek anchor did not bring the hoped benefits to the patients with a trapeziometacarpal arthritis.

Structure of the glycosyl-phosphatidylinositol membrane anchor of the Leishmania major promastigote surface protease.

In common with many other plasma membrane glycoproteins of eukaryotic origin, the promastigote surface protease (PSP) of the protozoan parasite Leishmania contains a glycosyl-phosphatidylinositol (GPI) membrane anchor. The GPI anchor of Leishmania major PSP was purified following proteolysis of the PSP and analyzed by two-dimensional 1H-1H NMR, compositional and methylation linkage analyses, chemical and enzymatic modifications, and amino acid sequencing. From these results, the structure of the GPI-containing peptide was found to be Asp-Gly-Gly-Asn-ethanolamine-PO4-6Man alpha 1-6Man alpha 1-4GlcN alpha 1-6myo-inositol-1-PO4-(1-alkyl-2-acyl-glycerol). The glycan structure is identical to the conserved glycan core regions of the GPI anchor of Trypanosoma brucei variant surface glycoprotein and rat brain Thy-1 antigen, supporting the notion that this portion of GPIs are highly conserved. The phosphatidylinositol moiety of the PSP anchor is unusual, containing a fully saturated, unbranched 1-O-alkyl chain (mainly C24:0) and a mixture of fully saturated unbranched 2-O-acyl chains (C12:0, C14:0, C16:0, and C18:0). This lipid composition differs significantly from those of the GPIs of T. brucei variant surface glycoprotein and mammalian erythrocyte acetylcholinesterase but is similar to that of a family of glycosylated phosphoinositides found uniquely in Leishmania.

Anchor or Accelerate – A Study on Cancer Cell Adhesion and Motility

Cell migration and adhesion to the extracellular matrix (ECM) are crucial in many biological and pathological processes such as morphogenesis, tissue repair, inflammatory responses, survival, and cancer. Cell-matrix adhesion is mediated by the integrin family of transmembrane receptors, which not only anchor cells to their surroundings, but also transmit bidirectional signalling at the cell surface and couple the ECM to the cytoskeleton. Another group of adhesion receptors are the syndecan proteoglycans, which engage the ECM and possess signalling activity in response to a variety of ligands. Cell migration is a complex process that requires spatial and temporal coordination of adhesion, cell contractility, intracellular traffic of integrins, and matrix turnover by matrix metalloproteinases (MMPs). Thus, integrins and syndecans, as well as MMPs, play essential roles in cancer cell migration and invasion. The understanding of the cooperation of syndecans and integrins was broadened in this thesis study. The results reveal that syndecan-1 functions in concert with 21 integrin in cell adhesion to collagen, whereas syndecan-4 is essential in 21 integrin-mediated matrix contraction. Finally, oncogenic K-Ras was shown to regulate 21 integrin, membrane-type 1 MMP, and syndecan-1 and -4 expression and their cooperation in cell invasion. Epithelial-mesenchymal transition (EMT) is fundamental during embryogenesis and organ development. Activation of EMT processes, including the upregulation of mesenchymal intermediate filament protein vimentin, has also been implicated in the acquisition of a malignant phenotype by epithelial cancer cells. Members of the protein kinase C (PKC) superfamily are involved in cell migration and various integrindependent cellular functions. One aim of this work was to shed light on the role of vimentin in the regulation of integrin traffic and cell motility. In addition, the mechanism by which vimentin participates in EMT was investigated. The results show that integrin recycling and motility are dependent on the PKC–mediated phosphorylation of vimentin. In addition, vimentin was found to be a positive regulator of EMT and regulate the expression of several migratory genes. Specifically, vimentin governs the expression of receptor tyrosine kinase Axl, which is implicated in tumour growth and metastasis. Taken together, the findings described in this thesis reveal novel aspects of the complex interplay between distinct cellular components: integrins, syndecans, and the vimentin cytoskeleton, which all contribute to the regulation of human cancer cell adhesion, migration, and invasion.

3-D reconstruction by extended depth-of-field in failure analysis - Case study I: Qualitative fractographic investigation of fractured bolts in a partial valve

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Anchor Areas to Improve Conservation and Increase Connectivity within the Brazilian "Mesopotamia of Biodiversity"

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Developing an Anchor system to enhance postural control

This article uses an anchor metaphor to explain the dynamic interplay between the human body's active uses of nonrigid tools to mediate information about its adjacent environment to enhance postural control. The author used an anchor system (ropes attached to varying weights resting on the floor) to test blindfolded adults who performed a restricted-balance task (30 s one-foot standing). Participants were tested while holding the anchors under a variety of weight conditions (125 g, 250 g, 500 g, and I kg) and again during a baseline condition (no anchors). When compared with the baseline condition, there was a significant reduction in the amount of body sway across the anchor conditions. The author found that mechanical support provided by the anchor system was secondary to its haptic exploratory function and that an individual can use the anchoring strategy with a dual purpose: for resting and for reorientation after intrinsic disruptions.

Intermittent use of an anchor system improves postural control in healthy older adults

Haptic information, provided by a non-rigid tool (i.e., an anchor system), can reduce body sway in individuals who perform a standing postural task. However, it was not known whether or not continuous use of the anchor system would improve postural control after its removal. Additionally, it was unclear as to whether or not frequency of use of the anchor system is related to improved control in older adults. The present study evaluated the effect of the prolonged use of the anchor system on postural control in healthy older individuals, at different frequencies of use, while they performed a postural control task (semi-tandem position). Participants were divided into three groups according to the frequency of the anchor system's use (0%, 50%, and 100%). Pre-practice phase (without anchor) was followed by a practice phase (they used the anchor system at the predefined frequency), and a post-practice phase (immediate and late-without anchor). All three groups showed a persistent effect 15. min after the end of the practice phase (immediate post-practice phase). However, only the 50% group showed a persistent effect in the late post-practice phase (24. h after finishing the practice phase). Older adults can improve their postural control by practicing the standing postural task, and use of the anchor system limited to half of their practice time can provide additional improvement in their postural control. © 2013 Elsevier B.V.

First principles investigations on the electronic structure of anchor groups on ZnO nanowires and surfaces

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Evaluation of the efficiencies of helical anchor plates in sand by centrifuge model tests

The uplift capacity of helical anchors normally increases with the number of helical plates. The rate of capacity gain is variable, considering that the disturbance caused by the anchor installation is generally more pronounced in the soil mass above the upper plates than above the lower plates, because the upper soil layers are penetrated more times. The present investigation examines the effect of the number of helices on the performance of helical anchors in sand, based on the results of centrifuge model tests. Uplift loading tests were performed on 12 different types of piles installed in two containers of dry sand prepared with different densities. The measured fractions of the uplift capacity related to each individual helical plate of multi-helix anchors were compared with the fractions predicted by the individual bearing method. The results of this investigation indicate that in double- and triple-helix anchors, the contributions of the second and third plate to the total anchor uplift capacity decreased with the increase of sand relative density and plate diameter. In addition, these experiments demonstrated that the variation of the anchor load-displacement behavior with the number of helices also depends on these parameters.

Finite element analysis of stress distribution in anchor teeth in surgically assisted rapid palatal expansion

The treatment of a transverse maxillary deficiency in skeletally mature individuals should include surgically assisted rapid palatal expansion. This study evaluated the distribution of stresses that affect the expander's anchor teeth using finite element analysis when the osteotomy is varied. Five virtual models were built and the surgically assisted rapid palatal expansion was simulated. Results showed tension on the lingual face of the teeth and alveolar bone, and compression on the buccal side of the alveolar bone. The subtotal Le Fort I osteotomy combined with intermaxillary suture osteotomy seemed to reduce the dissipation of tensions. Therefore, subtotal Le Fort I osteotomy without a step in the zygomaticomaxillary buttress, combined with intermaxillary suture osteotomy and pterygomaxillary disjunction may be the osteotomy of choice to reduce tensions on anchor teeth, which tend to move mesiobuccally (premolar) and distobuccally (molar)

A molecular approach towards tethered bilayer lipid membranes: synthesis and characterization of novel anchor lipids

Tethered bilayer lipid membranes (tBLMs) are a promising model system for the natural cell membrane. They consist of a lipid bilayer that is covalently coupled to a solid support via a spacer group. In this study, we developed a suitable approach to increase the submembrane space in tBLMs. The challenge is to create a membrane with a lower lipid density in order to increase the membrane fluidity, but to avoid defects that might appear due to an increase in the lateral space within the tethered monolayers. Therefore, various synthetic strategies and different monolayer preparation techniques were examined. Synthetical attempts to achieve a large ion reservoir were made in two directions: increasing the spacer length of the tether lipids and increasing the lateral distribution of the lipids in the monolayer. The first resulted in the synthesis of a small library of tether lipids (DPTT, DPHT and DPOT) characterized by 1H and 13C NMR, FD-MS, ATR, DSC and TGA. The synthetic strategy for their preparation includes synthesis of precursor with a double bond anchor that can be easily modified for different substrates (e.g. metal and metaloxide). Here, the double bond was modified into a thiol group suitable for gold surface. Another approach towards the preparation of homogeneous monolayers with decreased two-dimensional packing density was the synthesis of two novel anchor lipids: DPHDL and DDPTT. DPHDL is “self-diluted” tether lipid containing two lipoic anchor moieties. DDPTT has an extended lipophylic part that should lead to the preparation of diluted, leakage free proximal layers that will facilitate the completion of the bilayer. Our tool-box of tether lipids was completed with two fluorescent labeled lipid precursors with respectively one and two phytanyl chains in the hydrophobic region and a dansyl group as a fluorophore. The use of such fluorescently marked lipids is supposed to give additional information for the lipid distribution on the air-water interface. The Langmuir film balance was used to investigate the monolayer properties of four of the synthesized thiolated anchor lipids. The packing density and mixing behaviour were examined. The results have shown that mixing anchor with free lipids can homogeneously dilute the anchor lipid monolayers. Moreover, an increase in the hydrophylicity (PEG chain length) of the anchor lipids leads to a higher packing density. A decrease in the temperature results in a similar trend. However, increasing the number of phytanyl chains per lipid molecule is shown to decrease the packing density. LB-monolayers based on pure and mixed lipids in different ratio and transfer pressure were tested to form tBLMs with diluted inner layers. A combination of the LB-monolayer transfer with the solvent exchange method accomplished successfully the formation of tBLMs based on pure DPOT. Some preliminary investigations of the electrical sealing properties and protein incorporation of self-assembled DPOT and DDPTT-based tBLMs were conducted. The bilayer formation performed by solvent exchange resulted in membranes with high resistances and low capacitances. The appearance of space beneath the membrane is clearly visible in the impedance spectra expressed by a second RC element. The latter brings the conclusion that the longer spacer in DPOT and the bigger lateral space between the DDPTT molecules in the investigated systems essentially influence the electrical parameters of the membrane. Finally, we could show the functional incorporation of the small ion carrier valinomycin in both types of membranes.

Stability of the tunnel face reinforced by bolts under seepage flow conditions

Il presente lavoro tratta la stabilità del fronte di scavo, rinforzato con barre di consolidamento ed interessato da drenaggi in avanzamento, di gallerie sotto falda in rocce tenere o terreni. Tale studio è stato sviluppato dal progetto di Tesi attraverso l’analisi all’equilibrio limite che approssima il fronte di scavo con un rettangolo e considera un meccanismo di rottura composto da un cuneo, a tergo del fronte, caricato da un prisma. Il metodo descritto consente di tenere conto dell’effetto stabilizzante delle barre, mediante una distribuzione della pressione di supporto non uniforme. Nel caso di gallerie sotto falda, lo stesso metodo permette inoltre di considerare l’effetto destabilizzante dei gradienti idraulici. Sono state ricavate soluzioni analitiche per la valutazione della stabilità, ed implementate successivamente nel software di analisi numerica MATLAB. Dalle analisi condotte è emerso che il numero minimo di barre per garantire la stabilità del fronte di scavo è in molti casi elevato e risulta impossibile da porre in opera in terreni scarsamente coesivi o in gallerie sotto elevati battenti d’acqua. Per risolvere questa situazione si può prevedere l’inserimento di drenaggi in avanzamento, con lo scopo di diminuire i gradienti idraulici nei pressi del fronte della galleria. Il modello che descrive il nuovo andamento dei carichi idraulici, considerando la presenza di dreni, è stato realizzato con il software commerciale agli elementi finiti COMSOL. Una volta determinati gli andamenti dei carichi idraulici, sono stati condotti studi parametrici sull’effetto dei dreni combinato con gli elementi di rinforzo. Dopo tali analisi sono stati ricavati nomogrammi adimensionali che tengano conto della presenza contemporanea delle barre e dei dreni. Tali diagrammi costituiscono uno strumento utile e valido per la progettazione del rinforzo del fronte di scavo. Infine sono stati realizzati confronti fra casi di studio reali e risultati ottenuti dal modello.