Research notes : polymer concrete bridge deck overlays : Deschutes River Bridge (Biggs), Maupin Bridge (Maupin) : final report.

DOT National Transportation Integrated Search

1995-07-01

This report documents the construction and performance of two thin polymer concrete (with polyester/styrene resins) bridge deck overlays. The overlays were constructed in Biggs and Maupin, Oregon in June 1993. Construction of the overlays was less th...

Research notes : thin polymer overlays on bridge decks.

DOT National Transportation Integrated Search

2007-11-01

Winters in Oregon are hard on our bridges. Vehicles with studded tires gouge the pavement and abrade the deck surface. This abrasion often leads to reduced skid resistance. As the bridge deck ages, cracks often develop. Deicing chemicals used in wint...

Thick-to-Thin Filament Surface Distance Modulates Cross-Bridge Kinetics in Drosophila Flight Muscle

PubMed Central

Tanner, Bertrand C.W.; Farman, Gerrie P.; Irving, Thomas C.; Maughan, David W.; Palmer, Bradley M.; Miller, Mark S.

2012-01-01

The demembranated (skinned) muscle fiber preparation is widely used to investigate muscle contraction because the intracellular ionic conditions can be precisely controlled. However, plasma membrane removal results in a loss of osmotic regulation, causing abnormal hydration of the myofilament lattice and its proteins. We investigated the structural and functional consequences of varied myofilament lattice spacing and protein hydration on cross-bridge rates of force development and detachment in Drosophila melanogaster indirect flight muscle, using x-ray diffraction to compare the lattice spacing of dissected, osmotically compressed skinned fibers to native muscle fibers in living flies. Osmolytes of different sizes and exclusion properties (Dextran T-500 and T-10) were used to differentially alter lattice spacing and protein hydration. At in vivo lattice spacing, cross-bridge attachment time (ton) increased with higher osmotic pressures, consistent with a reduced cross-bridge detachment rate as myofilament protein hydration decreased. In contrast, in the swollen lattice, ton decreased with higher osmotic pressures. These divergent responses were reconciled using a structural model that predicts ton varies inversely with thick-to-thin filament surface distance, suggesting that cross-bridge rates of force development and detachment are modulated more by myofilament lattice geometry than protein hydration. Generalizing these findings, our results suggest that cross-bridge cycling rates slow as thick-to-thin filament surface distance decreases with sarcomere lengthening, and likewise, cross-bridge cycling rates increase during sarcomere shortening. Together, these structural changes may provide a mechanism for altering cross-bridge performance throughout a contraction-relaxation cycle. PMID:22995500

Thick-to-Thin Filament Surface Distance Modulates Cross-Bridge Kinetics in Drosophila Flight Muscle

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tanner, Bertrand C.W.; Farman, Gerrie P.; Irving, Thomas C.

2012-09-19

The demembranated (skinned) muscle fiber preparation is widely used to investigate muscle contraction because the intracellular ionic conditions can be precisely controlled. However, plasma membrane removal results in a loss of osmotic regulation, causing abnormal hydration of the myofilament lattice and its proteins. We investigated the structural and functional consequences of varied myofilament lattice spacing and protein hydration on cross-bridge rates of force development and detachment in Drosophila melanogaster indirect flight muscle, using x-ray diffraction to compare the lattice spacing of dissected, osmotically compressed skinned fibers to native muscle fibers in living flies. Osmolytes of different sizes and exclusion propertiesmore » (Dextran T-500 and T-10) were used to differentially alter lattice spacing and protein hydration. At in vivo lattice spacing, cross-bridge attachment time (t{sub on}) increased with higher osmotic pressures, consistent with a reduced cross-bridge detachment rate as myofilament protein hydration decreased. In contrast, in the swollen lattice, t{sub on} decreased with higher osmotic pressures. These divergent responses were reconciled using a structural model that predicts t{sub on} varies inversely with thick-to-thin filament surface distance, suggesting that cross-bridge rates of force development and detachment are modulated more by myofilament lattice geometry than protein hydration. Generalizing these findings, our results suggest that cross-bridge cycling rates slow as thick-to-thin filament surface distance decreases with sarcomere lengthening, and likewise, cross-bridge cycling rates increase during sarcomere shortening. Together, these structural changes may provide a mechanism for altering cross-bridge performance throughout a contraction-relaxation cycle.« less

Scanning electron microscopy of the trabecular meshwork: understanding the pathogenesis of primary angle closure glaucoma.

PubMed

Sihota, Ramanjit; Goyal, Amita; Kaur, Jasbir; Gupta, Viney; Nag, Tapas C

2012-01-01

To study ultrastructural changes of the trabecular meshwork in acute and chronic primary angle closure glaucoma (PACG) and primary open angle glaucoma (POAG) eyes by scanning electron microscopy. Twenty-one trabecular meshwork surgical specimens from consecutive glaucomatous eyes after a trabeculectomy and five postmortem corneoscleral specimens were fixed immediately in Karnovsky solution. The tissues were washed in 0.1 M phosphate buffer saline, post-fixed in 1% osmium tetraoxide, dehydrated in acetone series (30-100%), dried and mounted. Normal trabecular tissue showed well-defined, thin, cylindrical uveal trabecular beams with many large spaces, overlying flatter corneoscleral beams and numerous smaller spaces. In acute PACG eyes, the trabecular meshwork showed grossly swollen, irregular trabecular endothelial cells with intercellular and occasional basal separation with few spaces. Numerous activated macrophages, leucocytes and amorphous debris were present. Chronic PACG eyes had a few, thickened posterior uveal trabecular beams visible. A homogenous deposit covered the anterior uveal trabeculae and spaces. Converging, fan-shaped trabecular beam configuration corresponded to gonioscopic areas of peripheral anterior synechiae. In POAG eyes, anterior uveal trabecular beams were thin and strap-like, while those posteriorly were wide, with a homogenous deposit covering and bridging intertrabecular spaces, especially posteriorly. Underlying corneoscleral trabecular layers and spaces were visualized in some areas. In acute PACG a marked edema of the endothelium probably contributes for the acute and marked intraocular pressure (IOP) elevation. Chronically raised IOP in chronic PACG and POAG probably results, at least in part, from decreased aqueous outflow secondary to widening and fusion of adjacent trabecular beams, together with the homogenous deposit enmeshing trabecular beams and spaces.

The molecular basis of induction and formation of tunneling nanotubes.

PubMed

Kimura, Shunsuke; Hase, Koji; Ohno, Hiroshi

2013-04-01

Tunneling nanotubes (TNTs) and associated structures are recently recognized structures for intercellular communication. They are F-actin-containing thin protrusions of the plasma membrane of a cell and allow a direct physical connection to the plasma membranes of remote cells. TNTs and associated structures serve as mediators for intercellular transfer of organelles as well as membrane components and cytoplasmic molecules. Moreover, several pathogens have been shown to exploit these structures to spread among cells. Because of their contribution to normal cellular functions and importance in pathological conditions, studies on TNTs and related structures have accelerated over the past few years. These studies have revealed key molecules for their induction and/or formation; HIV Nef and M-Sec can induce the formation of TNTs in coordination with the remodeling of the actin cytoskeleton and vesicle trafficking.

Polymer concrete overlay on the Big Swan Creek Bridge : interim report no. 1--installation and initial condition of overlay.

DOT National Transportation Integrated Search

1984-01-01

The installation of a thin polymer concrete overlay on the Big Swan Creek Bridge provides further evidence that an overlay of low permeability can be soundly bonded to a concrete bridge deck by maintenance forces with a minimum of disruption to traff...

The effects of changes of water balance on the renal pelvic epithelium of the rat.

PubMed

Khorshid, M R; Moffat, D B

1975-01-01

The effects of changes of water balance on the renal pelvic epithelium of the rat. The fine structure of the various epithelia which line the renal pelvis was investigated in five hydropenic rats and five rats undergoing a water diuresis. In the former, the thin epithelium which covers the outer medulla showed dilated intercellular spaces and an increased number of cytoplasmic vacuoles whereas the intercellular spaces were tightly closed and there were few vacuoles in the diuretic rats. It was considered that these changes indicate an exchange of water and solute between pelvic urine and the outer since medulla they are similar to those occurring in epithelia elsewhere which are engaged in transport of salt or water. Similar but less marked changes were found in the papillary epithelium. Changes in the transitional epithelium were similar to those which have previously been described elsewhere in the urinary tract.

Immunogold Localization of the Citrus Exocortis Viroid-Induced Pathogenesis-Related Proteinase P69 in Tomato Leaves 1

PubMed Central

Vera, Pablo; Yago, José Hernández; Conejero, Vicente

1989-01-01

Citrus exocortis viroid induces in tomato plants (Lycopersicon esculentum) synthesis and accumulation of a pathogenesis-related protein (P69) previously reported to be a proteinase (Vera P, Conejero V [1988] Plant Physiol 87: 58-63). By immunogold/transmission electron microscopy, we have studied the distribution of this protein in thin sections of parenchymatous leaf tissue. The enzyme was present intra- and extracellularly. The intracellular location was limited to the vacuole and was always associated with engulfed cell material. When extracellularly located, the enzyme was associated with a dispersed, electron-dense material in the intercellular spaces. This latter location was confirmed after analysis of intercellular washing fluids obtained by vacuum infiltration of leaves. These observations provide new data for the understanding of viroid pathogenesis and the biological role of the pathogenesis-related proteinase P69. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 PMID:16666981

Strength and stiffness of reinforced yellow-poplar glued-laminated beams

Treesearch

R. Hernandez; R. C. Moody

In bridge applications, it is often necessary to minimize the depth of the bridge structure to provide for the required hydraulic opening or reduce the volume of approach fill. For bridges that utilize structural glued-laminated (glulam) timber beams as stringers, reinforcement using thin strips of pultruded E-glass-fiber-reinforced plastic (GFRP) composites may permit...

Self-stressed sandwich bridge decks.

DOT National Transportation Integrated Search

1971-01-01

Proposed is an entirely new type of bridge deck, consisting of an unreinforced lightweight concrete slab made of expanding cement sandwiched between two thin plates of steel. The expanding core serves to prestress the panel. Laboratory tests were con...

Production of footbridge with double curvature made of UHPC

NASA Astrophysics Data System (ADS)

Kolísko, J.; Čítek, D.; Tej, P.; Rydval, M.

2017-09-01

This article present a mix design, preparation and production of thin-walled footbridge made from UHPFRC. In this case an experimental pedestrian bridge was design and prepared. Bridge with span of 10 m and the clear width of 1.50 m designed as single-span bridge. Optimization of UHPFRC matrix and parameters of this material leads to the design of very thin structures. Total thickness of shell structure 30 - 45 mm. Bridge was cast as a prefabricated element in one piece. Self-compacting character of UHPFRC with high flowability allows the production of the final structure. Extensive research was done before production of footbridge. Experimental reached data were compared with extensive numerical analysis and the final design of structure and UHPFRC matrix were optimized in many details. Two versions of large scale mock-ups were casted and tested. According to the complexity of whole experiment a casting technology and production of formwork were tested and optimized many times.

Ultrafast single molecule technique for the study of force dependent kinetics and conformational changes of actin-protein interaction involved in mechanotransduction

NASA Astrophysics Data System (ADS)

Sergides, M.; Arbore, C.; Pavone, F. S.; Capitanio, M.

2018-02-01

Mechanical signals occurring at the interface between cell membrane and extracellular matrix and at intercellular junctions trigger biochemical signals that are fundamental for cell growth, development and regulation. Adaptor proteins, which link the cell membrane to the actin cytoskeleton, seem to partake in this process of mechanotransduction. In particular, catenins play a key role in intercellular junctions, where they act as a bridge between the cell membrane and actin. Studies suggest that α-catenin contains a domain that normally masks vinculin binding sites, which can become accessible after a conformational change induced by an external force. Here we demonstrate a single-molecule technique for investigating actin-protein interactions at different forces (up to 17 pN) with adequate temporal resolution (sub-ms). This system is based on the ultrafast force-clamp spectroscopy technique that has been recently developed by our group and is adapted to study and measure force-dependent kinetics of the catenin-actin interaction, as well as the amplitude of the expected conformational changes such as force-induced protein unfolding.

Thin film atomic hydrogen detectors

NASA Technical Reports Server (NTRS)

Gruber, C. L.

1977-01-01

Thin film and bead thermistor atomic surface recombination hydrogen detectors were investigated both experimentally and theoretically. Devices were constructed on a thin Mylar film substrate. Using suitable Wheatstone bridge techniques sensitivities of 80 microvolts/2x10 to the 13th power atoms/sec are attainable with response time constants on the order of 5 seconds.

Flow of a surfactant across a thin liquid film wetting a solid substrate

DOE Office of Scientific and Technical Information (OSTI.GOV)

Peng, J.B.; He, S.X.; Dutta, P.

1989-12-15

Transfer of a monolayer (valinomycin) between two Langmuir troughs via an interconnecting glass bridge'' has been observed when there is a difference between the monolayer pressures in the two troughs. Assuming that the transfer occurs over the surface of the thin film of water that wets the bridge,'' the transfer rate can be used to estimate the thickness of the liquid film. The thickness thus determined may be used to determine the retarded van der Waals (Hamaker) constant associated with the interaction of the film with the underlying substrate.

Acantholysis revisited: back to basics.

PubMed

Seshadri, Divya; Kumaran, M Sendhil; Kanwar, Amrinder J

2013-01-01

Acantholysis means loss of coherence between epidermal cells due to the breakdown of intercellular bridges. It is an important pathogenetic mechanism underlying various bullous disorders, particularly the pemphigus group, as well as many non-blistering disorders. Although a well-known concept, the student often has to refer to many sources to comprehend acantholysis completely. Thorough knowledge of this topic helps in clinching many diagnoses. The etiopathogenesis, classification, clinical signs, and laboratory demonstration of acantholysis are discussed in detail to help students build clear concepts. We have focused on various distinguishing points in different disorders for an easy grasp of the topic.

Proposal for monitoring concrete painting as a preventive maintenance tool (Abutments and pier caps).

DOT National Transportation Integrated Search

2017-07-01

One of the growing number of preventive bridge maintenance activities conducted by the Kentucky Transportation Cabinet (KYTC) is washing and applying thin film protective coatings to bridge abutments and piers. Previous work conducted by Kentucky Tra...

Formation of a cylindrical bridge in cell division

NASA Astrophysics Data System (ADS)

Citron, Daniel; Schmidt, Laura E.; Reichl, Elizabeth; Ren, Yixin; Robinson, Douglas; Zhang, Wendy W.

2007-11-01

In nature, the shape transition associated with the division of a mother cell into two daughter cells proceeds via a variety of routes. In the cylinder-thinning route, which has been observed in Dictyostelium and most animal cells, the mother cell first forms a broad bridge-like region, also known as a furrow, between two daughter cells. The furrow then rapidly evolves into a cylindrical bridge, which thins and eventually severs the mother cell into two. The fundamental mechanism underlying this division route is not understood. Recent experiments on Dictyostelium found that, while the cylinder-thinning route persists even when key actin cross-linking proteins are missing, it is disrupted by the removal of force-generating myosin-II proteins. Other measurements revealed that mutant cells lacking myosin-II have a much more uniform tension over the cell surface than wild-type cells. This suggests that tension variation may be important. Here we use a fluid model, previously shown to reproduce the thinning dynamics [Zhang & Robinson, PNAS 102, 7186 (2005)], to test this idea. Consistent with the experiments, the model shows that the cylinder formation process occurs regardless of the exact viscoelastic properties of the cell. In contrast to the experiments, a tension variation in the model hinders, rather then expedites, the cylinder formation.

Evaluation of thin epoxy system overlays for concrete bridge decks : interim report No. 1.

DOT National Transportation Integrated Search

1986-05-01

Four epoxy overlay systems were applied to concrete bridge decks in north Louisiana to evaluate their performance as skid resistant surfaces primarily and also as concrete sealers. Dural Flexolith, Poly-Carb Flexogrid, and Con/Chem Cono/Crete were pl...

Evaluation of thin epoxy system overlays for concrete bridge decks : final report.

DOT National Transportation Integrated Search

1991-12-01

Four overlay systems were applied to concrete bridge decks in north Louisiana in May 1995 to evaluate their performance as friction surface and also as concrete sealers. Dural Flexolith, Poly-Carb Flexogrid, and Con/Chem Con/Crete were placed on thre...

Tungsten bridge for the low energy ignition of explosive and energetic materials

DOEpatents

Benson, David A.; Bickes, Jr., Robert W.; Blewer, Robert S.

1990-01-01

A tungsten bridge device for the low energy ignition of explosive and energetic materials is disclosed. The device is fabricated on a silicon-on-sapphire substrate which has an insulating bridge element defined therein using standard integrated circuit fabrication techniques. Then, a thin layer of tungsten is selectively deposited on the silicon bridge layer using chemical vapor deposition techniques. Finally, conductive lands are deposited on each end of the tungsten bridge layer to form the device. It has been found that this device exhibits substantially shorter ignition times than standard metal bridges and foil igniting devices. In addition, substantially less energy is required to cause ignition of the tungsten bridge device of the present invention than is required for common metal bridges and foil devices used for the same purpose.

Surfing the wave, cycle, life history, and genes/proteins expressed by testicular germ cells. Part 4: intercellular bridges, mitochondria, nuclear envelope, apoptosis, ubiquitination, membrane/voltage-gated channels, methylation/acetylation, and transcription factors.

PubMed

Hermo, Louis; Pelletier, R-Marc; Cyr, Daniel G; Smith, Charles E

2010-04-01

As germ cells divide and differentiate from spermatogonia to spermatozoa, they share a number of structural and functional features that are common to all generations of germ cells and these features are discussed herein. Germ cells are linked to one another by large intercellular bridges which serve to move molecules and even large organelles from the cytoplasm of one cell to another. Mitochondria take on different shapes and features and topographical arrangements to accommodate their specific needs during spermatogenesis. The nuclear envelope and pore complex also undergo extensive modifications concomitant with the development of germ cell generations. Apoptosis is an event that is normally triggered by germ cells and involves many proteins. It occurs to limit the germ cell pool and acts as a quality control mechanism. The ubiquitin pathway comprises enzymes that ubiquitinate as well as deubiquitinate target proteins and this pathway is present and functional in germ cells. Germ cells express many proteins involved in water balance and pH control as well as voltage-gated ion channel movement. In the nucleus, proteins undergo epigenetic modifications which include methylation, acetylation, and phosphorylation, with each of these modifications signaling changes in chromatin structure. Germ cells contain specialized transcription complexes that coordinate the differentiation program of spermatogenesis, and there are many male germ cell-specific differences in the components of this machinery. All of the above features of germ cells will be discussed along with the specific proteins/genes and abnormalities to fertility related to each topic. Copyright 2009 Wiley-Liss, Inc.

Fine-tuning of actin dynamics by the HSPB8-BAG3 chaperone complex facilitates cytokinesis and contributes to its impact on cell division.

PubMed

Varlet, Alice Anaïs; Fuchs, Margit; Luthold, Carole; Lambert, Herman; Landry, Jacques; Lavoie, Josée N

2017-07-01

The small heat shock protein HSPB8 and its co-chaperone BAG3 are proposed to regulate cytoskeletal proteostasis in response to mechanical signaling in muscle cells. Here, we show that in dividing cells, the HSPB8-BAG3 complex is instrumental to the accurate disassembly of the actin-based contractile ring during cytokinesis, a process required to allow abscission of daughter cells. Silencing of HSPB8 markedly decreased the mitotic levels of BAG3 in HeLa cells, supporting its crucial role in BAG3 mitotic functions. Cells depleted of HSPB8 were delayed in cytokinesis, remained connected via a disorganized intercellular bridge, and exhibited increased incidence of nuclear abnormalities that result from failed cytokinesis (i.e., bi- and multi-nucleation). Such phenotypes were associated with abnormal accumulation of F-actin at the intercellular bridge of daughter cells at telophase. Remarkably, the actin sequestering drug latrunculin A, like the inhibitor of branched actin polymerization CK666, normalized F-actin during cytokinesis and restored proper cell division in HSPB8-depleted cells, implicating deregulated actin dynamics as a cause of abscission failure. Moreover, this HSPB8-dependent phenotype could be corrected by rapamycin, an autophagy-promoting drug, whereas it was mimicked by drugs impairing lysosomal function. Together, the results further support a role for the HSPB8-BAG3 chaperone complex in quality control of actin-based structure dynamics that are put under high tension, notably during cell cytokinesis. They expand a so-far under-appreciated connection between selective autophagy and cellular morphodynamics that guide cell division.

Mechanical Flexibility of Zinc Oxide Thin-Film Transistors Prepared by Transfer Printing Method

NASA Astrophysics Data System (ADS)

Eun, K. T.; Hwang, W. J.; Sharma, B. K.; Ahn, J. H.; Lee, Y. K.; Choa, S. H.

In the present study, we demonstrate the performance of Zinc oxide thin film transistors (ZnO TFTs) array subjected to the strain under high bending test and the reliability of TFTs was confirmed for the bending fatigue test of 2000 cycles. Initially, ZnO TFTs were fabricated on Si substrate and subsequently transferred on flexible PET substrate using transfer printing process. It was observed that when the bending radius reached ≥ 11 mm then cracks start to initiate first at SiO2 bridges, acting as interconnecting layers among individual TFT. Whatever the strain is applied to the devices, it is almost equivalently adopted by the SiO2 bridges, as they are relatively weak compared to rest of the part. The initial cracking of destructed SiO2 bridge leads to the secondary cracks to the ITO electrodes upon further increment of bending radius. Numerical simulation suggested that the strain of SiO2 layer reached to fracture level of 0.55% which was concentrated at the edge of SiO2 bridge layer. It also suggests that the round shape of SiO2 bridge can be more fruitful to compensate the stress concentration and to prevent failure of device.

Polymer concrete overlay on Beulah Road Bridge : interim report no. 1--installation and initial condition of overlay.

DOT National Transportation Integrated Search

1982-01-01

The installation of a thin polymer concrete overlay on the Beulah Road bridge demonstrates that an overlay of low permeability and high skid resistance can be successfully installed by maintenance forces with a minimum of disruption to traffic, appro...

Evaluation of the construction and performance of polymer concrete overlays on five bridges : interim report no. 1.

DOT National Transportation Integrated Search

1983-01-01

The installation of thin polymer concrete overlays on five bridges on I-85 near Williamsburg, Virginia, has demonstrated that an overlay of low permeability and high skid resistance can be successfully installed by a contractor with a minimum of disr...

Tunneling nanotubes: A versatile target for cancer therapy.

PubMed

Sahoo, Pragyaparamita; Jena, Soumya Ranjan; Samanta, Luna

2017-11-29

Currently Cancer is the leading cause of death worldwide. Malignancy or cancer is a class of diseases characterized by uncontrolled cell growth that eventually invade other tissues and dvelop secondary malignant growth at other sites by metastasis. Intercellular communication plays a major in cancer, particularly in the process of cell proliferation and coordination which in turn leads to tumor invasion, metastasis and development of resistance to therapy. Cells communicate among themselves in a variety of ways, namely, i) via gap junctions with adjacent cells, ii) via exosomes with nearby cells and iii) via chemical messengers with distant cells. Besides, cell - cell connection by tunneling nanotubes (TnTs) is recently gaining importance where intercellular components are transferred between cells. In general cell organelles like Golgi vesicle and mitochondria; and biomolecules like nucleic acids and proteins are transferred through these TnTs. These TnTs are long cytoplasmic extensions made up of actin that function as intercellular bridge and connect a wide verity of cell types. Malignant cells form TnTs with either another malignant cells or cells of the surrounding tumor matrix. These TnTs help in the process of initiation of tumor formation, its organization and propagation. The current review focuses on the role of TnTs mediated cell – cell signaling in cancer micro-environment. Drugs that inhibit TnT-formation such as metformin and everolimus can be targeted towards TnTs in the management of cancer growth, proliferation, tumor invasion and metastasis. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

Macrophage conditioned medium induced cellular network formation in MCF-7 cells through enhanced tunneling nanotube formation and tunneling nanotube mediated release of viable cytoplasmic fragments

DOE Office of Scientific and Technical Information (OSTI.GOV)

Patheja, Pooja, E-mail: pooja.patheja8@gmail.com; Homi Bhabha National Institute, Training School Complex, Anushaktinagar, Mumbai 400094, Maharashtra; Sahu, Khageswar

Infiltrating macrophages in tumor microenvironment, through their secreted cytokines and growth factors, regulate several processes of cancer progression such as cancer cell survival, proliferation, invasion, metastasis and angiogenesis. Recently, intercellular cytoplasmic bridges between cancer cells referred as tunneling nanotubes (TNTs) have been recognized as novel mode of intercellular communication between cancer cells. In this study, we investigated the effect of inflammatory mediators present in conditioned medium derived from macrophages on the formation of TNTs in breast adenocarcinoma cells MCF-7. Results show that treatment with macrophage conditioned medium (MφCM) not only enhanced TNT formation between cells but also stimulated the releasemore » of independently migrating viable cytoplasmic fragments, referred to as microplasts, from MCF-7 cells. Time lapse microscopy revealed that microplasts were released from parent cancer cells in extracellular space through formation of TNT-like structures. Mitochondria, vesicles and cytoplasm could be transferred from parent cell body to microplasts through connecting TNTs. The microplasts could also be resorbed into the parent cell body by retraction of the connecting TNTs. Microplast formation inhibited in presence cell migration inhibitor, cytochalasin-B. Notably by utilizing migratory machinery within microplasts, distantly located MCF-7 cells formed several TNT based intercellular connections, leading to formation of physically connected network of cells. Together, these results demonstrate novel role of TNTs in microplast formation, novel modes of TNT formation mediated by microplasts and stimulatory effect of MφCM on cellular network formation in MCF-7 cells mediated through enhanced TNT and microplast formation.« less

Acute Slices of Mice Testis Seminiferous Tubules Unveil Spontaneous and Synchronous Ca2+ Oscillations in Germ Cell Clusters1

PubMed Central

Sánchez-Cárdenas, Claudia; Guerrero, Adán; Treviño, Claudia Lydia; Hernández-Cruz, Arturo; Darszon, Alberto

2012-01-01

ABSTRACT Spermatogenic cell differentiation involves changes in the concentration of cytoplasmic Ca2+ ([Ca2+]i); however, very few studies exist on [Ca2+]i dynamics in these cells. Other tissues display Ca2+ oscillations involving multicellular functional arrangements. These phenomena have been studied in acute slice preparations that preserve tissue architecture and intercellular communications. Here we report the implementation of intracellular Ca2+ imaging in a sliced seminiferous tubule (SST) preparation to visualize [Ca2+]i changes of living germ cells in situ within the SST preparation. Ca2+ imaging revealed that a subpopulation of male germ cells display spontaneous [Ca2+]i fluctuations resulting from Ca2+ entry possibly throughout CaV3 channels. These [Ca2+]i fluctuation patterns are also present in single acutely dissociated germ cells, but they differ from those recorded from germ cells in the SST preparation. Often, spontaneous Ca2+ fluctuations of spermatogenic cells in the SST occur synchronously, so that clusters of cells can display Ca2+ oscillations for at least 10 min. Synchronous Ca2+ oscillations could be mediated by intercellular communication via gap junctions, although intercellular bridges could also be involved. We also observed an increase in [Ca2+]i after testosterone application, suggesting the presence of functional Sertoli cells in the SST. In summary, we believe that the SST preparation is suitable to explore the physiology of spermatogenic cells in their natural environment, within the seminiferous tubules, in particular Ca2+ signaling phenomena, functional cell-cell communication, and multicellular functional arrangements. PMID:22914313

Drosophila E-cadherin is required for the maintenance of ring canals anchoring to mechanically withstand tissue growth.

PubMed

Loyer, Nicolas; Kolotuev, Irina; Pinot, Mathieu; Le Borgne, Roland

2015-10-13

Intercellular bridges called "ring canals" (RCs) resulting from incomplete cytokinesis play an essential role in intercellular communication in somatic and germinal tissues. During Drosophila oogenesis, RCs connect the maturing oocyte to nurse cells supporting its growth. Despite numerous genetic screens aimed at identifying genes involved in RC biogenesis and maturation, how RCs anchor to the plasma membrane (PM) throughout development remains unexplained. In this study, we report that the clathrin adaptor protein 1 (AP-1) complex, although dispensable for the biogenesis of RCs, is required for the maintenance of the anchorage of RCs to the PM to withstand the increased membrane tension associated with the exponential tissue growth at the onset of vitellogenesis. Here we unravel the mechanisms by which AP-1 enables the maintenance of RCs' anchoring to the PM during size expansion. We show that AP-1 regulates the localization of the intercellular adhesion molecule E-cadherin and that loss of AP-1 causes the disappearance of the E-cadherin-containing adhesive clusters surrounding the RCs. E-cadherin itself is shown to be required for the maintenance of the RCs' anchorage, a function previously unrecognized because of functional compensation by N-cadherin. Scanning block-face EM combined with transmission EM analyses reveals the presence of interdigitated, actin- and Moesin-positive, microvilli-like structures wrapping the RCs. Thus, by modulating E-cadherin trafficking, we show that the sustained E-cadherin-dependent adhesion organizes the microvilli meshwork and ensures the proper attachment of RCs to the PM, thereby counteracting the increasing membrane tension induced by exponential tissue growth.

Drosophila E-cadherin is required for the maintenance of ring canals anchoring to mechanically withstand tissue growth

PubMed Central

Loyer, Nicolas; Kolotuev, Irina; Pinot, Mathieu; Le Borgne, Roland

2015-01-01

Intercellular bridges called “ring canals” (RCs) resulting from incomplete cytokinesis play an essential role in intercellular communication in somatic and germinal tissues. During Drosophila oogenesis, RCs connect the maturing oocyte to nurse cells supporting its growth. Despite numerous genetic screens aimed at identifying genes involved in RC biogenesis and maturation, how RCs anchor to the plasma membrane (PM) throughout development remains unexplained. In this study, we report that the clathrin adaptor protein 1 (AP-1) complex, although dispensable for the biogenesis of RCs, is required for the maintenance of the anchorage of RCs to the PM to withstand the increased membrane tension associated with the exponential tissue growth at the onset of vitellogenesis. Here we unravel the mechanisms by which AP-1 enables the maintenance of RCs’ anchoring to the PM during size expansion. We show that AP-1 regulates the localization of the intercellular adhesion molecule E-cadherin and that loss of AP-1 causes the disappearance of the E-cadherin–containing adhesive clusters surrounding the RCs. E-cadherin itself is shown to be required for the maintenance of the RCs’ anchorage, a function previously unrecognized because of functional compensation by N-cadherin. Scanning block-face EM combined with transmission EM analyses reveals the presence of interdigitated, actin- and Moesin-positive, microvilli-like structures wrapping the RCs. Thus, by modulating E-cadherin trafficking, we show that the sustained E-cadherin–dependent adhesion organizes the microvilli meshwork and ensures the proper attachment of RCs to the PM, thereby counteracting the increasing membrane tension induced by exponential tissue growth. PMID:26424451

Tungsten bridge for the low energy ignition of explosive and energetic materials

DOEpatents

Benson, D.A.; Bickes, R.W. Jr.; Blewer, R.S.

1990-12-11

A tungsten bridge device for the low energy ignition of explosive and energetic materials is disclosed. The device is fabricated on a silicon-on-sapphire substrate which has an insulating bridge element defined therein using standard integrated circuit fabrication techniques. Then, a thin layer of tungsten is selectively deposited on the silicon bridge layer using chemical vapor deposition techniques. Finally, conductive lands are deposited on each end of the tungsten bridge layer to form the device. It has been found that this device exhibits substantially shorter ignition times than standard metal bridges and foil igniting devices. In addition, substantially less energy is required to cause ignition of the tungsten bridge device of the present invention than is required for common metal bridges and foil devices used for the same purpose. 2 figs.

Lithium Oxysilicate Compounds Final Report.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Apblett, Christopher A.; Coyle, Jaclyn

In this study, the structure and composition of lithium silicate thin films deposited by RF magnetron co-sputtering is investigated. Five compositions ranging from Li2Si2O5 to Li8SiO6 were confirmed by inductively coupled plasma-optical emission spectroscopy (ICP-OES) and structure analysis on the evolution of non-bridging oxygens in the thin films was conducted with fourier transform infrared (FTIR) spectroscopy. It was found that non-bridging oxygens (NBOs) increased as the silicate network breaks apart with increasing lithium content which agrees with previous studies on lithium silicates. Thin film impurities were examined with x-ray photoelectron spectroscopy (XPS) and time of flight secondary ion mass spectroscopymore » (TOFSIMS) and traced back to target synthesis. This study utilizes a unique synthesis technique for lithium silicate thin films and can be referred to in future studies on the ionic conductivity of lithium silicates formed on the surface of silicon anodes in lithium ion batteries.« less

Space Paint

NASA Technical Reports Server (NTRS)

1976-01-01

Developed at Goddard, this improved inorganic paint may help protect coastal bridges subject to extreme corrosion from seawater spray. Potassium silicate formulated into a thin waterbase binder that sprays easily, adheres readily, and can be heavily loaded with zinc particles to provide uniform coverage in a single coat. Stanford Research Institute has measured an annual market in excess of $2 billion in painting highway bridges, utility pipelines, nuclear reactors, and railcar hoppers. Other suitable markets include offshore drilling facilities, railroad bridges, and ships.

Performance of thin bonded epoxy overlays on asphalt and concrete bridge deck surfaces.

DOT National Transportation Integrated Search

2014-06-01

This study is the evaluation of two thin bonded epoxy overlays: SafeLane (marketed by Cargill), and Flexogrid : (developed by PolyCarb). SafeLane is advertised as an anti-skid/anti-icing overlay that stores deicing chemicals for : release during wint...

Final report : Polymer concrete overlay on Beulah Road Bridge : condition of overlay after one year in service.

DOT National Transportation Integrated Search

1983-01-01

An evaluation of the thin polymer concrete overlay placed on the Beulah Road bridge indicates that the overlay is securely bonded to the base concrete and is providing low permeability and high skid resistance after 1 year of service life. The lane c...

Growth, patterning, and weak-link fabrication of superconducting YBa2Cu3O(7-x) thin films

NASA Astrophysics Data System (ADS)

Hilton, G. C.; Harris, E. B.; van Harlingen, D. J.

1988-09-01

Thin films of the high-temperature superconducting ceramic oxides have been grown, and techniques for fabricating weak-link structures have been investigated. Films of YBa2Cu3O(7-x) grown on SrTiO3 by a combination of dc magnetron sputtering and thermal evaporation from the three sources have been patterned into microbridges with widths down to 2 microns. Evidence is found that the bridges behave as arrays of Josephson-coupled superconducting islands. Further weak-link behavior is induced by in situ modification of the coupling by ion milling through the bridge.

Ground penetrating radar applied to rebar corrosion inspection

NASA Astrophysics Data System (ADS)

Eisenmann, David; Margetan, Frank; Chiou, Chien-Ping T.; Roberts, Ron; Wendt, Scott

2013-01-01

In this paper we investigate the use of ground penetrating radar (GPR) to detect corrosion-induced thinning of rebar in concrete bridge structures. We consider a simple pulse/echo amplitude-based inspection, positing that the backscattered response from a thinned rebar will be smaller than the similar response from a fully-intact rebar. Using a commercial 1600-MHz GPR system we demonstrate that, for laboratory specimens, backscattered amplitude measurements can detect a thinning loss of 50% in rebar diameter over a short length. GPR inspections on a highway bridge then identify several rebar with unexpectedly low amplitudes, possibly signaling thinning. To field a practical amplitude-based system for detecting thinned rebar, one must be able to quantify and assess the many factors that can potentially contribute to GPR signal amplitude variations. These include variability arising from the rebar itself (e.g., thinning) and from other factors (concrete properties, antenna orientation and liftoff, etc.). We report on early efforts to model the GPR instrument and the inspection process so as to assess such variability and to optimize inspections. This includes efforts to map the antenna radiation pattern, to predict how backscattered responses will vary with rebar size and location, and to assess detectability improvements via synthetic aperture focusing techniques (SAFT).

Flight loads measurements obtained from calibrated strain-gage bridges mounted externally on the skin of a low-aspect-ratio wing

NASA Technical Reports Server (NTRS)

Eckstrom, C. V.

1976-01-01

Flight-test measurements of wingloads (shear, bending moment, and torque) were obtained by means of strain-gage bridges mounted on the exterior surface of a low-aspect-ratio, thin, swept wing which had a structural skin, full-depth honeycomb core, sandwich construction. Details concerning the strain-gage bridges, the calibration procedures used, and the flight-test results are presented along with some pressure measurements and theoretical calculations for comparison purposes.

Epithelial cytoskeletal framework and nuclear matrix-intermediate filament scaffold: three-dimensional organization and protein composition.

PubMed

Fey, E G; Wan, K M; Penman, S

1984-06-01

Madin-Darby canine kidney (MDCK) cells grow as differentiated, epithelial colonies that display tissue-like organization. We examined the structural elements underlying the colony morphology in situ using three consecutive extractions that produce well-defined fractions for both microscopy and biochemical analysis. First, soluble proteins and phospholipid were removed with Triton X-100 in a physiological buffer. The resulting skeletal framework retained nuclei, dense cytoplasmic filament networks, intercellular junctional complexes, and apical microvillar structures. Scanning electron microscopy showed that the apical cell morphology is largely unaltered by detergent extraction. Residual desmosomes, as can be seen in thin sections, were also well-preserved. The skeletal framework was visualized in three dimensions as an unembedded whole mount that revealed the filament networks that were masked in Epon-embedded thin sections of the same preparation. The topography of cytoskeletal filaments was relatively constant throughout the epithelial sheet, particularly across intercellular borders. This ordering of epithelial skeletal filaments across contiguous cell boundaries was in sharp contrast to the more independent organization of networks in autonomous cells such as fibroblasts. Further extraction removed the proteins of the salt-labile cytoskeleton and the chromatin as separate fractions, and left the nuclear matrix-intermediate filament (NM-IF) scaffold. The NM-IF contained only 5% of total cellular protein, but whole mount transmission electron microscopy and immunofluorescence showed that this scaffold was organized as in the intact epithelium. Immunoblots demonstrate that vimentin, cytokeratins, desmosomal proteins, and a 52,000-mol-wt nuclear matrix protein were found almost exclusively in the NM-IF scaffold. Vimentin was largely perinuclear while the cytokeratins were localized at the cell borders. The 52,000-mol-wt nuclear matrix protein was confined to the chromatin-depleted matrix and the desmosomal proteins were observed in punctate polygonal arrays at intercellular junctions. The filaments of the NM-IF were seen to be interconnected, via the desmosomes, over the entire epithelial colony. The differentiated epithelial morphology was reflected in both the cytoskeletal framework and the NM-IF scaffold.

Epithelial cytoskeletal framework and nuclear matrix-intermediate filament scaffold: three-dimensional organization and protein composition

PubMed Central

1984-01-01

Madin-Darby canine kidney (MDCK) cells grow as differentiated, epithelial colonies that display tissue-like organization. We examined the structural elements underlying the colony morphology in situ using three consecutive extractions that produce well-defined fractions for both microscopy and biochemical analysis. First, soluble proteins and phospholipid were removed with Triton X-100 in a physiological buffer. The resulting skeletal framework retained nuclei, dense cytoplasmic filament networks, intercellular junctional complexes, and apical microvillar structures. Scanning electron microscopy showed that the apical cell morphology is largely unaltered by detergent extraction. Residual desmosomes, as can be seen in thin sections, were also well- preserved. The skeletal framework was visualized in three dimensions as an unembedded whole mount that revealed the filament networks that were masked in Epon-embedded thin sections of the same preparation. The topography of cytoskeletal filaments was relatively constant throughout the epithelial sheet, particularly across intercellular borders. This ordering of epithelial skeletal filaments across contiguous cell boundaries was in sharp contrast to the more independent organization of networks in autonomous cells such as fibroblasts. Further extraction removed the proteins of the salt-labile cytoskeleton and the chromatin as separate fractions, and left the nuclear matrix-intermediate filament (NM-IF) scaffold. The NM-IF contained only 5% of total cellular protein, but whole mount transmission electron microscopy and immunofluorescence showed that this scaffold was organized as in the intact epithelium. Immunoblots demonstrate that vimentin, cytokeratins, desmosomal proteins, and a 52,000-mol-wt nuclear matrix protein were found almost exclusively in the NM-IF scaffold. Vimentin was largely perinuclear while the cytokeratins were localized at the cell borders. The 52,000-mol-wt nuclear matrix protein was confined to the chromatin- depleted matrix and the desmosomal proteins were observed in punctate polygonal arrays at intercellular junctions. The filaments of the NM-IF were seen to be interconnected, via the desmosomes, over the entire epithelial colony. The differentiated epithelial morphology was reflected in both the cytoskeletal framework and the NM-IF scaffold. PMID:6202700

Extracellular Vesicles, Tunneling Nanotubes, and Cellular Interplay: Synergies and Missing Links

PubMed Central

Nawaz, Muhammad; Fatima, Farah

2017-01-01

The process of intercellular communication seems to have been a highly conserved evolutionary process. Higher eukaryotes use several means of intercellular communication to address both the changing physiological demands of the body and to fight against diseases. In recent years, there has been an increasing interest in understanding how cell-derived nanovesicles, known as extracellular vesicles (EVs), can function as normal paracrine mediators of intercellular communication, but can also elicit disease progression and may be used for innovative therapies. Over the last decade, a large body of evidence has accumulated to show that cells use cytoplasmic extensions comprising open-ended channels called tunneling nanotubes (TNTs) to connect cells at a long distance and facilitate the exchange of cytoplasmic material. TNTs are a different means of communication to classical gap junctions or cell fusions; since they are characterized by long distance bridging that transfers cytoplasmic organelles and intracellular vesicles between cells and represent the process of heteroplasmy. The role of EVs in cell communication is relatively well-understood, but how TNTs fit into this process is just emerging. The aim of this review is to describe the relationship between TNTs and EVs, and to discuss the synergies between these two crucial processes in the context of normal cellular cross-talk, physiological roles, modulation of immune responses, development of diseases, and their combinatory effects in tissue repair. At the present time this review appears to be the first summary of the implications of the overlapping roles of TNTs and EVs. We believe that a better appreciation of these parallel processes will improve our understanding on how these nanoscale conduits can be utilized as novel tools for targeted therapies. PMID:28770210

Macrophage conditioned medium induced cellular network formation in MCF-7 cells through enhanced tunneling nanotube formation and tunneling nanotube mediated release of viable cytoplasmic fragments.

PubMed

Patheja, Pooja; Sahu, Khageswar

2017-06-15

Infiltrating macrophages in tumor microenvironment, through their secreted cytokines and growth factors, regulate several processes of cancer progression such as cancer cell survival, proliferation, invasion, metastasis and angiogenesis. Recently, intercellular cytoplasmic bridges between cancer cells referred as tunneling nanotubes (TNTs) have been recognized as novel mode of intercellular communication between cancer cells. In this study, we investigated the effect of inflammatory mediators present in conditioned medium derived from macrophages on the formation of TNTs in breast adenocarcinoma cells MCF-7. Results show that treatment with macrophage conditioned medium (MɸCM) not only enhanced TNT formation between cells but also stimulated the release of independently migrating viable cytoplasmic fragments, referred to as microplasts, from MCF-7 cells. Time lapse microscopy revealed that microplasts were released from parent cancer cells in extracellular space through formation of TNT-like structures. Mitochondria, vesicles and cytoplasm could be transferred from parent cell body to microplasts through connecting TNTs. The microplasts could also be resorbed into the parent cell body by retraction of the connecting TNTs. Microplast formation inhibited in presence cell migration inhibitor, cytochalasin-B. Notably by utilizing migratory machinery within microplasts, distantly located MCF-7 cells formed several TNT based intercellular connections, leading to formation of physically connected network of cells. Together, these results demonstrate novel role of TNTs in microplast formation, novel modes of TNT formation mediated by microplasts and stimulatory effect of MɸCM on cellular network formation in MCF-7 cells mediated through enhanced TNT and microplast formation. Copyright © 2017 Elsevier Inc. All rights reserved.

Evolution and sign control of square-wave-like anisotropic magneto-resistance in spatially confined La{sub 0.3}Pr{sub 0.4}Ca{sub 0.3}MnO{sub 3}/LaAlO{sub 3}(001) manganite thin films

DOE Office of Scientific and Technical Information (OSTI.GOV)

Alagoz, H. S., E-mail: alagoz@ualberta.ca; Jeon, J.; Keating, S.

2016-04-14

We investigated magneto-transport properties of a compressively strained spatially confined La{sub 0.3}Pr{sub 0.4}Ca{sub 0.3}MnO{sub 3} (LPCMO) thin film micro-bridge deposited on LaAlO{sub 3}. Angular dependence of the magneto-resistance R(θ) of this bridge, where θ is the angle between the magnetic field and the current directions in the film plane, exhibits sharp positive and negative percolation jumps near T{sub MIT}. The sign and the magnitude of these jumps can be tuned using the magnetic field. Such behavior has not been observed in LPCMO micro-bridges subjected to tensile strain, indicating a correlation between the type of the lattice strain, the distribution ofmore » electronic domains, and the anisotropic magneto-resistance in spatially confined manganite systems.« less

Strengthening of Existing Bridge Structures for Shear and Bending with Carbon Textile-Reinforced Mortar.

PubMed

Herbrand, Martin; Adam, Viviane; Classen, Martin; Kueres, Dominik; Hegger, Josef

2017-09-19

Increasing traffic loads and changes in code provisions lead to deficits in shear and flexural capacity of many existing highway bridges. Therefore, a large number of structures are expected to require refurbishment and strengthening in the future. This projection is based on the current condition of many older road bridges. Different strengthening methods for bridges exist to extend their service life, all having specific advantages and disadvantages. By applying a thin layer of carbon textile-reinforced mortar (CTRM) to bridge deck slabs and the webs of pre-stressed concrete bridges, the fatigue and ultimate strength of these members can be increased significantly. The CTRM layer is a combination of a corrosion resistant carbon fiber reinforced polymer (CFRP) fabric and an efficient mortar. In this paper, the strengthening method and the experimental results obtained at RWTH Aachen University are presented.

Strengthening of Existing Bridge Structures for Shear and Bending with Carbon Textile-Reinforced Mortar

PubMed Central

Herbrand, Martin; Classen, Martin; Kueres, Dominik; Hegger, Josef

2017-01-01

Increasing traffic loads and changes in code provisions lead to deficits in shear and flexural capacity of many existing highway bridges. Therefore, a large number of structures are expected to require refurbishment and strengthening in the future. This projection is based on the current condition of many older road bridges. Different strengthening methods for bridges exist to extend their service life, all having specific advantages and disadvantages. By applying a thin layer of carbon textile-reinforced mortar (CTRM) to bridge deck slabs and the webs of pre-stressed concrete bridges, the fatigue and ultimate strength of these members can be increased significantly. The CTRM layer is a combination of a corrosion resistant carbon fiber reinforced polymer (CFRP) fabric and an efficient mortar. In this paper, the strengthening method and the experimental results obtained at RWTH Aachen University are presented. PMID:28925962

Fabrication and design of vanadium oxide microbolometer

NASA Astrophysics Data System (ADS)

Abdel-Rahman, M.; Al-Khalli, N.; Zia, M. F.; Alduraibi, M.; Ilahi, B.; Awad, E.; Debbar, N.

2017-02-01

Vanadium oxide (VxOy) multilayer sandwich structures previously studied by our group were found to yield a sensitive thermometer thin film material suitable for microbolometer applications. In this work, we aim to estimate the performance of a proposed air-bridge microbolometer configuration based on VxOy multilayer sandwich structure thermometer thin films. For this purpose, a microbolometer was fabricated on silicon (Si) substrate covered with a silicon nitride (Si3N4) insulating layer using VxOy thermometer thin film material. The fabricated microbolometer was patterned using electron-beam lithography and liftoff techniques and it was characterized in terms of its voltage repsonsivity (Rv), signal to noise ratio (SNR), noise equivalent power (NEP) and detectivity D*. A model was then developed by the aid of numerical optical/thermal simulations and experimentally measured parameters to estimate the performance of the microbolometer when fabricated in an air-bridge configuration. The estimated D* was found to be 1.55×107 cm.√Hz/ W.

Low polarity water, a novel transition species at the polyethylene-water interface.

PubMed

Kosower, Edward M; Borz, Galina

2015-10-14

The bridge between water repelling and water-attracting regions is recognized here as low polarity water, a novel "neutral" form of water; its identity as a dipole-dipole water dimer is supported by spectroscopic evidence of its presence in thin films of water on a polyethylene surface. High resolution (0.5 cm(-1)), low signal energies (Sg 100) and short scans (0.1 s) are used to ensure that all peaks are detected. Thin films may be trapped between two polyethylene windows, affirming the low polarity of such water; the spectra of the trapped films ("sandwich") are similar to those from a subtraction procedure. Use of the "sandwich" is a new and useful technique in surface studies. In general, intermediate forms might bridge incompatibility between different regimes, from sets of molecules (chemistry and physics) to sets of organisms (biology and sociology). Thin films of water on polyethylene also display strong and transient peaks of water oligomers, cyclic pentamers and cyclic hexamers (chair and boat), bicyclic hexamers (books 1 and 2) and tricyclic hexamers (prism) that have been previously identified in thin films of water on a silver halide surface.

Superconducting properties of NbN film, bridge and meanders

NASA Astrophysics Data System (ADS)

Joshi, Lalit M.; Verma, Apoorva; Gupta, Anurag; Rout, P. K.; Husale, Sudhir; Budhani, R. C.

2018-05-01

The transport properties of superconducting NbN nanostructures in the form of thin film, bridge of width (w) = 50 μm and three meanders of w = 500, 250 and 100 nm have been investigated by resistance (R) measurements in temperature (T) range = 2 -300 K and magnetic field (B) range = 0 - 7 Tesla. The nanostructuring was carried out using Focused Ion Beam (FIB) milling. Reduction of sample width results in significant changes in the normal and superconducting state properties. For instance, the observed metallic behavior in the thin film sample is lost and the normal state resistance increases drastically from 2.4 Ω to 418 kΩ for the 100 nm meander. In the superconducting state, the value of critical temperature Tc (upper critical field Bc2 at T = 0 K) reduces gradually with width reduction, it changes from 13.15 K (42.8 Tesla) in the case of thin film sample to 5.7 K (12.7 Tesla) for the 100 nm meander sample. The superconducting transitions are found to get broader for the bridge sample and the meanders additionally show low-temperature resistive tails. In case of all the samples with reduced width, the transition onsets are found to be rounded at surprisingly high values of T ˜ 25 K >> Tc. These results are discussed in terms of the possible effects of FIB processing and weak localization in our samples.

Nonlinear force-length relationship in the ADP-induced contraction of skeletal myofibrils.

PubMed

Shimamoto, Yuta; Kono, Fumiaki; Suzuki, Madoka; Ishiwata, Shin'ichi

2007-12-15

The regulatory mechanism of sarcomeric activity has not been fully clarified yet because of its complex and cooperative nature, which involves both Ca(2+) and cross-bridge binding to the thin filament. To reveal the mechanism of regulation mediated by the cross-bridges, separately from the effect of Ca(2+), we investigated the force-sarcomere length (SL) relationship in rabbit skeletal myofibrils (a single myofibril or a thin bundle) at SL > 2.2 microm in the absence of Ca(2+) at various levels of activation by exogenous MgADP (4-20 mM) in the presence of 1 mM MgATP. The individual SLs were measured by phase-contrast microscopy to confirm the homogeneity of the striation pattern of sarcomeres during activation. We found that at partial activation with 4-8 mM MgADP, the developed force nonlinearly depended on the length of overlap between the thick and the thin filaments; that is, contrary to the maximal activation, the maximal active force was generated at shorter overlap. Besides, the active force became larger, whereas this nonlinearity tended to weaken, with either an increase in [MgADP] or the lateral osmotic compression of the myofilament lattice induced by the addition of a macromolecular compound, dextran T-500. The model analysis, which takes into account the [MgADP]- and the lattice-spacing-dependent probability of cross-bridge formation, was successfully applied to account for the force-SL relationship observed at partial activation. These results strongly suggest that the cross-bridge works as a cooperative activator, the function of which is highly sensitive to as little as

Isotonic force modulates force redevelopment rate of intact frog muscle fibres: evidence for cross-bridge induced thin filament activation

PubMed Central

Vandenboom, Rene; Hannon, James D; Sieck, Gary C

2002-01-01

We tested the hypothesis that force-velocity history modulates thin filament activation, as assessed by the rate of force redevelopment after shortening (+dF/dtR). The influence of isotonic force on +dF/dtR was assessed by imposing uniform amplitude (2.55 to 2.15 μm sarcomere−1) but different speed releases to intact frog muscle fibres during fused tetani. Each release consisted of a contiguous ramp- and step-change in length. Ramp speed was changed from release to release to vary fibre shortening speed from 1.00 (2.76 ± 0.11 μm half-sarcomere−1 s−1) to 0.30 of maximum unloaded shortening velocity (Vu), thereby modulating isotonic force from 0 to 0.34 Fo, respectively. The step zeroed force and allowed the fibre to shorten unloaded for a brief period of time prior to force redevelopment. Although peak force redevelopment after different releases was similar, +dF/dtR increased by 81 ± 6% (P < 0.05) as fibre shortening speed was reduced from 1.00 Vu. The +dF/dtR after different releases was strongly correlated with the preceding isotonic force (r = 0.99, P < 0.001). Results from additional experiments showed that the slope of slack test plots produced by systematically increasing the step size that followed each ramp were similar. Thus, isotonic force did not influence Vu (mean: 2.84 ± 0.10 μm half-sarcomere−1 s−1, P < 0.05). We conclude that isotonic force modulates +dF/dtR independent of change in Vu, an outcome consistent with a cooperative influence of attached cross-bridges on thin filament activation that increases cross-bridge attachment rate without alteration to cross-bridge detachment rate. PMID:12205189

Identifying Molecular Targets for Chemoprevention in a Rat Model

DTIC Science & Technology

2007-06-01

accompanied by a reactive stromal proliferation resulting in a distinct thickening of the thin muscular layer surrounding individual glands. These...accompanied by reactive stromal proliferation, resulting in a distinct thickening of the thin muscular layer surrounding individual glands. These proliferations...epithelial cells forming solid bridges and circular apolar lumina. The lesions filled the glandular lumen but did not show distension with foci of

Design of a CMOS readout circuit on ultra-thin flexible silicon chip for printed strain gauges

NASA Astrophysics Data System (ADS)

Elsobky, Mourad; Mahsereci, Yigit; Keck, Jürgen; Richter, Harald; Burghartz, Joachim N.

2017-09-01

Flexible electronics represents an emerging technology with features enabling several new applications such as wearable electronics and bendable displays. Precise and high-performance sensors readout chips are crucial for high quality flexible electronic products. In this work, the design of a CMOS readout circuit for an array of printed strain gauges is presented. The ultra-thin readout chip and the printed sensors are combined on a thin Benzocyclobutene/Polyimide (BCB/PI) substrate to form a Hybrid System-in-Foil (HySiF), which is used as an electronic skin for robotic applications. Each strain gauge utilizes a Wheatstone bridge circuit, where four Aerosol Jet® printed meander-shaped resistors form a full-bridge topology. The readout chip amplifies the output voltage difference (about 5 mV full-scale swing) of the strain gauge. One challenge during the sensor interface circuit design is to compensate for the relatively large dc offset (about 30 mV at 1 mA) in the bridge output voltage so that the amplified signal span matches the input range of an analog-to-digital converter (ADC). The circuit design uses the 0. 5 µm mixed-signal GATEFORESTTM technology. In order to achieve the mechanical flexibility, the chip fabrication is based on either back thinned wafers or the ChipFilmTM technology, which enables the manufacturing of silicon chips with a thickness of about 20 µm. The implemented readout chip uses a supply of 5 V and includes a 5-bit digital-to-analog converter (DAC), a differential difference amplifier (DDA), and a 10-bit successive approximation register (SAR) ADC. The circuit is simulated across process, supply and temperature corners and the simulation results indicate excellent performance in terms of circuit stability and linearity.

Pulsed laser deposition to synthesize the bridge structure of artificial nacre: Comparison of nano- and femtosecond lasers

NASA Astrophysics Data System (ADS)

Melaibari, Ammar A.; Molian, Pal

2012-11-01

Nature offers inspiration to new adaptive technologies that allow us to build amazing shapes and structures such as nacre using synthetic materials. Consequently, we have designed a pulsed laser ablation manufacturing process involving thin film deposition and micro-machining to create hard/soft layered "brick-bridge-mortar" nacre of AlMgB14 (hard phase) with Ti (soft phase). In this paper, we report pulsed laser deposition (PLD) to mimic brick and bridge structures of natural nacre in AlMgB14. Particulate formation inherent in PLD is exploited to develop the bridge structure. Mechanical behavior analysis of the AlMgB14/Ti system revealed that the brick is to be 250 nm thick, 9 μm lateral dimensions while the bridge (particle) is to have a diameter of 500 nm for a performance equivalent to natural nacre. Both nanosecond (ns) and femtosecond (fs) pulsed lasers were employed for PLD in an iterative approach that involves varying pulse energy, pulse repetition rate, and target-to-substrate distance to achieve the desired brick and bridge characteristics. Scanning electron microscopy, x-ray photoelectron spectroscopy, and optical profilometer were used to evaluate the film thickness, particle size and density, stoichiometry, and surface roughness of thin films. Results indicated that both ns-pulsed and fs-pulsed lasers produce the desired nacre features. However, each laser may be chosen for different reasons: fs-pulsed laser is preferred for much shorter deposition time, better stoichiometry, uniform-sized particles, and uniform film thickness, while ns-pulsed laser is favored for industrial acceptance, reliability, ease of handling, and low cost.

Intercellular junctions between palisade nerve endings and outer root sheath cells of rat vellus hairs.

PubMed

Kaidoh, T; Inoué, T

2000-05-15

Hair follicles have a longitudinal set of sensory nerve endings called palisade nerve endings (PN). We examined the junctional structures between the PN and outer root sheath (ORS) cells of hair follicles in the rat external ear. Transmission electron microscopy of serial thin sections showed that the processes of the ORS cells penetrated the basal lamina of the hair follicle, forming intercellular junctions with the PN (PN-ORS junctions). Two types of junctions were found: junctions between nerve endings and ORS cells (N-ORS junctions) and those between Schwann cell processes and ORS cells (S-ORS junctions). The N-ORS junctions had two subtypes: 1) a short process or small eminence of the ORS cell was attached to the nerve ending (type I); or 2) a process of the ORS cell was invaginated into the nerve ending (type II). The S-ORS junctions also had two subtypes: 1) a short process or small eminence of the ORS cell was abutted on the Schwann cell process (type I); or 2) a process of the ORS cell was invaginated into the Schwann cell process (type II). Vesicles, coated pits, coated vesicles, and endosomes were sometimes seen in nerve endings, Schwann cells, and ORS cells near the junctions. Computer-aided reconstruction of the serial thin sections displayed the three-dimensional structure of these junctions. These results suggested that the PN-ORS junctions provided direct relationships between the PN and ORS in at least four different patterns. The discovery of these junctions shows the PN-ORS relationship to be closer than previously realized. We speculate that these junctions may have roles in attachment of the PN to the ORS, contributing to increases in the sensitivity of the PN, and in chemical signaling between the PN and ORS.

Estimation of Bridge Height over Water from Polarimetric SAR Image Data Using Mapping and Projection Algorithm and De-Orientation Theory

NASA Astrophysics Data System (ADS)

Wang, Haipeng; Xu, Feng; Jin, Ya-Qiu; Ouchi, Kazuo

An inversion method of bridge height over water by polarimetric synthetic aperture radar (SAR) is developed. A geometric ray description to illustrate scattering mechanism of a bridge over water surface is identified by polarimetric image analysis. Using the mapping and projecting algorithm, a polarimetric SAR image of a bridge model is first simulated and shows that scattering from a bridge over water can be identified by three strip lines corresponding to single-, double-, and triple-order scattering, respectively. A set of polarimetric parameters based on the de-orientation theory is applied to analysis of three types scattering, and the thinning-clustering algorithm and Hough transform are then employed to locate the image positions of these strip lines. These lines are used to invert the bridge height. Fully polarimetric image data of airborne Pi-SAR at X-band are applied to inversion of the height and width of the Naruto Bridge in Japan. Based on the same principle, this approach is also applicable to spaceborne ALOSPALSAR single-polarization data of the Eastern Ocean Bridge in China. The results show good feasibility to realize the bridge height inversion.

Apparatus for melt growth of crystalline semiconductor sheets

DOEpatents

Ciszek, Theodore F.; Hurd, Jeffery L.

1986-01-01

An economical method is presented for forming thin sheets of crystalline silicon suitable for use in a photovoltaic conversion cell by solidification from the liquid phase. Two spatially separated, generally coplanar filaments wettable by liquid silicon and joined together at the end by a bridge member are immersed in a silicon melt and then slowly withdrawn from the melt so that a silicon crystal is grown between the edge of the bridge and the filaments.

Book lung development in juveniles and adults of the cobweb spider, Parasteatoda tepidariorum C. L. Koch, 1841 (Araneomorphae, Theridiidae).

PubMed

Farley, Roger D

2018-03-01

Light and transmission electron microscopy were used to study the development of new book lung lamellae in juvenile and adult spiders (Parasteatoda tepidariorum). As hypothesized earlier in a study of embryos, mesenchyme cells dispersed throughout the opisthosoma (EMT) are a likely source of precursor epithelial cells (MET) for the new lamellae. The precursor cells in juveniles and adults continue many of the complex activities observed in embryos, e.g., migration, alignment, lumen formation, thinning, elongation, and secretion of the cuticle of air channel walls and trabeculae. The apicobasal polarity of precursor cells for new channels is apparently induced by the polarity pattern of precursor cells of channels produced earlier. Thus, new air and hemolymph channels extend and continue the alternating pattern of older channels. At sites more distant from the spiracle and atrium, new channels are usually produced by the mode II process (intracellular alignment and merging of vesicles). These air channels have bridging trabeculae and are quite stable in size throughout their length. At sites closer to the spiracle and atrium, new channels may be produced by mode I (coalescence of merocrine vesicle secretion). This raises the hypothesis that structural and functional differences in mode I and II channels and differing oxygen and fluid conditions with distance from the spiracle and atrium determine the mode of formation of new channels. Observations herein support an earlier hypothesis that there is some intercellular apical/apical and basal/basal affinity among the opposed surfaces of aligned precursor cells. This results in the alternating pattern of air channels at the apical and hemolymph channels at the basal cell surfaces. Copyright © 2018 The Author(s). Published by Elsevier Ltd.. All rights reserved.

Shearing of nanoscopic bridges in two-component thin liquid layers between chemically patterned walls.

PubMed

Hemming, C J; Patey, G N

2004-10-01

Bridge phases associated with a phase transition between two liquid phases occur when a two-component liquid mixture is confined between chemically patterned walls. In the bulk the liquid mixture with components A, B undergoes phase separation into an A-rich phase and a B-rich phase. The walls bear stripes attractive to A. In the bridge phase A-rich and B-rich regions alternate. Grand canonical Monte Carlo studies are performed with the alignment between stripes on opposite walls varied. Misalignment of the stripes places the nanoscopic liquid bridges under shear strain. The bridges exert a Hookean restoring force on the walls for small displacements from equilibrium. As the strain increases there are deviations from Hooke's law. Eventually there is an abrupt yielding of the bridges. Molecular dynamics simulations show the bridges form or disintegrate on time scales which are fast compared to wall motion and transport of molecules into or from the confined space. Some interesting possible applications of the phenomena are discussed. (c) 2004 American Institute of Physics

Extracellular vesicle communication pathways as regulatory targets of oncogenic transformation.

PubMed

Choi, Dongsic; Lee, Tae Hoon; Spinelli, Cristiana; Chennakrishnaiah, Shilpa; D'Asti, Esterina; Rak, Janusz

2017-07-01

Pathogenesis of human cancers bridges intracellular oncogenic driver events and their impact on intercellular communication. Among multiple mediators of this 'pathological connectivity' the role of extracellular vesicles (EVs) and their subsets (exosomes, ectosomes, oncosomes) is of particular interest for several reasons. The release of EVs from cancer cells represents a unique mechanism of regulated expulsion of bioactive molecules, a process that also mediates cell-to-cell transfer of lipids, proteins, and nucleic acids. Biological effects of these processes have been implicated in several aspects of cancer-related pathology, including tumour growth, invasion, angiogenesis, metastasis, immunity and thrombosis. Notably, the emerging evidence suggests that oncogenic mutations may impact several aspects of EV-mediated cell-cell communication including: (i) EV release rate and protein content; (ii) molecular composition of cancer EVs; (iii) the inclusion of oncogenic and mutant macromolecules in the EV cargo; (iv) EV-mediated release of genomic DNA; (v) deregulation of mechanisms responsible for EV biogenesis (vesiculome) and (vi) mechanisms of EV uptake by cancer cells. Intriguingly, EV-mediated intercellular transfer of mutant and oncogenic molecules between subpopulations of cancer cells, their indolent counterparts and stroma may exert profound biological effects that often resemble (but are not tantamount to) oncogenic transformation, including changes in cell growth, clonogenicity and angiogenic phenotype, or cause cell stress and death. However, several biological barriers likely curtail a permanent horizontal transformation of normal cells through EV-mediated mechanisms. The ongoing analysis and targeting of EV-mediated intercellular communication pathways can be viewed as a new therapeutic paradigm in cancer, while the analysis of oncogenic cargo contained in EVs released from cancer cells into biofluids is being developed for clinical use as a biomarker and companion diagnostics. Indeed, studies are underway to further explore the multiple links between molecular causality in cancer and various aspects of cellular vesiculation. Copyright © 2017 Elsevier Ltd. All rights reserved.

Polymer concrete overlay on SH-51, bridge deck

NASA Astrophysics Data System (ADS)

Borg, T. M.

1982-06-01

A thin resinous overlay was placed on a sound bridge deck in Oklahoma to evaluate its performance over one year using various physical tests. The evaluation shows how well the overlay protects the reinforcing steel from corrosion due to deicing salts. The steps leading to the construction of the overlay are detailed as well as the actual placing of the overlay. The results of various physical tests are reported for both before and after the overlay.

Method and apparatus for melt growth of crystalline semiconductor sheets

DOEpatents

Ciszek, T.F.; Hurd, J.L.

1981-02-25

An economical method is presented for forming thin sheets of crystalline silicon suitable for use in a photovoltaic conversion cell by solidification from the liquid phase. Two spatially separated, generally coplanar filaments wettable by liquid silicon and joined together at the end by a bridge member are immersed in a silicon melt and then slowly withdrawn from the melt so that a silicon crystal is grown between the edge of the bridge and the filaments.

Citrate bridges between mineral platelets in bone

PubMed Central

Davies, Erika; Müller, Karin H.; Wong, Wai Ching; Pickard, Chris J.; Reid, David G.; Skepper, Jeremy N.; Duer, Melinda J.

2014-01-01

We provide evidence that citrate anions bridge between mineral platelets in bone and hypothesize that their presence acts to maintain separate platelets with disordered regions between them rather than gradual transformations into larger, more ordered blocks of mineral. To assess this hypothesis, we take as a model for a citrate bridging between layers of calcium phosphate mineral a double salt octacalcium phosphate citrate (OCP-citrate). We use a combination of multinuclear solid-state NMR spectroscopy, powder X-ray diffraction, and first principles electronic structure calculations to propose a quantitative structure for this material, in which citrate anions reside in a hydrated layer, bridging between apatitic layers. To assess the relevance of such a structure in native bone mineral, we present for the first time, to our knowledge, 17O NMR data on bone and compare them with 17O NMR data for OCP-citrate and other calcium phosphate minerals relevant to bone. The proposed structural model that we deduce from this work for bone mineral is a layered structure with thin apatitic platelets sandwiched between OCP-citrate–like hydrated layers. Such a structure can explain a number of known structural features of bone mineral: the thin, plate-like morphology of mature bone mineral crystals, the presence of significant quantities of strongly bound water molecules, and the relatively high concentration of hydrogen phosphate as well as the maintenance of a disordered region between mineral platelets. PMID:24706850

Lipid composition of the stratum corneum and cutaneous water loss in birds along an aridity gradient.

PubMed

Champagne, Alex M; Muñoz-Garcia, Agustí; Shtayyeh, Tamer; Tieleman, B Irene; Hegemann, Arne; Clement, Michelle E; Williams, Joseph B

2012-12-15

Intercellular and covalently bound lipids within the stratum corneum (SC), the outermost layer of the epidermis, are the primary barrier to cutaneous water loss (CWL) in birds. We compared CWL and intercellular SC lipid composition in 20 species of birds from desert and mesic environments. Furthermore, we compared covalently bound lipids with CWL and intercellular lipids in the lark family (Alaudidae). We found that CWL increases in birds from more mesic environments, and this increase was related to changes in intercellular SC lipid composition. The most consistent pattern that emerged was a decrease in the relative amount of cerebrosides as CWL increased, a pattern that is counterintuitive based on studies of mammals with Gaucher disease. Although covalently bound lipids in larks did not correlate with CWL, we found that covalently bound cerebrosides correlated positively with intercellular cerebrosides and intercellular cholesterol ester, and intercellular cerebrosides correlated positively with covalently bound free fatty acids. Our results led us to propose a new model for the organization of lipids in the avian SC, in which the sugar moieties of cerebrosides lie outside of intercellular lipid layers, where they may interdigitate with adjacent intercellular cerebrosides or with covalently bound cerebrosides.

Signaling from the Podocyte Intercellular Junction to the Actin Cytoskeleton

PubMed Central

George, Britta; Holzman, Lawrence B.

2012-01-01

Observations of hereditary glomerular disease support the contention that podocyte intercellular junction proteins are essential for junction formation and maintenance. Genetic deletion of most of these podocyte intercellular junction proteins results in foot process effacement and proteinuria. This review focuses on the current understanding of molecular mechanisms by which podocyte intercellular junction proteins such as the Nephrin-Neph1-Podocin receptor complex coordinate cytoskeletal dynamics and thus intercellular junction formation, maintenance and injury-dependent remodeling. PMID:22958485

Cavitation of intercellular spaces is critical to establishment of hydraulic properties of compression wood of Chamaecyparis obtusa seedlings

PubMed Central

Nakaba, Satoshi; Hirai, Asami; Kudo, Kayo; Yamagishi, Yusuke; Yamane, Kenichi; Kuroda, Katsushi; Nugroho, Widyanto Dwi; Kitin, Peter; Funada, Ryo

2016-01-01

Background and Aims When the orientation of the stems of conifers departs from the vertical as a result of environmental influences, conifers form compression wood that results in restoration of verticality. It is well known that intercellular spaces are formed between tracheids in compression wood, but the function of these spaces remains to be clarified. In the present study, we evaluated the impact of these spaces in artificially induced compression wood in Chamaecyparis obtusa seedlings. Methods We monitored the presence or absence of liquid in the intercellular spaces of differentiating xylem by cryo-scanning electron microscopy. In addition, we analysed the relationship between intercellular spaces and the hydraulic properties of the compression wood. Key Results Initially, we detected small intercellular spaces with liquid in regions in which the profiles of tracheids were not rounded in transverse surfaces, indicating that the intercellular spaces had originally contained no gases. In the regions where tracheids had formed secondary walls, we found that some intercellular spaces had lost their liquid. Cavitation of intercellular spaces would affect hydraulic conductivity as a consequence of the induction of cavitation in neighbouring tracheids. Conclusions Our observations suggest that cavitation of intercellular spaces is the critical event that affects not only the functions of intercellular spaces but also the hydraulic properties of compression wood. PMID:26818592

Intercellular salicylic acid accumulation during compatible and incompatible Arabidopsis-Pseudomonas syringae interactions

PubMed Central

Wilson, Daniel C; Carella, Philip; Cameron, Robin K

2014-01-01

The phytohormone salicylic acid (SA) plays an important role in several disease resistance responses. During the Age-Related Resistance (ARR) response that occurs in mature Arabidopsis responding to Pseudomonas syringae pv tomato (Pst), SA accumulates in the intercellular space where it may act as an antimicrobial agent. Recently we measured intracellular and intercellular SA levels in young, ARR-incompetent plants responding to virulent and avirulent strains of Pst to determine if intercellular SA accumulation is a component of additional defense responses to Pst. In young plants virulent Pst suppressed both intra- and intercellular SA accumulation in a coronatine-dependent manner. In contrast, high levels of intra- and intercellular SA accumulated in response to avirulent Pst. Our results support the idea that SA accumulation in the intercellular space is an important component of multiple defense responses. Future research will include understanding how mature plants counteract the effects of coronatine during the ARR response. PMID:25763618

Human cardiac telocytes: 3D imaging by FIB-SEM tomography

PubMed Central

Cretoiu, D; Hummel, E; Zimmermann, H; Gherghiceanu, M; Popescu, L M

2014-01-01

Telocyte (TC) is a newly identified type of cell in the cardiac interstitium (www.telocytes.com). TCs are described by classical transmission electron microscopy as cells with very thin and long telopodes (Tps; cellular prolongations) having podoms (dilations) and podomers (very thin segments). TCs’ three-dimensional (3D) morphology is still unknown. Cardiac TCs seem to be particularly involved in long and short distance intercellular signalling and, therefore, their 3D architecture is important for understanding their spatial connections. Using focused ion beam scanning electron microscopy (FIB-SEM) we show, for the first time, the whole ultrastructural anatomy of cardiac TCs. 3D reconstruction of cardiac TCs by FIB-SEM tomography confirms that they have long, narrow but flattened (ribbon-like) telopodes, with humps generated by the podoms. FIB-SEM tomography also confirms the network made by TCs in the cardiac interstitium through adherens junctions. This study provides the first FIB-SEM tomography of a human cell type. PMID:25327290

Enzyme-assisted growth of nacreous CaCO3/polymer hybrid nanolaminates via the formation of mineral bridges

NASA Astrophysics Data System (ADS)

Yeom, Bongjun; Char, Kookheon

2016-06-01

Laminated nanostructures in nacre have been adopted as models in the fabrication of strong, tough synthetic nanocomposites. However, the utilization of CaCO3 biominerals in these composites is limited by the complexity of the synthesis method for nanosized biominerals. In this study, we use the enzymatic reaction of urease to generate a nanoscale CaCO3 thin film to prepare CaCO3/polymer hybrid nanolaminates. Additional layers of CaCO3 thin film are consecutively grown over the base CaCO3 layer with the intercalation of organic layers. The morphology and crystallinity of the added CaCO3 layers depend strongly on the thickness of the organic layer coated on the underlying CaCO3 layer. When the organic layer is less than 20 nm thick, the amorphous CaCO3 layer is spontaneously transformed into crystalline calcite layer during the growth process. We also observe crystalline continuity between adjacent CaCO3 layers through interconnecting mineral bridges. The formation of these mineral bridges is crucial to the epitaxial growth of CaCO3 layers, similar to the formation of natural nacre.

Stabilization of electrically conducting capillary bridges using feedback control of radial electrostatic stresses and the shapes of extended bridges

NASA Astrophysics Data System (ADS)

Marr-Lyon, Mark J.; Thiessen, David B.; Blonigen, Florian J.; Marston, Philip L.

2000-05-01

Electrically conducting, cylindrical liquid bridges in a density-matched, electrically insulating bath were stabilized beyond the Rayleigh-Plateau (RP) limit using electrostatic stresses applied by concentric ring electrodes. A circular liquid cylinder of length L and radius R in real or simulated zero gravity becomes unstable when the slenderness S=L/2R exceeds π. The initial instability involves the growth of the so-called (2, 0) mode of the bridge in which one side becomes thin and the other side rotund. A mode-sensing optical system detects the growth of the (2, 0) mode and an analog feedback system applies the appropriate voltages to a pair of concentric ring electrodes positioned near the ends of the bridge in order to counter the growth of the (2, 0) mode and prevent breakup of the bridge. The conducting bridge is formed between metal disks which are grounded. Three feedback algorithms were tested and each found capable of stabilizing a bridge well beyond the RP limit. All three algorithms stabilized bridges having S as great as 4.3 and the extended bridges broke immediately when feedback was terminated. One algorithm was suitable for stabilization approaching S=4.493… where the (3, 0) mode is predicted to become unstable for cylindrical bridges. For that algorithm the equilibrium shapes of bridges that were slightly under or over inflated corresponded to solutions of the Young-Laplace equation with negligible electrostatic stresses. The electrical conductivity of the bridge liquid need not be large. The conductivity was associated with salt added to the aqueous bridge liquid.

Investigation of Intercellular Salicylic Acid Accumulation during Compatible and Incompatible Arabidopsis-Pseudomonas syringae Interactions Using a Fast Neutron-Generated Mutant Allele of EDS5 Identified by Genetic Mapping and Whole-Genome Sequencing

PubMed Central

Catana, Vasile; Golding, Brian; Weretilnyk, Elizabeth A.; Cameron, Robin K.

2014-01-01

A whole-genome sequencing technique developed to identify fast neutron-induced deletion mutations revealed that iap1-1 is a new allele of EDS5 (eds5-5). RPS2-AvrRpt2-initiated effector-triggered immunity (ETI) was compromised in iap1-1/eds5-5 with respect to in planta bacterial levels and the hypersensitive response, while intra- and intercellular free salicylic acid (SA) accumulation was greatly reduced, suggesting that SA contributes as both an intracellular signaling molecule and an antimicrobial agent in the intercellular space during ETI. During the compatible interaction between wild-type Col-0 and virulent Pseudomonas syringae pv. tomato (Pst), little intercellular free SA accumulated, which led to the hypothesis that Pst suppresses intercellular SA accumulation. When Col-0 was inoculated with a coronatine-deficient strain of Pst, high levels of intercellular SA accumulation were observed, suggesting that Pst suppresses intercellular SA accumulation using its phytotoxin coronatine. This work suggests that accumulation of SA in the intercellular space is an important component of basal/PAMP-triggered immunity as well as ETI to pathogens that colonize the intercellular space. PMID:24594657

Cavitation of intercellular spaces is critical to establishment of hydraulic properties of compression wood of Chamaecyparis obtusa seedlings.

PubMed

Nakaba, Satoshi; Hirai, Asami; Kudo, Kayo; Yamagishi, Yusuke; Yamane, Kenichi; Kuroda, Katsushi; Nugroho, Widyanto Dwi; Kitin, Peter; Funada, Ryo

2016-03-01

When the orientation of the stems of conifers departs from the vertical as a result of environmental influences, conifers form compression wood that results in restoration of verticality. It is well known that intercellular spaces are formed between tracheids in compression wood, but the function of these spaces remains to be clarified. In the present study, we evaluated the impact of these spaces in artificially induced compression wood in Chamaecyparis obtusa seedlings. We monitored the presence or absence of liquid in the intercellular spaces of differentiating xylem by cryo-scanning electron microscopy. In addition, we analysed the relationship between intercellular spaces and the hydraulic properties of the compression wood. Initially, we detected small intercellular spaces with liquid in regions in which the profiles of tracheids were not rounded in transverse surfaces, indicating that the intercellular spaces had originally contained no gases. In the regions where tracheids had formed secondary walls, we found that some intercellular spaces had lost their liquid. Cavitation of intercellular spaces would affect hydraulic conductivity as a consequence of the induction of cavitation in neighbouring tracheids. Our observations suggest that cavitation of intercellular spaces is the critical event that affects not only the functions of intercellular spaces but also the hydraulic properties of compression wood. © The Author 2016. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Appendix C : SR-72 tests.

DOT National Transportation Integrated Search

2013-03-01

The Florida highway system includes some of the earliest (circa 1950s) pretensioned : concrete bridges in the United States. Shear capacity of Floridas early pretensioned girders is of : interest because the early designs had thin webs and only li...

Cardiac myofilaments: mechanics and regulation

NASA Technical Reports Server (NTRS)

de Tombe, Pieter P.; Bers, D. M. (Principal Investigator)

2003-01-01

The mechanical properties of the cardiac myofilament are an important determinant of pump function of the heart. This report is focused on the regulation of myofilament function in cardiac muscle. Calcium ions form the trigger that induces activation of the thin filament which, in turn, allows for cross-bridge formation, ATP hydrolysis, and force development. The structure and protein-protein interactions of the cardiac sarcomere that are responsible for these processes will be reviewed. The molecular mechanism that underlies myofilament activation is incompletely understood. Recent experimental approaches have been employed to unravel the mechanism and regulation of myofilament mechanics and energetics by activator calcium and sarcomere length, as well as contractile protein phosphorylation mediated by protein kinase A. Central to these studies is the question whether such factors impact on muscle function simply by altering thin filament activation state, or whether modulation of cross-bridge cycling also plays a part in the responses of muscle to these stimuli.

Ultrasonic measurements of thin zinc layers on concrete

NASA Astrophysics Data System (ADS)

Jansen, Henri; Brooks, Bill; Nguyen, Vinh; Koretsky, Milo

2008-05-01

In order to protect bridges at the coast from corrosion, a thin layer (approximately 0.5 mm) of zinc is sprayed on the concrete of the bridge. When this zinc layer is electrically connected to the reinforcing steel (rebar) and placed at a positive potential with respect to the rebar, oxidation is favored at the zinc layer and reduced at the rebar. The resulting protection of the rebar fails when the zinc layer delaminates from the concrete or when the zinc oxidation product layer becomes too thick. We have used ultrasonic detection to investigate the properties of the zinc layer. This method has been applied very successfully in the semiconductor industry. We present the details of the method and the expected response. Unfortunately, we are not able to measure changes in the zinc layer, because either the frequency we use (10-20 MHz) is too low, or scattering in the concrete is a dominant effect.

Intercellular nanotubes: insights from imaging studies and beyond

PubMed Central

Hurtig, Johan; Chiu, Daniel T.; Önfelt, Björn

2017-01-01

Cell-cell communication is critical to the development, maintenance, and function of multicellular organisms. Classical mechanisms for intercellular communication include secretion of molecules into the extracellular space and transport of small molecules through gap junctions. Recent reports suggest that cells also can communicate over long distances via a network of transient intercellular nanotubes. Such nanotubes have been shown to mediate intercellular transfer of organelles as well as membrane components and cytoplasmic molecules. Moreover, intercellular nanotubes have been observed in vivo and have been shown to enhance the transmission of pathogens such as human immunodeficiency virus (HIV)-1 and prions in vitro. These studies indicate that intercellular nanotubes may play a role both in normal physiology and in disease. PMID:20166114

AC electric field induced dielectrophoretic assembly behavior of gold nanoparticles in a wide frequency range

NASA Astrophysics Data System (ADS)

Liu, Weiyu; Wang, Chunhui; Ding, Haitao; Shao, Jinyou; Ding, Yucheng

2016-05-01

In this work, we focus on frequency-dependence of pearl chain formations (PCF) of gold nanoparticles driven by AC dielectrophoresis (DEP), especially in a low field-frequency range, where induced double-layer charging effect at ideally polarizable surfaces on particle DEP behavior and surrounding liquid motion need not be negligible. As field frequency varies, grown features of DEP assembly structures ranging from low-frequency non-bridged gap to high-frequency single gold nanoparticle-made nanowires bridging the electrodes are demonstrated experimentally. Specifically, at 10 kHz, a kind of novel channel-like structure with parallel opposing banks is formed at the center of interelectrode gap. In stark contrast, at 1 MHz, thin PCF with diameter of 100 nm is created along the shortest distance of the isolation spacing. Moreover, a particular conductive path of nanoparticle chains is produced at 1 MHz in a DEP device embedded with multiple floating electrodes. A theoretical framework taking into account field-induced double-layer polarization at both the particle/electrolyte and electrode/electrolyte interface is developed to correlate these experimental observations with induced-charge electrokinetic (ICEK) phenomenon. And a RC circuit model is helpful in accounting for the formation of this particular non-bridged channel-like structure induced by a low-frequency AC voltage. As compared to thin PCF formed at high field frequency that effectively short circuits the electrode pair, though it is difficult for complete PCF bridging to occur at low frequency, the non-bridged conducting microstructure has potential to further miniaturize the size of electrode gap fabricated by standard micromachining process and may find useful application in biochemical sensing.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Martz, Joseph Christopher; Spearing, Dane Robert; Labouriau, Andrea

Microclad is a composite material consisting of a thin copper coating applied on a single side over a Kapton substrate. Kapton is the commercial designator for polyimide supplied by DuPont. Microclad is a key material in detonator manufacture and function. Detonators which utilize Microclad function when a large current applied through a thin bridge etched into the copper produces a plasma, accelerating a Kapton flyer into an explosive (PETN) pellet. The geometry and properties of the Microclad are a critical element of this process.

Nanoporous thin film additives to improve precast concrete construction of transportation facilities.

DOT National Transportation Integrated Search

2011-12-01

The national transportation network contains a significant number of highway and railway bridges. This research is intended to transform the use of precast/pre-stressed materials in the transportation infrastructure. Specifically it examines how nano...

Sensing sheet: the response of full-bridge strain sensors to thermal variations for detecting and characterizing cracks

NASA Astrophysics Data System (ADS)

Tung, S.-T.; Glisic, B.

2016-12-01

Sensing sheets based on large-area electronics consist of a dense array of unit strain sensors. This new technology has potential for becoming an effective and affordable monitoring tool that can identify, localize and quantify surface damage in structures. This research contributes to their development by investigating the response of full-bridge unit strain sensors to thermal variations. Overall, this investigation quantifies the effects of temperature on thin-film full-bridge strain sensors monitoring uncracked and cracked concrete. Additionally, an empirical formula is developed to estimate crack width given an observed strain change and a measured temperature change. This research led to the understanding of the behavior of full-bridge strain sensors installed on cracked concrete and exposed to temperature variations. It proves the concept of the sensing sheet and its suitability for application in environments with variable temperature.

Self-healing patterns in ferromagnetic-superconducting hybrids

DOE Office of Scientific and Technical Information (OSTI.GOV)

Vlasko-Vlasov, V. K.; Palacious, E.; Rosenmann, D.

We study magnetic flux dynamic effects in a superconducting bridge with thin soft magnetic stripes placed either on top or under the bridge. Voltage-current (VI) measurements reveal that the edges of magnetic stripes oriented transvers or along the bridge introduce channels or barriers for vortex motion, resulting in the decrease or increase of the critical current, respectively. We demonstrate a remarkable self-healing effect whereby the magnetic pinning strength for the longitudinal stripes increases with current. The self-field of the current polarizes the magnetic stripes along their width, which enhances the stray fields at their edges and creates a dynamic vortexmore » pinning landscape to impede vortex flow. Our results highlight new strategies to engineer adaptive pinning topologies in superconducting-ferromagnetic hybrids.« less

Molecular diodes and ultra-thin organic rectifying junctions: Au-S-CnH2n-Q3CNQ and TCNQ derivatives.

PubMed

Ashwell, Geoffrey J; Moczko, Katarzyna; Sujka, Marta; Chwialkowska, Anna; Hermann High, L R; Sandman, Daniel J

2007-02-28

Attempts to obtain derivatives of the molecular diode, 2-{4-[1-cyano-2-(1-(omega-acetylsulfanylalkyl)-1H-quinolin-4-ylidene)-ethylidene]-cyclohexa-2,5-dienylidene}-malonitrile [1, CH(3)CO-S-C(n)H(2n)-Q3CNQ], from either 2,3,5,6-tetrafluoro-7,7,8,8-tetracyano-p-quinodimethane (TCNQF(4)) or 2,3,5,6-tetramethyl-7,7,8,8-tetracyano-p-quinodimethane (TMTCNQ) result in ring closure via the cyano group of the pi-bridge and yield di-substituted analogues: 2-{2,3,5,6-tetrafluoro-4-[6-(10-acetylsulfanyldecyl)-3-(1-(10-acetylsulfanyldecyl)-1H-quinolin-4-ylidenemethyl)-6H-benzo[f][1,7]naphthyridin-2-ylidene]-cyclohexa-2,5-dienylidene}-malonitrile (2a) and the 2,3,5,6-tetramethyl derivative (2b). Self-assembled monolayers (SAMs) of these donor-(pi-bridge)-acceptor molecular diodes exhibit asymmetric current-voltage characteristics with electron flow at forward bias from the top contact to surface C(CN)(2) groups. Comparison is made with I-V curves from ultra-thin films of an organic rectifying junction in which TCNQ(-) is electron-donating and a donor-(sigma-bridge)-acceptor diode in which TCNQ degrees is electron-accepting.

Polymer concrete overlay test program : Lebanon Ditch Bridge : final report.

DOT National Transportation Integrated Search

1983-01-01

This report presents information on the installation of a thin polymer concrete overlay and the evaluation of its durability after a 15-month in-service period. The project was performed by the Oregon State Highway Division under the sponsorship of t...

Electrically Tunable Integrated Thin-Film Magnetoelectric Resonators

DOE Office of Scientific and Technical Information (OSTI.GOV)

El-Ghazaly, Amal; Evans, Joseph T.; Sato, Noriyuki

Magnetoelectrics have attracted much attention for their ability to control magnetic behavior electrically and electrical behavior magnetically. This feature provides numerous benefits to electronic systems and can potentially serve as the bridge needed to integrate magnetic devices into mainstream electronics. This natural next step is pursued and thin-film integrated magnetoelectric devices are produced for radio-frequency (RF) electronics. The first fully integrated, thin-film magnetoelectric modulators for tunable RF electronics are presented. Moreover, these devices provide electric field control of magnetic permeability in order to change the phase velocity and resonance frequency of coplanar waveguides. During this study, the various thin-film materialmore » phenomena, trade-offs, and integration considerations for composite magnetoelectrics are analyzed and discussed. The fabricated devices achieve reversible tunability of the resonance frequency, characterized by a remarkable converse magnetoelectric coupling coefficient of up to 24 mG cm V -1 using just thin films. Based on this work, suggestions are given for additional optimizations of future designs that will maximize the thin-film magnetoelectric interactions.« less

A generalized theory of thin film growth

NASA Astrophysics Data System (ADS)

Du, Feng; Huang, Hanchen

2018-03-01

This paper reports a theory of thin film growth that is generalized for arbitrary incidence angle during physical vapor deposition in two dimensions. The accompanying kinetic Monte Carlo simulations serve as verification. A special theory already exists for thin film growth with zero incidence angle, and another theory also exists for nanorod growth with a glancing angle. The theory in this report serves as a bridge to describe the transition from thin film growth to nanorod growth. In particular, this theory gives two critical conditions in analytical form of critical coverage, ΘI and ΘII. The first critical condition defines the onset when crystal growth or step dynamics stops following the wedding cake model for thin film growth. The second critical condition defines the onset when multiple-layer surface steps form to enable nanorod growth. Further, this theory also reveals a critical incidence angle, below which nanorod growth is impossible. The critical coverages, together with the critical incidence angle, defines a phase diagram of thin growth versus nanorod growth.

Electrically Tunable Integrated Thin-Film Magnetoelectric Resonators

DOE PAGES

El-Ghazaly, Amal; Evans, Joseph T.; Sato, Noriyuki; ...

2017-06-14

Magnetoelectrics have attracted much attention for their ability to control magnetic behavior electrically and electrical behavior magnetically. This feature provides numerous benefits to electronic systems and can potentially serve as the bridge needed to integrate magnetic devices into mainstream electronics. This natural next step is pursued and thin-film integrated magnetoelectric devices are produced for radio-frequency (RF) electronics. The first fully integrated, thin-film magnetoelectric modulators for tunable RF electronics are presented. Moreover, these devices provide electric field control of magnetic permeability in order to change the phase velocity and resonance frequency of coplanar waveguides. During this study, the various thin-film materialmore » phenomena, trade-offs, and integration considerations for composite magnetoelectrics are analyzed and discussed. The fabricated devices achieve reversible tunability of the resonance frequency, characterized by a remarkable converse magnetoelectric coupling coefficient of up to 24 mG cm V -1 using just thin films. Based on this work, suggestions are given for additional optimizations of future designs that will maximize the thin-film magnetoelectric interactions.« less

Variations in gap junctional intercellular communication and connexin expression in fibroblasts derived from keloid and hypertrophic scars.

PubMed

Lu, Feng; Gao, JianHua; Ogawa, Rei; Hyakusoku, Hiko

2007-03-01

Expression of connexins and other constituent proteins of gap junctions along with gap junctional intercellular communication are involved in cellular development and differentiation processes. In addition, an increasing number of hereditary skin disorders appear to be linked to connexins. Therefore, in this report, the authors studied in vitro gap junctional intercellular communication function and connexin expression in fibroblasts derived from keloid and hypertrophic scar patients. Fibroblasts harvested from each of six keloid and hypertrophic scar patients were used for this study. Gap junctional intercellular communication function was investigated using the gap fluorescence recovery after photobleaching method, and expression of connexin proteins was studied using quantitative confocal microscopic analyses. Compared with normal skin, a decreased level of gap junctional intercellular communication was seen in fibroblasts derived from hypertrophic scar tissue, whereas an extremely low gap junctional intercellular communication level was detected in fibroblasts derived from keloid tissue. We also detected little connexin 43 (Cx43) protein localized in fibroblasts derived from keloids. Moreover, Cx43 protein levels were much lower in fibroblasts derived from hypertrophic scars than in those derived from normal skin. The authors' data suggest that the loss of gap junctional intercellular communication and connexin expression may affect intercellular recognition and thus break the proliferation and apoptosis balance in fibroblasts derived from keloid and hypertrophic scar tissue.

Large-scale models reveal the two-component mechanics of striated muscle.

PubMed

Jarosch, Robert

2008-12-01

This paper provides a comprehensive explanation of striated muscle mechanics and contraction on the basis of filament rotations. Helical proteins, particularly the coiled-coils of tropomyosin, myosin and alpha-actinin, shorten their H-bonds cooperatively and produce torque and filament rotations when the Coulombic net-charge repulsion of their highly charged side-chains is diminished by interaction with ions. The classical "two-component model" of active muscle differentiated a "contractile component" which stretches the "series elastic component" during force production. The contractile components are the helically shaped thin filaments of muscle that shorten the sarcomeres by clockwise drilling into the myosin cross-bridges with torque decrease (= force-deficit). Muscle stretch means drawing out the thin filament helices off the cross-bridges under passive counterclockwise rotation with torque increase (= stretch activation). Since each thin filament is anchored by four elastic alpha-actinin Z-filaments (provided with force-regulating sites for Ca(2+) binding), the thin filament rotations change the torsional twist of the four Z-filaments as the "series elastic components". Large scale models simulate the changes of structure and force in the Z-band by the different Z-filament twisting stages A, B, C, D, E, F and G. Stage D corresponds to the isometric state. The basic phenomena of muscle physiology, i. e. latency relaxation, Fenn-effect, the force-velocity relation, the length-tension relation, unexplained energy, shortening heat, the Huxley-Simmons phases, etc. are explained and interpreted with the help of the model experiments.

Large-scale Models Reveal the Two-component Mechanics of Striated Muscle

PubMed Central

Jarosch, Robert

2008-01-01

This paper provides a comprehensive explanation of striated muscle mechanics and contraction on the basis of filament rotations. Helical proteins, particularly the coiled-coils of tropomyosin, myosin and α-actinin, shorten their H-bonds cooperatively and produce torque and filament rotations when the Coulombic net-charge repulsion of their highly charged side-chains is diminished by interaction with ions. The classical “two-component model” of active muscle differentiated a “contractile component” which stretches the “series elastic component” during force production. The contractile components are the helically shaped thin filaments of muscle that shorten the sarcomeres by clockwise drilling into the myosin cross-bridges with torque decrease (= force-deficit). Muscle stretch means drawing out the thin filament helices off the cross-bridges under passive counterclockwise rotation with torque increase (= stretch activation). Since each thin filament is anchored by four elastic α-actinin Z-filaments (provided with force-regulating sites for Ca2+ binding), the thin filament rotations change the torsional twist of the four Z-filaments as the “series elastic components”. Large scale models simulate the changes of structure and force in the Z-band by the different Z-filament twisting stages A, B, C, D, E, F and G. Stage D corresponds to the isometric state. The basic phenomena of muscle physiology, i. e. latency relaxation, Fenn-effect, the force-velocity relation, the length-tension relation, unexplained energy, shortening heat, the Huxley-Simmons phases, etc. are explained and interpreted with the help of the model experiments. PMID:19330099

Biomolecule detection using wheatstone bridge giant magnetoresistance (GMR) sensors based on CoFeB spin-valve thin film

NASA Astrophysics Data System (ADS)

Elda Swastika, P.; Antarnusa, G.; Suharyadi, E.; Kato, T.; Iwata, S.

2018-04-01

A potential wheatstone bridge giant magnetoresistance (GMR) biosensor have been successfully developed for biomolecule detection. [IrMn(10 nm)/CoFe(3 nm)/Cu(2.2 nm)/CoFeB(10 nm)] spin-valve structure has been chosen as the magnetic sensing surface, showing a magnetoresistance (MR) of 6% fabricated by DC magnetron sputtering method. The Fe3O4 magnetic nanoparticles used as biomolecular labels (nanotags) was synthesized by co-precipitation method, exhibiting soft magnetic behavior with saturation magnetization (Ms), remanent magnetization (Mr) and coercivity (Hc) is 77.2 emu/g, 7.8 emu/g and 51 Oe, respectively. The X-ray diffraction (XRD) patterns and transmission electron microscopy (TEM) images showed that Fe3O4 was well crystallized and grew in their inverse spinel structure, highly uniform morphology with an average grain size was about 20 nm. Fe3O4 was coated with polyethylene-glycol (PEG)-4000 for surface functionalization. Detection of biomolecule such as formalin, gelatin from bovine-skin and porcine-skin were dispersed in ethanol at room temperature. Induction would cause a shift in output voltage with a minimum delta output voltage (ΔV) 4.937 mV (10%) for formalin detection, 2.268 mV (7%) for bovine-skin gelatin and 2.943 mV (7%) for porcine-skin gelatin detection. The ΔV of the wheatstone bridge in real-time measurement decrease by increase in biomolecules concentration. The change of ΔV with various concentration of biomolecule indicates that the spin-valve thin film with wheatstone-bridge circuit is potential as a biosensor.

Carbon-bridged oligo(p-phenylenevinylene)s for photostable and broadly tunable, solution-processable thin film organic lasers

PubMed Central

Morales-Vidal, Marta; Boj, Pedro G.; Villalvilla, José M.; Quintana, José A.; Yan, Qifan; Lin, Nai-Ti; Zhu, Xiaozhang; Ruangsupapichat, Nopporn; Casado, Juan; Tsuji, Hayato; Nakamura, Eiichi; Díaz-García, María A.

2015-01-01

Thin film organic lasers represent a new generation of inexpensive, mechanically flexible devices for spectroscopy, optical communications and sensing. For this purpose, it is desired to develop highly efficient, stable, wavelength-tunable and solution-processable organic laser materials. Here we report that carbon-bridged oligo(p-phenylenevinylene)s serve as optimal materials combining all these properties simultaneously at the level required for applications by demonstrating amplified spontaneous emission and distributed feedback laser devices. A series of six compounds, with the repeating unit from 1 to 6, doped into polystyrene films undergo amplified spontaneous emission from 385 to 585 nm with remarkably low threshold and high net gain coefficients, as well as high photostability. The fabricated lasers show narrow linewidth (<0.13 nm) single mode emission at very low thresholds (0.7 kW cm−2), long operational lifetimes (>105 pump pulses for oligomers with three to six repeating units) and wavelength tunability across the visible spectrum (408–591 nm). PMID:26416643

Role of Al2O3 thin layer on improving the resistive switching properties of Ta5Si3-based conductive bridge random accesses memory device

NASA Astrophysics Data System (ADS)

Kumar, Dayanand; Aluguri, Rakesh; Chand, Umesh; Tseng, Tseung-Yuen

2018-04-01

Ta5Si3-based conductive bridge random access memory (CBRAM) devices have been investigated to improve their resistive switching characteristics for their application in future nonvolatile memory technology. Changes in the switching characteristics by the addition of a thin Al2O3 layer of different thicknesses at the bottom electrode interface of a Ta5Si3-based CBRAM devices have been studied. The double-layer device with a 1 nm Al2O3 layer has shown improved resistive switching characteristics over the single layer one with a high on/off resistance ratio of 102, high endurance of more than 104 cycles, and good retention for more than 105 s at the temperature of 130 °C. The higher thermal conductivity of Al2O3 over Ta5Si3 has been attributed to the enhanced switching properties of the double-layer devices.

A Bottle-around-a-Ship Method To Generate Hollow Thin-Shelled Particles Containing Encapsulated Iron Species with Application to the Environmental Decontamination of Chlorinated Compounds.

PubMed

Su, Yang; Wang, Yingqing; Owoseni, Olasehinde; Zhang, Yueheng; Gamliel, David Pierce; Valla, Julia A; McPherson, Gary L; John, Vijay T

2018-04-25

Thin-shelled hollow silica particles are synthesized using an aerosol-based process where the concentration of a silica precursor tetraethyl orthosilicate (TEOS) determines the shell thickness. The synthesis involves a novel concept of the salt bridging of an iron salt, FeCl 3 , to a cationic surfactant, cetyltrimethylammonium bromide (CTAB), which modulates the templating effect of the surfactant on silica porosity. The salt bridging leads to a sequestration of the surfactant in the interior of the droplet with the formation of a dense silica shell around the organic material. Subsequent calcination consistently results in hollow particles with encapsulated iron oxides. Control of the TEOS levels leads to the generation of ultrathin-shelled (∼10 nm) particles which become susceptible to rupture upon exposure to ultrasound. The dense silica shell that is formed is impervious to entry of chemical species. Mesoporosity is restored to the shell through desilication and reassembly, again using CTAB as a template. The mesoporous-shelled hollow particles show good reactivity toward the reductive dichlorination of trichloroethylene (TCE), indicating access of TCE to the particle interior. The ordered mesoporous thin-shelled particles containing active iron species are viable systems for chemical reaction and catalysis.

Human cardiac telocytes: 3D imaging by FIB-SEM tomography.

PubMed

Cretoiu, D; Hummel, E; Zimmermann, H; Gherghiceanu, M; Popescu, L M

2014-11-01

Telocyte (TC) is a newly identified type of cell in the cardiac interstitium (www.telocytes.com). TCs are described by classical transmission electron microscopy as cells with very thin and long telopodes (Tps; cellular prolongations) having podoms (dilations) and podomers (very thin segments). TCs' three-dimensional (3D) morphology is still unknown. Cardiac TCs seem to be particularly involved in long and short distance intercellular signalling and, therefore, their 3D architecture is important for understanding their spatial connections. Using focused ion beam scanning electron microscopy (FIB-SEM) we show, for the first time, the whole ultrastructural anatomy of cardiac TCs. 3D reconstruction of cardiac TCs by FIB-SEM tomography confirms that they have long, narrow but flattened (ribbon-like) telopodes, with humps generated by the podoms. FIB-SEM tomography also confirms the network made by TCs in the cardiac interstitium through adherens junctions. This study provides the first FIB-SEM tomography of a human cell type. © 2014 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Blood-Borne Macrophage-Neural Cell Interactions Hitchhike Endosome Networks for Cell-Based Nanozyme Brain Delivery

PubMed Central

Haney, Matthew J.; Suresh, Poornima; Zhao, Yuling; Kanmogne, Georgette D.; Kadiu, Irena; Sokolsky-Papkov, Marina; Klyachko, Natalia L.; Mosley, R. Lee; Kabanov, Alexander V.; Gendelman, Howard E.; Batrakova, Elena V.

2012-01-01

Background Macrophage carried nanoformulated catalase (“nanozyme”) attenuates neuroinflammation and protects nigrostriatal neurons from 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine intoxication. This is facilitated by effective enzyme transfer from blood borne macrophages to adjacent endothelial cells and neurons leading to the decomposition of reactive oxygen species. Methods We now examine the intra- and intercellular trafficking mechanisms of nanozymes. Results In macrophages, nanozymes are internalized mainly by clathrin mediated endocytosis then traffic to recycling endosomes. The enzyme is subsequently released in exosomes facilitated by bridging conduits. Nanozyme transfer from macrophages to adjacent cells by endocytosis-independent mechanisms diffusing broadly throughout the recipient cells. In contrast, macrophage-free nanozymes are localized in lysosomes following endocytic entry. Conclusion Facilitated transfer of nanozyme from cell to cell can improve neuroprotection against oxidative stress commonly seen during neurodegenerative disease processes. PMID:22236307

Review article: mitogen-activated protein kinases in chronic intestinal inflammation - targeting ancient pathways to treat modern diseases.

PubMed

Waetzig, G H; Schreiber, S

2003-07-01

Conventional treatment of chronic inflammatory disorders, including inflammatory bowel diseases, employs broad-range anti-inflammatory drugs. In order to reduce the side-effects and increase the efficacy of treatment, several strategies have been developed in the last decade to interfere with intercellular and intracellular inflammatory signalling processes. The highly conserved mitogen-activated protein kinase pathways regulate most cellular processes, particularly defence mechanisms such as stress reactions and inflammation. In this review, we provide an overview of the current knowledge of the specificity and interconnection of mitogen-activated protein kinase pathways, their functions in the gut immune system and published and ongoing studies on the role of mitogen-activated protein kinases in inflammatory bowel disease. The development of mitogen-activated protein kinase inhibitors and their use for the therapy of inflammatory disorders is a paradigm of the successful bridging of the gap between basic research and clinical practice.

Ectopic decidua and metastatic squamous carcinoma: presentation in a single pelvic lymph node.

PubMed

Cobb, C J

1988-06-01

The presence of ectopic decidua in pelvic lymph nodes from patients with squamous carcinoma of the cervix makes evaluation for metastatic disease difficult due to the light microscopic similarity between decidua and sheets of squamous epithelial cells. A patient is present in whom decidualized endometriosis was intimately associated with metastatic moderately differentiate squamous carcinoma in a single pelvic lymph node. This phenomenon afforded an excellent opportunity to study the unique morphologic features that distinguish these two entities. A prior report of this kind was not found. In the absence of obvious squamous differentiation (i.e., intercellular bridges, dyskeratosis, and keratin "pearl" formation), as is frequently the case with squamous carcinoma of the cervix, the light microscopic features that are most useful in distinguishing squamous carcinoma from decidua include the presence of well-defined nests of cohesive cells, nuclear hyperchromasia, and cellular pleomorphism.

Keratin-lipid structural organization in the corneous layer of snake.

PubMed

Ripamonti, Alberto; Alibardi, Lorenzo; Falini, Giuseppe; Fermani, Simona; Gazzano, Massimo

2009-12-01

The shed epidermis (molt) of snakes comprises four distinct layers. The upper two layers, here considered as beta-layer, contain essentially beta-keratin. The following layer, known as mesos-layer, is similar to the human stratum corneum, and is formed by thin cells surrounded by intercellular lipids. The latter layer mainly contains alpha-keratin. In this study, the molecular assemblies of proteins and lipids contained in these layers have been analyzed in the scale of two species of snakes, the elapid Tiger snake (TS, Notechis scutatus) and the viperid Gabon viper (GV, Bitis gabonica). Scanning X-ray micro-diffraction, FTIR and Raman spectroscopies, thermal analysis, and scanning electron microscopy experiments confirm the presence of the three layers in the GV skin scale. Conversely, in the TS molt a typical alpha-keratin layer appears to be absent. In the latter, experimental data suggest the presence of two domains similar to those found in the lipid intercellular matrix of stratum corneum. X-ray diffraction data also allow to determine the relative orientation of keratins and lipids. The keratin fibrils are randomly oriented inside the layers parallel to the surface of scales while the lipids are organized in lamellar structures having aliphatic chains normal to the scale surface. The high ordered lipid organization in the mature mesos layer probably increases its effectiveness in limiting water-loss.

Electronic modules easily separated from heat sink

NASA Technical Reports Server (NTRS)

1965-01-01

Metal heat sink and electronic modules bonded to a thermal bridge can be easily cleaved for removal of the modules for replacement or repair. A thin film of grease between a fluorocarbon polymer film on the metal heat sink and an adhesive film on the modules acts as the cleavage plane.

Method and system for making integrated solid-state fire-sets and detonators

DOEpatents

O'Brien, Dennis W.; Druce, Robert L.; Johnson, Gary W.; Vogtlin, George E.; Barbee, Jr., Troy W.; Lee, Ronald S.

1998-01-01

A slapper detonator comprises a solid-state high-voltage capacitor, a low-jitter dielectric breakdown switch and trigger circuitry, a detonator transmission line, an exploding foil bridge, and a flier material. All these components are fabricated in a single solid-state device using thin film deposition techniques.

Surface-geophysical techniques used to detect existing and infilled scour holes near bridge piers

USGS Publications Warehouse

Placzek, Gary; Haeni, F.P.

1995-01-01

Surface-geophysical techniques were used with a position-recording system to study riverbed scour near bridge piers. From May 1989 to May 1993. Fathometers, fixed- and swept-frequency con- tinuous seismic-reflection profiling (CSP) systems, and a ground-penetrating radar (GPR) system were used with a laser-positioning system to measure the depth and extent of existing and infilled scour holes near bridge piers. Equipment was purchased commercially and modified when necessary to interface the components and (or) to improve their performance. Three 200-kHz black-and-white chart- recording Fathometers produced profiles of the riverbed that included existing scour holes and exposed pier footings. The Fathometers were used in conjunction with other geophysical techniques to help interpret the geophysical data. A 20-kHz color Fathometer delineated scour-hole geometry and, in some cases, the thickness of fill material in the hole. The signal provided subbottom information as deep as 10 ft in fine-grained materials and resolved layers of fill material as thin as 1 foot thick. Fixed-frequency and swept-frequency CSP systems were evaluated. The fixed-frequency system used a 3.5-, 7.0-, or 14-kHz signal. The 3.5-kHz signal pene- trated up to 50 ft of fine-grained material and resolved layers as thin as 2.5-ft thick. The 14-kHz signal penetrated up to 20 ft of fine-grained material and resolved layers as thin as 1-ft thick. The swept-frequency systems used a signal that swept from 2- to 16-kHz. With this system, up to 50 ft of penetration was achieved, and fill material as thin as 1 ft was resolved. Scour-hole geometry, exposed pier footings, and fill thickness in scour holes were detected with both CSP systems. The GPR system used an 80-, 100-, or 300-megahertz signal. The technique produced records in water up to 15 ft deep that had a specific conductance less than 200x11ms/cm. The 100-MHz signal penetrated up to 40 ft of resistive granular material and resolved layers as thin as 2-ft thick. Scour-hole geometry, the thickness of fill material in scour holes, and riverbed deposition were detected using this technique. Processing techniques were applied after data collection to assist with the interpretation of the data. Data were transferred from the color Fathometer, CSP, and GPR systems to a personal computer, and a commercially available software package designed to process GPR data was used to process the GPR and CSP data. Digital filtering, predictive-deconvolution, and migration algorithms were applied to some of the data. The processed data were displayed and printed as color amplitude or wiggle-trace plots. These processing methods eased and improved the interpretation of some of the data, but some interference from side echoes from bridge piers and multiple reflections remained in the data. The surface-geophysical techniques were applied at six bridge sites in Connecticut. Each site had different water depths, specific conductance, and riverbed materials. Existing and infilled scour holes, exposed pier footings, and riverbed deposition were detected by the surveys. The interpretations of the geophysical data were confirmed by comparing the data with lithologic and (or) probing data.

Tunneling from the past horizon

NASA Astrophysics Data System (ADS)

Kang, Subeom; Yeom, Dong-han

2018-04-01

We investigate a tunneling and emission process of a thin-shell from a Schwarzschild black hole, where the shell was initially located beyond the Einstein-Rosen bridge and finally appears at the right side of the Penrose diagram. In order to obtain such a solution, we should assume that the areal radius of the black hole horizon increases after the tunneling. Hence, there is a parameter range such that the tunneling rate is exponentially enhanced, rather than suppressed. We may have two interpretations regarding this. First, such a tunneling process from the past horizon is improbable by physical reasons; second, such a tunneling is possible in principle, but in order to obtain a stable Einstein-Rosen bridge, one needs to restrict the parameter spaces. If such a process is allowed, this can be a nonperturbative contribution to Einstein-Rosen bridges as well as eternal black holes.

Maintenance of Air in Intercellular Spaces of Plants

PubMed Central

Woolley, Joseph T.

1983-01-01

Although air-filled intercellular spaces are necessary and ubiquitous in higher plants, little attention has been paid to the possible mechanisms by which these spaces are kept from being flooded. The most likely mechanism is that the living plant cell may maintain a hydrophobic monolayer on the surfaces of adjacent intercellular spaces. The existence of `apparent free space' in cell walls and the fact that detergent solutions do not enter the intercellular spaces argue against this hypothesis. It is concluded that the mechanism by which these important air spaces are maintained is still unknown. Images Fig. 1 Fig. 2 PMID:16663150

Release of Applied Mechanical Loading Stimulates Intercellular Calcium Waves in Drosophila Wing Discs.

PubMed

Narciso, Cody E; Contento, Nicholas M; Storey, Thomas J; Hoelzle, David J; Zartman, Jeremiah J

2017-07-25

Mechanical forces are critical but poorly understood inputs for organogenesis and wound healing. Calcium ions (Ca 2+ ) are critical second messengers in cells for integrating environmental and mechanical cues, but the regulation of Ca 2+ signaling is poorly understood in developing epithelial tissues. Here we report a chip-based regulated environment for microorgans that enables systematic investigations of the crosstalk between an organ's mechanical stress environment and biochemical signaling under genetic and chemical perturbations. This method enabled us to define the essential conditions for generating organ-scale intercellular Ca 2+ waves in Drosophila wing discs that are also observed in vivo during organ development. We discovered that mechanically induced intercellular Ca 2+ waves require fly extract growth serum as a chemical stimulus. Using the chip-based regulated environment for microorgans, we demonstrate that not the initial application but instead the release of mechanical loading is sufficient, but not necessary, to initiate intercellular Ca 2+ waves. The Ca 2+ response depends on the prestress intercellular Ca 2+ activity and not on the magnitude or duration of the mechanical stimulation applied. Mechanically induced intercellular Ca 2+ waves rely on IP 3 R-mediated Ca 2+ -induced Ca 2+ release and propagation through gap junctions. Thus, intercellular Ca 2+ waves in developing epithelia may be a consequence of stress dissipation during organ growth. Copyright © 2017 Biophysical Society. Published by Elsevier Inc. All rights reserved.

INHIBITION OF GAP JUNCTIONAL INTERCELLULAR COMMUNICATION BY PERFLUORINATED COMPOUNDS IN RAT LIVER AND DOLPHIN KIDNEY EPITHELIAL CELL LINES IN VITRO AND SPRAGUE-DAWLEY RATS IN VIVO

EPA Science Inventory

Abstract

Gap Junctional Intercellular Communication (GJIC) is the major pathway of intercellular signal transduction, and is, thus, important for normal cell growth and function. Recent studies have revealed a global distribution of some perfluorinated organic compounds e...

The vascular plant-pathogenic bacterium Ralstonia solanacearum produces biofilms required for its virulence on the surfaces of tomato cells adjacent to intercellular spaces.

PubMed

Mori, Yuka; Inoue, Kanako; Ikeda, Kenichi; Nakayashiki, Hitoshi; Higashimoto, Chikaki; Ohnishi, Kouhei; Kiba, Akinori; Hikichi, Yasufumi

2016-08-01

The mechanism of colonization of intercellular spaces by the soil-borne and vascular plant-pathogenic bacterium Ralstonia solanacearum strain OE1-1 after invasion into host plants remains unclear. To analyse the behaviour of OE1-1 cells in intercellular spaces, tomato leaves with the lower epidermis layers excised after infiltration with OE1-1 were observed under a scanning electron microscope. OE1-1 cells formed microcolonies on the surfaces of tomato cells adjacent to intercellular spaces, and then aggregated surrounded by an extracellular matrix, forming mature biofilm structures. Furthermore, OE1-1 cells produced mushroom-type biofilms when incubated in fluids of apoplasts including intercellular spaces, but not xylem fluids from tomato plants. This is the first report of biofilm formation by R. solanacearum on host plant cells after invasion into intercellular spaces and mushroom-type biofilms produced by R. solanacearum in vitro. Sugar application led to enhanced biofilm formation by OE1-1. Mutation of lecM encoding a lectin, RS-IIL, which reportedly exhibits affinity for these sugars, led to a significant decrease in biofilm formation. Colonization in intercellular spaces was significantly decreased in the lecM mutant, leading to a loss of virulence on tomato plants. Complementation of the lecM mutant with native lecM resulted in the recovery of mushroom-type biofilms and virulence on tomato plants. Together, our findings indicate that OE1-1 produces mature biofilms on the surfaces of tomato cells after invasion into intercellular spaces. RS-IIL may contribute to biofilm formation by OE1-1, which is required for OE1-1 virulence. © 2015 BSPP AND JOHN WILEY & SONS LTD.

Oesophageal mucosal intercellular space diameter and reflux pattern in childhood erosive and non-erosive reflux disease.

PubMed

Mancini, Valentina; Ribolsi, Mentore; Gentile, Massimo; de'Angelis, Gianluigi; Bizzarri, Barbara; Lindley, Keith J; Cucchiara, Salvatore; Cicala, Michele; Borrelli, Osvaldo

2012-12-01

We sought to compare intercellular space diameter in children with non-erosive and erosive reflux disease, and a control group. We also aimed to characterize the reflux pattern in erosive and non-erosive reflux disease patients, and to explore the relationship between intercellular space diameter values and reflux parameters. Twenty-four children with non-erosive reflux disease, 20 with erosive reflux disease, and 10 controls were prospectively studied. All patients and controls underwent upper endoscopy. Biopsies were taken at 2-3 cm above the Z-line, and intercellular space diameter was measured using transmission electron microscopy. Non-erosive and erosive reflux disease patients underwent impedance-pH monitoring. Mean intercellular space diameter values were significantly higher in both non-erosive (0.9 ± 0.2 μm) and erosive reflux disease (1 ± 0.2 μm) compared to controls (0.5 ± 0.2 μm, p<0.01). No difference was found between the two patient groups. Acid exposure time, the number of acid, weakly acidic and weakly alkaline reflux events did not differ between the two patient groups. No difference was found in the mean intercellular space diameter between non-erosive reflux disease children with and without abnormal acid exposure time (1 ± 0.3 vs. 0.9 ± 0.2 μm). No correlation was found between any reflux parameter and intercellular space diameter values. Dilated intercellular space diameter seems to be a useful and objective marker of oesophageal damage in paediatric gastro-oesophageal reflux disease, regardless of acid exposure. In childhood, different gastro-oesophageal reflux disease phenotypes cannot be discriminated on the basis of reflux pattern. Copyright © 2012 Editrice Gastroenterologica Italiana S.r.l. Published by Elsevier Ltd. All rights reserved.

Intercellular and systemic spread of RNA and RNAi in plants.

PubMed

Nazim Uddin, Mohammad; Kim, Jae-Yean

2013-01-01

Plants possess dynamic networks of intercellular communication that are crucial for plant development and physiology. In plants, intercellular communication involves a combination of ligand-receptor-based apoplasmic signaling, and plasmodesmata and phloem-mediated symplasmic signaling. The intercellular trafficking of macromolecules, including RNAs and proteins, has emerged as a novel mechanism of intercellular communication in plants. Various forms of regulatory RNAs move over distinct cellular boundaries through plasmodesmata and phloem. This plant-specific, non-cell-autonomous RNA trafficking network is also involved in development, nutrient homeostasis, gene silencing, pathogen defense, and many other physiological processes. However, the mechanism underlying macromolecular trafficking in plants remains poorly understood. Current progress made in RNA trafficking research and its biological relevance to plant development will be summarized. Diverse plant regulatory mechanisms of cell-to-cell and systemic long-distance transport of RNAs, including mRNAs, viral RNAs, and small RNAs, will also be discussed. Copyright © 2013 John Wiley & Sons, Ltd.

Estimating intercellular surface tension by laser-induced cell fusion.

PubMed

Fujita, Masashi; Onami, Shuichi

2011-12-01

Intercellular surface tension is a key variable in understanding cellular mechanics. However, conventional methods are not well suited for measuring the absolute magnitude of intercellular surface tension because these methods require determination of the effective viscosity of the whole cell, a quantity that is difficult to measure. In this study, we present a novel method for estimating the intercellular surface tension at single-cell resolution. This method exploits the cytoplasmic flow that accompanies laser-induced cell fusion when the pressure difference between cells is large. Because the cytoplasmic viscosity can be measured using well-established technology, this method can be used to estimate the absolute magnitudes of tension. We applied this method to two-cell-stage embryos of the nematode Caenorhabditis elegans and estimated the intercellular surface tension to be in the 30-90 µN m(-1) range. Our estimate was in close agreement with cell-medium surface tensions measured at single-cell resolution.

Effects of fabrication procedures and weld melt-through on fatigue resistance of orthotropic steel deck welds.

DOT National Transportation Integrated Search

2008-12-01

A common practice for the fabrication of orthotropic bridge deck in the US involves using 80% partial-joint-penetration groove welds (PJP) to join : closed ribs to a deck plate. Avoiding weld melt-through with the thin rib plate may be difficult to a...

Method and system for making integrated solid-state fire-sets and detonators

DOEpatents

O`Brien, D.W.; Druce, R.L.; Johnson, G.W.; Vogtlin, G.E.; Barbee, T.W. Jr.; Lee, R.S.

1998-03-24

A slapper detonator comprises a solid-state high-voltage capacitor, a low-jitter dielectric breakdown switch and trigger circuitry, a detonator transmission line, an exploding foil bridge, and a flier material. All these components are fabricated in a single solid-state device using thin film deposition techniques. 13 figs.

Epitaxial growth of (001)-oriented Ba{sub 0.5}Sr{sub 0.5}TiO{sub 3} thin films on a-plane sapphire with an MgO/ZnO bridge layer

DOE Office of Scientific and Technical Information (OSTI.GOV)

Xiao Bo; Liu Hongrui; Avrutin, Vitaliy

2009-11-23

High quality (001)-oriented Ba{sub 0.5}Sr{sub 0.5}TiO{sub 3} (BST) thin films have been grown on a-plane sapphire (1120) by rf magnetron sputtering using a double bridge layer consisting of (0001)-oriented ZnO (50 nm) and (001)-oriented MgO (10 nm) prepared by plasma-assisted molecular beam epitaxy. X-ray diffraction revealed the formation of three sets of in-plane BST domains, offset from one another by 30 deg., which is consistent with the in-plane symmetry of the MgO layer observed by in situ reflective high electron energy diffraction. The in-plane epitaxial relationship of BST, MgO, and ZnO has been determined to be BST [110]//MgO [110]//ZnO [1120]more » and BST [110]/MgO [110]//ZnO [1100]. Capacitance-voltage measurements performed on BST coplanar interdigitated capacitor structures revealed a high dielectric tunability of up to 84% at 1 MHz.« less

Mechanical Properties of Respiratory Muscles

PubMed Central

Sieck, Gary C.; Ferreira, Leonardo F.; Reid, Michael B.; Mantilla, Carlos B.

2014-01-01

Striated respiratory muscles are necessary for lung ventilation and to maintain the patency of the upper airway. The basic structural and functional properties of respiratory muscles are similar to those of other striated muscles (both skeletal and cardiac). The sarcomere is the fundamental organizational unit of striated muscles and sarcomeric proteins underlie the passive and active mechanical properties of muscle fibers. In this respect, the functional categorization of different fiber types provides a conceptual framework to understand the physiological properties of respiratory muscles. Within the sarcomere, the interaction between the thick and thin filaments at the level of cross-bridges provides the elementary unit of force generation and contraction. Key to an understanding of the unique functional differences across muscle fiber types are differences in cross-bridge recruitment and cycling that relate to the expression of different myosin heavy chain isoforms in the thick filament. The active mechanical properties of muscle fibers are characterized by the relationship between myoplasmic Ca2+ and cross-bridge recruitment, force generation and sarcomere length (also cross-bridge recruitment), external load and shortening velocity (cross-bridge cycling rate), and cross-bridge cycling rate and ATP consumption. Passive mechanical properties are also important reflecting viscoelastic elements within sarcomeres as well as the extracellular matrix. Conditions that affect respiratory muscle performance may have a range of underlying pathophysiological causes, but their manifestations will depend on their impact on these basic elemental structures. PMID:24265238

Innexin-3 forms connexin-like intercellular channels.

PubMed

Landesman, Y; White, T W; Starich, T A; Shaw, J E; Goodenough, D A; Paul, D L

1999-07-01

Innexins comprise a large family of genes that are believed to encode invertebrate gap junction channel-forming proteins. However, only two Drosophila innexins have been directly tested for the ability to form intercellular channels and only one of those was active. Here we tested the ability of Caenorhabditis elegans family members INX-3 and EAT-5 to form intercellular channels between paired Xenopus oocytes. We show that expression of INX-3 but not EAT-5, induces electrical coupling between the oocyte pairs. In addition, analysis of INX-3 voltage and pH gating reveals a striking degree of conservation in the functional properties of connexin and innnexin channels. These data strongly support the idea that innexin genes encode intercellular channels.

Intercellular ultrafast Ca2+ wave in vascular smooth muscle cells: numerical and experimental study

NASA Astrophysics Data System (ADS)

Quijano, J. C.; Raynaud, F.; Nguyen, D.; Piacentini, N.; Meister, J. J.

2016-08-01

Vascular smooth muscle cells exhibit intercellular Ca2+ waves in response to local mechanical or KCl stimulation. Recently, a new type of intercellular Ca2+ wave was observed in vitro in a linear arrangement of smooth muscle cells. The intercellular wave was denominated ultrafast Ca2+ wave and it was suggested to be the result of the interplay between membrane potential and Ca2+ dynamics which depended on influx of extracellular Ca2+, cell membrane depolarization and its intercel- lular propagation. In the present study we measured experimentally the conduction velocity of the membrane depolarization and performed simulations of the ultrafast Ca2+ wave along coupled smooth muscle cells. Numerical results reproduced a wide spectrum of experimental observations, including Ca2+ wave velocity, electrotonic membrane depolarization along the network, effects of inhibitors and independence of the Ca2+ wave speed on the intracellular stores. The numerical data also provided new physiological insights suggesting ranges of crucial model parameters that may be altered experimentally and that could significantly affect wave kinetics allowing the modulation of the wave characteristics experimentally. Numerical and experimental results supported the hypothesis that the propagation of membrane depolarization acts as an intercellular messenger mediating intercellular ultrafast Ca2+ waves in smooth muscle cells.

A Bridge between Two Important Problems in Optics and Electrostatics

ERIC Educational Resources Information Center

Capelli, R.; Pozzi, G.

2008-01-01

It is shown how the same physically appealing method can be applied to find analytic solutions for two difficult and apparently unrelated problems in optics and electrostatics. They are: (i) the diffraction of a plane wave at a perfectly conducting thin half-plane and (ii) the electrostatic field associated with a parallel array of stripes held at…

Tunneling Nanotubes as a Novel Route of Cell-to-Cell Spread of Herpesviruses.

PubMed

Panasiuk, Mirosława; Rychłowski, Michał; Derewońko, Natalia; Bieńkowska-Szewczyk, Krystyna

2018-05-15

Various types of intercellular connections that are essential for communication between cells are often utilized by pathogens. Recently, a new type of cellular connection, consisting of long, thin, actin-rich membrane extensions named tunneling nanotubes (TNTs), has been shown to play an important role in cell-to-cell spread of HIV and influenza virus. In the present report, we show that TNTs are frequently formed by cells infected by an alphaherpesvirus, bovine herpesvirus 1 (BoHV-1). Viral proteins, such as envelope glycoprotein E (gE), capsid protein VP26, and tegument protein Us3, as well as cellular organelles (mitochondria) were detected by immunofluorescence and live-cell imaging of nanotubes formed by bovine primary fibroblasts and oropharynx cells (KOP cells). Time-lapse confocal studies of live cells infected with fluorescently labeled viruses showed that viral particles were transmitted via TNTs. This transfer also occurred in the presence of neutralizing antibodies, which prevented free entry of BoHV-1. We conclude that TNT formation contributes to successful cell-to-cell spread of BoHV-1 and demonstrate for the first time the participation of membrane nanotubes in intercellular transfer of a herpesvirus in live cells. IMPORTANCE Efficient transmission of viral particles between cells is an important factor in successful infection by herpesviruses. Herpesviruses can spread by the free-entry mode or direct cell-to-cell transfer via cell junctions and long extensions of neuronal cells. In this report, we show for the first time that an alphaherpesvirus can also spread between various types of cells using tunneling nanotubes, intercellular connections that are utilized by HIV and other viruses. Live-cell monitoring revealed that viral transmission occurs between the cells of the same type as well as between epithelial cells and fibroblasts. This newly discovered route of herpesviruses spread may contribute to efficient transmission despite the presence of host immune responses, especially after reactivation from latency that developed after primary infection. Long-range communication provided by TNTs may facilitate the spread of herpesviruses between many tissues and organs of an infected organism. Copyright © 2018 American Society for Microbiology.

Characterization of ultrathin SOI film and application to short channel MOSFETs.

PubMed

Tang, Xiaohui; Reckinger, Nicolas; Larrieu, Guilhem; Dubois, Emmanuel; Flandre, Denis; Raskin, Jean-Pierre; Nysten, Bernard; Jonas, Alain M; Bayot, Vincent

2008-04-23

In this study, a very dilute solution (NH(4)OH:H(2)O(2):H(2)O 1:8:64 mixture) was employed to reduce the thickness of commercially available SOI wafers down to 3 nm. The etch rate is precisely controlled at 0.11 Å s(-1) based on the self-limited etching speed of the solution. The thickness uniformity of the thin film, evaluated by spectroscopic ellipsometry and by high-resolution x-ray reflectivity, remains constant through the thinning process. Moreover, the film roughness, analyzed by atomic force microscopy, slightly improves during the thinning process. The residual stress in the thin film is much smaller than that obtained by sacrificial oxidation. Mobility, measured by means of a bridge-type Hall bar on 15 nm film, is not significantly reduced compared to the value of bulk silicon. Finally, the thinned SOI wafers were used to fabricate Schottky-barrier metal-oxide-semiconductor field-effect transistors with a gate length down to 30 nm, featuring state-of-the-art current drive performance.

Characteristics research of pressure sensor based on nanopolysilicon thin films resistors

NASA Astrophysics Data System (ADS)

Zhao, Xiaofeng; Li, Dandan; Wen, Dianzhong

2017-10-01

To further improve the sensitivity temperature characteristics of pressure sensor, a kind of pressure sensor taking nanopolysilicon thin films as piezoresistors is proposed in this paper. On the basis of the microstructure analysis by X-ray diffraction (XRD) and scanning electron microscope (SEM) tests, the preparing process of nanopolysilicon thin films is optimized. The effects of film thickness and annealing temperature on the micro-structure of nanopolysilicon thin films were studied, respectively. In order to realize the measurement of external pressure, four nanopolysilicon thin films resistors were arranged at the edges of square silicon diaphragm connected to a Wheatstone bridge, and the chip of the sensor was designed and fabricated on a 〈100〉 orientation silicon wafer by microelectromechanical system (MEMS) technology. Experimental result shows that when I = 6.80 mA, the sensitivity of the sensor PS-1 is 0.308 mV/kPa, and the temperature coefficient of sensitivity (TCS) is about -1742 ppm/∘C in the range of -40-140∘C. It is possible to obviously improve the sensitivity temperature characteristics of pressure sensor by the proposed sensors.

Disproportionate loss of thin filaments in human soleus muscle after 17-day bed rest

NASA Technical Reports Server (NTRS)

Riley, D. A.; Bain, J. L.; Thompson, J. L.; Fitts, R. H.; Widrick, J. J.; Trappe, S. W.; Trappe, T. A.; Costill, D. L.

1998-01-01

Previously we reported that, after 17-day bed rest unloading of 8 humans, soleus slow fibers atrophied and exhibited increased velocity of shortening without fast myosin expression. The present ultrastructural study examined fibers from the same muscle biopsies to determine whether decreased myofilament packing density accounted for the observed speeding. Quantitation was by computer-assisted morphometry of electron micrographs. Filament densities were normalized for sarcomere length, because density depends directly on length. Thick filament density was unchanged by bed rest. Thin filaments/microm2 decreased 16-23%. Glycogen filled the I band sites vacated by filaments. The percentage decrease in thin filaments (Y) correlated significantly (P < 0.05) with the percentage increase in velocity (X), (Y = 0.1X + 20%, R2 = 0.62). An interpretation is that fewer filaments increases thick to thin filament spacing and causes earlier cross-bridge detachment and faster cycling. Increased velocity helps maintain power (force x velocity) as atrophy lowers force. Atrophic muscles may be prone to sarcomere reloading damage because force/microm2 was near normal, and force per thin filament increased an estimated 30%.

Single crystalline thin films as a novel class of electrocatalysts

DOE PAGES

Snyder, Joshua; Markovic, Nenad; Stamenkovic, Vojislav

2013-01-01

The ubiquitous use of single crystal metal electrodes has garnered invaluable insight into the relationship between surface atomic structure and functional electrochemical properties. But, the sensitivity of their electrochemical response to surface orientation and the amount of precious metal required can limit their use. We present here a generally applicable procedure for producing thin metal films with a large proportion of atomically flat (111) terraces without the use of an epitaxial template. Thermal annealing in a controlled atmosphere induces long-range ordering of magnetron sputtered thin metal films deposited on an amorphous substrate. The ordering transition in these thin metal filmsmore » yields characteristic (111) electrochemical signatures with minimal amount of material and provides an adequate replacement for oriented bulk single crystals. Our procedure can be generalized towards a novel class of practical multimetallic thin film based electrocatalysts with tunable near-surface compositional profile and morphology. Annealing of atomically corrugated sputtered thin film Pt-alloy catalysts yields an atomically smooth structure with highly crystalline, (111)-like ordered and Pt segregated surface that displays superior functional properties, bridging the gap between extended/bulk surfaces and nanoscale systems.« less

Theory and practical considerations of multilayer dielectric thin-film stacks in Ag-coated hollow waveguides.

PubMed

Bledt, Carlos M; Melzer, Jeffrey E; Harrington, James A

2014-02-01

This analysis explores the theory and design of dielectric multilayer reflection-enhancing thin film stacks based on high and low refractive index alternating layers of cadmium sulfide (CdS) and lead sulfide (PbS) on silver (Ag)-coated hollow glass waveguides (HGWs) for low loss transmission at midinfrared wavelengths. The fundamentals for determining propagation losses in such multilayer thin-film-coated Ag hollow waveguides is thoroughly discussed, and forms the basis for further theoretical analysis presented in this study. The effects on propagation loss resulting from several key parameters of these multilayer thin film stacks is further explored in order to bridge the gap between results predicted through calculation under ideal conditions and deviations from such ideal models that often arise in practice. In particular, the effects on loss due to the number of dielectric thin film layers deposited, deviation from ideal individual layer thicknesses, and surface roughness related scattering losses are presented and thoroughly investigated. Through such extensive theoretical analysis the level of understanding of the underlying loss mechanisms of multilayer thin-film Ag-coated HGWs is greatly advanced, considerably increasing the potential practical development of next-generation ultralow-loss mid-IR Ag/multilayer dielectric-coated HGWs.

Effect of Ag doping and annealing on thermoelectric properties of PbTe

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bala, Manju, E-mail: Manjubala474@gmail.com; Tripathi, T. S.; Avasthi, D. K.

2015-06-24

The present study reveals that annealing Ag doped PbTe thin films enhance thermoelectric properties. Phase formation was identified by using X-ray diffraction measurement. Annealing increases the crystallinity of both undoped and Ag doped PbTe. Electrical resistivity and thermoelectric power measurements are done using four probe and bridge method respectively. The increase in thermoelectric power of Ag doped PbTe is 29 % in comparison to undoped PbTe and it further increases to 34 % after annealing at 250{sup o} C for 1 hour whereas thermoelectric power increases by 14 % on annealing undoped PbTe thin films at same temperature.

Tunneling nanotube (TNT)-mediated neuron-to neuron transfer of pathological Tau protein assemblies.

PubMed

Tardivel, Meryem; Bégard, Séverine; Bousset, Luc; Dujardin, Simon; Coens, Audrey; Melki, Ronald; Buée, Luc; Colin, Morvane

2016-11-04

A given cell makes exchanges with its neighbors through a variety of means ranging from diffusible factors to vesicles. Cells use also tunneling nanotubes (TNTs), filamentous-actin-containing membranous structures that bridge and connect cells. First described in immune cells, TNTs facilitate HIV-1 transfer and are found in various cell types, including neurons. We show that the microtubule-associated protein Tau, a key player in Alzheimer's disease, is a bona fide constituent of TNTs. This is important because Tau appears beside filamentous actin and myosin 10 as a specific marker of these fine protrusions of membranes and cytosol that are difficult to visualize. Furthermore, we observed that exogenous Tau species increase the number of TNTs established between primary neurons, thereby facilitating the intercellular transfer of Tau fibrils. In conclusion, Tau may contribute to the formation and function of the highly dynamic TNTs that may be involved in the prion-like propagation of Tau assemblies.

Oxidation of F-actin controls the terminal steps of cytokinesis

PubMed Central

Frémont, Stéphane; Hammich, Hussein; Bai, Jian; Wioland, Hugo; Klinkert, Kerstin; Rocancourt, Murielle; Kikuti, Carlos; Stroebel, David; Romet-Lemonne, Guillaume; Pylypenko, Olena; Houdusse, Anne; Echard, Arnaud

2017-01-01

Cytokinetic abscission, the terminal step of cell division, crucially depends on the local constriction of ESCRT-III helices after cytoskeleton disassembly. While the microtubules of the intercellular bridge are cut by the ESCRT-associated enzyme Spastin, the mechanism that clears F-actin at the abscission site is unknown. Here we show that oxidation-mediated depolymerization of actin by the redox enzyme MICAL1 is key for ESCRT-III recruitment and successful abscission. MICAL1 is recruited to the abscission site by the Rab35 GTPase through a direct interaction with a flat three-helix domain found in MICAL1 C terminus. Mechanistically, in vitro assays on single actin filaments demonstrate that MICAL1 is activated by Rab35. Moreover, in our experimental conditions, MICAL1 does not act as a severing enzyme, as initially thought, but instead induces F-actin depolymerization from both ends. Our work reveals an unexpected role for oxidoreduction in triggering local actin depolymerization to control a fundamental step of cell division. PMID:28230050

Activation of acid-sensing ion channels by localized proton transient reveals their role in proton signaling.

PubMed

Zeng, Wei-Zheng; Liu, Di-Shi; Liu, Lu; She, Liang; Wu, Long-Jun; Xu, Tian-Le

2015-09-15

Extracellular transients of pH alterations likely mediate signal transduction in the nervous system. Neuronal acid-sensing ion channels (ASICs) act as sensors for extracellular protons, but the mechanism underlying ASIC activation remains largely unknown. Here, we show that, following activation of a light-activated proton pump, Archaerhodopsin-3 (Arch), proton transients induced ASIC currents in both neurons and HEK293T cells co-expressing ASIC1a channels. Using chimera proteins that bridge Arch and ASIC1a by a glycine/serine linker, we found that successful coupling occurred within 15 nm distance. Furthermore, two-cell sniffer patch recording revealed that regulated release of protons through either Arch or voltage-gated proton channel Hv1 activated neighbouring cells expressing ASIC1a channels. Finally, computational modelling predicted the peak proton concentration at the intercellular interface to be at pH 6.7, which is acidic enough to activate ASICs in vivo. Our results highlight the pathophysiological role of proton signalling in the nervous system.

Bridging the divide

PubMed Central

McLean, Peter F; Cooley, Lynn

2014-01-01

Ring canals are made from arrested cleavage furrows, and provide direct cytoplasmic connections among sibling cells. They are well documented for their participation in Drosophila oogenesis, but little is known about their role in several somatic tissues in which they are also found. Using a variety of genetic tools in live and fixed tissue, we recently demonstrated that rapid intercellular exchange occurs through somatic ring canals by diffusion, and presented evidence that ring canals permit equilibration of protein among transcriptionally mosaic cells. We also used a novel combination of markers to evaluate the extent of protein movement within and across mitotic clones in follicle cells and imaginal discs, providing evidence of robust movement of GFP between the 2 sides of mitotic clones and frequently into non-recombined cells. These data suggest that, depending on the experimental setup and proteins of interest, inter-clonal diffusion of protein may alter the interpretation of clonal data in follicle cells. Here, we discuss these results and provide additional insight into the impact of ring canals in Drosophila somatic tissues. PMID:24406334

Ultrastructural Aspects of the Prenatal Bovine Ovary Differentiation with a Special Focus on the Interstitial Cells.

PubMed

Kenngott, R A-M; Scholz, W; Sinowatz, F

2016-10-01

The aim of this investigation was to study the ultrastructural features during the development of fetal bovine ovaries (crown rump length ranging from 11.4 to 94.0 cm). An interesting observation was the occurrence of big elongated cells containing a variety of electron dense granules and light homogenous vacuoles/bodies. They were located between the stroma cells surrounding the germ cell cord ends, adjacent to the first formed primordial follicles, typically situated near blood vessels. ER alpha and ER beta receptor positive cells could be detected in the same regions by means of immunohistochemistry. Intercellular bridges linked the germ cells nests oogonia. Germ cell cords consisted of centrally located, large, pale oogonia, surrounded by elongated somatic cells with very long cytoplasm extensions. Primordial follicles with flat pale follicular cells could be observed on the inner end of the cords. Extrusions of the outer nuclear membrane could often been recognised in voluminous oocytes. © 2016 Blackwell Verlag GmbH.

Visualization of predentine matrix components and endocytic structures in rat incisor odontoblasts with tannic acid.

PubMed

Goldberg, M; Septier, D

1989-12-01

Rat incisor odontoblasts and predentine fixed with tannic acid-glutaraldehyde-osmium tetroxide (Tago) were compared with those obtained by prior incubation in tannic acid-Ringer before conventional fixation with glutaraldehyde-osmium-tetroxide (Tari) The Tago method allowed visualization of complex glycoconjugates along the plasma membrane, in the pericellular spaces and in the intercellular predentine matrix. The non-collagenous proteins, proteoglycans and lipids were seen as granules and thin filaments located between the collagen fibers and at their surface. The collagen fibers themselves were also stained. The Tari method which was used to visualize exocytosis, mainly revealed endocytosis in the form of large intracellular vacuoles containing tannic acid and stained proteoglycans. It is suggested that tannic acid-Ringer incubation prior to fixation increases the endocytosis of the matrix components, which acculumates in these large vesicles.

Porcine Reproductive and Respiratory Syndrome Virus Utilizes Nanotubes for Intercellular Spread

PubMed Central

Guo, Rui; Katz, Benjamin B.; Tomich, John M.; Gallagher, Tom

2016-01-01

ABSTRACT Intercellular nanotube connections have been identified as an alternative pathway for cellular spreading of certain viruses. In cells infected with porcine reproductive and respiratory syndrome virus (PRRSV), nanotubes were observed connecting two distant cells with contiguous membranes, with the core infectious viral machinery (viral RNA, certain replicases, and certain structural proteins) present in/on the intercellular nanotubes. Live-cell movies tracked the intercellular transport of a recombinant PRRSV that expressed green fluorescent protein (GFP)-tagged nsp2. In MARC-145 cells expressing PRRSV receptors, GFP-nsp2 moved from one cell to another through nanotubes in the presence of virus-neutralizing antibodies. Intercellular transport of viral proteins did not require the PRRSV receptor as it was observed in receptor-negative HEK-293T cells after transfection with an infectious clone of GFP-PRRSV. In addition, GFP-nsp2 was detected in HEK-293T cells cocultured with recombinant PRRSV-infected MARC-145 cells. The intercellular nanotubes contained filamentous actin (F-actin) with myosin-associated motor proteins. The F-actin and myosin IIA were identified as coprecipitates with PRRSV nsp1β, nsp2, nsp2TF, nsp4, nsp7-nsp8, GP5, and N proteins. Drugs inhibiting actin polymerization or myosin IIA activation prevented nanotube formation and viral clusters in virus-infected cells. These data lead us to propose that PRRSV utilizes the host cell cytoskeletal machinery inside nanotubes for efficient cell-to-cell spread. This form of virus transport represents an alternative pathway for virus spread, which is resistant to the host humoral immune response. IMPORTANCE Extracellular virus particles transmit infection between organisms, but within infected hosts intercellular infection can be spread by additional mechanisms. In this study, we describe an alternative pathway for intercellular transmission of PRRSV in which the virus uses nanotube connections to transport infectious viral RNA, certain replicases, and certain structural proteins to neighboring cells. This process involves interaction of viral proteins with cytoskeletal proteins that form the nanotube connections. Intercellular viral spread through nanotubes allows the virus to escape the neutralizing antibody response and may contribute to the pathogenesis of viral infections. The development of strategies that interfere with this process could be critical in preventing the spread of viral infection. PMID:26984724

Cell remodeling and subtilase gene expression in the actinorhizal plant Discaria trinervis highlight host orchestration of intercellular Frankia colonization.

PubMed

Fournier, Joëlle; Imanishi, Leandro; Chabaud, Mireille; Abdou-Pavy, Iltaf; Genre, Andrea; Brichet, Lukas; Lascano, Hernán Ramiro; Muñoz, Nacira; Vayssières, Alice; Pirolles, Elodie; Brottier, Laurent; Gherbi, Hassen; Hocher, Valérie; Svistoonoff, Sergio; Barker, David G; Wall, Luis G

2018-05-23

Nitrogen-fixing filamentous Frankia colonize the root tissues of its actinorhizal host Discaria trinervis via an exclusively intercellular pathway. Here we present studies aimed at uncovering mechanisms associated with this little-researched mode of root entry, and in particular the extent to which the host plant is an active partner during this process. Detailed characterization of the expression patterns of infection-associated actinorhizal host genes has provided valuable tools to identify intercellular infection sites, thus allowing in vivo confocal microscopic studies of the early stages of Frankia colonization. The subtilisin-like serine protease gene Dt12, as well as its Casuarina glauca homolog Cg12, are specifically expressed at sites of Frankia intercellular colonization of D. trinervis outer root tissues. This is accompanied by nucleo-cytoplasmic reorganization in the adjacent host cells and major remodeling of the intercellular apoplastic compartment. These findings lead us to propose that the actinorhizal host plays a major role in modifying both the size and composition of the intercellular apoplast in order to accommodate the filamentous microsymbiont. The implications of these findings are discussed in the light of the analogies that can be made with the orchestrating role of host legumes during intracellular root hair colonization by nitrogen-fixing rhizobia. © 2018 The Authors New Phytologist © 2018 New Phytologist Trust.

The p.Cys169Tyr variant of connexin 26 is not a polymorphism

PubMed Central

Zonta, Francesco; Girotto, Giorgia; Buratto, Damiano; Crispino, Giulia; Morgan, Anna; Abdulhadi, Khalid; Alkowari, Moza; Badii, Ramin; Gasparini, Paolo; Mammano, Fabio

2015-01-01

Mutations in the GJB2 gene, which encodes the gap junction protein connexin 26 (Cx26), are the primary cause of hereditary prelingual hearing impairment. Here, the p.Cys169Tyr missense mutation of Cx26 (Cx26C169Y), previously classified as a polymorphism, has been identified as causative of severe hearing loss in two Qatari families. We have analyzed the effect of this mutation using a combination of confocal immunofluorescence microscopy and molecular dynamics simulations. At the cellular level, our results show that the mutant protein fails to form junctional channels in HeLa transfectants despite being correctly targeted to the plasma membrane. At the molecular level, this effect can be accounted for by disruption of the disulfide bridge that Cys169 forms with Cys64 in the wild-type structure (Cx26WT). The lack of the disulfide bridge in the Cx26C169Y protein causes a spatial rearrangement of two important residues, Asn176 and Thr177. In the Cx26WT protein, these residues play a crucial role in the intra-molecular interactions that permit the formation of an intercellular channel by the head-to-head docking of two opposing hemichannels resident in the plasma membrane of adjacent cells. Our results elucidate the molecular pathogenesis of hereditary hearing loss due to the connexin mutation and facilitate the understanding of its role in both healthy and affected individuals. PMID:25628337

Altered myocardial force generation in end-stage human heart failure.

PubMed

Papp, Zoltán; van der Velden, Jolanda; Borbély, Attila; Édes, István; Stienen, Ger J M

2014-12-01

This study aimed to elucidate the molecular background of increased Ca 2+ sensitivity of force production in cardiomyocytes of end-stage human heart failure. Ca 2+ -activated isometric force and the cross-bridge specific rate of force redevelopment (k tr ) were determined in Triton-skinned myocytes from end-stage failing and non-failing donor hearts. Measurements (control: pH 7.2, 0 mM inorganic phosphate (P i )) were performed under test conditions that probed either the Ca 2+ -regulatory function of the thin filaments (pH 6.5), the kinetics of the actin-myosin cross-bridge cycle (10 mM P i ), or both (pH 6.5, 10 mM P i ). The control maximal Ca 2+ -activated force (F o ) and k trmax did not differ between failing and non-failing myocytes. At submaximal [Ca 2+ ], however, both force and k tr were higher in failing than in donor myocytes. The difference in the Ca 2+ sensitivities of force production was preserved when the thin filament regulatory function was perturbed by acidosis (pH 6.5) but was abolished by cross-bridge modulation (i.e. by P i ) both at pH 7.2 and at pH 6.5. P i induced a larger reduction in force but a smaller increase in k tr in the failing myocytes than in the non-failing myocytes at submaximal [Ca 2+ ]. The enhanced P i sensitivity of the actin-myosin interaction suggests that the P i release step of the actin-myosin cross-bridge cycle is modified during end-stage human heart failure. This might be of functional importance when P i accumulates (e.g. during cardiac ischaemia). Moreover, this alteration can influence cardiac energetics and the clinical efficacy of sarcomere targeted agents in human heart failure. © 2015 The Authors. ESC Heart Failure published by John Wiley & Sons Ltd on behalf of the European Society of Cardiology.

Activation Dependence of Stretch Activation in Mouse Skinned Myocardium: Implications for Ventricular Function

PubMed Central

Stelzer, Julian E.; Larsson, Lars; Fitzsimons, Daniel P.; Moss, Richard L.

2006-01-01

Recent evidence suggests that ventricular ejection is partly powered by a delayed development of force, i.e., stretch activation, in regions of the ventricular wall due to stretch resulting from torsional twist of the ventricle around the apex-to-base axis. Given the potential importance of stretch activation in cardiac function, we characterized the stretch activation response and its Ca2+ dependence in murine skinned myocardium at 22°C in solutions of varying Ca2+ concentrations. Stretch activation was induced by suddenly imposing a stretch of 0.5–2.5% of initial length to the isometrically contracting muscle and then holding the muscle at the new length. The force response to stretch was multiphasic: force initially increased in proportion to the amount of stretch, reached a peak, and then declined to a minimum before redeveloping to a new steady level. This last phase of the response is the delayed force characteristic of myocardial stretch activation and is presumably due to increased attachment of cross-bridges as a consequence of stretch. The amplitude and rate of stretch activation varied with Ca2+ concentration and more specifically with the level of isometric force prior to the stretch. Since myocardial force is regulated both by Ca2+ binding to troponin-C and cross-bridge binding to thin filaments, we explored the role of cross-bridge binding in the stretch activation response using NEM-S1, a strong-binding, non-force–generating derivative of myosin subfragment 1. NEM-S1 treatment at submaximal Ca2+-activated isometric forces significantly accelerated the rate of the stretch activation response and reduced its amplitude. These data show that the rate and amplitude of myocardial stretch activation vary with the level of activation and that stretch activation involves cooperative binding of cross-bridges to the thin filament. Such a mechanism would contribute to increased systolic ejection in response to increased delivery of activator Ca2+ during excitation–contraction coupling. PMID:16446502

Highly electrically conductive layered carbon derived from polydopamine and its functions in SnO2-based lithium ion battery anodes.

PubMed

Kong, Junhua; Yee, Wu Aik; Yang, Liping; Wei, Yuefan; Phua, Si Lei; Ong, Hock Guan; Ang, Jia Ming; Li, Xu; Lu, Xuehong

2012-10-25

Thin carbonized polydopamine (C-PDA) coatings are found to have similar structures and electrical conductivities to those of multilayered graphene doped with heteroatoms. Greatly enhanced electrochemical properties are achieved with C-PDA-coated SnO(2) nanoparticles where the coating functions as a mechanical buffer layer and conducting bridge.

Moving contact lines in partial wetting: bridging the gap across the scales

NASA Astrophysics Data System (ADS)

Pahlavan, Amir; Cueto-Felgueroso, Luis; McKinley, Gareth; Juanes, Ruben

2017-11-01

The spreading and dewetting of liquid films on solid substrates is a common phenomenon in nature and industry from a snail secreting a mucosal film to printing and coating processes. A quantitative description of these phenomena, however, requires a detailed understanding of the flow physics at the nanoscale as the intermolecular interactions become important close to the contact line. Classical hydrodynamic theory describes wetting as an interplay between viscous and interfacial forces, neglecting the intermolecular interactions, leading to a paradox known as the moving contact line singularity. By contrast, molecular kinetic theory describes wetting as an activated process, neglecting the bulk hydrodynamics in the spreading viscous fluid film altogether. Here, we show that our recently developed model for thin liquid films in partial wetting, which properly incorporates the role of van der Waals interactions in a thin spreading fluid layer into a height-dependent surface tension, bridges the gap between these two approaches and leads to a unified framework for the description of wetting phenomena. We further use our model to investigate the instability and dewetting of nanometric liquid films, and show that it brings theoretical predictions closer to experimental observations.

Effect of sound on gap-junction-based intercellular signaling: Calcium waves under acoustic irradiation.

PubMed

Deymier, P A; Swinteck, N; Runge, K; Deymier-Black, A; Hoying, J B

2015-01-01

We present a previously unrecognized effect of sound waves on gap-junction-based intercellular signaling such as in biological tissues composed of endothelial cells. We suggest that sound irradiation may, through temporal and spatial modulation of cell-to-cell conductance, create intercellular calcium waves with unidirectional signal propagation associated with nonconventional topologies. Nonreciprocity in calcium wave propagation induced by sound wave irradiation is demonstrated in the case of a linear and a nonlinear reaction-diffusion model. This demonstration should be applicable to other types of gap-junction-based intercellular signals, and it is thought that it should be of help in interpreting a broad range of biological phenomena associated with the beneficial therapeutic effects of sound irradiation and possibly the harmful effects of sound waves on health.

Domain wall effects on the magnetoresistance in epitaxial nanostructures

NASA Astrophysics Data System (ADS)

Perason, David; Zambano, Antonio; Lukaszew, R. Alejandra

2004-03-01

It has been postulated that adiabatic magneto-transport in sufficiently small contacts in nano-bridges should exhibit significant magnetoresistance at room temperature.[1] In order to study this phenomenon we have patterned nano-bridges on epitaxial ferromagnetic thin films using e-beam lithography. We have tested (001) and (111) oriented Ni films, as well as (001) and (011) CrO2 films. We have patterned bridges with different geometric orientations with respect to the crystallographic axes in the samples. We will show magnetic force microscopy images of the devices and will compare them with OOMMF simulations of the magnetization dynamics during reversal. The magneto-transport was studied using 4-point probe with AC current and Lock-In techniques. We notice here that the size of the observed effect is inversely proportional to the width of the nano-contact. The typical MR observed was 1-2at room temperature. We will show a comparison of MR effects observed in the various films and geometries described. [1]. P. Bruno, Phys. Rev. Lett. 83, 2425, (1999).

Optical properties of paint-on resins for shade modification of crown and bridge resins--light transmittance characteristics--.

PubMed

Arikawa, Hiroyuki; Kanie, Takahito; Fujii, Koichi; Homma, Tetsuya; Takahashi, Hideo; Ban, Seiji

2003-09-01

The purpose of this study was to examine the light transmittance characteristics of the paint-on resins for shade modification. Three shades of paint-on resin, one crown and bridge resin, and human enamel were used. Specimens with four different thicknesses (75-150 microm) were prepared. The light transmittances including its wavelength distribution and diffusion characteristics were measured. The color values and the color differences among thicknesses of specimens were also determined. The light transmittance values of the paint-on resins ranged from 60.3% to 88.3% at 100 microm thickness, which were lower or nearly equal in comparison with the crown and bridge resin and enamel. Although differences in the wavelength distribution of transmittance among materials were found at lower wavelengths, all materials showed similar diffusion characteristics. The thin layer of paint-on resin effectively changed the color of restorative resin. The paint-on resin may be an effective material for the modification of the color appearance matching required.

[Thin filament elasticity and its role in the muscle contraction].

PubMed

Skubiszak, L

2006-01-01

The available experimental methods do not allow one to establish unambiguously the molecular structural events during muscle contraction. To resolve the existing controversies, I have devised an unconventional original computer program. The new approach allows the reconstruction of the hexagonal lattice of the sarcomere for different muscle states and verification of the structure by comparison of the calculated Fourier spectra with the real diffraction patterns. Previously, by the use of this approach, the real structure of a myosin filament from vertebrate striated muscle has been reconstructed (http://zope.ibib.waw.pl/pspk). In this work, a reconstruction for the thin filament is presented for three states: relaxed, after activation, and during contraction. Good consistency of the calculated Fourier spectra with the real diffraction patterns available in the literature suggests that the thin filament, due to flexibility, plays an active part in muscle contraction, as myosin cross-bridges do.

Oscillations in a Sunspot with Light Bridges

NASA Astrophysics Data System (ADS)

Yuan, Ding; Nakariakov, Valery M.; Huang, Zhenghua; Li, Bo; Su, Jiangtao; Yan, Yihua; Tan, Baolin

2014-09-01

The Solar Optical Telescope on board Hinode observed a sunspot (AR 11836) with two light bridges (LBs) on 2013 August 31. We analyzed a two-hour Ca II H emission intensity data set and detected strong five-minute oscillation power on both LBs and in the inner penumbra. The time-distance plot reveals that the five-minute oscillation phase does not vary significantly along the thin bridge, indicating that the oscillations are likely to originate from underneath it. The slit taken along the central axis of the wide LB exhibits a standing wave feature. However, at the center of the wide bridge, the five-minute oscillation power is found to be stronger than at its sides. Moreover, the time-distance plot across the wide bridge exhibits a herringbone pattern that indicates a counter-stream of two running waves, which originated at the bridge's sides. Thus, the five-minute oscillations on the wide bridge also resemble the properties of running penumbral waves. The five-minute oscillations are suppressed in the umbra, while the three-minute oscillations occupy all three cores of the sunspot's umbra, separated by the LBs. The three-minute oscillations were found to be in phase at both sides of the LBs. This may indicate that either LBs do not affect umbral oscillations, or that umbral oscillations at different umbral cores share the same source. It also indicates that LBs are rather shallow objects situated in the upper part of the umbra. We found that umbral flashes (UFs) follow the life cycles of umbral oscillations with much larger amplitudes. They cannot propagate across LBs. UFs dominate the three-minute oscillation power within each core; however, they do not disrupt the phase of umbral oscillation.

Oscillations in a sunspot with light bridges

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yuan, Ding; Su, Jiangtao; Yan, Yihua

2014-09-01

The Solar Optical Telescope on board Hinode observed a sunspot (AR 11836) with two light bridges (LBs) on 2013 August 31. We analyzed a two-hour Ca II H emission intensity data set and detected strong five-minute oscillation power on both LBs and in the inner penumbra. The time-distance plot reveals that the five-minute oscillation phase does not vary significantly along the thin bridge, indicating that the oscillations are likely to originate from underneath it. The slit taken along the central axis of the wide LB exhibits a standing wave feature. However, at the center of the wide bridge, the five-minutemore » oscillation power is found to be stronger than at its sides. Moreover, the time-distance plot across the wide bridge exhibits a herringbone pattern that indicates a counter-stream of two running waves, which originated at the bridge's sides. Thus, the five-minute oscillations on the wide bridge also resemble the properties of running penumbral waves. The five-minute oscillations are suppressed in the umbra, while the three-minute oscillations occupy all three cores of the sunspot's umbra, separated by the LBs. The three-minute oscillations were found to be in phase at both sides of the LBs. This may indicate that either LBs do not affect umbral oscillations, or that umbral oscillations at different umbral cores share the same source. It also indicates that LBs are rather shallow objects situated in the upper part of the umbra. We found that umbral flashes (UFs) follow the life cycles of umbral oscillations with much larger amplitudes. They cannot propagate across LBs. UFs dominate the three-minute oscillation power within each core; however, they do not disrupt the phase of umbral oscillation.« less

Cancer and intercellular cooperation

PubMed Central

Dieli, Anna Maria

2017-01-01

The major transitions approach in evolutionary biology has shown that the intercellular cooperation that characterizes multicellular organisms would never have emerged without some kind of multilevel selection. Relying on this view, the Evolutionary Somatic view of cancer considers cancer as a breakdown of intercellular cooperation and as a loss of the balance between selection processes that take place at different levels of organization (particularly single cell and individual organism). This seems an elegant unifying framework for healthy organism, carcinogenesis, tumour proliferation, metastasis and other phenomena such as ageing. However, the gene-centric version of Darwinian evolution, which is often adopted in cancer research, runs into empirical problems: proto-tumoural and tumoural features in precancerous cells that would undergo ‘natural selection’ have proved hard to demonstrate; cells are radically context-dependent, and some stages of cancer are poorly related to genetic change. Recent perspectives propose that breakdown of intercellular cooperation could depend on ‘fields’ and other higher-level phenomena, and could be even mutations independent. Indeed, the field would be the context, allowing (or preventing) genetic mutations to undergo an intra-organism process analogous to natural selection. The complexities surrounding somatic evolution call for integration between multiple incomplete frameworks for interpreting intercellular cooperation and its pathologies. PMID:29134064

Biomaterials-Based Electronics: Polymers and Interfaces for Biology and Medicine

PubMed Central

Muskovich, Meredith; Bettinger, Christopher J.

2012-01-01

Advanced polymeric biomaterials continue to serve as a cornerstone of new medical technologies and therapies. The vast majority of these materials, both natural and synthetic, interact with biological matter without direct electronic communication. However, biological systems have evolved to synthesize and employ naturally-derived materials for the generation and modulation of electrical potentials, voltage gradients, and ion flows. Bioelectric phenomena can be interpreted as potent signaling cues for intra- and inter-cellular communication. These cues can serve as a gateway to link synthetic devices with biological systems. This progress report will provide an update on advances in the application of electronically active biomaterials for use in organic electronics and bio-interfaces. Specific focus will be granted to the use of natural and synthetic biological materials as integral components in technologies such as thin film electronics, in vitro cell culture models, and implantable medical devices. Future perspectives and emerging challenges will also be highlighted. PMID:23184740

Switch from intracellular to intercellular invasion during water stress-tolerant legume nodulation

PubMed Central

Goormachtig, Sofie; Capoen, Ward; James, Euan K.; Holsters, Marcelle

2004-01-01

Rhizobia colonize their legume hosts by different modes of entry while initiating symbiotic nitrogen fixation. Most legumes are invaded via growing root hairs by the root hair-curl mechanism, which involves epidermal cell responses. However, invasion of a number of tropical legumes happens through fissures at lateral root bases by cortical, intercellular crack entry. In the semiaquatic Sesbania rostrata, the bacteria entered via root hair curls under nonflooding conditions. Upon flooding, root hair growth was prevented, invasion on accessible root hairs was inhibited, and intercellular invasion was recruited. The plant hormone ethylene was involved in these processes. The occurrence of both invasion pathways on the same host plant enabled a comparison to be made of the structural requirements for the perception of nodulation factors, which were more stringent for the epidermal root hair invasion than for the cortical intercellular invasion at lateral root bases. PMID:15079070

Asymmetric homotypic interactions of the atypical cadherin Flamingo mediate intercellular polarity signaling

PubMed Central

Chen, Wei-Shen; Antic, Dragana; Matis, Maja; Logan, Catriona Y.; Povelones, Michael; Anderson, Graham; Nusse, Roel; Axelrod, Jeffrey D.

2008-01-01

Acquisition of planar cell polarity (PCP) in epithelia involves intercellular communication, during which cells align their polarity with that of their neighbors. The transmembrane proteins Frizzled (Fz) and Van Gogh (Vang) are essential components of the intercellular communication mechanism, as loss of either strongly perturbs the polarity of neighboring cells. How Fz and Vang communicate polarity information between neighboring cells is poorly understood. The atypical cadherin, Flamingo (Fmi), is implicated in this process, yet whether Fmi acts permissively as a scaffold, or instructively as a signal is unclear. Here, we provide evidence that Fmi functions instructively to mediate Fz-Vang intercellular signal relay, recruiting Fz and Vang to opposite sides of cell boundaries. We propose that two functional forms of Fmi, one of which is induced by and physically interacts with Fz, form cadherin homodimers that signal bidirectionally and asymmetrically, instructing unequal responses in adjacent cell membranes to establish molecular asymmetry. PMID:18555784

THE EFFECT OF SMOOTH MUSCLE ON THE INTERCELLULAR SPACES IN TOAD URINARY BLADDER

PubMed Central

DiBona, Donald R.; Civan, Mortimer M.

1970-01-01

Phase microscopy of toad urinary bladder has demonstrated that vasopressin can cause an enlargement of the epithelial intercellular spaces under conditions of no net transfer of water or sodium. The suggestion that this phenomenon is linked to the hormone's action as a smooth muscle relaxant has been tested and verified with the use of other agents effecting smooth muscle: atropine and adenine compounds (relaxants), K+ and acetylcholine (contractants). Furthermore, it was possible to reduce the size and number of intercellular spaces, relative to a control, while increasing the rate of osmotic water flow. A method for quantifying these results has been developed and shows that they are, indeed, significant. It is concluded, therefore, that the configuration of intercellular spaces is not a reliable index of water flow across this epithelium and that such a morphologic-physiologic relationship is tenuous in any epithelium supported by a submucosa rich in smooth muscle. PMID:4915450

Thin glass based packaging and photonic single-mode waveguide integration by ion-exchange technology on board and module level

NASA Astrophysics Data System (ADS)

Brusberg, Lars; Lang, Günter; Schröder, Henning

2011-01-01

The proposed novel packaging approach merges micro-system packaging and glass integrated optics. It provides 3D optical single-mode intra system links to bridge the gap between novel photonic integrated circuits and the glass fibers for inter system interconnects. We introduce our hybrid 3D photonic packaging approach based on thin glass substrates with planar integrated optical single-mode waveguides for fiber-to-chip and chip-to-chip links. Optical mirrors and lenses provide optical mode matching for photonic IC assemblies and optical fiber interconnects. Thin glass is commercially available in panel and wafer formats and characterizes excellent optical and high-frequency properties as reviewed in the paper. That makes it perfect for micro-system packaging. The adopted planar waveguide process based on ion-exchange technology is capable for high-volume manufacturing. This ion-exchange process and the optical propagation are described in detail for thin glass substrates. An extensive characterization of all basic circuit elements like straight and curved waveguides, couplers and crosses proves the low attenuation of the optical circuit elements.

Depressed Frank-Starling mechanism in the left ventricular muscle of the knock-in mouse model of dilated cardiomyopathy with troponin T deletion mutation ΔK210.

PubMed

Inoue, Takahiro; Kobirumaki-Shimozawa, Fuyu; Kagemoto, Tatsuya; Fujii, Teruyuki; Terui, Takako; Kusakari, Yoichiro; Hongo, Kenichi; Morimoto, Sachio; Ohtsuki, Iwao; Hashimoto, Kazuhiro; Fukuda, Norio

2013-10-01

It has been reported that the Frank-Starling mechanism is coordinately regulated in cardiac muscle via thin filament "on-off" equilibrium and titin-based lattice spacing changes. In the present study, we tested the hypothesis that the deletion mutation ΔK210 in the cardiac troponin T gene shifts the equilibrium toward the "off" state and accordingly attenuate the sarcomere length (SL) dependence of active force production, via reduced cross-bridge formation. Confocal imaging in isolated hearts revealed that the cardiomyocytes were enlarged, especially in the longitudinal direction, in ΔK210 hearts, with striation patterns similar to those in wild type (WT) hearts, suggesting that the number of sarcomeres is increased in cardiomyocytes but the sarcomere length remains unaltered. For analysis of the SL dependence of active force, skinned muscle preparations were obtained from the left ventricle of WT and knock-in (ΔK210) mice. An increase in SL from 1.90 to 2.20μm shifted the mid-point (pCa50) of the force-pCa curve leftward by ~0.21pCa units in WT preparations. In ΔK210 muscles, Ca(2+) sensitivity was lower by ~0.37pCa units, and the SL-dependent shift of pCa50, i.e., ΔpCa50, was less pronounced (~0.11pCa units), with and without protein kinase A treatment. The rate of active force redevelopment was lower in ΔK210 preparations than in WT preparations, showing blunted thin filament cooperative activation. An increase in thin filament cooperative activation upon an increase in the fraction of strongly bound cross-bridges by MgADP increased ΔpCa50 to ~0.21pCa units. The depressed Frank-Starling mechanism in ΔK210 hearts is the result of a reduction in thin filament cooperative activation. © 2013.

Intercellular Diffusion of a Fluorescent Sucrose Analog via the Septal Junctions in a Filamentous Cyanobacterium

PubMed Central

Nürnberg, Dennis J.; Mariscal, Vicente; Bornikoel, Jan; Nieves-Morión, Mercedes; Krauß, Norbert; Herrero, Antonia

2015-01-01

ABSTRACT Many filamentous cyanobacteria produce specialized nitrogen-fixing cells called heterocysts, which are located at semiregular intervals along the filament with about 10 to 20 photosynthetic vegetative cells in between. Nitrogen fixation in these complex multicellular bacteria depends on metabolite exchange between the two cell types, with the heterocysts supplying combined-nitrogen compounds but dependent on the vegetative cells for photosynthetically produced carbon compounds. Here, we used a fluorescent tracer to probe intercellular metabolite exchange in the filamentous heterocyst-forming cyanobacterium Anabaena sp. strain PCC 7120. We show that esculin, a fluorescent sucrose analog, is incorporated by a sucrose import system into the cytoplasm of Anabaena cells. The cytoplasmic esculin is rapidly and reversibly exchanged across vegetative-vegetative and vegetative-heterocyst cell junctions. Our measurements reveal the kinetics of esculin exchange and also show that intercellular metabolic communication is lost in a significant fraction of older heterocysts. SepJ, FraC, and FraD are proteins located at the intercellular septa and are suggested to form structures analogous to gap junctions. We show that a ΔsepJ ΔfraC ΔfraD triple mutant shows an altered septum structure with thinner septa but a denser peptidoglycan layer. Intercellular diffusion of esculin and fluorescein derivatives is impaired in this mutant, which also shows a greatly reduced frequency of nanopores in the intercellular septal cross walls. These findings suggest that FraC, FraD, and SepJ are important for the formation of junctional structures that constitute the major pathway for feeding heterocysts with sucrose. PMID:25784700

Germ cell cluster organization and oogenesis in the tardigrade Dactylobiotus parthenogeneticus Bertolani, 1982 (Eutardigrada, Murrayidae).

PubMed

Poprawa, Izabela; Hyra, Marta; Rost-Roszkowska, Magdalena Maria

2015-07-01

Germ cell cluster organization and the process of oogenesis in Dactylobiotus parthenogeneticus have been described using transmission electron microscopy and light microscopy. The reproductive system of D. parthenogeneticus is composed of a single, sac-like, meroistic ovary and a single oviduct that opens into the cloaca. Two zones can be distinguished in the ovary: a small germarium that is filled with oogonia and a vitellarium that is filled with germ cell clusters. The germ cell cluster, which has the form of a modified rosette, consists of eight cells that are interconnected by stable cytoplasmic bridges. The cell that has the highest number of stable cytoplasmic bridges (four bridges) finally develops into the oocyte, while the remaining cells become trophocytes. Vitellogenesis of a mixed type occurs in D. parthenogeneticus. One part of the yolk material is produced inside the oocyte (autosynthesis), while the second part is synthesized in the trophocytes and transported to the oocyte through the cytoplasmic bridges. The eggs are covered with two envelopes: a thin vitelline envelope and a three-layered chorion. The surface of the chorion forms small conical processes, the shape of which is characteristic for the species that was examined. In our paper, we present the first report on the rosette type of germ cell clusters in Parachela.

Structural and functional aspects of the myosin essential light chain in cardiac muscle contraction

DOE Office of Scientific and Technical Information (OSTI.GOV)

Muthu, Priya; Wang, Li; Yuan, Chen-Ching

2012-04-02

The myosin essential light chain (ELC) is a structural component of the actomyosin cross-bridge, but its function is poorly understood, especially the role of the cardiac specific N-terminal extension in modulating actomyosin interaction. Here, we generated transgenic (Tg) mice expressing the A57G (alanine to glycine) mutation in the cardiac ELC known to cause familial hypertrophic cardiomyopathy (FHC). The function of the ELC N-terminal extension was investigated with the Tg-{Delta}43 mouse model, whose myocardium expresses a truncated ELC. Low-angle X-ray diffraction studies on papillary muscle fibers in rigor revealed a decreased interfilament spacing ({approx} 1.5 nm) and no alterations in cross-bridgemore » mass distribution in Tg-A57G mice compared to Tg-WT, expressing the full-length nonmutated ELC. The truncation mutation showed a 1.3-fold increase in I{sub 1,1}/I{sub 1,0}, indicating a shift of cross-bridge mass from the thick filament backbone toward the thin filaments. Mechanical studies demonstrated increased stiffness in Tg-A57G muscle fibers compared to Tg-WT or Tg-{Delta}43. The equilibrium constant for the cross-bridge force generation step was smallest in Tg-{Delta}43. These results support an important role for the N-terminal ELC extension in prepositioning the cross-bridge for optimal force production. Subtle changes in the ELC sequence were sufficient to alter cross-bridge properties and lead to pathological phenotypes.« less

Patterning of wound-induced intercellular Ca2+ flashes in a developing epithelium

NASA Astrophysics Data System (ADS)

Narciso, Cody; Wu, Qinfeng; Brodskiy, Pavel; Garston, George; Baker, Ruth; Fletcher, Alexander; Zartman, Jeremiah

2015-10-01

Differential mechanical force distributions are increasingly recognized to provide important feedback into the control of an organ’s final size and shape. As a second messenger that integrates and relays mechanical information to the cell, calcium ions (Ca2+) are a prime candidate for providing important information on both the overall mechanical state of the tissue and resulting behavior at the individual-cell level during development. Still, how the spatiotemporal properties of Ca2+ transients reflect the underlying mechanical characteristics of tissues is still poorly understood. Here we use an established model system of an epithelial tissue, the Drosophila wing imaginal disc, to investigate how tissue properties impact the propagation of Ca2+ transients induced by laser ablation. The resulting intercellular Ca2+ flash is found to be mediated by inositol 1,4,5-trisphosphate and depends on gap junction communication. Further, we find that intercellular Ca2+ transients show spatially non-uniform characteristics across the proximal-distal axis of the larval wing imaginal disc, which exhibit a gradient in cell size and anisotropy. A computational model of Ca2+ transients is employed to identify the principle factors explaining the spatiotemporal patterning dynamics of intercellular Ca2+ flashes. The relative Ca2+ flash anisotropy is principally explained by local cell shape anisotropy. Further, Ca2+ velocities are relatively uniform throughout the wing disc, irrespective of cell size or anisotropy. This can be explained by the opposing effects of cell diameter and cell elongation on intercellular Ca2+ propagation. Thus, intercellular Ca2+ transients follow lines of mechanical tension at velocities that are largely independent of tissue heterogeneity and reflect the mechanical state of the underlying tissue.

Stimulating the Release of Exosomes Increases the Intercellular Transfer of Prions.

PubMed

Guo, Belinda B; Bellingham, Shayne A; Hill, Andrew F

2016-03-04

Exosomes are small extracellular vesicles released by cells and play important roles in intercellular communication and pathogen transfer. Exosomes have been implicated in several neurodegenerative diseases, including prion disease and Alzheimer disease. Prion disease arises upon misfolding of the normal cellular prion protein, PrP(C), into the disease-associated isoform, PrP(Sc). The disease has a unique transmissible etiology, and exosomes represent a novel and efficient method for prion transmission. The precise mechanism by which prions are transmitted from cell to cell remains to be fully elucidated, although three hypotheses have been proposed: direct cell-cell contact, tunneling nanotubes, and exosomes. Given the reported presence of exosomes in biological fluids and in the lipid and nucleic acid contents of exosomes, these vesicles represent an ideal mechanism for encapsulating prions and potential cofactors to facilitate prion transmission. This study investigates the relationship between exosome release and intercellular prion dissemination. Stimulation of exosome release through treatment with an ionophore, monensin, revealed a corresponding increase in intercellular transfer of prion infectivity. Conversely, inhibition of exosome release using GW4869 to target the neutral sphingomyelinase pathway induced a decrease in intercellular prion transmission. Further examination of the effect of monensin on PrP conversion revealed that monensin also alters the conformational stability of PrP(C), leading to increased generation of proteinase K-resistant prion protein. The findings presented here provide support for a positive relationship between exosome release and intercellular transfer of prion infectivity, highlighting an integral role for exosomes in facilitating the unique transmissible nature of prions. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

Intercellular communications-redox interactions in radiation toxicity; potential targets for radiation mitigation.

PubMed

Farhood, Bagher; Goradel, Nasser Hashemi; Mortezaee, Keywan; Khanlarkhani, Neda; Salehi, Ensieh; Nashtaei, Maryam Shabani; Shabeeb, Dheyauldeen; Musa, Ahmed Eleojo; Fallah, Hengameh; Najafi, Masoud

2018-06-17

Nowadays, using ionizing radiation (IR) is necessary for clinical, agricultural, nuclear energy or industrial applications. Accidental exposure to IR after a radiation terror or disaster poses a threat to human. In contrast to the old dogma of radiation toxicity, several experiments during the last two recent decades have revealed that intercellular signaling and communications play a key role in this procedure. Elevated level of cytokines and other intercellular signals increase oxidative damage and inflammatory responses via reduction/oxidation interactions (redox system). Intercellular signals induce production of free radicals and inflammatory mediators by some intermediate enzymes such as cyclooxygenase-2 (COX-2), nitric oxide synthase (NOS), NADPH oxidase, and also via triggering mitochondrial ROS. Furthermore, these signals facilitate cell to cell contact and increasing cell toxicity via cohort effect. Nitric oxide is a free radical with ability to act as an intercellular signal that induce DNA damage and changes in some signaling pathways in irradiated as well as non-irradiated adjacent cells. Targeting of these mediators by some anti-inflammatory agents or via antioxidants such as mitochondrial ROS scavengers opens a window to mitigate radiation toxicity after an accidental exposure. Experiments which have been done so far suggests that some cytokines such as IL-1β, TNF-α, TGF-β, IL-4 and IL-13 are some interesting targets that depend on irradiated organs and may help mitigate radiation toxicity. Moreover, animal experiments in recent years indicated that targeting of toll like receptors (TLRs) may be more useful for radioprotection and mitigation. In this review, we aimed to describe the role of intercellular interactions in oxidative injury, inflammation, cell death and killing effects of IR. Moreover, we described evidence on potential mitigation of radiation injury via targeting of these mediators.

Mass flow sensor utilizing a resistance bridge

NASA Technical Reports Server (NTRS)

Fralick, Gustave C. (Inventor); Hwang, Danny P. (Inventor); Wrbanek, John D. (Inventor)

2004-01-01

A mass flow sensor to be mounted within a duct and measures the mass flow of a fluid stream moving through the duct. The sensor is an elongated thin quartz substrate having a plurality of platinum strips extending in a parallel relationship on the strip, with certain of the strips being resistors connected to an excitation voltage. The resistors form the legs of a Wheatstone bridge. The resistors are spaced a sufficient distance inwardly from the leading and trailing edges of the substrate to lie within the velocity recovery region so that the measured flow is the same as the actual upstream flow. The resistor strips extend at least half-way through the fluid stream to include a substantial part of the velocity profile of the stream. Certain of the resistors detect a change in temperature as the fluid stream moves across the substrate to provide an output signal from the Wheatstone bridge which is representative of the fluid flow. A heater is located in the midst of the resistor array to heat the air as it passes over the array.

Cross-regulation between Aurora B and Citron kinase controls midbody architecture in cytokinesis

PubMed Central

McKenzie, Callum; Bassi, Zuni I.; Debski, Janusz; Gottardo, Marco; Callaini, Giuliano; Dadlez, Michal; D'Avino, Pier Paolo

2016-01-01

Cytokinesis culminates in the final separation, or abscission, of the two daughter cells at the end of cell division. Abscission relies on an organelle, the midbody, which forms at the intercellular bridge and is composed of various proteins arranged in a precise stereotypic pattern. The molecular mechanisms controlling midbody organization and function, however, are obscure. Here we show that proper midbody architecture requires cross-regulation between two cell division kinases, Citron kinase (CIT-K) and Aurora B, the kinase component of the chromosomal passenger complex (CPC). CIT-K interacts directly with three CPC components and is required for proper midbody architecture and the orderly arrangement of midbody proteins, including the CPC. In addition, we show that CIT-K promotes Aurora B activity through phosphorylation of the INCENP CPC subunit at the TSS motif. In turn, Aurora B controls CIT-K localization and association with its central spindle partners through phosphorylation of CIT-K's coiled coil domain. Our results identify, for the first time, a cross-regulatory mechanism between two kinases during cytokinesis, which is crucial for establishing the stereotyped organization of midbody proteins. PMID:27009191

CGGBP1 is a nuclear and midbody protein regulating abscission

DOE Office of Scientific and Technical Information (OSTI.GOV)

Singh, Umashankar, E-mail: umashankar.singh@genpat.uu.se; Westermark, Bengt

2011-01-15

Abscission marks the completion of cell division and its failure is associated with delayed cytokinesis and even tetraploidization. Aberrant abscission and consequential ploidy changes can underlie various diseases including cancer. Midbody, a transient structure formed in the intercellular bridge during telophase, contains several proteins including Aurora kinase B (AURKB), which participate in abscission. We report here an unexpected expression pattern and function of the transcription repressor protein CGG triplet repeat-binding protein 1 (CGGBP1), in normal human fibroblasts. We show that CGGBP1, a chromatin-associated protein, trans-localizes to spindle midzone and midbodies in a manner similar to that of AURKB. CGGBP1 depletionmore » resulted in a cell cycle block at G2, characterized by failure of cells to undergo mitosis and also reduced entry into S phase. Consistent with its presence in the midbodies, live microscopy showed that CGGBP1 deficiency caused mitotic failure at abscission resulting in tetraploidy, which could be rescued by CGGBP1 overexpression. These results show that CGGBP1 is a bona fide midbody protein required for normal abscission and mitosis in general.« less

Inter-Cellular Exchange of Cellular Components via VE-Cadherin-Dependent Trans-Endocytosis

PubMed Central

Sakurai, Takashi; Woolls, Melissa J.; Jin, Suk-Won

2014-01-01

Cell-cell communications typically involve receptor-mediated signaling initiated by soluble or cell-bound ligands. Here, we report a unique mode of endocytosis: proteins originating from cell-cell junctions and cytosolic cellular components from the neighboring cell are internalized, leading to direct exchange of cellular components between two adjacent endothelial cells. VE-cadherins form transcellular bridges between two endothelial cells that are the basis of adherence junctions. At such adherens junction sites, we observed the movement of the entire VE-cadherin molecule from one endothelial cell into the other with junctional and cytoplasmic components. This phenomenon, here termed trans-endocytosis, requires the establishment of a VE-cadherin homodimer in trans with internalization proceeding in a Rac1-, and actomyosin-dependent manner. Importantly, the trans-endocytosis is not dependent on any known endocytic pathway including clathrin-dependent endocytosis, macropinocytosis or phagocytosis. This novel form of cell-cell communications, leading to a direct exchange of cellular components, was observed in 2D and 3D-cultured endothelial cells as well as in the developing zebrafish vasculature. PMID:24603875

The midbody ring scaffolds the abscission machinery in the absence of midbody microtubules

PubMed Central

Green, Rebecca A.; Mayers, Jonathan R.; Wang, Shaohe; Lewellyn, Lindsay; Desai, Arshad; Audhya, Anjon

2013-01-01

Abscission completes cytokinesis to form the two daughter cells. Although abscission could be organized from the inside out by the microtubule-based midbody or from the outside in by the contractile ring–derived midbody ring, it is assumed that midbody microtubules scaffold the abscission machinery. In this paper, we assess the contribution of midbody microtubules versus the midbody ring in the Caenorhabditis elegans embryo. We show that abscission occurs in two stages. First, the cytoplasm in the daughter cells becomes isolated, coincident with formation of the intercellular bridge; proper progression through this stage required the septins (a midbody ring component) but not the membrane-remodeling endosomal sorting complex required for transport (ESCRT) machinery. Second, the midbody and midbody ring are released into a specific daughter cell during the subsequent cell division; this stage required the septins and the ESCRT machinery. Surprisingly, midbody microtubules were dispensable for both stages. These results delineate distinct steps during abscission and highlight the central role of the midbody ring, rather than midbody microtubules, in their execution. PMID:24217623

Activation of acid-sensing ion channels by localized proton transient reveals their role in proton signaling

PubMed Central

Zeng, Wei-Zheng; Liu, Di-Shi; Liu, Lu; She, Liang; Wu, Long-Jun; Xu, Tian-Le

2015-01-01

Extracellular transients of pH alterations likely mediate signal transduction in the nervous system. Neuronal acid-sensing ion channels (ASICs) act as sensors for extracellular protons, but the mechanism underlying ASIC activation remains largely unknown. Here, we show that, following activation of a light-activated proton pump, Archaerhodopsin-3 (Arch), proton transients induced ASIC currents in both neurons and HEK293T cells co-expressing ASIC1a channels. Using chimera proteins that bridge Arch and ASIC1a by a glycine/serine linker, we found that successful coupling occurred within 15 nm distance. Furthermore, two-cell sniffer patch recording revealed that regulated release of protons through either Arch or voltage-gated proton channel Hv1 activated neighbouring cells expressing ASIC1a channels. Finally, computational modelling predicted the peak proton concentration at the intercellular interface to be at pH 6.7, which is acidic enough to activate ASICs in vivo. Our results highlight the pathophysiological role of proton signalling in the nervous system. PMID:26370138

Clustering and negative feedback by endocytosis in planar cell polarity signaling is modulated by ubiquitinylation of prickle.

PubMed

Cho, Bomsoo; Pierre-Louis, Gandhy; Sagner, Andreas; Eaton, Suzanne; Axelrod, Jeffrey D

2015-05-01

The core components of the planar cell polarity (PCP) signaling system, including both transmembrane and peripheral membrane associated proteins, form asymmetric complexes that bridge apical intercellular junctions. While these can assemble in either orientation, coordinated cell polarization requires the enrichment of complexes of a given orientation at specific junctions. This might occur by both positive and negative feedback between oppositely oriented complexes, and requires the peripheral membrane associated PCP components. However, the molecular mechanisms underlying feedback are not understood. We find that the E3 ubiquitin ligase complex Cullin1(Cul1)/SkpA/Supernumerary limbs(Slimb) regulates the stability of one of the peripheral membrane components, Prickle (Pk). Excess Pk disrupts PCP feedback and prevents asymmetry. We show that Pk participates in negative feedback by mediating internalization of PCP complexes containing the transmembrane components Van Gogh (Vang) and Flamingo (Fmi), and that internalization is activated by oppositely oriented complexes within clusters. Pk also participates in positive feedback through an unknown mechanism promoting clustering. Our results therefore identify a molecular mechanism underlying generation of asymmetry in PCP signaling.

Inheritance of the Golgi Apparatus and Cytokinesis Are Controlled by Degradation of GBF1.

PubMed

Magliozzi, Roberto; Carrero, Zunamys I; Low, Teck Yew; Yuniati, Laurensia; Valdes-Quezada, Christian; Kruiswijk, Flore; van Wijk, Koen; Heck, Albert J R; Jackson, Catherine L; Guardavaccaro, Daniele

2018-06-12

Although much is known about how chromosome segregation is coupled to cell division, how intracellular organelles partition during mitotic division is poorly understood. We report that the phosphorylation-dependent degradation of the ARFGEF GBF1 regulates organelle trafficking during cell division. We show that, in mitosis, GBF1 is phosphorylated on Ser292 and Ser297 by casein kinase-2 allowing recognition by the F-box protein βTrCP. GBF1 interaction with βTrCP recruits GBF1 to the SCF βTrCP ubiquitin ligase complex, triggering its degradation. Phosphorylation and degradation of GBF1 occur along microtubules at the intercellular bridge of telophase cells and are required for Golgi membrane positioning and postmitotic Golgi reformation. Indeed, expression of a non-degradable GBF1 mutant inhibits the transport of the Golgi cluster adjacent to the midbody toward the Golgi twin positioned next to the centrosome and results in defective Golgi reassembly and cytokinesis failure. These findings define a mechanism that controls postmitotic Golgi reassembly and inheritance. Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

Structural dynamics of F-actin: I. Changes in the C terminus.

PubMed

Orlova, A; Egelman, E H

1995-02-03

The biochemical properties of G-actin, and the kinetics of polymerization of G-actin into F-actin, are dependent upon whether Mg2+ or Ca2+ is bound at the high-affinity metal-binding site in actin. Three-dimensional reconstructions from electron micrographs show that a bridge of density, that we interpret as arising from a major shift of the C terminus, exists between the two strands of the filament in Ca(2+)-actin that is absent in Mg(2+)-actin. This bridge is also absent in models of F-actin built from an atomic structure of G-Ca(2+)-actin. The cleavage of the DNase I-binding loop in actin between residues 42 and 43, with the non-covalent association of the 42 cleaved residues with the remainder of the actin, induces an even larger bridge of density between the two strands. When the bridge is absent, the two C-terminal residues in F-actin are easily cleaved by trypsin, while these residues become increasingly resistant to tryptic cleavage as the bridge becomes more prominent. Conversely, cleavage of the two C-terminal residues leads to a conformational change in the DNase I-binding loop. Since both the DNase I-binding loop and the metal-binding site are quite distant from the C terminus, large allosteric effects must exist in F-actin. The conformational change in F-actin that results from the creation of this bridge may be induced by myosin binding, since this movement generates changes in actin's diffraction that are very similar to the changes in the muscle X-ray pattern during activation that are associated with the binding of myosin to the thin filament.

Reverse actin sliding triggers strong myosin binding that moves tropomyosin

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bekyarova, T.I.; Reedy, M.C.; Baumann, B.A.J.

2008-09-03

Actin/myosin interactions in vertebrate striated muscles are believed to be regulated by the 'steric blocking' mechanism whereby the binding of calcium to the troponin complex allows tropomyosin (TM) to change position on actin, acting as a molecular switch that blocks or allows myosin heads to interact with actin. Movement of TM during activation is initiated by interaction of Ca{sup 2+} with troponin, then completed by further displacement by strong binding cross-bridges. We report x-ray evidence that TM in insect flight muscle (IFM) moves in a manner consistent with the steric blocking mechanism. We find that both isometric contraction, at highmore » [Ca{sup 2+}], and stretch activation, at lower [Ca{sup 2+}], develop similarly high x-ray intensities on the IFM fourth actin layer line because of TM movement, coinciding with x-ray signals of strong-binding cross-bridge attachment to helically favored 'actin target zones.' Vanadate (Vi), a phosphate analog that inhibits active cross-bridge cycling, abolishes all active force in IFM, allowing high [Ca{sup 2+}] to elicit initial TM movement without cross-bridge attachment or other changes from relaxed structure. However, when stretched in high [Ca{sup 2+}], Vi-'paralyzed' fibers produce force substantially above passive response at pCa {approx} 9, concurrent with full conversion from resting to active x-ray pattern, including x-ray signals of cross-bridge strong-binding and TM movement. This argues that myosin heads can be recruited as strong-binding 'brakes' by backward-sliding, calcium-activated thin filaments, and are as effective in moving TM as actively force-producing cross-bridges. Such recruitment of myosin as brakes may be the major mechanism resisting extension during lengthening contractions.« less

Changes in corneal biometry and the associated histology in rhesus monkeys wearing orthokeratology contact lenses.

PubMed

Ding, Hui; Pu, Aijun; He, Hong; Xie, Ruo Zhong; Yang, Jun; Liao, Aiping; Gao, Shaohui; Zhong, Xingwu

2012-08-01

This study compared the effect of orthokeratology (OK) lens wear on corneal topography and microstructure between the 1-night and 30-night wearers to predict any adverse responses of the eyes to long-term OK lens wear. Nine rhesus monkeys with one eye wearing an OK lens and the fellow eye wearing a rigid gas permeable (RGP) lens were used in this study. The lenses were worn for 11 hours, with the measurement of corneal curvature and thickness and axial components of the eye at 0 night (n = 9), 1 night (n = 9), and 30 nights (n = 6). Histology was performed at 1 and 30 nights of lens wear. Corneal surface power in the 3-mm central region was reduced in the OK eye when compared with the RGP eye (P < 0.05). Central corneal thickness was only reduced in 30 nights of OK lens wear (P < 0.05). Under light microscopy, the 1-night OK or RGP eyes showed normal morphology in all layers of the cornea. The 30-night OK eyes showed a thinned central corneal epithelium and a thickened but less stratified paracentral corneal epithelium when compared with the 30-night RGP eyes. Under electron microscopy, intercellular junctions of corneal epithelium in the 30-night OK eyes were much looser than those in the 1-night OK eyes. However, the density and morphology of hemidesmosomes were similar between the OK and RGP eyes at 30 nights of lens wear. Overnight OK can change the corneal curvature through anterior corneal remodeling, resulting in an effective correction of myopia. However, a decrease in intercellular junctions during long-term wear may compromise the functions of the corneal epithelium as a border mechanical barrier.

Prediction of water loss and viscoelastic deformation of apple tissue using a multiscale model.

PubMed

Aregawi, Wondwosen A; Abera, Metadel K; Fanta, Solomon W; Verboven, Pieter; Nicolai, Bart

2014-11-19

A two-dimensional multiscale water transport and mechanical model was developed to predict the water loss and deformation of apple tissue (Malus × domestica Borkh. cv. 'Jonagold') during dehydration. At the macroscopic level, a continuum approach was used to construct a coupled water transport and mechanical model. Water transport in the tissue was simulated using a phenomenological approach using Fick's second law of diffusion. Mechanical deformation due to shrinkage was based on a structural mechanics model consisting of two parts: Yeoh strain energy functions to account for non-linearity and Maxwell's rheological model of visco-elasticity. Apparent parameters of the macroscale model were computed from a microscale model. The latter accounted for water exchange between different microscopic structures of the tissue (intercellular space, the cell wall network and cytoplasm) using transport laws with the water potential as the driving force for water exchange between different compartments of tissue. The microscale deformation mechanics were computed using a model where the cells were represented as a closed thin walled structure. The predicted apparent water transport properties of apple cortex tissue from the microscale model showed good agreement with the experimentally measured values. Deviations between calculated and measured mechanical properties of apple tissue were observed at strains larger than 3%, and were attributed to differences in water transport behavior between the experimental compression tests and the simulated dehydration-deformation behavior. Tissue dehydration and deformation in the high relative humidity range ( > 97% RH) could, however, be accurately predicted by the multiscale model. The multiscale model helped to understand the dynamics of the dehydration process and the importance of the different microstructural compartments (intercellular space, cell wall, membrane and cytoplasm) for water transport and mechanical deformation.

An intercellular polyamine transfer via gap junctions regulates proliferation and response to stress in epithelial cells

PubMed Central

Desforges, Bénédicte; Curmi, Patrick A.; Bounedjah, Ouissame; Nakib, Samir; Hamon, Loic; De Bandt, Jean-Pascal; Pastré, David

2013-01-01

In the organism, quiescent epithelial cells have the potential to resume cycling as a result of various stimuli, including wound healing or oxidative stress. Because quiescent cells have a low polyamine level, resuming their growth requires an increase of their intracellular polyamine levels via de novo polyamine synthesis or their uptake from plasma. Another alternative, explored here, is an intercellular exchange with polyamine-rich cycling cells via gap junctions. We show that polyamines promote gap junction communication between proliferating cells by promoting dynamical microtubule plus ends at the cell periphery and thus allow polyamine exchange between cells. In this way, cycling cells favor regrowth in adjacent cells deprived of polyamines. In addition, intercellular interactions mediated by polyamines can coordinate the translational response to oxidative stress through the formation of stress granules. Some putative in vivo consequences of polyamine-mediated intercellular interactions are also discussed regarding cancer invasiveness and tissue regeneration. PMID:23515223

Host and viral RNA-binding proteins involved in membrane targeting, replication and intercellular movement of plant RNA virus genomes

PubMed Central

Hyodo, Kiwamu; Kaido, Masanori; Okuno, Tetsuro

2014-01-01

Many plant viruses have positive-strand RNA [(+)RNA] as their genome. Therefore, it is not surprising that RNA-binding proteins (RBPs) play important roles during (+)RNA virus infection in host plants. Increasing evidence demonstrates that viral and host RBPs play critical roles in multiple steps of the viral life cycle, including translation and replication of viral genomic RNAs, and their intra- and intercellular movement. Although studies focusing on the RNA-binding activities of viral and host proteins, and their associations with membrane targeting, and intercellular movement of viral genomes have been limited to a few viruses, these studies have provided important insights into the molecular mechanisms underlying the replication and movement of viral genomic RNAs. In this review, we briefly overview the currently defined roles of viral and host RBPs whose RNA-binding activity have been confirmed experimentally in association with their membrane targeting, and intercellular movement of plant RNA virus genomes. PMID:25071804

Molecular Engineering for Enhanced Charge Transfer in Thin-Film Photoanode.

PubMed

Kim, Jeong Soo; Kim, Byung-Man; Kim, Un-Young; Shin, HyeonOh; Nam, Jung Seung; Roh, Deok-Ho; Park, Jun-Hyeok; Kwon, Tae-Hyuk

2017-10-11

We developed three types of dithieno[3,2-b;2',3'-d]thiophene (DTT)-based organic sensitizers for high-performance thin photoactive TiO 2 films and investigated the simple but powerful molecular engineering of different types of bonding between the triarylamine electron donor and the conjugated DTT π-bridge by the introduction of single, double, and triple bonds. As a result, with only 1.3 μm transparent and 2.5-μm TiO 2 scattering layers, the triple-bond sensitizer (T-DAHTDTT) shows the highest power conversion efficiency (η = 8.4%; V OC = 0.73 V, J SC = 15.4 mA·cm -2 , and FF = 0.75) in an iodine electrolyte system under one solar illumination (AM 1.5, 1000 W·m -2 ), followed by the single-bond sensitizer (S-DAHTDTT) (η = 7.6%) and the double-bond sensitizer (D-DAHTDTT) (η = 6.4%). We suggest that the superior performance of T-DAHTDTT comes from enhanced intramolecular charge transfer (ICT) induced by the triple bond. Consequently, T-DAHTDTT exhibits the most active photoelectron injection and charge transport on a TiO 2 film during operation, which leads to the highest photocurrent density among the systems studied. We analyzed these correlations mainly in terms of charge injection efficiency, level of photocharge storage, and charge-transport kinetics. This study suggests that the molecular engineering of a triple bond between the electron donor and the π-bridge of a sensitizer increases the performance of dye-sensitized solar cell (DSC) with a thin photoactive film by enhancing not only J SC through improved ICT but also V OC through the evenly distributed sensitizer surface coverage.

Coronary Heart Disease Alters Intercellular Communication by Modifying Microparticle-Mediated MicroRNA Transport

PubMed Central

Finn, Nnenna A.; Eapen, Danny; Manocha, Pankaj; Kassem, Hatem Al; Lassegue, Bernard; Ghasemzadeh, Nima; Quyyumi, Arshed; Searles, Charles D.

2013-01-01

Coronary heart disease (CHD) is characterized by abnormal intercellular communication and circulating microRNAs (miRNAs) are likely involved in this process. Here, we show that CHD was associated with changes in the transport of circulating miRNA, particularly decreased miRNA enrichment in microparticles (MPs). Additionally, MPs from CHD patients were less efficient at transferring miRNA to cultured HUVECs, which correlated with their diminished capacity to bind developmental endothelial locus-1 (Del-1). In summary, CHD was associated with distinct changes in circulating miRNA transport and these changes may contribute to the abnormal intercellular communication that underlies CHD initiation and progression. PMID:24042051

Wafer-scale growth of VO2 thin films using a combinatorial approach

PubMed Central

Zhang, Hai-Tian; Zhang, Lei; Mukherjee, Debangshu; Zheng, Yuan-Xia; Haislmaier, Ryan C.; Alem, Nasim; Engel-Herbert, Roman

2015-01-01

Transition metal oxides offer functional properties beyond conventional semiconductors. Bridging the gap between the fundamental research frontier in oxide electronics and their realization in commercial devices demands a wafer-scale growth approach for high-quality transition metal oxide thin films. Such a method requires excellent control over the transition metal valence state to avoid performance deterioration, which has been proved challenging. Here we present a scalable growth approach that enables a precise valence state control. By creating an oxygen activity gradient across the wafer, a continuous valence state library is established to directly identify the optimal growth condition. Single-crystalline VO2 thin films have been grown on wafer scale, exhibiting more than four orders of magnitude change in resistivity across the metal-to-insulator transition. It is demonstrated that ‘electronic grade' transition metal oxide films can be realized on a large scale using a combinatorial growth approach, which can be extended to other multivalent oxide systems. PMID:26450653

Connexin 32 and its derived homotypic gap junctional intercellular communication inhibit the migration and invasion of transfected HeLa cells via enhancement of intercellular adhesion.

PubMed

Yang, Jie; Liu, Bing; Wang, Qin; Yuan, Dongdong; Hong, Xiaoting; Yang, Yan; Tao, Liang

2011-01-01

The effects of connexin (Cx) and its derived homotypic gap junctional intercellular communication (GJIC) between tumor cells on the invasion of metastatic cancers and the underlying mechanisms remain unclear. In this study, we investigated the influence of Cx32 and the homotypic GJIC mediated by this Cx on the migration, invasion and intercellular adhesion of transfected HeLa cells. The expression of Cx32 significantly increased cell adhesion and inhibited migration and invasion. The inhibition of GJIC by oleamide, a widely used GJIC inhibitor, reduced the enhanced adhesion and partly reversed the decreased migration and invasion that had been induced by Cx32 expression. Blockage of the p38 and extracellular signal-regulated kinase 1 and 2 mitogen-activated protein kinase (ERK1/2 MAPKs) pathways using their specific inhibitors attenuated the effects of Cx32, but not those of GJIC, on cell adhesion, migration and invasion. These results indicate that the homotypic GJIC mediated by Cx32, as well as the Cx itself, inhibit cell migration and invasion, most likely through the elevation of intercellular adhesion. The suppressive effect of Cx32 on the migration and invasion of cancer cells, but not that of its derived homotypic GJIC, partly depends on the activation of the p38 and the ERK1/2 MAPKs pathways.

Structural Changes in Isometrically Contracting Insect Flight Muscle Trapped following a Mechanical Perturbation

PubMed Central

Wu, Shenping; Liu, Jun; Perz-Edwards, Robert J.; Tregear, Richard T.; Winkler, Hanspeter; Franzini-Armstrong, Clara; Sasaki, Hiroyuki; Goldman, Yale E.; Reedy, Michael K.; Taylor, Kenneth A.

2012-01-01

The application of rapidly applied length steps to actively contracting muscle is a classic method for synchronizing the response of myosin cross-bridges so that the average response of the ensemble can be measured. Alternatively, electron tomography (ET) is a technique that can report the structure of the individual members of the ensemble. We probed the structure of active myosin motors (cross-bridges) by applying 0.5% changes in length (either a stretch or a release) within 2 ms to isometrically contracting insect flight muscle (IFM) fibers followed after 5–6 ms by rapid freezing against a liquid helium cooled copper mirror. ET of freeze-substituted fibers, embedded and thin-sectioned, provides 3-D cross-bridge images, sorted by multivariate data analysis into ∼40 classes, distinct in average structure, population size and lattice distribution. Individual actin subunits are resolved facilitating quasi-atomic modeling of each class average to determine its binding strength (weak or strong) to actin. ∼98% of strong-binding acto-myosin attachments present after a length perturbation are confined to “target zones” of only two actin subunits located exactly midway between successive troponin complexes along each long-pitch helical repeat of actin. Significant changes in the types, distribution and structure of actin-myosin attachments occurred in a manner consistent with the mechanical transients. Most dramatic is near disappearance, after either length perturbation, of a class of weak-binding cross-bridges, attached within the target zone, that are highly likely to be precursors of strong-binding cross-bridges. These weak-binding cross-bridges were originally observed in isometrically contracting IFM. Their disappearance following a quick stretch or release can be explained by a recent kinetic model for muscle contraction, as behaviour consistent with their identification as precursors of strong-binding cross-bridges. The results provide a detailed model for contraction in IFM that may be applicable to contraction in other types of muscle. PMID:22761792

EXPOSURE OF CULTURED MYOCYTES TO ZINC RESULTS IN ALTERED BEAT RATE AND INTERCELLULAR COMMUNICATION.

EPA Science Inventory

Exposure of cultured myocytes to zinc results in altered beat rate and intercellular communication

Graff, Donald W, Devlin, Robert B, Brackhan, Joseph A, Muller-Borer, Barbara J, Bowman, Jill S, Cascio, Wayne E.

Exposure to ambient air pollution particulate matter (...

Piezoelectric micromachined ultrasonic transducers based on PZT thin films.

PubMed

Muralt, Paul; Ledermann, Nicolas; Baborowski, Jacek; Barzegar, Abdolghaffar; Gentil, Sandrine; Belgacem, Brahim; Petitgrand, Sylvain; Bosseboeuf, Alain; Setter, Nava

2005-12-01

This paper describes fabrication and characterization results of piezoelectric micromachined ultrasonic transducers (pMUTs) based on 2-microm-thick Pb(Zr0.53Ti0.47O3) (PZT) thin films. The applied structures are circular plates held at four bridges, thus partially unclamped. A simple analytical model for the fully clamped structure is used as a reference to optimize design parameters such as thickness relations and electrodes, and to provide approximate predictions for coupling coefficients related to previously determined thin film properties. The best coupling coefficient was achieved with a 270-microm plate and amounted to kappa2 = 5.3%. This value compares well with the calculated value based on measured small signal dielectric (epsilon = 1050) and piezoelectric (e3l,f = 15 Cm(-2)) properties of the PZT thin film at 100 kV/cm dc bias. The resonances show relatively large Q-factors, which can be partially explained by the small diameters as compared to the sound wavelength in air and in the test liquid (Fluorinert 77). A transmit-receive experiment with two quasi-identical pMUTs was performed showing significant signal transmission up to a distance of 20 cm in air and 2 cm in the test liquid.

Guard cell photosynthesis is critical for stomatal turgor production, yet does not directly mediate CO2- and ABA-induced stomatal closing

PubMed Central

Azoulay-Shemer, Tamar; Palomares, Axxell; Bagheri, Andish; Israelsson-Nordstrom, Maria; Engineer, Cawas B.; Bargmann, Bastiaan O.R.; Stephan, Aaron B.; Schroeder, Julian I.

2015-01-01

SUMMARY Stomata mediate gas exchange between the inter-cellular spaces of leaves and the atmosphere. CO2 levels in leaves (Ci) are determined by respiration, photosynthesis, stomatal conductance and atmospheric [CO2]. [CO2] in leaves mediates stomatal movements. The role of guard-cell photosynthesis in stomatal conductance responses is a matter of debate, and genetic approaches are needed. We have generated transgenic Arabidopsis plants that are chlorophyll-deficient in guard cells only, expressing a constitutively active chlorophyllase in a guard-cell specific enhancer trap-line. Our data show that more than 90% of guard cells were chlorophyll-deficient. Interestingly, approximately ~ 45% of stomata had an unusual, previously not-described, morphology of thin-shaped chlorophyll-less stomata. Nevertheless, stomatal size, stomatal index, plant morphology, and whole-leaf photosynthetic parameters (PSII, qP, qN, FV′/FM′) were comparable to wild-type plants. Time-resolved intact leaf gas exchange analyses showed a reduction in stomatal conductance and carbon assimilation rates of the transgenic plants. Normalization of CO2 responses showed that stomata of transgenic plants respond to [CO2] shifts. Detailed stomatal aperture measurements of normal kidney-shaped stomata, which lack chlorophyll, showed stomatal closing responses to [CO2] elevation and abscisic acid (ABA), while thin-shaped stomata were continuously closed. Our present findings show that stomatal movement responses to [CO2] and ABA are functional in guard cells that lack chlorophyll. These data suggest that guard-cell CO2 and ABA signal transduction are not directly modulated by guard-cell photosynthesis/electron transport. Moreover, the finding that chlorophyll-less stomata cause a “deflated” thin-shaped phenotype, suggests that photosynthesis in guard cells is critical for energization and guard-cell turgor production. PMID:26096271

THE FINE STRUCTURE OF MEISSNER's TOUCH CORPUSCLES OF HUMAN FINGERS

PubMed Central

Cauna, Nikolajs; Ross, Leonard L.

1960-01-01

Thin slices of the finger pads of six individuals were fixed in buffered 1 per cent osmic acid, embedded in deaerated, nitrogenated methacrylate, and cut into thin sections for electron microscopic study. Before embedding, the slices were trimmed so as to include several digital tactile corpuscles. Some thin sections were stained in 10 per cent aqueous phosphotungstic acid solution. The principal part of Meissner's corpuscle is made up of flattened laminar cells stretching across the corpuscle in irregular layers. The perinuclear cytoplasm of these cells contains numerous small mitochondria, a sparse granular endoplasmic reticulum, and a large number of small vesicles. Nerve fibers enter the side or base of the corpuscle, lose their myelin sheaths, and follow a meandering course between the laminar cell plates. The nerve endings enter into a close appositional relationship with the flattened portions of the laminar cells. In some areas the apposed axolemma and cell membranes are slightly thickened with small vesicles located along the cell membrane or on both surfaces. These regions are interpreted as synapses. The most prominent feature of the nerve endings is an extraordinary accumulation of small mitochondria which vary in size and internal density. The nerve endings also contain vacuoles, groups of dense concentric membranes, and small dense vesicles of irregular distribution. The laminar cells are separated from one another by a dense intercellular substance of uniform thickness which also envelops the entire corpuscle. This material contains randomly oriented collagen fibers and fine fibrils bound together by a dense material at nodal points recurring at regular intervals of approximately 120 mµ. These findings are discussed in relation to the problems of the function of Meissner's corpuscle, neural material loss and replacement, and the presence of synapses. PMID:13691669

Dissociative electron attachment to DNA-diamine thin films: Impact of the DNA close environment on the OH− and O− decay channels

PubMed Central

Boulanouar, Omar; Fromm, Michel; Mavon, Christophe; Cloutier, Pierre; Sanche, Léon

2013-01-01

We measure the desorption of anions stimulated by the impact of 0–20 eV electrons on highly uniform thin films of plasmid DNA-diaminopropane. The results are accurately correlated with film thickness and composition by AFM and XPS measurements, respectively. Resonant structures in the H−, O−, and OH− yield functions are attributed to the decay of transient anions into the dissociative electron attachment (DEA) channel. The diamine induces ammonium-phosphate bridges along the DNA backbone, which suppresses the DEA O− channel and in counter-part increases considerably the desorption of OH−. The close environment of the phosphate groups may therefore play an important role in modulating the rate and type of DNA damages induced by low energy electrons. PMID:23927286

Suction and cohesion demise in desaturating granular medium

NASA Astrophysics Data System (ADS)

Hueckel, T.; Mielniczuk, B.; El-Youssoufi, S. M.

2017-12-01

Continuum mechanics for unsaturated soils is based on the assumption of a one-to-one relationship betwee saturation degree and suction represented by the characteristic curve. Such curve commonly shows exceedingly high values of suction at saturation decreasing below 10%. We have performed a series of experiments on physical micro-structural models of 8-, 5, 4, 3, and 2-grain assemblies filled with water forming capillary, funicular and pendular bridges. Dynamic variables characterizing the evolution include: Laplace pressure, surface tension force, total intergralular force, contact angle and contact perimeter length. The Laplace pressure was calculated from the directly measured curvatures of interface surface for 2-grain bridges, and estimated from tomography stills for 3 grain bridges. The initial negative Laplace pressure (suction) as well as total intergranular force increase modestly at the begining of evaporation, but undergo an unstable decrease at the advanced stage, often with a jump in the force known as a Haines jumps since 1925. Laplace pressure turns into positive values prior to rupture for 2-grain bodies. For 3-grain bridges there is never an exceedingly high intergranular force of suction, reported in macro-scale experiments. For multiple-grain bodies there are two types of instabilities, depending on densitiy of the assembly and the Gaussian curvature (GC): at positive GC points it is thin-sheet instability, while at negative GC points instability is linked with air entry fingers, all associated with the split of assemblies into smaller isolated funicular, and eventually pendular bodies. The multi-grain bridges instabilities are linked to material drying cracking, the instabilities in 2 grain systems mean eventual loss of cohesion.

Superconducting noise bolometer with microwave bias and readout for array applications

NASA Astrophysics Data System (ADS)

Kuzmin, A. A.; Semenov, A. D.; Shitov, S. V.; Merker, M.; Wuensch, S. H.; Ustinov, A. V.; Siegel, M.

2017-07-01

We present a superconducting noise bolometer for terahertz radiation, which is suitable for large-format arrays. It is based on an antenna-coupled superconducting micro-bridge embedded in a high-quality factor superconducting resonator for a microwave bias and readout with frequency-division multiplexing in the GHz range. The micro-bridge is kept below its critical temperature and biased with a microwave current of slightly lower amplitude than the critical current of the micro-bridge. The response of the detector is the rate of superconducting fluctuations, which depends exponentially on the concentration of quasiparticles in the micro-bridge. Excess quasiparticles are generated by an incident THz signal. Since the quasiparticle lifetime increases exponentially at lower operation temperature, the noise equivalent power rapidly decreases. This approach allows for large arrays of noise bolometers operating above 1 K with sensitivity, limited by 300-K background noise. Moreover, the response of the bolometer always dominates the noise of the readout due to relatively large amplitude of the bias current. We performed a feasibility study on a proof-of-concept device with a 1.0 × 0.5 μm2 micro-bridge from a 9-nm thin Nb film on a sapphire substrate. Having a critical temperature of 5.8 K, it operates at 4.2 K and is biased at the frequency 5.6 GHz. For the quasioptical input at 0.65 THz, we measured the noise equivalent power ≈3 × 10-12 W/Hz1/2, which is close to expectations for this particular device in the noise-response regime.

Coronary heart disease alters intercellular communication by modifying microparticle-mediated microRNA transport.

PubMed

Finn, Nnenna A; Eapen, Danny; Manocha, Pankaj; Al Kassem, Hatem; Lassegue, Bernard; Ghasemzadeh, Nima; Quyyumi, Arshed; Searles, Charles D

2013-11-01

Coronary heart disease (CHD) is characterized by abnormal intercellular communication and circulating microRNAs (miRNAs) are likely involved in this process. Here, we show that CHD was associated with changes in the transport of circulating miRNA, particularly decreased miRNA enrichment in microparticles (MPs). Additionally, MPs from CHD patients were less efficient at transferring miRNA to cultured HUVECs, which correlated with their diminished capacity to bind developmental endothelial locus-1 (Del-1). In summary, CHD was associated with distinct changes in circulating miRNA transport and these changes may contribute to the abnormal intercellular communication that underlies CHD initiation and progression. Published by Elsevier B.V.

Antibody immobilization using pneumatic spray: comparison with the avidin-biotin bridge immobilization method.

PubMed

Figueroa, Jhon; Magaña, Sonia; Lim, Daniel V; Schlaf, Rudy

2012-12-14

The formation of a thin antibody film on a glass surface using pneumatic spray was investigated as a potential immobilization technique for capturing pathogenic targets. Goat-Escherichia coli O157:H7 IgG films were made by pneumatic spray and compared against the avidin-biotin bridge immobilized films by assaying with green fluorescent protein (GFP) transformed E. coli O157:H7 cells and fluorescent reporter antibodies. Functionality, stability, and immobilization of the films were tested. The pneumatic spray films had lower fluorescence intensity values than the avidin-biotin bridge films but resulted in similar detection for E. coli O157:H7 at 10(5)-10(7)cells/ml sample concentrations with no detection of non-E. coli O157:H7 strains. Both methods also resulted in similar percent capture efficiencies. The results demonstrated that immobilization of antibody via pneumatic spray did not render the antibody non-functional and produced stable antibody films. The amount of time necessary for immobilization of the antibody was reduced significantly from 24h for the avidin-biotin bridge to 7 min using the pneumatic spray technique, with additional benefits of greatly reduced use of materials and chemicals. The pneumatic spray technique promises to be an alternative for the immobilization of antibodies on glass slides for capturing pathogenic targets and use in biosensor type devices. Copyright © 2012. Published by Elsevier B.V.

MicroRNA Intercellular Transfer and Bioelectrical Regulation of Model Multicellular Ensembles by the Gap Junction Connectivity.

PubMed

Cervera, Javier; Meseguer, Salvador; Mafe, Salvador

2017-08-17

We have studied theoretically the microRNA (miRNA) intercellular transfer through voltage-gated gap junctions in terms of a biophysically grounded system of coupled differential equations. Instead of modeling a specific system, we use a general approach describing the interplay between the genetic mechanisms and the single-cell electric potentials. The dynamics of the multicellular ensemble are simulated under different conditions including spatially inhomogeneous transcription rates and local intercellular transfer of miRNAs. These processes result in spatiotemporal changes of miRNA, mRNA, and ion channel protein concentrations that eventually modify the bioelectrical states of small multicellular domains because of the ensemble average nature of the electrical potential. The simulations allow a qualitative understanding of the context-dependent nature of the effects observed when specific signaling molecules are transferred through gap junctions. The results suggest that an efficient miRNA intercellular transfer could permit the spatiotemporal control of small cellular domains by the conversion of single-cell genetic and bioelectric states into multicellular states regulated by the gap junction interconnectivity.

A Study of Intercellular Spaces in the Rabbit Jejunum during Acute Volume Expansion and after Treatment with Cholera Toxin

PubMed Central

DiBona, Donald R.; Chen, Lincoln C.; Sharp, Geoffrey W. G.

1974-01-01

The effects of acute volume expansion and of intraluminal administration of cholera toxin have been examined in rabbit jejunum. Acute volume expansion was shown to reverse the normal reabsorptive flux of water and cause significant fluid secretion. Phase and electronmicroscopic examination of the jejunal epithelium showed that marked distension of the intercellular spaces had occurred. Examination of the jejunal epithelium after treatment with cholera toxin showed that, in association with high rates of fluid secretion, the intercellular spaces were extremely small and lateral membranes of adjacent cells were in close apposition to one another. Thus the mechanisms of fluid secretion in these two situations would appear to be quite different. The secretion associated with volume expansion, and accompanied by a rise in venous pressure and bullous deformations of terminal junctions, could well be due to hydrostatic pressure applied through intercellular channels. The secretion of cholera appears to be unrelated to hydrostatic pressure and is more likely due to body-to-lumen active ion transport. Images PMID:4596506

Kinetic Measurements Reveal Enhanced Protein-Protein Interactions at Intercellular Junctions

PubMed Central

Shashikanth, Nitesh; Kisting, Meridith A.; Leckband, Deborah E.

2016-01-01

The binding properties of adhesion proteins are typically quantified from measurements with soluble fragments, under conditions that differ radically from the confined microenvironment of membrane bound proteins in adhesion zones. Using classical cadherin as a model adhesion protein, we tested the postulate that confinement within quasi two-dimensional intercellular gaps exposes weak protein interactions that are not detected in solution binding assays. Micropipette-based measurements of cadherin-mediated, cell-cell binding kinetics identified a unique kinetic signature that reflects both adhesive (trans) bonds between cadherins on opposing cells and lateral (cis) interactions between cadherins on the same cell. In solution, proposed lateral interactions were not detected, even at high cadherin concentrations. Mutations postulated to disrupt lateral cadherin association altered the kinetic signatures, but did not affect the adhesive (trans) binding affinity. Perturbed kinetics further coincided with altered cadherin distributions at junctions, wound healing dynamics, and paracellular permeability. Intercellular binding kinetics thus revealed cadherin interactions that occur within confined, intermembrane gaps but not in solution. Findings further demonstrate the impact of these revealed interactions on the organization and function of intercellular junctions. PMID:27009566

Molecular Diffusion through Cyanobacterial Septal Junctions.

PubMed

Nieves-Morión, Mercedes; Mullineaux, Conrad W; Flores, Enrique

2017-01-03

Heterocyst-forming cyanobacteria grow as filaments in which intercellular molecular exchange takes place. During the differentiation of N 2 -fixing heterocysts, regulators are transferred between cells. In the diazotrophic filament, vegetative cells that fix CO 2 through oxygenic photosynthesis provide the heterocysts with reduced carbon and heterocysts provide the vegetative cells with fixed nitrogen. Intercellular molecular transfer has been traced with fluorescent markers, including calcein, 5-carboxyfluorescein, and the sucrose analogue esculin, which are observed to move down their concentration gradient. In this work, we used fluorescence recovery after photobleaching (FRAP) assays in the model heterocyst-forming cyanobacterium Anabaena sp. strain PCC 7120 to measure the temperature dependence of intercellular transfer of fluorescent markers. We find that the transfer rate constants are directly proportional to the absolute temperature. This indicates that the "septal junctions" (formerly known as "microplasmodesmata") linking the cells in the filament allow molecular exchange by simple diffusion, without any activated intermediate state. This constitutes a novel mechanism for molecular transfer across the bacterial cytoplasmic membrane, in addition to previously characterized mechanisms for active transport and facilitated diffusion. Cyanobacterial septal junctions are functionally analogous to the gap junctions of metazoans. Although bacteria are frequently considered just as unicellular organisms, there are bacteria that behave as true multicellular organisms. The heterocyst-forming cyanobacteria grow as filaments in which cells communicate. Intercellular molecular exchange is thought to be mediated by septal junctions. Here, we show that intercellular transfer of fluorescent markers in the cyanobacterial filament has the physical properties of simple diffusion. Thus, cyanobacterial septal junctions are functionally analogous to metazoan gap junctions, although their molecular components appear unrelated. Like metazoan gap junctions, the septal junctions of cyanobacteria allow the rapid intercellular exchange of small molecules, without stringent selectivity. Our finding expands the repertoire of mechanisms for molecular transfer across the plasma membrane in prokaryotes. Copyright © 2017 Nieves-Morión et al.

Biomaterials-based electronics: polymers and interfaces for biology and medicine.

PubMed

Muskovich, Meredith; Bettinger, Christopher J

2012-05-01

Advanced polymeric biomaterials continue to serve as a cornerstone for new medical technologies and therapies. The vast majority of these materials, both natural and synthetic, interact with biological matter in the absence of direct electronic communication. However, biological systems have evolved to synthesize and utilize naturally-derived materials for the generation and modulation of electrical potentials, voltage gradients, and ion flows. Bioelectric phenomena can be translated into potent signaling cues for intra- and inter-cellular communication. These cues can serve as a gateway to link synthetic devices with biological systems. This progress report will provide an update on advances in the application of electronically active biomaterials for use in organic electronics and bio-interfaces. Specific focus will be granted to covering technologies where natural and synthetic biological materials serve as integral components such as thin film electronics, in vitro cell culture models, and implantable medical devices. Future perspectives and emerging challenges will also be highlighted. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Cell Connections by Tunneling Nanotubes: Effects of Mitochondrial Trafficking on Target Cell Metabolism, Homeostasis, and Response to Therapy

PubMed Central

2017-01-01

Intercellular communications play a major role in tissue homeostasis and responses to external cues. Novel structures for this communication have recently been described. These tunneling nanotubes (TNTs) consist of thin-extended membrane protrusions that connect cells together. TNTs allow the cell-to-cell transfer of various cellular components, including proteins, RNAs, viruses, and organelles, such as mitochondria. Mesenchymal stem cells (MSCs) are both naturally present and recruited to many different tissues where their interaction with resident cells via secreted factors has been largely documented. Their immunosuppressive and repairing capacities constitute the basis for many current clinical trials. MSCs recruited to the tumor microenvironment also play an important role in tumor progression and resistance to therapy. MSCs are now the focus of intense scrutiny due to their capacity to form TNTs and transfer mitochondria to target cells, either in normal physiological or in pathological conditions, leading to changes in cell energy metabolism and functions, as described in this review. PMID:28659978

Coalescence of Drops of a Power-law Fluid

NASA Astrophysics Data System (ADS)

Kamat, Pritish; Thete, Sumeet; Basaran, Osman

2014-11-01

Drop coalescence is crucial in a host of industrial, household, and natural processes that involve dispersions. Coalescence is a rate-controlling process in breaking emulsions and strongly influences drop-size-distributions in sprays. In a continuum approach, coalescence begins by the formation of a microscopic, non-slender bridge connecting the two drops. Indefinitely large axial curvature at the neck results in local lowering of pressure that drives fluid from the bulk of the drops toward the neck, thereby causing the bridge radius r (t) and height z (t) to increase in time t. The coalescence of Newtonian drops in air has heretofore been thoroughly studied. Here, we extend these earlier studies by analyzing the coalescence of drops of power-law fluids because many fluids encountered in real applications, including cosmetic creams, shampoos, grease, and paint, exhibit power-law (deformation-rate thinning) rheology. On account of the non-slender geometry of the liquid bridge connecting the two drops (z << r) , we analyze the resulting free surface flow problem by numerical simulation. Among other results, we present and discuss the nature of flows and scaling behaviors for r and z as functions of the initial viscosity and power-law index (0 < n <= 1) .

Intercellular communication in Arabidopsis thaliana pollen discovered via AHG3 transcript movement from the vegetative cell to sperm

USDA-ARS?s Scientific Manuscript database

An Arabidopsis pollen grain (male gametophyte) consists of three cells: the vegetative cell, which forms the pollen tube, and two sperm cells enclosed within the vegetative cell. It is still unclear if there is intercellular communication between the vegetative cell and the sperm cells. Here we show...

Thin film strain transducer

NASA Technical Reports Server (NTRS)

Rand, J. L. (Inventor)

1984-01-01

A strain transducer system and process for making the same is disclosed. A beryllium copper ring having four strain gages is electrically connected in Wheatstone bridge fashion to the output instrumentation. Tabs are bonded to a balloon or like surface with strain on the surface causing bending of a ring which provides an electrical signal through the gages proportional to the surface strain. A photographic pattern of a one half ring segment as placed on a sheet of beryllium copper for chem-mill etch formation is illustrated.

Research on pressure sensors for biomedical instruments

NASA Technical Reports Server (NTRS)

Angell, J. B.

1975-01-01

The development of a piezo-resistive pressure transducer is discussed suitable for recording pressures typically encountered in biomedical applications. The pressure transducer consists of a thin silicon diaphragm containing four strain-sensitive resistors, and is fabricated using silicon monolithic integrated-circuit technology. The pressure transducers can be as small as 0.7 mm outer diameter, and are, as a result, suitable for mounting at the tip of a catheter. Pressure-induced stress in the diaphragm is sensed by the resistors, which are interconnected to form a Wheatstone bridge.

Modeling Self-Heating Effects in Nanoscale Devices

NASA Astrophysics Data System (ADS)

Raleva, K.; Shaik, A. R.; Vasileska, D.; Goodnick, S. M.

2017-08-01

Accurate thermal modeling and the design of microelectronic devices and thin film structures at the micro- and nanoscales poses a challenge to electrical engineers who are less familiar with the basic concepts and ideas in sub-continuum heat transport. This book aims to bridge that gap. Efficient heat removal methods are necessary to increase device performance and device reliability. The authors provide readers with a combination of nanoscale experimental techniques and accurate modeling methods that must be employed in order to determine a device's temperature profile.

1976 Navy Study on Superconductive Electronics, August 2-13, 1976, Naval Postgraduate School, Monterey, California

DTIC Science & Technology

1976-01-01

excluded from this Study except as points of reference. The conclusions identify a number of relevant functional technology items which are of...SQUID 20 4.2 4.3 The RF SQUID and its associated circuitry and operating characteristics 22 Bulk point contact SQUID formed by...a point contact, a constriction or microbridge in a thin film ("Dayem Bridge"), an S-N-S junction (in which pairs move through a "normal" metal

Fundamental transport mechanisms, fabrication and potential applications of nanoporous atomically thin membranes.

PubMed

Wang, Luda; Boutilier, Michael S H; Kidambi, Piran R; Jang, Doojoon; Hadjiconstantinou, Nicolas G; Karnik, Rohit

2017-06-06

Graphene and other two-dimensional materials offer a new approach to controlling mass transport at the nanoscale. These materials can sustain nanoscale pores in their rigid lattices and due to their minimum possible material thickness, high mechanical strength and chemical robustness, they could be used to address persistent challenges in membrane separations. Here we discuss theoretical and experimental developments in the emerging field of nanoporous atomically thin membranes, focusing on the fundamental mechanisms of gas- and liquid-phase transport, membrane fabrication techniques and advances towards practical application. We highlight potential functional characteristics of the membranes and discuss applications where they are expected to offer advantages. Finally, we outline the major scientific questions and technological challenges that need to be addressed to bridge the gap from theoretical simulations and proof-of-concept experiments to real-world applications.

Variable temperature superconducting microscope

NASA Astrophysics Data System (ADS)

Cheng, Bo; Yeh, W. J.

2000-03-01

We have developed and tested a promising type of superconducting quantum interference device (SQUID) microscope, which can be used to detect vortex motion and can operate in magnetic fields over a large temperature range. The system utilizes a single-loop coupling transformer, consisting of a patterned high Tc superconducting thin film. At one end of the transformer, a 20 μm diam detecting loop is placed close to the sample. At the other end, a large loop is coupled to a NbTi coil, which is connected to a low Tc SQUID sensor. Transformers in a variety of sizes have been tested and calibrated. The results show that the system is capable of detecting the motion of a single vortex. We have used the microscope to study the behavior of moving vortices at various positions in a YBa2Cu3O7 thin film bridge.

Intrinsic superconducting transport properties of ultra-thin Fe1+ y Te0.6Se0.4 microbridges

NASA Astrophysics Data System (ADS)

Sun, HanCong; Lv, YangYang; Lu, DaChuan; Yang, ZhiBao; Zhou, XianJing; Hao, LuYao; Xing, XiangZhuo; Zou, Wei; Li, Jun; Shi, ZhiXiang; Xu, WeiWei; Wang, HuaBing; Wu, PeiHeng

2017-11-01

We investigated the superconducting properties of Fe1+ y Te0.6Se0.4 single-crystalline microbridges with a width of 4 μm and thicknesses ranging from 20.8 to 136.2 nm. The temperature-dependent in-plane resistance of the bridges exhibited a type of metal-insulator transition in the normal state. The critical current density ( J c) of the microbridge with a thickness of 136.2 nm was 82.3 kA/cm2 at 3K and reached 105 kA/cm2 after extrapolation to T = 0 K. The current versus voltage characteristics of the microbridges showed a Josephson-like behavior with an obvious hysteresis. These results demonstrate the potential application of ultra-thin Fe-based microbridges in superconducting electronic devices such as bolometric detectors.

Mechanistic understanding of cellular level of water in plant-based food material

NASA Astrophysics Data System (ADS)

Khan, Md. Imran H.; Kumar, C.; Karim, M. A.

2017-06-01

Understanding of water distribution in plant-based food material is crucial for developing an accurate heat and mass transfer drying model. Generally, in plant-based food tissue, water is distributed in three different spaces namely, intercellular water, intracellular water, and cell wall water. For hygroscopic material, these three types of water transport should be considered for actual understanding of heat and mass transfer during drying. However, there is limited study dedicated to the investigation of the moisture distribution in a different cellular environment in the plant-based food material. Therefore, the aim of the present study was to investigate the proportion of intercellular water, intracellular water, and cell wall water inside the plant-based food material. During this study, experiments were performed for two different plant-based food tissues namely, eggplant and potato tissue using 1H-NMR-T2 relaxometry. Various types of water component were calculated by using multicomponent fits of the T2 relaxation curves. The experimental result showed that in potato tissue 80-82% water exist in intracellular space; 10-13% water in intercellular space and only 4-6% water exist in the cell wall space. In eggplant tissue, 90-93% water in intracellular space, 4-6% water exists in intercellular space and the remaining percentage of water is recognized as cell wall water. The investigated results quantify different types of water in plant-based food tissue. The highest proportion of water exists in intracellular spaces. Therefore, it is necessary to include different transport mechanism for intracellular, intercellular and cell wall water during modelling of heat and mass transfer during drying.

Intercellular interaction mechanisms for the origination of blast crisis in chronic myeloid leukemia

PubMed Central

Sachs, Rainer; Johnsson, Kerstin; Hahnfeldt, Philip; Luo, Janet; Chen, Allen; Hlatky, Lynn

2011-01-01

Chronic myeloid leukemia (CML) is characterized by a specific chromosome translocation, and its pathobiology is considered comparatively well understood. Thus, quantitative analysis of CML and its progression to blast crisis may help elucidate general mechanisms of carcinogenesis and cancer progression. Hitherto it has been widely postulated that CML blast crisis originates mainly via cell-autonomous mechanisms such as secondary mutations or genomic instability, rather than by intercellular interactions. However, recent results suggest that intercellular interactions play an important role in carcinogenesis. In this study, we analyzed alternative mechanisms, including pairwise intercellular interactions, for CML blast crisis origination. A quantitative, mechanistic cell population dynamics model was employed. This model used recent data on imatinib-treated CML; it also used earlier clinical data, not previously incorporated into current mathematical CML/imatinib models. With the pre-imatinib data, which include results on many more blast crises, we obtained evidence that the driving mechanism for blast crisis origination is intercellular interaction between specific cell types. Assuming leukemic-normal interactions resulted in a statistically significant improvement over assuming either cell-autonomous mechanisms or interactions between leukemic cells. This conclusion was robust with regard to changes in the model’s adjustable parameters. Application of the results to patients treated with imatinib suggests that imatinib may act not only on malignant blast precursors, but also, to a limited degree, on the malignant blasts themselves. Major Findings A comprehensive mechanistic model gives evidence that the main driving mechanisms for CML blast crisis origination are interactions between leukemic and normal cells. PMID:21487044

Subsets of ATP-sensitive potassium channel (KATP) inhibitors increase gap junctional intercellular communication in metastatic cancer cell lines independent of SUR expression

USDA-ARS?s Scientific Manuscript database

Gap junctional intercellular communication (GJIC) is a process whereby cells share molecules and nutrients with each other by physical contact through cell membrane pores. In tumor cells, GJIC is often altered, suggesting that this process may be important in the context of cancer. Certain ion chan...

Increase in the adhesion molecule P-selectin in endothelium overlying atherosclerotic plaques. Coexpression with intercellular adhesion molecule-1.

PubMed Central

Johnson-Tidey, R. R.; McGregor, J. L.; Taylor, P. R.; Poston, R. N.

1994-01-01

P-selectin (GMP-140) is an adhesion molecule present within endothelial cells that is rapidly translocated to the cell membrane upon activation, where it mediates endothelial-leukocyte interactions. Immunohistochemical analysis of human atherosclerotic plaques has shown strong expression of P-selectin by the endothelium overlying active atherosclerotic plaques. P-selectin is not, however, detected in normal arterial endothelium or in endothelium overlying inactive fibrous plaques. Color image analysis was used to quantitate the degree of P-selectin expression in the endothelium and demonstrates a statistically significant increase in P-selectin expression by atherosclerotic endothelial cells. Double immunofluorescence shows that some of this P-selectin is expressed on the luminal surface of the endothelial cells. Previous work has demonstrated a significant up-regulation in the expression of the intercellular adhesion molecule-1 in atherosclerotic endothelium and a study on the expression of intercellular adhesion molecule-1 and P-selectin in atherosclerosis shows a highly positive correlation. These results suggest that the selective and cooperative expression of P-selectin and intercellular adhesion molecule-1 may be involved in the recruitment of monocytes into sites of atherosclerosis. Images Figure 1 Figure 3 Figure 4 Figure 5 PMID:7513951

Transient inter-cellular polymeric linker.

PubMed

Ong, Siew-Min; He, Lijuan; Thuy Linh, Nguyen Thi; Tee, Yee-Han; Arooz, Talha; Tang, Guping; Tan, Choon-Hong; Yu, Hanry

2007-09-01

Three-dimensional (3D) tissue-engineered constructs with bio-mimicry cell-cell and cell-matrix interactions are useful in regenerative medicine. In cell-dense and matrix-poor tissues of the internal organs, cells support one another via cell-cell interactions, supplemented by small amount of the extra-cellular matrices (ECM) secreted by the cells. Here we connect HepG2 cells directly but transiently with inter-cellular polymeric linker to facilitate cell-cell interaction and aggregation. The linker consists of a non-toxic low molecular-weight polyethyleneimine (PEI) backbone conjugated with multiple hydrazide groups that can aggregate cells within 30 min by reacting with the aldehyde handles on the chemically modified cell-surface glycoproteins. The cells in the cellular aggregates proliferated; and maintained the cortical actin distribution of the 3D cell morphology while non-aggregated cells died over 7 days of suspension culture. The aggregates lost distinguishable cell-cell boundaries within 3 days; and the ECM fibers became visible around cells from day 3 onwards while the inter-cellular polymeric linker disappeared from the cell surfaces over time. The transient inter-cellular polymeric linker can be useful for forming 3D cellular and tissue constructs without bulk biomaterials or extensive network of engineered ECM for various applications.

Analysis of gap junctional intercellular communications using a dielectrophoresis-based microchip.

PubMed

Tellez-Gabriel, Marta; Charrier, Céline; Brounais-Le Royer, Bénédicte; Mullard, Mathilde; Brown, Hannah K; Verrecchia, Franck; Heymann, Dominique

2017-03-01

Gap junctions are transmembrane structures that directly connect the cytoplasm of adjacent cells, making intercellular communications possible. It has been shown that the behaviour of several tumours - such as bone tumours - is related to gap junction intercellular communications (GJIC). Several methodologies are available for studying GJIC, based on measuring different parameters that are useful for multiple applications, such as the study of carcinogenesis for example. These methods nevertheless have several limitations. The present manuscript describes the setting up of a dielectrophoresis (DEP)-based lab-on-a-chip platform for the real-time study of Gap Junctional Intercellular Communication between osteosarcoma cells and the main cells accessible to their microenvironment. We conclude that using the DEParray technology for the GJIC assessment has several advantages comparing to current techniques. This methodology is less harmful for cells integrity; cells can be recovered after interaction to make further molecular analysis; it is possible to study GJIC in real time; we can promote cell interactions using up to five different populations. The setting up of this new methodology overcomes several difficulties to perform experiments for solving questions about GJIC process that we are not able to do with current technics. Copyright © 2017 Elsevier GmbH. All rights reserved.

Intercellular signaling pathways active during intervertebral disc growth, differentiation, and aging.

PubMed

Dahia, Chitra Lekha; Mahoney, Eric J; Durrani, Atiq A; Wylie, Christopher

2009-03-01

Intervertebral discs at different postnatal ages were assessed for active intercellular signaling pathways. To generate a spatial and temporal map of the signaling pathways active in the postnatal intervertebral disc (IVD). The postnatal IVD is a complex structure, consisting of 3 histologically distinct components, the nucleus pulposus, fibrous anulus fibrosus, and endplate. These differentiate and grow during the first 9 weeks of age in the mouse. Identification of the major signaling pathways active during and after the growth and differentiation period will allow functional analysis using mouse genetics and identify targets for therapy for individual components of the disc. Antibodies specific for individual cell signaling pathways were used on cryostat sections of IVD at different postnatal ages to identify which components of the IVD were responding to major classes of intercellular signal, including sonic hedgehog, Wnt, TGFbeta, FGF, and BMPs. We present a spatial/temporal map of these signaling pathways during growth, differentiation, and aging of the disc. During growth and differentiation of the disc, its different components respond at different times to different intercellular signaling ligands. Most of these are dramatically downregulated at the end of disc growth.

Calcium spikes, waves and oscillations in a large, patterned epithelial tissue

PubMed Central

Balaji, Ramya; Bielmeier, Christina; Harz, Hartmann; Bates, Jack; Stadler, Cornelia; Hildebrand, Alexander; Classen, Anne-Kathrin

2017-01-01

While calcium signaling in excitable cells, such as muscle or neurons, is extensively characterized, calcium signaling in epithelial tissues is little understood. Specifically, the range of intercellular calcium signaling patterns elicited by tightly coupled epithelial cells and their function in the regulation of epithelial characteristics are little explored. We found that in Drosophila imaginal discs, a widely studied epithelial model organ, complex spatiotemporal calcium dynamics occur. We describe patterns that include intercellular waves traversing large tissue domains in striking oscillatory patterns as well as spikes confined to local domains of neighboring cells. The spatiotemporal characteristics of intercellular waves and oscillations arise as emergent properties of calcium mobilization within a sheet of gap-junction coupled cells and are influenced by cell size and environmental history. While the in vivo function of spikes, waves and oscillations requires further characterization, our genetic experiments suggest that core calcium signaling components guide actomyosin organization. Our study thus suggests a possible role for calcium signaling in epithelia but importantly, introduces a model epithelium enabling the dissection of cellular mechanisms supporting the initiation, transmission and regeneration of long-range intercellular calcium waves and the emergence of oscillations in a highly coupled multicellular sheet. PMID:28218282

GAL4 transactivation-based assay for the detection of selective intercellular protein movement.

PubMed

Kumar, Dhinesh; Chen, Huan; Rim, Yeonggil; Kim, Jae-Yean

2015-01-01

Several plant proteins function as intercellular messenger to specify cell fate and coordinate plant development. Such intercellular communication can be achieved by direct, selective, or nonselective (diffusion-based) trafficking through plasmodesmata (PD), the symplasmic membrane-lined nanochannels adjoining two cells. A trichome rescue trafficking assay was reported to allow the detection of protein movement in Arabidopsis leaf tissue using transgenic gene expression. Here, we provide a protocol to dissect the mode of intercellular protein movement in Arabidopsis root. This assay system involves a root ground tissue-specific GAL4/UAS transactivation expression system in combination with fluorescent reporter proteins. In this system, mCherry, a red fluorescent protein, can move cell to cell via diffusion, while mCherry-H2B is tightly cell autonomous. Thus, a protein fused to mCherry-H2B that can move out from the site of synthesis likely contains a selective trafficking signal to impart a cell-to-cell gain-of-trafficking function to the cell-autonomous mCherry-H2B. This approach can be adapted to investigate the cell-to-cell trafficking properties of any protein of interest.

'Til Eph do us part': intercellular signaling via Eph receptors and ephrin ligands guides cerebral cortical development from birth through maturation.

PubMed

North, Hilary A; Clifford, Meredith A; Donoghue, Maria J

2013-08-01

Eph receptors, the largest family of surface-bound receptor tyrosine kinases and their ligands, the ephrins, mediate a wide variety of cellular interactions in most organ systems throughout both development and maturity. In the forming cerebral cortex, Eph family members are broadly and dynamically expressed in particular sets of cortical cells at discrete times. Here, we review the known functions of Eph-mediated intercellular signaling in the generation of progenitors, the migration of maturing cells, the differentiation of neurons, the formation of functional connections, and the choice between life and death during corticogenesis. In synthesizing these results, we posit a signaling paradigm in which cortical cells maintain a life history of Eph-mediated intercellular interactions that guides subsequent cellular decision-making.

Evolution and failure of liquid bridges between grains due to evaporation and due to extension

NASA Astrophysics Data System (ADS)

Hueckel, T.; Mielniczuk, B.; Said El Youssoufi, M.

2012-04-01

Evolution and rupture of liquid bridges between glass spheres during liquid evaporation and during mechanical extension was examined. The latter type of the tests has been widely studied, while a number of pertinent measurements during transient evaporation have not yet been reported. Also the resultant total capillary forces were measured and geometrical characteristics (curvature radii)were recorded with a photo camera and high-speed camera and subsequently digitalized. The obtained results reveal substantial differences in geometry of liquid bridges during extension and evaporation. On the other hand, evaporation and extension of liquid bridgelead to a similar qualitative response in terms of the pressure within the liquid bridge, starting with a significant suction, which initially somewhat increases during evaporation to reach a maximum, followed by a rapid monotonic decrease until zero, to become a sizable positive pressure prior to rupture. Extension same pattern is followed, except that there is no initial suction increase. Hence, in both cases, rupture consistently occurs at a positive fluid pressure. The pressure evolution is a simple resultant of the evolution of radii of curvature, with the neck radius becoming smaller than meridian radius. In terms of resultant capillary force, as the area of the bridge cross-section decreases with the square of the neck radius, the pressure difference is almost entirely negative, in part also due to surface tension component. Nevertheless, the suction decreases nearly monotonically during both processes. Rupture during evaporation of the bridges occurs most abruptly for larger separations, as early as after 25% volume evaporated. It is seen as a bifurcation of the geometry of equilibrium, as demonstrated on a movie with 27, 000 shots per second. The evolution of a bridge between three spheres exhibits a centrally located thin film instability with a circular hole growing within 1/3000th of a second. All these findings have an enormous impact on the mechanics of unsaturated media, as they determine the conditions and criteria for the loss of the "sand-castle" cohesion effect in the final phase of soil drying.

ALIX and ESCRT-III Coordinately Control Cytokinetic Abscission during Germline Stem Cell Division In Vivo

PubMed Central

Eikenes, Åsmund H.; Malerød, Lene; Christensen, Anette Lie; Steen, Chloé B.; Mathieu, Juliette; Nezis, Ioannis P.; Liestøl, Knut; Huynh, Jean-René; Stenmark, Harald; Haglund, Kaisa

2015-01-01

Abscission is the final step of cytokinesis that involves the cleavage of the intercellular bridge connecting the two daughter cells. Recent studies have given novel insight into the spatiotemporal regulation and molecular mechanisms controlling abscission in cultured yeast and human cells. The mechanisms of abscission in living metazoan tissues are however not well understood. Here we show that ALIX and the ESCRT-III component Shrub are required for completion of abscission during Drosophila female germline stem cell (fGSC) division. Loss of ALIX or Shrub function in fGSCs leads to delayed abscission and the consequent formation of stem cysts in which chains of daughter cells remain interconnected to the fGSC via midbody rings and fusome. We demonstrate that ALIX and Shrub interact and that they co-localize at midbody rings and midbodies during cytokinetic abscission in fGSCs. Mechanistically, we show that the direct interaction between ALIX and Shrub is required to ensure cytokinesis completion with normal kinetics in fGSCs. We conclude that ALIX and ESCRT-III coordinately control abscission in Drosophila fGSCs and that their complex formation is required for accurate abscission timing in GSCs in vivo. PMID:25635693

Manipulation of intestinal epithelial cell function by the cell contact-dependent type III secretion systems of Vibrio parahaemolyticus

PubMed Central

O'Boyle, Nicky; Boyd, Aoife

2013-01-01

Vibrio parahaemolyticus elicits gastroenteritis by deploying Type III Secretion Systems (TTSS) to deliver effector proteins into epithelial cells of the human intestinal tract. The bacteria must adhere to the human cells to allow colonization and operation of the TTSS translocation apparatus bridging the bacterium and the host cell. This article first reviews recent advances in identifying the molecules responsible for intercellular adherence. V. parahaemolyticus possesses two TTSS, each of which delivers an exclusive set of effectors and mediates unique effects on the host cell. TTSS effectors primarily target and alter the activation status of host cell signaling proteins, thereby bringing about changes in the regulation of cellular behavior. TTSS1 is responsible for the cytotoxicity of V. parahaemolyticus, while TTSS2 is necessary for the enterotoxicity of the pathogen. Recent publications have elucidated the function of several TTSS effectors and their importance in the virulence of the bacterium. This review will explore the ability of the TTSS to manipulate activities of human intestinal cells and how this modification of cell function favors bacterial colonization and persistence of V. parahaemolyticus in the host. PMID:24455490

Motor activity of centromere-associated protein-E contributes to its localization at the center of the midbody to regulate cytokinetic abscission

PubMed Central

Ohashi, Akihiro; Ohori, Momoko; Iwai, Kenichi

2016-01-01

Accurate control of cytokinesis is critical for genomic stability to complete high-fidelity transmission of genetic material to the next generation. A number of proteins accumulate in the intercellular bridge (midbody) during cytokinesis, and the dynamics of these proteins are temporally and spatially orchestrated to complete the process. In this study, we demonstrated that localization of centromere-associated protein-E (CENP-E) at the midbody is involved in cytokinetic abscission. The motor activity of CENP-E and the C-terminal midbody localization domain, which includes amino acids 2659–2666 (RYFDNSSL), are involved in the anchoring of CENP-E to the center of the midbody. Furthermore, CENP-E motor activity contributes to the accumulation of protein regulator of cytokinesis 1 (PRC1) in the midbody during cytokinesis. Midbody localization of PRC1 is critical to the antiparallel microtubule structure and recruitment of other midbody-associated proteins. Therefore, CENP-E motor activity appears to play important roles in the organization of these proteins to complete cytokinetic abscission. Our findings will be helpful for understanding how each step of cytokinesis is regulated to complete cytokinetic abscission. PMID:27835888

Intercellular diffusion of a fluorescent sucrose analog via the septal junctions in a filamentous cyanobacterium.

PubMed

Nürnberg, Dennis J; Mariscal, Vicente; Bornikoel, Jan; Nieves-Morión, Mercedes; Krauß, Norbert; Herrero, Antonia; Maldener, Iris; Flores, Enrique; Mullineaux, Conrad W

2015-03-17

Many filamentous cyanobacteria produce specialized nitrogen-fixing cells called heterocysts, which are located at semiregular intervals along the filament with about 10 to 20 photosynthetic vegetative cells in between. Nitrogen fixation in these complex multicellular bacteria depends on metabolite exchange between the two cell types, with the heterocysts supplying combined-nitrogen compounds but dependent on the vegetative cells for photosynthetically produced carbon compounds. Here, we used a fluorescent tracer to probe intercellular metabolite exchange in the filamentous heterocyst-forming cyanobacterium Anabaena sp. strain PCC 7120. We show that esculin, a fluorescent sucrose analog, is incorporated by a sucrose import system into the cytoplasm of Anabaena cells. The cytoplasmic esculin is rapidly and reversibly exchanged across vegetative-vegetative and vegetative-heterocyst cell junctions. Our measurements reveal the kinetics of esculin exchange and also show that intercellular metabolic communication is lost in a significant fraction of older heterocysts. SepJ, FraC, and FraD are proteins located at the intercellular septa and are suggested to form structures analogous to gap junctions. We show that a ΔsepJ ΔfraC ΔfraD triple mutant shows an altered septum structure with thinner septa but a denser peptidoglycan layer. Intercellular diffusion of esculin and fluorescein derivatives is impaired in this mutant, which also shows a greatly reduced frequency of nanopores in the intercellular septal cross walls. These findings suggest that FraC, FraD, and SepJ are important for the formation of junctional structures that constitute the major pathway for feeding heterocysts with sucrose. Anabaena and its relatives are filamentous cyanobacteria that exhibit a sophisticated form of prokaryotic multicellularity, with the formation of differentiated cell types, including normal photosynthetic cells and specialized nitrogen-fixing cells called heterocysts. The question of how heterocysts communicate and exchange metabolites with other cells in the filament is key to understanding this form of bacterial multicellularity. Here we provide the first information on the intercellular exchange of a physiologically important molecule, sucrose. We show that a fluorescent sucrose analog can be imported into the Anabaena cytoplasm by a sucrose import system. Once in the cytoplasm, it is rapidly and reversibly exchanged among all of the cells in the filament by diffusion across the septal junctions. Photosynthetically produced sucrose likely follows the same route from cytoplasm to cytoplasm. We identify some of the septal proteins involved in sucrose exchange, and our results indicate that these proteins form structures functionally analogous to metazoan gap junctions. Copyright © 2015 Nürnberg et al.

Three distinct suppressors of RNA silencing encoded by a 20-kb viral RNA genome

NASA Astrophysics Data System (ADS)

Lu, Rui; Folimonov, Alexey; Shintaku, Michael; Li, Wan-Xiang; Falk, Bryce W.; Dawson, William O.; Ding, Shou-Wei

2004-11-01

Viral infection in both plant and invertebrate hosts requires a virus-encoded function to block the RNA silencing antiviral defense. Here, we report the identification and characterization of three distinct suppressors of RNA silencing encoded by the 20-kb plus-strand RNA genome of citrus tristeza virus (CTV). When introduced by genetic crosses into plants carrying a silencing transgene, both p20 and p23, but not coat protein (CP), restored expression of the transgene. Although none of the CTV proteins prevented DNA methylation of the transgene, export of the silencing signal (capable of mediating intercellular silencing spread) was detected only from the F1 plants expressing p23 and not from the CP- or p20-expressing F1 plants, demonstrating suppression of intercellular silencing by CP and p20 but not by p23. Thus, intracellular and intercellular silencing are each targeted by a CTV protein, whereas the third, p20, inhibits silencing at both levels. Notably, CP suppresses intercellular silencing without interfering with intracellular silencing. The novel property of CP suggests a mechanism distinct to p20 and all of the other viral suppressors known to interfere with intercellular silencing and that this class of viral suppressors may not be consistently identified by Agrobacterium coinfiltration because it also induces RNA silencing against the infiltrated suppressor transgene. Our analyses reveal a sophisticated viral counter-defense strategy that targets the silencing antiviral pathway at multiple steps and may be essential for protecting CTV with such a large RNA genome from antiviral silencing in the perennial tree host. RNA interference | citrus tristeza virus | virus synergy | antiviral immunity

Intercellular Transfer of a Soluble Viral Superantigen

PubMed Central

Reilly, Melissa; Mix, Denise; Reilly, Andrew A.; Yang Ye, Xiang; Winslow, Gary M.

2000-01-01

Mouse mammary tumor virus (MMTV) superantigens (vSAgs) can undergo intercellular transfer in vivo and in vitro such that a vSAg can be presented to T cells by major histocompatibility complex (MHC) class II proteins on antigen-presenting cells (APCs) that do not express the superantigen. This process may allow T-cell activation to occur prior to viral infection. Consistent with these findings, vSAg produced by Chinese hamster ovary (CHO) cells was readily transferred to class II IE and IA (H-2k and H-2d) proteins on a B-cell lymphoma or mouse splenocytes. Fixed class II-expressing acceptor cells were used to demonstrate that the vSAg, but not the class II proteins, underwent intercellular transfer, indicating that vSAg binding to class II MHC could occur directly at the cell surface. Intercellular transfer also occurred efficiently to splenocytes from endogenous retrovirus-free mice, indicating that other proviral proteins were not involved. Presentation of vSAg7 produced by a class II-negative, furin protease-deficient CHO variant (FD11) was unsuccessful, indicating that proteolytic processing was a requisite event and that proteolytic activity could not be provided by an endoprotease on the acceptor APC. Furthermore, vSAg presentation was effected using cell-free supernatant from class II-negative, vSAg-positive cells, indicating that a soluble molecule, most likely produced by proteolytic processing, was sufficient to stimulate T cells. Because the membrane-proximal endoproteolytic cleavage site in the vSAg (residues 68 to 71) was not necessary for intercellular transfer, the data support the notion that the carboxy-terminal endoproteolytic cleavage product is an active vSAg moiety. PMID:10954523

Dissociative electron attachment to DNA-diamine thin films: Impact of the DNA close environment on the OH{sup −} and O{sup −} decay channels

DOE Office of Scientific and Technical Information (OSTI.GOV)

Boulanouar, Omar; Fromm, Michel; Mavon, Christophe

We measure the desorption of anions stimulated by the impact of 0–20 eV electrons on highly uniform thin films of plasmid DNA-diaminopropane. The results are accurately correlated with film thickness and composition by AFM and XPS measurements, respectively. Resonant structures in the H{sup −}, O{sup −}, and OH{sup −} yield functions are attributed to the decay of transient anions into the dissociative electron attachment (DEA) channel. The diamine induces ammonium-phosphate bridges along the DNA backbone, which suppresses the DEA O{sup −} channel and in counter-part increases considerably the desorption of OH{sup −}. The close environment of the phosphate groups maymore » therefore play an important role in modulating the rate and type of DNA damages induced by low energy electrons.« less

Fabrication of periodical surface structures by picosecond laser irradiation of carbon thin films: transformation of amorphous carbon in nanographite

NASA Astrophysics Data System (ADS)

Popescu, C.; Dorcioman, G.; Bita, B.; Besleaga, C.; Zgura, I.; Himcinschi, C.; Popescu, A. C.

2016-12-01

Thin films of carbon were synthesized by ns pulsed laser deposition in vacuum on silicon substrates, starting from graphite targets. Further on, the films were irradiated with a picosecond laser source emitting in visible at 532 nm. After tuning of laser parameters, we obtained a film surface covered by laser induced periodical surface structures (LIPSS). They were investigated by optical, scanning electron and atomic force microscopy. It was observed that changing the irradiation angle influences the LIPSS covered area. At high magnification it was revealed that the LIPSS pattern was quite complex, being composed of other small LIPSS islands, interconnected by bridges of nanoparticles. Raman spectra for the non-irradiated carbon films were typical for a-C type of diamond-like carbon, while the LIPSS spectra were characteristic to nano-graphite. The pristine carbon film was hydrophilic, while the LIPSS covered film surface was hydrophobic.

Artificial gravity field, astrophysical analogues, and topological phase transitions in strained topological semimetals

NASA Astrophysics Data System (ADS)

Yu, Zhiming; Guan, Shan; Yao, Yugui; Yang, Shengyuan

Effective gravity and gauge fields are emergent properties intrinsic for low-energy quasiparticles in topological semimetals. Here, taking two Dirac semimetals as examples, we demonstrate that applied lattice strain can generate warped spacetime, with fascinating analogues in astrophysics. Particularly, we study the possibility of simulating black-hole/white-hole event horizons and gravitational lensing effect. Furthermore, we discover strain-induced topological phase transitions, both in the bulk materials and in their thin films. Especially in thin films, the transition between the quantum spin Hall and the trivial insulating phases can be achieved by a small strain, naturally leading to the proposition of a novel piezo-topological transistor device. Our result not only bridges multiple disciplines, revealing topological semimetals as a unique table-top platform for exploring interesting phenomena in astrophysics and general relativity; it also suggests realistic materials and methods to achieve controlled topological phase transitions with great potential for device applications.

Intercellular signalling in Stigmatella aurantiaca.

PubMed

Plaga, W; Ulrich, S H

1999-12-01

The myxobacterium Stigmatella aurantiaca is a prokaryotic model used to study intercellular signalling and the genetic determination of morphogenesis. Signalling factors and genes required for the generation of the elaborate multicellular fruiting body are to be identified. Recently, the structure of stigmolone, which is the pheromone necessary for fruiting body formation, was elucidated, and genes involved in development were characterised. Progress has also been made in the genetic accessibility of S. aurantiaca.

A transwell assay that excludes exosomes for assessment of tunneling nanotube-mediated intercellular communication.

PubMed

Thayanithy, Venugopal; O'Hare, Patrick; Wong, Phillip; Zhao, Xianda; Steer, Clifford J; Subramanian, Subbaya; Lou, Emil

2017-11-13

Tunneling nanotubes (TNTs) are naturally-occurring filamentous actin-based membranous extensions that form across a wide spectrum of mammalian cell types to facilitate long-range intercellular communication. Valid assays are needed to accurately assess the downstream effects of TNT-mediated transfer of cellular signals in vitro. We recently reported a modified transwell assay system designed to test the effects of intercellular transfer of a therapeutic oncolytic virus, and viral-activated drugs, between cells via TNTs. The objective of the current study was to demonstrate validation of this in vitro approach as a new method for effectively excluding diffusible forms of long- and close-range intercellular transfer of intracytoplasmic cargo, including exosomes/microvesicles and gap junctions in order to isolate TNT-selective cell communication. We designed several steps to effectively reduce or eliminate diffusion and long-range transfer via these extracellular vesicles, and used Nanoparticle Tracking Analysis to quantify exosomes following implementation of these steps. The experimental approach outlined here effectively reduced exosome trafficking by >95%; further use of heparin to block exosome uptake by putative recipient cells further impeded transfer of these extracellular vesicles. This validated assay incorporates several steps that can be taken to quantifiably control for extracellular vesicles in order to perform studies focused on TNT-selective communication.

Role of auxin during intercellular infection of Discaria trinervis by Frankia

PubMed Central

Imanishi, Leandro; Perrine-Walker, Francine M.; Ndour, Adama; Vayssières, Alice; Conejero, Genevieve; Lucas, Mikaël; Champion, Antony; Laplaze, Laurent; Wall, Luis; Svistoonoff, Sergio

2014-01-01

Nitrogen-fixing nodules induced by Frankia in the actinorhizal plant Discaria trinervis result from a primitive intercellular root invasion pathway that does not involve root hair deformation and infection threads. Here, we analyzed the role of auxin in this intercellular infection pathway at the molecular level and compared it with our previous work in the intracellular infected actinorhizal plant Casuarina glauca. Immunolocalisation experiments showed that auxin accumulated in Frankia-infected cells in both systems. We then characterized the expression of auxin transporters in D. trinervis nodules. No activation of the heterologous CgAUX1 promoter was detected in infected cells in D. trinervis. These results were confirmed with the endogenous D. trinervis gene, DtAUX1. However, DtAUX1 was expressed in the nodule meristem. Consistently, transgenic D. trinervis plants containing the auxin response marker DR5:VENUS showed expression of the reporter gene in the meristem. Immunolocalisation experiments using an antibody against the auxin efflux carrier PIN1, revealed the presence of this transporter in the plasma membrane of infected cells. Finally, we used in silico cellular models to analyse auxin fluxes in D. trinervis nodules. Our results point to the existence of divergent roles of auxin in intercellularly- and intracellularly-infected actinorhizal plants, an ancestral infection pathways leading to root nodule symbioses. PMID:25191330

Cellulose and pectin localization in roots of mycorrhizalAllium porrum: labelling continuity between host cell wall and interfacial material.

PubMed

Bonfante-Fasolo, P; Vian, B; Perotto, S; Faccio, A; Knox, J P

1990-03-01

Two different types of contacts (or interfaces) exist between the plant host and the fungus during the vesicular-arbuscular mycorrhizal symbiosis, depending on whether the fungus is intercellular or intracellular. In the first case, the walls of the partners are in contact, while in the second case the fungal wall is separated from the host cytoplasm by the invaginated host plasmamembrane and by an interfacial material. In order to verify the origin of the interfacial material, affinity techniques which allow identification in situ of cell-wall components, were used. Cellobiohydrolase (CBH I) that binds to cellulose and a monoclonal antibody (JIM 5) that reacts with pectic components were tested on roots ofAllium porrum L. (leek) colonized byGlomus versiforme (Karst.) Berch. Both probes gave a labelling specific for the host cell wall, but each probe labelled over specific and distinct areas. The CBH I-colloidal gold complex heavily labelled the thick epidermal cell walls, whereas JIM 5 only labelled this area weakly. Labelling of the hypodermis was mostly on intercellular material after treatment with JIM 5 and only on the wall when CBH I was used. Suberin bands found on the radial walls were never labelled. Cortical cells were mostly labelled on the middle lamella with JIM 5 and on the wall with CBH I. Gold granules from the two probes were found in interfacial material both near the point where the fungus enters the cell and around the thin hyphae penetrating deep into the cell. The ultrastructural observations demonstrate that cellulose and pectic components have different but complementary distributions in the walls of root cells involved in the mycorrhizal symbiosis. These components show a similar distribution in the interfacial material laid down around the vesicular-arbuscular mycorrhizal fungus indicating that the interfacial material is of host origin.

Wireless Power Transmission to Organic Light Emitting Diode Lighting Panel with Magnetically Coupled Resonator

NASA Astrophysics Data System (ADS)

Kim, Yong-Hae; Han, Jun-Han; Kang, Seung-Youl; Cheon, Sanghoon; Lee, Myung-Lae; Ahn, Seong-Deok; Zyung, Taehyoung; Lee, Jeong-Ik; Moon, Jaehyun; Chu, Hye Yong

2012-09-01

We are successful to lit the organic light emitting diode (OLED) lighting panel through the magnetically coupled wireless power transmission technology. For the wireless power transmission, we used the operation frequency 932 kHz, specially designed double spiral type transmitter, small and thin receiver on the four layered printed circuit board, and schottky diodes for the full bridge rectifier. Our white OLED is a hybrid type, in which phosphorescent and fluorescent organics are used together to generate stable white color. The total efficiency of power transmission is around 72%.

Precise deformation measurement of prestressed concrete beam during a strain test using the combination of intersection photogrammetry and micro-network measurement

NASA Astrophysics Data System (ADS)

Urban, Rudolf; Braun, Jaroslav; Štroner, Martin

2015-05-01

The prestressed thin-walled concrete elements enable the bridge a relatively large span. These structures are advantageous in economic and environmental way due to their thickness and lower consumption of materials. The bending moments can be effectively influenced by using the pre-stress. The experiment was done to monitor deformation of the under load. During the experiment the discrete points were monitored. To determine a large number of points, the intersection photogrammetry combined with precise micro-network were chosen. Keywords:

Circumferential skin folds in a child: a case of Michelin tire baby syndrome.

PubMed

Palit, Aparna; Inamadar, Arun C

2007-01-01

A six-month-old girl who presented with dermatitis was found to have multiple, symmetric, deep, gyrate skin folds involving her trunk and similar circumferential lesions on her extremities since birth. She had a characteristic round face with hypertelorism, depressed nasal bridge, thin, down-turned vermillion border of upper lip and short neck. Skin biopsy demonstrated increased smooth muscle fibers in the deeper dermis. A diagnosis of Michelin tire baby syndrome was made. Clinical features, histopathology, differential diagnosis and prognosis of this rare disorder have been discussed.

Materials Design Through Chemical Control of Precursors

DTIC Science & Technology

2008-08-22

Ligand Sets,” H. M. El-Kaderi, M.J. Heeg, and C.H. Winter, Polyhedron 2006, 25, 224-234. 7. “Atomic Layer Deposition of Ga2O3 Films from a...poster presentation. 4. “Atomic Layer Deposition (ALD) of High Quality Ga2O3 Thin Films from a Dimeric Dialkylamido-Bridged Gallium Complex,” C. L...growth of Ga2O3 films from a new metalorganic precursor (“Atomic Layer Deposition of Ga2O3 Films from a 4 Dialkylamido-Based Precursor,” C. L

Fully integrated carbon nanotube composite thin film strain sensors on flexible substrates for structural health monitoring

NASA Astrophysics Data System (ADS)

Burton, A. R.; Lynch, J. P.; Kurata, M.; Law, K. H.

2017-09-01

Multifunctional thin film materials have opened many opportunities for novel sensing strategies for structural health monitoring. While past work has established methods of optimizing multifunctional materials to exhibit sensing properties, comparatively less work has focused on their integration into fully functional sensing systems capable of being deployed in the field. This study focuses on the advancement of a scalable fabrication process for the integration of multifunctional thin films into a fully integrated sensing system. This is achieved through the development of an optimized fabrication process that can create a broad range of sensing systems using multifunctional materials. A layer-by-layer deposited multifunctional composite consisting of single walled carbon nanotubes (SWNT) in a polyvinyl alcohol and polysodium-4-styrene sulfonate matrix are incorporated with a lithography process to produce a fully integrated sensing system deposited on a flexible substrate. To illustrate the process, a strain sensing platform consisting of a patterned SWNT-composite thin film as a strain-sensitive element within an amplified Wheatstone bridge sensing circuit is presented. Strain sensing is selected because it presents many of the design and processing challenges that are core to patterning multifunctional thin film materials into sensing systems. Strain sensors fabricated on a flexible polyimide substrate are experimentally tested under cyclic loading using standard four-point bending coupons and a partial-scale steel frame assembly under lateral loading. The study reveals the material process is highly repeatable to produce fully integrated strain sensors with linearity and sensitivity exceeding 0.99 and 5 {{V}}/{ε }, respectively. The thin film strain sensors are robust and are capable of high strain measurements beyond 3000 μ {ε }.

Intrinsically disordered proteins aggregate at fungal cell-to-cell channels and regulate intercellular connectivity.

PubMed

Lai, Julian; Koh, Chuan Hock; Tjota, Monika; Pieuchot, Laurent; Raman, Vignesh; Chandrababu, Karthik Balakrishna; Yang, Daiwen; Wong, Limsoon; Jedd, Gregory

2012-09-25

Like animals and plants, multicellular fungi possess cell-to-cell channels (septal pores) that allow intercellular communication and transport. Here, using a combination of MS of Woronin body-associated proteins and a bioinformatics approach that identifies related proteins based on composition and character, we identify 17 septal pore-associated (SPA) proteins that localize to the septal pore in rings and pore-centered foci. SPA proteins are not homologous at the primary sequence level but share overall physical properties with intrinsically disordered proteins. Some SPA proteins form aggregates at the septal pore, and in vitro assembly assays suggest aggregation through a nonamyloidal mechanism involving mainly α-helical and disordered structures. SPA loss-of-function phenotypes include excessive septation, septal pore degeneration, and uncontrolled Woronin body activation. Together, our data identify the septal pore as a complex subcellular compartment and focal point for the assembly of unstructured proteins controlling diverse aspects of intercellular connectivity.

Tunneling nanotubes spread fibrillar α-synuclein by intercellular trafficking of lysosomes.

PubMed

Abounit, Saïda; Bousset, Luc; Loria, Frida; Zhu, Seng; de Chaumont, Fabrice; Pieri, Laura; Olivo-Marin, Jean-Christophe; Melki, Ronald; Zurzolo, Chiara

2016-10-04

Synucleinopathies such as Parkinson's disease are characterized by the pathological deposition of misfolded α-synuclein aggregates into inclusions throughout the central and peripheral nervous system. Mounting evidence suggests that intercellular propagation of α-synuclein aggregates may contribute to the neuropathology; however, the mechanism by which spread occurs is not fully understood. By using quantitative fluorescence microscopy with co-cultured neurons, here we show that α-synuclein fibrils efficiently transfer from donor to acceptor cells through tunneling nanotubes (TNTs) inside lysosomal vesicles. Following transfer through TNTs, α-synuclein fibrils are able to seed soluble α-synuclein aggregation in the cytosol of acceptor cells. We propose that donor cells overloaded with α-synuclein aggregates in lysosomes dispose of this material by hijacking TNT-mediated intercellular trafficking. Our findings thus reveal a possible novel role of TNTs and lysosomes in the progression of synucleinopathies. © 2016 The Authors.

Cell-to-cell communication in plants, animals, and fungi: a comparative review.

PubMed

Bloemendal, Sandra; Kück, Ulrich

2013-01-01

Cell-to-cell communication is a prerequisite for differentiation and development in multicellular organisms. This communication has to be tightly regulated to ensure that cellular components such as organelles, macromolecules, hormones, or viruses leave the cell in a precisely organized way. During evolution, plants, animals, and fungi have developed similar ways of responding to this biological challenge. For example, in higher plants, plasmodesmata connect adjacent cells and allow communication to regulate differentiation and development. In animals, two main general structures that enable short- and long-range intercellular communication are known, namely gap junctions and tunneling nanotubes, respectively. Finally, filamentous fungi have also developed specialized structures called septal pores that allow intercellular communication via cytoplasmic flow. This review summarizes the underlying mechanisms for intercellular communication in these three eukaryotic groups and discusses its consequences for the regulation of differentiation and developmental processes.

Cellular level robotic surgery: Nanodissection of intermediate filaments in live keratinocytes.

PubMed

Yang, Ruiguo; Song, Bo; Sun, Zhiyong; Lai, King Wai Chiu; Fung, Carmen Kar Man; Patterson, Kevin C; Seiffert-Sinha, Kristina; Sinha, Animesh A; Xi, Ning

2015-01-01

We present the nanosurgery on the cytoskeleton of live cells using AFM based nanorobotics to achieve adhesiolysis and mimic the effect of pathophysiological modulation of intercellular adhesion. Nanosurgery successfully severs the intermediate filament bundles and disrupts cell-cell adhesion similar to the desmosomal protein disassembly in autoimmune disease, or the cationic modulation of desmosome formation. Our nanomechanical analysis revealed that adhesion loss results in a decrease in cellular stiffness in both cases of biochemical modulation of the desmosome junctions and mechanical disruption of intercellular adhesion, supporting the notion that intercellular adhesion through intermediate filaments anchors the cell structure as focal adhesion does and that intermediate filaments are integral components in cell mechanical integrity. The surgical process could potentially help reveal the mechanism of autoimmune pathology-induced cell-cell adhesion loss as well as its related pathways that lead to cell apoptosis. Copyright © 2015 Elsevier Inc. All rights reserved.

Mobile microRNAs hit the target.

PubMed

Gursanscky, Nial R; Searle, Iain R; Carroll, Bernard J

2011-11-01

MicroRNAs (miRNAs) are negative regulators of gene expression in eukaryotic organisms, whereas small interfering RNAs (siRNAs) guide host-cell defence against viruses, transposons and transgenes. A key issue in plant biology is whether miRNAs act only in cells in which they are formed, or if, like siRNAs, they also function after passive diffusion or active transportation into other cells. Recent reports show that miRNAs are indeed able to move between plant cells to direct developmental programming of gene expression. In both leaf and root development, miRNAs establish intercellular gradients of gene expression that are essential for cell and tissue differentiation. Gradients in gene expression also play crucial roles in animal development, and there is strong evidence for intercellular movement of miRNAs in animals. Thus, intercellular movement of miRNAs may be crucial to animal developmental biology as well as plants. © 2011 John Wiley & Sons A/S.

Inter-Cellular Forces Orchestrate Contact Inhibition of Locomotion

PubMed Central

Davis, John R.; Luchici, Andrei; Mosis, Fuad; Thackery, James; Salazar, Jesus A.; Mao, Yanlan; Dunn, Graham A.; Betz, Timo; Miodownik, Mark; Stramer, Brian M.

2015-01-01

Summary Contact inhibition of locomotion (CIL) is a multifaceted process that causes many cell types to repel each other upon collision. During development, this seemingly uncoordinated reaction is a critical driver of cellular dispersion within embryonic tissues. Here, we show that Drosophila hemocytes require a precisely orchestrated CIL response for their developmental dispersal. Hemocyte collision and subsequent repulsion involves a stereotyped sequence of kinematic stages that are modulated by global changes in cytoskeletal dynamics. Tracking actin retrograde flow within hemocytes in vivo reveals synchronous reorganization of colliding actin networks through engagement of an inter-cellular adhesion. This inter-cellular actin-clutch leads to a subsequent build-up in lamellar tension, triggering the development of a transient stress fiber, which orchestrates cellular repulsion. Our findings reveal that the physical coupling of the flowing actin networks during CIL acts as a mechanotransducer, allowing cells to haptically sense each other and coordinate their behaviors. PMID:25799385

Cell-to-cell communication in plants, animals, and fungi: a comparative review

NASA Astrophysics Data System (ADS)

Bloemendal, Sandra; Kück, Ulrich

2013-01-01

Cell-to-cell communication is a prerequisite for differentiation and development in multicellular organisms. This communication has to be tightly regulated to ensure that cellular components such as organelles, macromolecules, hormones, or viruses leave the cell in a precisely organized way. During evolution, plants, animals, and fungi have developed similar ways of responding to this biological challenge. For example, in higher plants, plasmodesmata connect adjacent cells and allow communication to regulate differentiation and development. In animals, two main general structures that enable short- and long-range intercellular communication are known, namely gap junctions and tunneling nanotubes, respectively. Finally, filamentous fungi have also developed specialized structures called septal pores that allow intercellular communication via cytoplasmic flow. This review summarizes the underlying mechanisms for intercellular communication in these three eukaryotic groups and discusses its consequences for the regulation of differentiation and developmental processes.

Actin cable dynamics and Rho/Rock orchestrate a polarized cytoskeletal architecture in the early steps of assembling a stratified epithelium.

PubMed

Vaezi, Alec; Bauer, Christoph; Vasioukhin, Valeri; Fuchs, Elaine

2002-09-01

To enable stratification and barrier function, the epidermis must permit self-renewal while maintaining adhesive connections. By generating K14-GFP-actin mice to monitor actin dynamics in cultured primary keratinocytes, we uncovered a role for the actin cytoskeleton in establishing cellular organization. During epidermal sheet formation, a polarized network of nascent intercellular junctions and radial actin cables assemble in the apical plane of the monolayer. These actin fibers anchor to a central actin-myosin network, creating a tension-based plane of cytoskeleton across the apical surface of the sheet. Movement of the sheet surface relative to its base expands the zone of intercellular overlap, catalyzing new sites for nascent intercellular junctions. This polarized cytoskeleton is dependent upon alpha-catenin, Rho, and Rock, and its regulation may be important for wound healing and/or stratification, where coordinated tissue movements are involved.

Relative Roles of Gap Junction Channels and Cytoplasm in Cell-to-Cell Diffusion of Fluorescent Tracers

NASA Astrophysics Data System (ADS)

Safranyos, Richard G. A.; Caveney, Stanley; Miller, James G.; Petersen, Nils O.

1987-04-01

Intercellular (tissue) diffusion of molecules requires cytoplasmic diffusion and diffusion through gap junctional (or cell-to-cell) channels. The rates of tissue and cytoplasmic diffusion of fluorescent tracers, expressed as an effective diffusion coefficient, De, and a cytoplasmic diffusion coefficient, Dcyt, have been measured among the developing epidermal cells of a larval beetle, Tenebrio molitor L., to determine the contribution of the junctional channels to intercellular diffusion. Tracer diffusion was measured by injecting fluorescent tracers into cells and quantitating the rate of subsequent spread into adjacent cells. Cytoplasmic diffusion was determined by fluorescence photobleaching. These experiments show that gap junctional channels constitute approximately 70-80% of the total cell-to-cell resistance to the diffusion of organic tracers at high concentrations in this tissue. At low concentrations, however, the binding of tracer to cytoplasm slows down the cytoplasmic diffusion, which may limit intercellular diffusion.

Ultraflexible nanostructures and implications for future nanorobots

NASA Astrophysics Data System (ADS)

Cohn, Robert W.; Panchapakesan, Balaji

2016-05-01

Several high aspect ratio nanostructures have been made by capillary force directed self-assembly including polymeric nanofiber air-bridges, trampoline-like membranes, microsphere-beaded nanofibers, and intermetallic nanoneedles. Arrays of polymer air-bridges form in seconds by simply hand brushing a bead of polymeric liquid over an array of micropillars. The domination of capillary force that is thinning unstable capillary bridges leads to uniform arrays of nanofiber air-bridges. Similarly, arrays of vertically oriented Ag2Ga nanoneedles have been formed by dipping silvercoated arrays of pyramidal silicon into melted gallium. Force-displacement measurements of these structures are presented. These nanostructures, especially when compressively or torsionally buckled, have extremely low stiffnesses, motion due to thermal fluctuations that is relatively easily detected, and the ability to move great distances for very small changes in applied force. Nanofibers with bead-on-a-string structure, where the beads are micron diameter and loaded with magnetic iron oxide (maghemite), are shown to be simply viewable under optical microscopes, have micronewton/ m stiffness, and have ultralow torsional stiffnesses enabling the bead to be rotated numerous revolutions without breaking. Combination of these high aspect ratio structures with stretched elastomers offer interesting possibilities for robotic actuation and locomotion. Polydimethylsiloxane loaded with nanomaterials, e.g. nanotubes, graphene or MoS2, can be efficiently heated with directed light. Heating produces considerable force through the thermoelastic effect, and this force can be used for continuous translation or to trigger reversible elastic buckling of the nanostructures. The remote stimulation of motion with light provides a possible mechanism for producing cooperative behavior between swarms of semiautonomous nanorobots.

Triboluminesence multifunctional cementitious composites with in situ damage sensing capability

NASA Astrophysics Data System (ADS)

Olawale, David O.; Dickens, Tarik; Uddin, Mohammed J.; Okoli, Okenwa O.

2012-04-01

Structural health monitoring of civil infrastructure systems like concrete bridges and dams has become critical because of the aging and overloading of these CIS. Most of the available SHM methods are not in-situ and can be very expensive. The triboluminescence multifunctional cementitious composites (TMCC) have in-built crack detection mechanism that can enable bridge engineers to monitor and detect abnormal crack formation in concrete structures so that timely corrective action can be taken to prevent costly or catastrophic failures. This article reports the fabrication process and test result of the flexural characterization of the TMCC. Accelerated durability test indicated that the 0.5 ZnS:Mn/Epoxy weight fraction ITOF sensor configuration to be more desirable in terms of durability. The alkaline environment at the highest temperature investigated (45 °C) resulted in significant reduction in the mean glass transition and storage moduli of the tested ITOF thin films. Further work is ongoing to correlate the TL response of the TMCC with damage, particularly crack opening.

Thin film mechanics

NASA Astrophysics Data System (ADS)

Cooper, Ryan C.

This doctoral thesis details the methods of determining mechanical properties of two classes of novel thin films suspended two-dimensional crystals and electron beam irradiated microfilms of polydimethylsiloxane (PDMS). Thin films are used in a variety of surface coatings to alter the opto-electronic properties or increase the wear or corrosion resistance and are ideal for micro- and nanoelectromechanical system fabrication. One of the challenges in fabricating thin films is the introduction of strains which can arise due to application techniques, geometrical conformation, or other spurious conditions. Chapters 2-4 focus on two dimensional materials. This is the intrinsic limit of thin films-being constrained to one atomic or molecular unit of thickness. These materials have mechanical, electrical, and optical properties ideal for micro- and nanoelectromechanical systems with truly novel device functionality. As such, the breadth of applications that can benefit from a treatise on two dimensional film mechanics is reason enough for exploration. This study explores the anomylously high strength of two dimensional materials. Furthermore, this work also aims to bridge four main gaps in the understanding of material science: bridging the gap between ab initio calculations and finite element analysis, bridging the gap between ab initio calculations and experimental results, nanoscale to microscale, and microscale to mesoscale. A nonlinear elasticity model is used to determine the necessary elastic constants to define the strain-energy density function for finite strain. Then, ab initio calculations-density functional theory-is used to calculate the nonlinear elastic response. Chapter 2 focuses on validating this methodology with atomic force microscope nanoindentation on molybdenum disulfide. Chapter 3 explores the convergence criteria of three density functional theory solvers to further verify the numerical calculations. Chapter 4 then uses this model to investigate the role of grain boundaries on the strength of chemical vapor deposited graphene. The results from these studies suggest that two dimensional films have remarkably high strength-reaching the intrinsic limit of molecular bonds. Chapter 5 explores the viscoelastic properties of heterogeneous polydimethylsiloxane (PDMS) microfilms through dynamic nanoindentation. PDMS microfilms are irradiated with an electron beam creating a 3 m-thick film with an increased cross-link density. The change in mechanical properties of PDMS due to thermal history and accelerator have been explored by a variety of tests, but the effect of electron beam irradiation is still unknown. The resulting structure is a stiff microfilm embedded in a soft rubber with some transformational strain induced by the cross-linking volume changes. Chapter 5 employs a combination of dynamic nanoindentation and finite element analysis to determine the change in stiffness as a function of electron beam irradiation. The experimental results are compared to the literature. The results of these experimental and numerical techniques provide exciting opportu- nities in future research. Two dimensional materials and flexible thin films are exciting materials for novel applications with new form factors, such as flexible electronics and microfluidic devices. The results herein indicate that you can accurately model the strength of two dimsensional materials and that these materials are robust against nanoscale defects. The results also reveal local variation of mechanical properties in PDMS microfilms. This allows one to design substrates that flex with varying amounts of strain on the surface. Combining the mechanics of two dimensional materials with that of a locally irradiated PDMS film could achieve a new class of flexible microelectromechanical systems. Large-scale growth of two dimensional materials will be structurally robust-even in the presence of nanostructural defects-and PDMS microfilms can be irradiated to vary strain of the electromechanical systems. These systems could be designed to investigate electromechanical coupling in two dimensional films or for a substitute to traditional silicon microdevices. (Abstract shortened by UMI.)

Guard cell photosynthesis is critical for stomatal turgor production, yet does not directly mediate CO2 - and ABA-induced stomatal closing.

PubMed

Azoulay-Shemer, Tamar; Palomares, Axxell; Bagheri, Andisheh; Israelsson-Nordstrom, Maria; Engineer, Cawas B; Bargmann, Bastiaan O R; Stephan, Aaron B; Schroeder, Julian I

2015-08-01

Stomata mediate gas exchange between the inter-cellular spaces of leaves and the atmosphere. CO2 levels in leaves (Ci) are determined by respiration, photosynthesis, stomatal conductance and atmospheric [CO2 ]. [CO2 ] in leaves mediates stomatal movements. The role of guard cell photosynthesis in stomatal conductance responses is a matter of debate, and genetic approaches are needed. We have generated transgenic Arabidopsis plants that are chlorophyll-deficient in guard cells only, expressing a constitutively active chlorophyllase in a guard cell specific enhancer trap line. Our data show that more than 90% of guard cells were chlorophyll-deficient. Interestingly, approximately 45% of stomata had an unusual, previously not-described, morphology of thin-shaped chlorophyll-less stomata. Nevertheless, stomatal size, stomatal index, plant morphology, and whole-leaf photosynthetic parameters (PSII, qP, qN, FV '/FM' ) were comparable with wild-type plants. Time-resolved intact leaf gas-exchange analyses showed a reduction in stomatal conductance and CO2 -assimilation rates of the transgenic plants. Normalization of CO2 responses showed that stomata of transgenic plants respond to [CO2 ] shifts. Detailed stomatal aperture measurements of normal kidney-shaped stomata, which lack chlorophyll, showed stomatal closing responses to [CO2 ] elevation and abscisic acid (ABA), while thin-shaped stomata were continuously closed. Our present findings show that stomatal movement responses to [CO2 ] and ABA are functional in guard cells that lack chlorophyll. These data suggest that guard cell CO2 and ABA signal transduction are not directly modulated by guard cell photosynthesis/electron transport. Moreover, the finding that chlorophyll-less stomata cause a 'deflated' thin-shaped phenotype, suggests that photosynthesis in guard cells is critical for energization and guard cell turgor production. © 2015 The Authors The Plant Journal © 2015 John Wiley & Sons Ltd.

Experimental Performance of a Micromachined Heat Flux Sensor

NASA Technical Reports Server (NTRS)

Stefanescu, S.; DeAnna, R. G.; Mehregany, M.

1998-01-01

Steady-state and frequency response calibration of a microfabricated heat-flux sensor have been completed. This sensor is batch fabricated using standard, micromachining techniques, allowing both miniaturization and the ability to create arrays of sensors and their corresponding interconnects. Both high-frequency and spatial response is desired, so the sensors are both thin and of small cross-sectional area. Thin-film, temperature-sensitive resistors are used as the active gauge elements. Two sensor configurations are investigated: (1) a Wheatstone-bridge using four resistors; and (2) a simple, two-resistor design. In each design, one resistor (or pair) is covered by a thin layer (5000 A) thermal barrier; the other resistor (or pair) is covered by a thick (5 microns) thermal barrier. The active area of a single resistor is 360 microns by 360 microns; the total gauge area is 1.5 mm square. The resistors are made of 2000 A-thick metal; and the entire gauge is fabricated on a 25 microns-thick flexible, polyimide substrate. Heat flux through the surface changes the temperature of the resistors and produces a corresponding change in resistance. Sensors were calibrated using two radiation heat sources: (1) a furnace for steady-state, and (2) a light and chopper for frequency response.

Temperature-Dependent Lithium-Ion Diffusion and Activation Energy of Li1.2Co0.13Ni0.13Mn0.54O2 Thin-Film Cathode at Nanoscale by Using Electrochemical Strain Microscopy.

PubMed

Yang, Shan; Yan, Binggong; Wu, Jiaxiong; Lu, Li; Zeng, Kaiyang

2017-04-26

This paper presents the in situ mapping of temperature-dependent lithium-ion diffusion at the nanometer level in thin film Li 1.2 Co 0.13 Ni 0.13 Mn 0.54 O 2 cathode using electrochemical strain microscopy. The thin-film Li 1.2 Co 0.13 Ni 0.13 Mn 0.54 O 2 cathode exhibits higher lithium-ion diffusivities with increasing temperature, which explains the higher capacity observed in the lithium-ion batteries with a Li-rich cathode at elevated temperature. In addition, the activation energy for lithium-ion diffusion can be extracted in an Arrhenius-type plot at the level of grain structure with the assumption that the ionic movement is diffusion controlled. Compared with the grain interiors, the grain boundaries show relatively lower activation energy; hence, it is the preferred diffusion path for lithium ions. This study has bridged the gap between atomistic calculations and traditional macroscopic experiments, showing direct evidence as well as mechanisms for ionic diffusion for Li-rich cathode material.

Structural comparison of Ag-Ge-S bulk glasses and thin films

NASA Astrophysics Data System (ADS)

Wang, Fei; Jain, Mukul; Dunn, Porter; de Leo, Carter; Boolchand, Punit

2007-03-01

Ternary glasses of composition (GeS3)1-xAgx (x=0.1 and 0.2) are studied in form of bulk and thin films. Bulk glasses are synthesized and examined in Raman scattering and SEM. Raman scattering results of bulk glasses show that with increasing x, an increasing fraction of the Ag additive enters the base glass as Ag^+ with S^-anions serving to form thiogermanate species with one, two and three non-bridging S^- species. SEM measurements of the bulk glass show the material is intrinsically phase separated. White colored islands are observed distributed in a dark base. The EDS measurements show islands are Ag rich and the base is relatively Ag deficient. The Ag rich islands are expected to be mainly glassy phase Ag2S. Thin films of same compositions are fabricated using thermal evaporation. Films are evaporated following two different procedures to prevent the material from spitting. One method was preheating outgas and the other method was using tungsten mesh wrapped boats. The stoichiometry and molecular structure of films under each procedure are analyzed by Raman scattering and SEM to be compared with bulk glasses.

Ultrastructural study of the semicircular canal cells of the frog Rana esculenta.

PubMed

Oudar, O; Ferrary, E; Feldmann, G

1988-03-01

The ultrastructure of the nonsensory cells (dark cells, transitional cells, and undifferentiated cells) of the frog semicircular canal was studied by using transmission electron microscopy in an attempt to correlate the structure with the functions of these epithelial cells. All the nonsensory cells were linked by tight junctions and desmosomes; this suggested that there is little paracellular ionic transport from perilymph to endolymph. In the dark cell epithelium, the apical intercellular spaces were dilated; in the basal part, numerous basolateral plasma membrane infoldings, containing mitochondria, delimited electron-lucent spaces. The undifferentiated cells and the transitional cells were devoid of any basal membrane infolding. Surrounding the semicircular canal, very flattened and interdigitated mesothelial cells constituted a thin multilayer tissue which limited the perilymphatic space. The morphological aspect of the dark cells suggests that they may play a role in the secretion and/or in the reabsorption of endolymph, which bathes the apical pole of these cells. The undifferentiated and transitional cells can play a role in the maintenance of the endolymphatic ionic composition because of their apical tight junctions and desmosomes.

Serum deprivation induces glucose response and intercellular coupling in human pancreatic adenocarcinoma PANC-1 cells.

PubMed

Hiram-Bab, Sahar; Shapira, Yuval; Gershengorn, Marvin C; Oron, Yoram

2012-03-01

This study aimed to investigate whether the previously described differentiating islet-like aggregates of human pancreatic adenocarcinoma cells (PANC-1) develop glucose response and exhibit intercellular communication. Fura 2-loaded PANC-1 cells in serum-free medium were assayed for changes in cytosolic free calcium ([Ca]i) induced by depolarization, tolbutamide inhibition of K(ATP) channels, or glucose. Dye transfer, assayed by confocal microscopy or by FACS, was used to detect intercellular communication. Changes in messenger RNA (mRNA) expression of genes of interest were assessed by quantitative real-time polymerase chain reaction. Proliferation was assayed by the MTT method. Serum-deprived PANC-1 cell aggregates developed [Ca]i response to KCl, tolbutamide, or glucose. These responses were accompanied by 5-fold increase in glucokinase mRNA level and, to a lesser extent, of mRNAs for K(ATP) and L-type calcium channels, as well as increase in mRNA levels of glucagon and somatostatin. Trypsin, a proteinase-activated receptor 2 agonist previously shown to enhance aggregation, modestly improved [Ca]i response to glucose. Glucose-induced coordinated [Ca]i oscillations and dye transfer demonstrated the emergence of intercellular communication. These findings suggest that PANC-1 cells, a pancreatic adenocarcinoma cell line, can be induced to express a differentiated phenotype in which cells exhibit response to glucose and form a functional syncytium similar to those observed in pancreatic islets.

Identification of infection- and defense-related genes via a dynamic host-pathogen interaction network using a Candida albicans-zebrafish infection model.

PubMed

Kuo, Zong-Yu; Chuang, Yung-Jen; Chao, Chun-Cheih; Liu, Fu-Chen; Lan, Chung-Yu; Chen, Bor-Sen

2013-01-01

Candida albicans infections and candidiasis are difficult to treat and create very serious therapeutic challenges. In this study, based on interactive time profile microarray data of C. albicans and zebrafish during infection, the infection-related protein-protein interaction (PPI) networks of the two species and the intercellular PPI network between host and pathogen were simultaneously constructed by a dynamic interaction model, modeled as an integrated network consisting of intercellular invasion and cellular defense processes during infection. The signal transduction pathways in regulating morphogenesis and hyphal growth of C. albicans were further investigated based on significant interactions found in the intercellular PPI network. Two cellular networks were also developed corresponding to the different infection stages (adhesion and invasion), and then compared with each other to identify proteins from which we can gain more insight into the pathogenic role of hyphal development in the C. albicans infection process. Important defense-related proteins in zebrafish were predicted using the same approach. The hyphal growth PPI network, zebrafish PPI network and host-pathogen intercellular PPI network were combined to form an integrated infectious PPI network that helps us understand the systematic mechanisms underlying the pathogenicity of C. albicans and the immune response of the host, and may help improve medical therapies and facilitate the development of new antifungal drugs. Copyright © 2013 S. Karger AG, Basel.

[The Functional Role of Exosomes in Cancer Biology and Their Potential as Biomarkers and Therapeutic Targets of Cancer].

PubMed

Naito, Yutaka; Yoshioka, Yusuke; Ochiya, Takahiro

2015-06-01

Intercellular communication plays an important role in the regulation of various cellular events. In particular, cancer cells and the surrounding cells communicate with each other, and this intercellular communication triggers cancer initiation and progression through the secretion of molecules, including growth factors and cytokines. Recent advances in cancer biology have indicated that small membrane vesicles, termed exosomes, also serve as regulatory agents in intercellular communications. Exosomes contain functional cellular components, including proteins and microRNAs (miRNAs), and they transfer these components to recipient cells. This exosome-mediated intercellular communication leads to increased growth, invasion, and metastasis of cancer. Thus, researchers regard exosomes as important cues to understanding the molecular mechanisms of cancer biology. Indeed, several lines of evidence have demonstrated that exosomes can explain multiple aspects of cancer biology. In addition, increasing evidence suggests that exosomes and their specific molecules are also attractive for use as biomarkers and therapeutic targets in cancer. Recent reports showed the efficacy of a novel diagnosis by detecting component molecules of cancer-derived exosomes, including miRNAs and membrane proteins. Furthermore, clinical trials that test the application of exosomes for cancer therapy have already been reported. From these points of view, we will summarize experimental data that support the role of exosomes in cancer progression and the potential of exosomes for use in novel diagnostic and therapeutic approaches for cancer.

Rescue of Notch signaling in cells incapable of GDP-L-fucose synthesis by gap junction transfer of GDP-L-fucose in Drosophila.

PubMed

Ayukawa, Tomonori; Matsumoto, Kenjiroo; Ishikawa, Hiroyuki O; Ishio, Akira; Yamakawa, Tomoko; Aoyama, Naoki; Suzuki, Takuya; Matsuno, Kenji

2012-09-18

Notch (N) is a transmembrane receptor that mediates cell-cell interactions to determine many cell-fate decisions. N contains EGF-like repeats, many of which have an O-fucose glycan modification that regulates N-ligand binding. This modification requires GDP-L-fucose as a donor of fucose. The GDP-L-fucose biosynthetic pathways are well understood, including the de novo pathway, which depends on GDP-mannose 4,6 dehydratase (Gmd) and GDP-4-keto-6-deoxy-D-mannose 3,5-epimerase/4-reductase (Gmer). However, the potential for intercellularly supplied GDP-L-fucose and the molecular basis of such transportation have not been explored in depth. To address these points, we studied the genetic effects of mutating Gmd and Gmer on fucose modifications in Drosophila. We found that these mutants functioned cell-nonautonomously, and that GDP-L-fucose was supplied intercellularly through gap junctions composed of Innexin-2. GDP-L-fucose was not supplied through body fluids from different isolated organs, indicating that the intercellular distribution of GDP-L-fucose is restricted within a given organ. Moreover, the gap junction-mediated supply of GDP-L-fucose was sufficient to support the fucosylation of N-glycans and the O-fucosylation of the N EGF-like repeats. Our results indicate that intercellular delivery is a metabolic pathway for nucleotide sugars in live animals under certain circumstances.

Spam1-associated transmission ratio distortion in mice: Elucidating the mechanism

PubMed Central

Martin-DeLeon, Patricia A; Zhang, Hong; Morales, Carlos R; Zhao, Yutong; Rulon, Michelle; Barnoski, Barry L; Chen, Hong; Galileo, Deni S

2005-01-01

Background While transmission ratio distortion, TRD, (a deviation from Mendelian ratio) is extensive in humans and well-documented in mice, the underlying mechanisms are unknown. Our earlier studies on carriers of spontaneous mutations of mouse Sperm Adhesion Molecule 1 (Spam1) suggested that TRD results from biochemically different sperm, due to a lack of transcript sharing through the intercellular cytoplasmic bridges of spermatids. These bridges usually allow transcript sharing among genetically different spermatids which develop into biochemically and functionally equivalent sperm. Objectives The goals of the study were to provide support for the lack of sharing (LOS) hypothesis, using transgene and null carriers of Spam1, and to determine the mechanism of Spam1-associated TRD. Methods Carriers of Spam1-Hyal5 BAC transgenes were mated with wild-type female mice and the progeny analyzed for TRD by PCR genotyping. Sperm from transgene and Spam1 null carriers were analyzed using flow cytometry and immunocytochemistry to detect quantities of Spam1 and/or Hyal5. Transgene-bearing sperm with Spam1 overexpression were detected by fluorescence in situ hybridization. In wild-type animals, EM studies of in situ transcript hybridization of testis sections and Northern analysis of biochemically fractionated testicular RNA were performed to localize Spam1 transcript. Finally, AU-rich motifs identified in the 3' UTR of Spam1 RNA were assayed by UV cross-linking to determine their ability to interact with testicular RNA binding proteins. Results The Tg8 line of transgene carriers had a significant (P < 0.001) TRD, due to reduced fertilizing ability of transgene-bearing sperm. These sperm retained large cytoplasmic droplets engorged with overexpressed Spam1 or Hyal5 protein. Caudal sperm from transgene carriers and caput sperm of null carriers showed a bimodal distribution of Spam1, indicating that the sperm in a male were biochemically different with respect to Spam1 quantities. Spam1 RNA was absent from the bridges, associated exclusively with the ER, and was shown to be anchored to the cytoskeleton. This compartmentalization of the transcript, mediated by cytoskeletal binding, occurs via protein interactions with 3' UTR AU-rich sequences that are likely involved in its stabilization. Conclusion We provide strong support for the LOS hypothesis, and have elucidated the mechanism of Spam1-associated TRD. PMID:16092963

Message-adjusted network (MAN) hypothesis in gastro-entero-pancreatic (GEP) endocrine system.

PubMed

Aykan, N Faruk

2007-01-01

Several types of communication coordinate body functions to maintain homeostasis. Clarifying intercellular communication systems is as important as intracellular signal mechanisms. In this study, we propose an intercellular network model to establish novel targets in GEP-endocrine system, based on up-to-date information from medical publications. As materials, two physiologic events which are Pavlov's sham-feeding assay and bicarbonate secretion into the duodenum from pancreas were explored by new biologic data from the literature. Major key words used in Pub-Med were modes of regulations (autocrine, paracrine, endocrine, neurocrine, juxtacrine, lumencrine), GEP cells, hormones, peptides and neuro-transmitters. In these two examples of physiologic events, we can design a model of network to clarify transmission of a message. When we take a simple, unique message, we can observe a complete intercellular network. In our examples, these messages are "food is coming" and "hydrogen ions are increasing" in human language (humanese). We need to find molecular counterparts of these unique messages in cell language (cellese). In this network (message-adjusted network; MAN), message is an input which can affect the physiologic equilibrium, mission is an output to improve the disequilibrium and aim is always maintenance of homeostasis. If we orientate to a transmission of a unique message we can distinguish that different cells use different chemical messengers in different modes of regulations to transmit the same message. This study also supports Shannon's information theory and cell language theories such as von Neumann-Patte principles. After human genome project (HU-GO) and protein organisations (HU-PO), finding true messages and the establishment of their networks (in our model HU-MAN project) can be a novel and exciting field in cell biology. We established an intercellular network model to understand intercellular communication in the physiology of GEP endocrine system. This model could help to explain complex physiologic events as well as to generate new treatment concepts.

Physics of lumen growth.

PubMed

Dasgupta, Sabyasachi; Gupta, Kapish; Zhang, Yue; Viasnoff, Virgile; Prost, Jacques

2018-05-22

We model the dynamics of formation of intercellular secretory lumens. Using conservation laws, we quantitatively study the balance between paracellular leaks and the build-up of osmotic pressure in the lumen. Our model predicts a critical pumping threshold to expand stable lumens. Consistently with experimental observations in bile canaliculi, the model also describes a transition between a monotonous and oscillatory regime during luminogenesis as a function of ion and water transport parameters. We finally discuss the possible importance of regulation of paracellular leaks in intercellular tubulogenesis.

Regulation of Yersina pestis Virulence by AI-2 Mediated Quorum Sensing

DOE Office of Scientific and Technical Information (OSTI.GOV)

Segelke, B; Hok, S; Lao, V

The proposed research was motivated by an interest in understanding Y. pestis virulence mechanisms and bacteria cell-cell communication. It is expected that a greater understanding of virulence mechanisms will ultimately lead to biothreat countermeasures and novel therapeutics. Y. pestis is the etiological agent of plague, the most devastating disease in human history. Y. pestis infection has a high mortality rate and a short incubation before mortality. There is no widely available and effective vaccine for Y. pestis and multi-drug resistant strains are emerging. Y. pestis is a recognized biothreat agent based on the wide distribution of the bacteria in researchmore » laboratories around the world and on the knowledge that methods exist to produce and aerosolize large amounts of bacteria. We hypothesized that cell-cell communication via signaling molecules, or quorum sensing, by Y. pestis is important for the regulation of virulence factor gene expression during host invasion, though a causative link had never been established. Quorum sensing is a mode of intercellular communication which enables orchestration of gene expression for many bacteria as a function of population density and available evidence suggests there may be a link between quorum sensing and regulation of Y. pesits virulence. Several pathogenic bacteria have been shown to regulate expression of virulence factor genes, including genes encoding type III secretion, via quorum sensing. The Y. pestis genome encodes several cell-cell signaling pathways and the interaction of at least three of these are thought to be involved in one or more modes of host invasion. Furthermore, Y. pestis gene expression array studies carried out at LLNL have established a correlation between expression of known virulence factors and genes involved in processing of the AI-2 quorum sensing signal. This was a basic research project that was intended to provide new insights into bacterial intercellular communication and how it is used to regulate virulence in Y. pestis. It is known that many bacteria use intercellular signaling molecules to orchestrate gene expression and cellular function. A fair amount is known about production and uptake of signaling molecules, but very little is known about how intercellular signaling regulates other pathways. Although several studies demonstrate that intercellular signaling plays a role in regulating virulence in other pathogens, the link between signaling and regulation of virulence has not been established. Very little work had been done directly with Y. pestis intercellular signaling apart from the work carried out at LLNL. The research we proposed was intended to both establish a causative link between AI-2 intercellular signaling and regulation of virulence in Y. pestis and elucidate the fate of the AI-2 signaling molecule after it is taken up and processed by Y. pestis. Elucidating the fate of AI-2 was expected to lead directly to the understanding of how AI-2 signal processing regulates other pathways as well as provide new insights in this direction.« less

Anillin acts as a bifunctional linker coordinating midbody ring biogenesis during cytokinesis

PubMed Central

Kechad, Amel; Jananji, Silvana; Ruella, Yvonne; Hickson, Gilles R. X.

2013-01-01

Summary Animal cell cytokinesis proceeds via constriction of an actomyosin-based contractile ring (CR) [1, 2]. Upon reaching a diameter of ~1 μm [3], a midbody ring (MR) forms to stabilize the intercellular bridge until abscission [4-6]. How MR formation is coupled to CR closure and how plasma membrane anchoring is maintained at this key transition is unknown. Time-lapse microscopy of Drosophila S2 cells depleted of the scaffold protein, Anillin [7-9], revealed that Anillin is required for complete closure of the CR and formation of the MR. Truncation analysis revealed that Anillin N-termini connected with the actomyosin CR and supported formation of stable MR-like structures, but these could not maintain anchoring of the plasma membrane. Conversely, Anillin C-termini failed to connect with the CR or MR but recruited the septin, Peanut, to ectopic structures at the equatorial cortex. Peanut depletion mimicked truncation of the Anillin C-terminus, resulting in MR-like structures that failed to anchor the membrane. These data demonstrate that Anillin coordinates the transition from CR to MR, and that it does so by linking two distinct cortical cytoskeletal elements. One apparently acts as the core structural template for MR assembly, while the other ensures stable anchoring of the plasma membrane beyond the CR stage. PMID:22226749

Aging Enhances Indirect Flight Muscle Fiber Performance yet Decreases Flight Ability in Drosophila

DOE Office of Scientific and Technical Information (OSTI.GOV)

Miller, Mark S.; Lekkas, Panagiotis; Braddock, Joan M.

2008-10-02

We investigated the effects of aging on Drosophila melanogaster indirect flight muscle from the whole organism to the actomyosin cross-bridge. Median-aged (49-day-old) flies were flight impaired, had normal myofilament number and packing, barely longer sarcomeres, and slight mitochondrial deterioration compared with young (3-day-old) flies. Old (56-day-old) flies were unable to beat their wings, had deteriorated ultrastructure with severe mitochondrial damage, and their skinned fibers failed to activate with calcium. Small-amplitude sinusoidal length perturbation analysis showed median-aged indirect flight muscle fibers developed greater than twice the isometric force and power output of young fibers, yet cross-bridge kinetics were similar. Large increasesmore » in elastic and viscous moduli amplitude under active, passive, and rigor conditions suggest that median-aged fibers become stiffer longitudinally. Small-angle x-ray diffraction indicates that myosin heads move increasingly toward the thin filament with age, accounting for the increased transverse stiffness via cross-bridge formation. We propose that the observed protein composition changes in the connecting filaments, which anchor the thick filaments to the Z-disk, produce compensatory increases in longitudinal stiffness, isometric tension, power and actomyosin interaction in aging indirect flight muscle. We also speculate that a lack of MgATP due to damaged mitochondria accounts for the decreased flight performance.« less

Intrinsically disordered proteins aggregate at fungal cell-to-cell channels and regulate intercellular connectivity

PubMed Central

Lai, Julian; Koh, Chuan Hock; Tjota, Monika; Pieuchot, Laurent; Raman, Vignesh; Chandrababu, Karthik Balakrishna; Yang, Daiwen; Wong, Limsoon; Jedd, Gregory

2012-01-01

Like animals and plants, multicellular fungi possess cell-to-cell channels (septal pores) that allow intercellular communication and transport. Here, using a combination of MS of Woronin body-associated proteins and a bioinformatics approach that identifies related proteins based on composition and character, we identify 17 septal pore-associated (SPA) proteins that localize to the septal pore in rings and pore-centered foci. SPA proteins are not homologous at the primary sequence level but share overall physical properties with intrinsically disordered proteins. Some SPA proteins form aggregates at the septal pore, and in vitro assembly assays suggest aggregation through a nonamyloidal mechanism involving mainly α-helical and disordered structures. SPA loss-of-function phenotypes include excessive septation, septal pore degeneration, and uncontrolled Woronin body activation. Together, our data identify the septal pore as a complex subcellular compartment and focal point for the assembly of unstructured proteins controlling diverse aspects of intercellular connectivity. PMID:22955885

Drosophila wing imaginal discs respond to mechanical injury via slow InsP3R-mediated intercellular calcium waves

NASA Astrophysics Data System (ADS)

Restrepo, Simon; Basler, Konrad

2016-08-01

Calcium signalling is a highly versatile cellular communication system that modulates basic functions such as cell contractility, essential steps of animal development such as fertilization and higher-order processes such as memory. We probed the function of calcium signalling in Drosophila wing imaginal discs through a combination of ex vivo and in vivo imaging and genetic analysis. Here we discover that wing discs display slow, long-range intercellular calcium waves (ICWs) when mechanically stressed in vivo or cultured ex vivo. These slow imaginal disc intercellular calcium waves (SIDICs) are mediated by the inositol-3-phosphate receptor, the endoplasmic reticulum (ER) calcium pump SERCA and the key gap junction component Inx2. The knockdown of genes required for SIDIC formation and propagation negatively affects wing disc recovery after mechanical injury. Our results reveal a role for ICWs in wing disc homoeostasis and highlight the utility of the wing disc as a model for calcium signalling studies.

siRNA-based Analysis of the Abrogation of the Protective Function of Membrane-associated Catalase of Tumor Cells.

PubMed

Bauer, Georg

2017-02-01

Tumor cells, in contrast to non-malignant cells, show sustained expression of membrane-associated NADPH oxidase-1 and therefore generate extracellular superoxide anions and their dismutation product H 2 O 2 In order to prevent intercellular reactive oxygen species/reactive nitrogen species (ROS/RNS)-dependent apoptosis-inducing signaling, tumor cells need to express membrane-associated catalase that interferes with HOCl and nitric oxide/peroxynitrite signaling. Catalase is attached to tumor cells through the activity of transglutaminase-2 and is prevented from superoxide anion-dependent inhibition through coexpression of membrane-associated superoxide dismutase. Therefore, specific inhibition of membrane-associated catalase should reactivate intercellular ROS/RNS-dependent apoptosis-inducing signaling. These processes are analyzed here through small interfering RNA-mediated knockdown of essential signaling compounds. This allows to establish a rather comprehensive picture of intercellular ROS/RNS signaling that may be instrumental for future therapeutic approaches. Copyright© 2017, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.

PI3K/Akt signaling is involved in the disruption of gap junctional communication caused by v-Src and TNF-α.

PubMed

Ito, Satoko; Hyodo, Toshinori; Hasegawa, Hitoki; Yuan, Hong; Hamaguchi, Michinari; Senga, Takeshi

2010-09-17

Gap junctional communication, which is mediated by the connexin protein family, is essential for the maintenance of normal tissue function and homeostasis. Loss of intercellular communication results in a failure to coordinately regulate cellular functions, and it can facilitate tumorigenesis. Expression of oncogenes and stimulation with cytokines has been shown to suppress intercellular communication; however, the exact mechanism by which intercellular communication is disrupted by these factors remains uncertain. In this report, we show that Akt is essential for the disruption of gap junctional communication in v-Src-transformed cells. In addition, inhibition of Akt restores gap junctional communication after it is suppressed by TNF-α signaling. Furthermore, we demonstrate that the expression of a constitutively active form of Akt1, but not of Akt2 or Akt3, is sufficient to suppress gap junctional communication. Our results clearly define Akt1 as one of the critical regulators of gap junctional communication. Copyright © 2010 Elsevier Inc. All rights reserved.

Tunneling nanotubes promote intercellular mitochondria transfer followed by increased invasiveness in bladder cancer cells

PubMed Central

Lu, Jinjin; Zheng, Xiufen; Li, Fan; Yu, Yang; Chen, Zhong; Liu, Zheng; Wang, Zhihua; Xu, Hua; Yang, Weimin

2017-01-01

Intercellular transfer of organelles via tunneling nanotubes (TNTs) is a novel means of cell-to-cell communication. Here we demonstrate the existence of TNTs between co-cultured RT4 and T24 bladder cancer cells using light microscopy, fluorescence imaging, and scanning electron microscopy (SEM). Spontaneous unidirectional transfer of mitochondria from T24 to RT4 cells was detected using fluorescence imaging and flow cytometry. The distribution of mitochondria migrated from T24 cells was in good agreement with the original mitochondria in RT4 cells, which may imply mitochondrial fusion. We detected cytoskeleton reconstruction in RT4-Mito-T24 cells by observing F-actin redistribution. Akt, mTOR, and their downstream mediators were activated and increased. The resultant increase in the invasiveness of bladder cancer cells was detected in vitro and in vivo. These data indicate that TNTs promote intercellular mitochondrial transfer between heterogeneous cells, followed by an increase in the invasiveness of bladder cancer cells. PMID:28107184

CO₂ processing and hydration of fruit and vegetable tissues by clathrate hydrate formation.

PubMed

Takeya, Satoshi; Nakano, Kohei; Thammawong, Manasikan; Umeda, Hiroki; Yoneyama, Akio; Takeda, Tohoru; Hyodo, Kazuyuki; Matsuo, Seiji

2016-08-15

CO2 hydrate can be used to preserve fresh fruits and vegetables, and its application could contribute to the processing of carbonated frozen food. We investigated water transformation in the frozen tissue of fresh grape samples upon CO2 treatment at 2-3 MPa and 3°C for up to 46 h. Frozen fresh bean, radish, eggplant and cucumber samples were also investigated for comparison. X-ray diffraction indicated that after undergoing CO2 treatment for several hours, structure I CO2 hydrate formed within the grape tissue. Phase-contrast X-ray imaging using the diffraction-enhanced imaging technique revealed the presence of CO2 hydrate within the intercellular spaces of these tissues. The carbonated produce became effervescent because of the dissociation of CO2 hydrate through the intercellular space, especially above the melting point of ice. In addition, suppressed metabolic activity resulting from CO2 hydrate formation, which inhibits water and nutrient transport through intercellular space, can be expected. Copyright © 2016 Elsevier Ltd. All rights reserved.

Inter-cellular forces orchestrate contact inhibition of locomotion.

PubMed

Davis, John R; Luchici, Andrei; Mosis, Fuad; Thackery, James; Salazar, Jesus A; Mao, Yanlan; Dunn, Graham A; Betz, Timo; Miodownik, Mark; Stramer, Brian M

2015-04-09

Contact inhibition of locomotion (CIL) is a multifaceted process that causes many cell types to repel each other upon collision. During development, this seemingly uncoordinated reaction is a critical driver of cellular dispersion within embryonic tissues. Here, we show that Drosophila hemocytes require a precisely orchestrated CIL response for their developmental dispersal. Hemocyte collision and subsequent repulsion involves a stereotyped sequence of kinematic stages that are modulated by global changes in cytoskeletal dynamics. Tracking actin retrograde flow within hemocytes in vivo reveals synchronous reorganization of colliding actin networks through engagement of an inter-cellular adhesion. This inter-cellular actin-clutch leads to a subsequent build-up in lamellar tension, triggering the development of a transient stress fiber, which orchestrates cellular repulsion. Our findings reveal that the physical coupling of the flowing actin networks during CIL acts as a mechanotransducer, allowing cells to haptically sense each other and coordinate their behaviors. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

Novel microscopy-based screening method reveals regulators of contact-dependent intercellular transfer

PubMed Central

Michael Frei, Dominik; Hodneland, Erlend; Rios-Mondragon, Ivan; Burtey, Anne; Neumann, Beate; Bulkescher, Jutta; Schölermann, Julia; Pepperkok, Rainer; Gerdes, Hans-Hermann; Kögel, Tanja

2015-01-01

Contact-dependent intercellular transfer (codeIT) of cellular constituents can have functional consequences for recipient cells, such as enhanced survival and drug resistance. Pathogenic viruses, prions and bacteria can also utilize this mechanism to spread to adjacent cells and potentially evade immune detection. However, little is known about the molecular mechanism underlying this intercellular transfer process. Here, we present a novel microscopy-based screening method to identify regulators and cargo of codeIT. Single donor cells, carrying fluorescently labelled endocytic organelles or proteins, are co-cultured with excess acceptor cells. CodeIT is quantified by confocal microscopy and image analysis in 3D, preserving spatial information. An siRNA-based screening using this method revealed the involvement of several myosins and small GTPases as codeIT regulators. Our data indicates that cellular protrusions and tubular recycling endosomes are important for codeIT. We automated image acquisition and analysis to facilitate large-scale chemical and genetic screening efforts to identify key regulators of codeIT. PMID:26271723

An ultramicroscopic study on rigor mortis.

PubMed

Suzuki, T

1976-01-01

Gastrocnemius muscles taken from decapitated mice at various intervals after death and from mice killed by 2,4-dinitrophenol or mono-iodoacetic acid injection to induce rigor mortis soon after death, were observed by electron microscopy. The prominent appearance of many fine cross striations in the myofibrils (occurring about every 400 A) was considered to be characteristic of rigor mortis. These striations were caused by minute granules studded along the surfaces of both thick and thin filaments and appeared to be the bridges connecting the 2 kinds of filaments and accounted for the hardness and rigidity of the muscle.

ADMiER-ing thin but complex fluids

NASA Astrophysics Data System (ADS)

McDonnell, Amarin G.; Bhattacharjee, Pradipto K.; Pan, Sharadwata; Hill, David; Danquah, Michael K.; Friend, James R.; Yeo, Leslie Y.; Prabhakar, Ranganathan

2011-12-01

The Acoustics Driven Microfluidic Extensional Rheometer (ADMiER) utilises micro litre volumes of liquid, with viscosities as low as that of water, to create valid and observable extensional flows, liquid bridges that pinch off due to capillary forces in this case. ADMiER allows the study fluids that have been beyond conventional methods and also study more subtle fluid properties. We can observe polymeric fluids with solvent viscosities far below those previously testable, accentuating elastic effects. Also, it has enabled the testing of aqueous solutions of living motile particles, which significantly change fluid properties, opening up the potential for diagnostic applications.

Silicon on insulator self-aligned transistors

DOEpatents

McCarthy, Anthony M.

2003-11-18

A method for fabricating thin-film single-crystal silicon-on-insulator (SOI) self-aligned transistors. Standard processing of silicon substrates is used to fabricate the transistors. Physical spaces, between the source and gate, and the drain and gate, introduced by etching the polysilicon gate material, are used to provide connecting implants (bridges) which allow the transistor to perform normally. After completion of the silicon substrate processing, the silicon wafer is bonded to an insulator (glass) substrate, and the silicon substrate is removed leaving the transistors on the insulator (glass) substrate. Transistors fabricated by this method may be utilized, for example, in flat panel displays, etc.

Serum Deprivation Induces Glucose Response and Intercellular Coupling in Human Pancreatic Adenocarcinoma PANC-1 Cells

PubMed Central

Hiram-Bab, Sahar; Shapira, Yuval; Gershengorn, Marvin C.; Oron, Yoram

2012-01-01

Objective This study aimed to investigate whether the previously described differentiating islet-like aggregates of human pancreatic adenocarcinoma cells (PANC-1) develop glucose response and exhibit intercellular communication. Methods Fura 2–loaded PANC-1 cells in serum-free medium were assayed for changes in cytosolic free calcium ([Ca]i) induced by depolarization, tolbutamide inhibition of K(ATP) channels, or glucose. Dye transfer, assayed by confocal microscopy or by FACS, was used to detect intercellular communication. Changes in messenger RNA (mRNA) expression of genes of interest were assessed by quantitative real-time polymerase chain reaction. Proliferation was assayed by the MTT method. Results Serum-deprived PANC-1 cell aggregates developed [Ca]i response to KCl, tolbutamide, or glucose. These responses were accompanied by 5-fold increase in glucokinase mRNA level and, to a lesser extent, of mRNAs for K(ATP) and L-type calcium channels, as well as increase in mRNA levels of glucagon and somatostatin. Trypsin, a proteinase-activated receptor 2 agonist previously shown to enhance aggregation, modestly improved [Ca]i response to glucose. Glucose-induced coordinated [Ca]i oscillations and dye transfer demonstrated the emergence of intercellular communication. Conclusions These findings suggest that PANC-1 cells, a pancreatic adenocarcinoma cell line, can be induced to express a differentiated phenotype, in which cells exhibit response to glucose and form a functional syncytium similar to those observed in pancreatic islets. PMID:22129530

Destruction of the hepatocyte junction by intercellular invasion of Leptospira causes jaundice in a hamster model of Weil's disease

PubMed Central

Miyahara, Satoshi; Saito, Mitsumasa; Kanemaru, Takaaki; Villanueva, Sharon Y A M; Gloriani, Nina G; Yoshida, Shin-ichi

2014-01-01

Weil's disease, the most severe form of leptospirosis, is characterized by jaundice, haemorrhage and renal failure. The mechanisms of jaundice caused by pathogenic Leptospira remain unclear. We therefore aimed to elucidate the mechanisms by integrating histopathological changes with serum biochemical abnormalities during the development of jaundice in a hamster model of Weil's disease. In this work, we obtained three-dimensional images of infected hamster livers using scanning electron microscope together with freeze-cracking and cross-cutting methods for sample preparation. The images displayed the corkscrew-shaped bacteria, which infiltrated the Disse's space, migrated between hepatocytes, detached the intercellular junctions and disrupted the bile canaliculi. Destruction of bile canaliculi coincided with the elevation of conjugated bilirubin, aspartate transaminase and alkaline phosphatase levels in serum, whereas serum alanine transaminase and γ-glutamyl transpeptidase levels increased slightly, but not significantly. We also found in ex vivo experiments that pathogenic, but not non-pathogenic leptospires, tend to adhere to the perijunctional region of hepatocyte couplets isolated from hamsters and initiate invasion of the intercellular junction within 1 h after co-incubation. Our results suggest that pathogenic leptospires invade the intercellular junctions of host hepatocytes, and this invasion contributes in the disruption of the junction. Subsequently, bile leaks from bile canaliculi and jaundice occurs immediately. Our findings revealed not only a novel pathogenicity of leptospires, but also a novel mechanism of jaundice induced by bacterial infection. PMID:24945433

Intercellular communication via gap junctions affected by mechanical load in the bovine annulus fibrosus.

PubMed

Desrochers, Jane; Duncan, Neil A

2014-01-01

Cells in the intervertebral disc, as in other connective tissues including tendon, ligament and bone, form interconnected cellular networks that are linked via functional gap junctions. These cellular networks may be necessary to affect a coordinated response to mechanical and environmental stimuli. Using confocal microscopy with fluorescence recovery after photobleaching methods, we explored the in situ strain environment of the outer annulus of an intact bovine disc and the effect of high-level flexion on gap junction signalling. The in situ strain environment in the extracellular matrix of the outer annulus under high flexion load was observed to be non-uniform with the extensive cellular processes remaining crimped sometimes at flexion angles greater than 25°. A significant transient disruption of intercellular communication via functional gap junctions was measured after 10 and 20 min under high flexion load. This study illustrates that in healthy annulus fibrosus tissue, high mechanical loads can impede the functioning of the gap junctions. Future studies will explore more complex loading conditions to determine whether losses in intercellular communication can be permanent and whether gap junctions in aged and degenerated tissues become more susceptible to load. The current research suggests that cellular structures such as gap junctions and intercellular networks, as well as other cell-cell and cell-matrix interconnections, need to be considered in computational models in order to fully understand how macroscale mechanical signals are transmitted across scales to the microscale and ultimately into a cellular biosynthetic response in collagenous tissues.

Analysis of the Gap Junction-dependent Transfer of miRNA with 3D-FRAP Microscopy.

PubMed

Lemcke, Heiko; Voronina, Natalia; Steinhoff, Gustav; David, Robert

2017-06-19

Small antisense RNAs, like miRNA and siRNA, play an important role in cellular physiology and pathology and, moreover, can be used as therapeutic agents in the treatment of several diseases. The development of new, innovative strategies for miRNA/siRNA therapy is based on an extensive knowledge of the underlying mechanisms. Recent data suggest that small RNAs are exchanged between cells in a gap junction-dependent manner, thereby inducing gene regulatory effects in the recipient cell. Molecular biological techniques and flow cytometric analysis are commonly used to study the intercellular exchange of miRNA. However, these methods do not provide high temporal resolution, which is necessary when studying the gap junctional flux of molecules. Therefore, to investigate the impact of miRNA/siRNA as intercellular signaling molecules, novel tools are needed that will allow for the analysis of these small RNAs at the cellular level. The present protocol describes the application of three-dimensional fluorescence recovery after photobleaching (3D-FRAP) microscopy to elucidating the gap junction-dependent exchange of miRNA molecules between cardiac cells. Importantly, this straightforward and non-invasive live-cell imaging approach allows for the visualization and quantification of the gap junctional shuttling of fluorescently labeled small RNAs in real time, with high spatio-temporal resolution. The data obtained by 3D-FRAP confirm a novel pathway of intercellular gene regulation, where small RNAs act as signaling molecules within the intercellular network.

Theoretical simulations of protective thin film Fabry-Pérot filters for integrated optical elements of diode pumped alkali lasers (DPAL)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Quarrie, L., E-mail: Lindsay.Quarrie@l-3com.com, E-mail: lindsay.o.quarrie@gmail.com; Air Force Research Laboratory, AFRL/RDLC Laser CoE, 3550 Aberdeen Avenue SE, Kirtland AFB, NM 87117-5776

The lifetime of Diode-Pumped Alkali Lasers (DPALs) is limited by damage initiated by reaction of the glass envelope of its gain medium with rubidium vapor. Rubidium is absorbed into the glass and the rubidium cations diffuse through the glass structure, breaking bridging Si-O bonds. A damage-resistant thin film was developed enhancing high-optical transmission at natural rubidium resonance input and output laser beam wavelengths of 780 nm and 795 nm, while protecting the optical windows of the gain cell in a DPAL. The methodology developed here can be readily modified for simulation of expected transmission performance at input pump and outputmore » laser wavelengths using different combination of thin film materials in a DPAL. High coupling efficiency of the light through the gas cell was accomplished by matching the air-glass and glass-gas interfaces at the appropriate wavelengths using a dielectric stack of high and low index of refraction materials selected to work at the laser energies and protected from the alkali metal vapor in the gain cell. Thin films as oxides of aluminum, zirconium, tantalum, and silicon were selected allowing the creation of Fabry-Perot optical filters on the optical windows achieving close to 100% laser transmission in a solid optic combination of window and highly reflective mirror. This approach allows for the development of a new whole solid optic laser.« less

Radiation resistance of a gamma-ray irradiated nonlinear optic chromophore

NASA Astrophysics Data System (ADS)

Zhang, Cheng; Taylor, Edward W.

2009-11-01

The radiation resistance of organic electro-optic and optoelectronic materials for space applications is receiving increased attention. An earlier investigation reported that guest-host poled polymer EO modulator devices composed of a phenyltetraene bridge-type chromophore in amorphous polycarbonate (CLD/APC) did not exhibit a decrease in EO response (i.e., an increase in modulation-switching voltage- Vπ) following irradiation by low dose [10-160 krad(Si)] 60Co gamma-rays. To provide further evidences to the observed radiation stability, the post-irradiation responses of 60Co gamma-rays on CLD1/APC thin films are examined by various chemical and spectroscopic methods including: a solubility test, thin-layer chromatography, proton nuclear magnetic resonance spectroscopy, UV-vis absorption, and infra-red absorption. The results indicate that CLD1 and APC did not decompose under gamma-ray irradiation at dose levels ranging from 66-274 krad(Si) and from 61-154 krad(Si), respectively which support the previously reported data.

Free Surface Flows and Extensional Rheology of Polymer Solutions

NASA Astrophysics Data System (ADS)

Dinic, Jelena; Jimenez, Leidy Nallely; Biagioli, Madeleine; Estrada, Alexandro; Sharma, Vivek

Free-surface flows - jetting, spraying, atomization during fuel injection, roller-coating, gravure printing, several microfluidic drop/particle formation techniques, and screen-printing - all involve the formation of axisymmetric fluid elements that spontaneously break into droplets by a surface-tension-driven instability. The growth of the capillary-driven instability and pinch-off dynamics are dictated by a complex interplay of inertial, viscous and capillary stresses for simple fluids. Additional contributions by elasticity, extensibility and extensional viscosity play a role for complex fluids. We show that visualization and analysis of capillary-driven thinning and pinch-off dynamics of the columnar neck in an asymmetric liquid bridge created by dripping-onto-substrate (DoS) can be used for characterizing the extensional rheology of complex fluids. Using a wide variety of complex fluids, we show the measurement of the extensional relaxation time, extensional viscosity, power-law index and shear viscosity. Lastly, we elucidate how polymer composition, flexibility, and molecular weight determine the thinning and pinch-off dynamics of polymeric complex fluids.

The ZnO-FET Biosensor for Cardiac Troponin I

NASA Astrophysics Data System (ADS)

Fathil, M. F. M.; Arshad, M. K. Md; Nuzaihan, M. N. M.; Gopinath, Subash C. B.; Ruslinda, A. R.; Hashim, U.

2018-03-01

This paper investigates the influence of substrate-gate coupling on the ZnO-FET biosensor’s sensitivity for detection of cardiac troponin I (cTnI), a ‘gold standard’ biomarker for acute myocardial infarction (AMI). The FET-based device with introduction of substrate-gate coupling on p-type silicon-on-insulator (SOI) substrate is fabricated using conventional lithography processes. An n-type zinc oxide (ZnO) thin film deposited via electron-beam evaporator is used as transducer for bridging the source and drain regions. Surface modifications via functionalization with 3-aminopropyltriethoxysilane (APTES) and glutaraldehyde (GA) as chemical linkers, followed by immobilization of cTnI monoclonal antibody (MAb-cTnI) as bio-receptor on the ZnO thin film allow different concentration of cTnI detection with high selectivity. The device’s sensitivity increases up to 9 %·(g/ml)-1 with the increase of the substrate-gate voltage (VSG) up to -10 V at very low limit of detection (LOD) down to 1.6 fg/ml.

Transition from Reconstruction toward Thin Film on the (110) Surface of Strontium Titanate

PubMed Central

2016-01-01

The surfaces of metal oxides often are reconstructed with a geometry and composition that is considerably different from a simple termination of the bulk. Such structures can also be viewed as ultrathin films, epitaxed on a substrate. Here, the reconstructions of the SrTiO3 (110) surface are studied combining scanning tunneling microscopy (STM), transmission electron diffraction, and X-ray absorption spectroscopy (XAS), and analyzed with density functional theory calculations. Whereas SrTiO3 (110) invariably terminates with an overlayer of titania, with increasing density its structure switches from n × 1 to 2 × n. At the same time the coordination of the Ti atoms changes from a network of corner-sharing tetrahedra to a double layer of edge-shared octahedra with bridging units of octahedrally coordinated strontium. This transition from the n × 1 to 2 × n reconstructions is a transition from a pseudomorphically stabilized tetrahedral network toward an octahedral titania thin film with stress-relief from octahedral strontia units at the surface. PMID:26954064

Molecular and Subcellular-Scale Modeling of Nucleotide Diffusion in the Cardiac Myofilament Lattice

PubMed Central

Kekenes-Huskey, Peter M.; Liao, Tao; Gillette, Andrew K.; Hake, Johan E.; Zhang, Yongjie; Michailova, Anushka P.; McCulloch, Andrew D.; McCammon, J. Andrew

2013-01-01

Contractile function of cardiac cells is driven by the sliding displacement of myofilaments powered by the cycling myosin crossbridges. Critical to this process is the availability of ATP, which myosin hydrolyzes during the cross-bridge cycle. The diffusion of adenine nucleotides through the myofilament lattice has been shown to be anisotropic, with slower radial diffusion perpendicular to the filament axis relative to parallel, and is attributed to the periodic hexagonal arrangement of the thin (actin) and thick (myosin) filaments. We investigated whether atomistic-resolution details of myofilament proteins can refine coarse-grain estimates of diffusional anisotropy for adenine nucleotides in the cardiac myofibril, using homogenization theory and atomistic thin filament models from the Protein Data Bank. Our results demonstrate considerable anisotropy in ATP and ADP diffusion constants that is consistent with experimental measurements and dependent on lattice spacing and myofilament overlap. A reaction-diffusion model of the half-sarcomere further suggests that diffusional anisotropy may lead to modest adenine nucleotide gradients in the myoplasm under physiological conditions. PMID:24209858

Seasonal and interannual evolution of Jakobshavn Isbrae, Greenland from a 2008-2015 high-res DEM and velocity time series

NASA Astrophysics Data System (ADS)

Shean, D. E.; Joughin, I.; Smith, B.; Floricioiu, D.

2015-12-01

Greenland's large marine-terminating outlet glaciers have displayed marked retreat, speedup, and thinning in recent decades. Jakobshavn Isbrae, one of Greenland's largest outlet glaciers, has retreated ~15 km, accelerated ~150%, and thinned ~200 m since the early 1990s. Here, we present a comprehensive analysis of high-resolution elevation (~2-5 m/px) and velocity (~100 m/px) time series with dense temporal coverage (daily-monthly). The Jakobshavn DEM time series consists of >70 WorldView-1/2/3 stereo DEMs and >11 TanDEM-X DEMs spanning 2008-2015. Complementary point elevation data from Operation IceBridge (ATM, LVIS), pre-IceBridge ATM flights, and ICESat-1 GLAS extend the surface elevation record to 1999 and provide essential absolute control data, enabling sub-meter horizontal/vertical accuracy for gridded DEMs. Velocity data are primarily derived from TerraSAR-X/TanDEM-X image pairs with 11-day interval from 2009-2015. These elevation and velocity data capture outlet glacier evolution with unprecedented detail during the post-ICESat era. The lower trunk of Jakobshavn displays significant seasonal velocity variations, with recent rates of ~8 km/yr during winter and >17 km/yr during summer. DEM data show corresponding seasonal elevation changes of -30 to -45 m in summer and +15 to +20 m in winter, with decreasing magnitude upstream. Seasonal discharge varies from ~30-35 Gt/yr in winter to ~45-55 Gt/yr in summer, and we integrate these measurements for improved long-term mass-balance estimates. Recent interannual trends show increased discharge, velocity, and thinning (-15 to -20 m/yr), which is consistent with long-term altimetry records. The DEM time series also reveal new details about calving front and mélange evolution during the seasonal cycle. Similar time series are available for Kangerdlugssuaq and Helheim Glaciers. These observations are improving our understanding of outlet glacier dynamics, while complementing ongoing efforts to constrain estimates for ice-sheet mass balance and present/future sea level rise contributions.

Cell contact as an independent factor modulating cardiac myocyte hypertrophy and survival in long-term primary culture

NASA Technical Reports Server (NTRS)

Clark, W. A.; Decker, M. L.; Behnke-Barclay, M.; Janes, D. M.; Decker, R. S.

1998-01-01

Cardiac myocytes maintained in cell culture develop hypertrophy both in response to mechanical loading as well as to receptor-mediated signaling mechanisms. However, it has been shown that the hypertrophic response to these stimuli may be modulated through effects of intercellular contact achieved by maintaining cells at different plating densities. In this study, we show that the myocyte plating density affects not only the hypertrophic response and features of the differentiated phenotype of isolated adult myocytes, but also plays a significant role influencing myocyte survival in vitro. The native rod-shaped phenotype of freshly isolated adult myocytes persists in an environment which minimizes myocyte attachment and spreading on the substratum. However, these conditions are not optimal for long-term maintenance of cultured adult cardiac myocytes. Conditions which promote myocyte attachment and spreading on the substratum, on the other hand, also promote the re-establishment of new intercellular contacts between myocytes. These contacts appear to play a significant role in the development of spontaneous activity, which enhances the redevelopment of highly differentiated contractile, junctional, and sarcoplasmic reticulum structures in the cultured adult cardiomyocyte. Although it has previously been shown that adult cardiac myocytes are typically quiescent in culture, the addition of beta-adrenergic agonists stimulates beating and myocyte hypertrophy, and thereby serves to increase the level of intercellular contact as well. However, in densely-plated cultures with intrinsically high levels of intercellular contact, spontaneous contractile activity develops without the addition of beta-adrenergic agonists. In this study, we compare the function, morphology, and natural history of adult feline cardiomyocytes which have been maintained in cultures with different levels of intercellular contact, with and without the addition of beta-adrenergic agonists. Intercellular contact, communication, and transmission of contractile forces between myocytes appears to play a primary role in remodeling the 2-dimensional cell layer into a parallel alignment of elongated myocytes with highly developed intercalated disk-like junctions. This highly differentiated state is very stable, and cultures which achieve this state exhibit significantly greater longevity than more sparsely plated myocytes. These myocytes typically continue beating, and survive from 6 to more than 12 weeks in culture. When this level of contact and differentiation are not achieved, even among beta-adrenergic stimulated myocytes, contractile activity is not sustained, myofibrils atrophy, there is little or no development of junctional complexes, and the period of myocyte viability is typically no more than 5 weeks in vitro.

Polygonal crack patterns by drying thin films under quasi-two-dimensional confinement

NASA Astrophysics Data System (ADS)

Ma, Xiaolei; Lowensohn, Janna; Burton, Justin

Cracks patterns such as T/Y junction cracks in dried mud are ubiquitous in nature. Although the conditions for cracking in solids is well-known, cracks in colloidal and granular systems are more complex. Here we report the formations of polygonal cracks by drying thin films of corn starch ( 10 μm in diameter) under quasi-2D confinement. We find there are two drying stages before the films are completely dried. Initially, a compaction front invades throughout the film. Then, a second drying stage ''percolates'' throughout the film with a characteristic branching pattern, leading to a dense packing of particles connected by liquid capillary bridges. Finally, polygonal cracks appear as the remaining liquid dries. The same drying kinetics occur for films with different thickness, h, except that fractal-like fracture patterns form in thin films, where the thickness is comparable to the particle size, while polygons form in thick films with many layers of particles. We also find that the average area of the polygons, A, in fully dried films scales with the thickness, A hβ , where β 1 . 5 , and the prefactor depends on the initial packing fraction of the suspension. This form is consistent with a simple energy balance criterion for crack formation.

Is titin a 'winding filament'? A new twist on muscle contraction.

PubMed

Nishikawa, Kiisa C; Monroy, Jenna A; Uyeno, Theodore E; Yeo, Sang Hoon; Pai, Dinesh K; Lindstedt, Stan L

2012-03-07

Recent studies have demonstrated a role for the elastic protein titin in active muscle, but the mechanisms by which titin plays this role remain to be elucidated. In active muscle, Ca(2+)-binding has been shown to increase titin stiffness, but the observed increase is too small to explain the increased stiffness of parallel elastic elements upon muscle activation. We propose a 'winding filament' mechanism for titin's role in active muscle. First, we hypothesize that Ca(2+)-dependent binding of titin's N2A region to thin filaments increases titin stiffness by preventing low-force straightening of proximal immunoglobulin domains that occurs during passive stretch. This mechanism explains the difference in length dependence of force between skeletal myofibrils and cardiac myocytes. Second, we hypothesize that cross-bridges serve not only as motors that pull thin filaments towards the M-line, but also as rotors that wind titin on the thin filaments, storing elastic potential energy in PEVK during force development and active stretch. Energy stored during force development can be recovered during active shortening. The winding filament hypothesis accounts for force enhancement during stretch and force depression during shortening, and provides testable predictions that will encourage new directions for research on mechanisms of muscle contraction.

Is titin a ‘winding filament’? A new twist on muscle contraction

PubMed Central

Nishikawa, Kiisa C.; Monroy, Jenna A.; Uyeno, Theodore E.; Yeo, Sang Hoon; Pai, Dinesh K.; Lindstedt, Stan L.

2012-01-01

Recent studies have demonstrated a role for the elastic protein titin in active muscle, but the mechanisms by which titin plays this role remain to be elucidated. In active muscle, Ca2+-binding has been shown to increase titin stiffness, but the observed increase is too small to explain the increased stiffness of parallel elastic elements upon muscle activation. We propose a ‘winding filament’ mechanism for titin's role in active muscle. First, we hypothesize that Ca2+-dependent binding of titin's N2A region to thin filaments increases titin stiffness by preventing low-force straightening of proximal immunoglobulin domains that occurs during passive stretch. This mechanism explains the difference in length dependence of force between skeletal myofibrils and cardiac myocytes. Second, we hypothesize that cross-bridges serve not only as motors that pull thin filaments towards the M-line, but also as rotors that wind titin on the thin filaments, storing elastic potential energy in PEVK during force development and active stretch. Energy stored during force development can be recovered during active shortening. The winding filament hypothesis accounts for force enhancement during stretch and force depression during shortening, and provides testable predictions that will encourage new directions for research on mechanisms of muscle contraction. PMID:21900329

Design and Experimentation with Sandwich Microstructure for Catalytic Combustion-Type Gas Sensors

PubMed Central

Gu, Jun-Tao; Zhang, Yong-De; Jiang, Jin-Gang

2014-01-01

The traditional handmade catalytic combustion gas sensor has some problems such as a pairing difficulty, poor consistency, high power consumption, and not being interchangeable. To address these issues, integrated double catalytic combustion of alcohol gas sensor was designed and manufactured using silicon micro-electro-mechanical systems (MEMS) technology. The temperature field of the sensor is analyzed using the ANSYS finite element analysis method. In this work, the silicon oxide-PECVD-oxidation technique is used to manufacture a SiO2-Si3N2-SiO2 microstructure carrier with a sandwich structure, while wet etching silicon is used to form a beam structure to reduce the heat consumption. Thin-film technology is adopted to manufacture the platinum-film sensitive resistance. Nano Al2O3-ZrO-ThO is coated to format the sensor carrier, and the sensitive unit is dipped in a Pt-Pd catalyst solution to form the catalytic sensitive bridge arm. Meanwhile the uncoated catalyst carrier is considered as the reference unit, realizing an integrated chip based on a micro double bridge and forming sensors. The lines of the Pt thin-film resistance have been observed with an electronic microscope. The compensation of the sensitive material carriers and compensation materials have been analyzed using an energy spectrum. The results show that the alcohol sensor can detect a volume fraction between 0 and 4,500 × 10−6 and has good linear output characteristic. The temperature ranges from −20 to +40 °C. The humidity ranges from 30% to 85% RH. The zero output of the sensor is less than ±2.0% FS. The power consumption is ≤0.2 W, and both the response and recovery time are approximately 20 s. PMID:24625742

An ionic-chemical-mechanical model for muscle contraction.

PubMed

Manning, Gerald S

2016-12-01

The dynamic process underlying muscle contraction is the parallel sliding of thin actin filaments along an immobile thick myosin fiber powered by oar-like movements of protruding myosin cross bridges (myosin heads). The free energy for functioning of the myosin nanomotor comes from the hydrolysis of ATP bound to the myosin heads. The unit step of translational movement is based on a mechanical-chemical cycle involving ATP binding to myosin, hydrolysis of the bound ATP with ultimate release of the hydrolysis products, stress-generating conformational changes in the myosin cross bridge, and relief of built-up stress in the myosin power stroke. The cycle is regulated by a transition between weak and strong actin-myosin binding affinities. The dissociation of the weakly bound complex by addition of salt indicates the electrostatic basis for the weak affinity, while structural studies demonstrate that electrostatic interactions among negatively charged amino acid residues of actin and positively charged residues of myosin are involved in the strong binding interface. We therefore conjecture that intermediate states of increasing actin-myosin engagement during the weak-to-strong binding transition also involve electrostatic interactions. Methods of polymer solution physics have shown that the thin actin filament can be regarded in some of its aspects as a net negatively charged polyelectrolyte. Here we employ polyelectrolyte theory to suggest how actin-myosin electrostatic interactions might be of significance in the intermediate stages of binding, ensuring an engaged power stroke of the myosin motor that transmits force to the actin filament, and preventing the motor from getting stuck in a metastable pre-power stroke state. We provide electrostatic force estimates that are in the pN range known to operate in the cycle. © 2016 Wiley Periodicals, Inc.

Discrete element modeling of microstructure of nacre

NASA Astrophysics Data System (ADS)

Chandler, Mei Qiang; Cheng, Jing-Ru C.

2018-04-01

The microstructure of nacre consists of polygon-shaped aragonite mineral tablets bonded by very thin layers of organic materials and is organized in a brick-mortar morphology. In this research, the discrete element method was utilized to model this structure. The aragonite mineral tablets were modeled with three-dimensional polygon particles generated by the Voronoi tessellation method to represent the Voronoi-like patterns of mineral tablets assembly observed in experiments. The organic matrix was modeled with a group of spring elements. The constitutive relations of the spring elements were inspired from the experimental results of organic molecules from the literature. The mineral bridges were modeled with simple elastic bonds with the parameters based on experimental data from the literature. The bulk stress-strain responses from the models agreed well with experimental results. The model results show that the mineral bridges play important roles in providing the stiffness and yield strength for the nacre, while the organic matrix in providing the ductility for the nacre. This work demonstrated the suitability of particle methods for modeling microstructures of nacre.

Increasing the endogenous NO level causes catalase inactivation and reactivation of intercellular apoptosis signaling specifically in tumor cells

PubMed Central

Bauer, Georg

2015-01-01

Tumor cells generate extracellular superoxide anions and are protected against intercellular apoptosis-inducing HOCl- and NO/peroxynitrite signaling through the expression of membrane-associated catalase. This enzyme decomposes H2O2 and thus prevents HOCl synthesis. It efficiently interferes with NO/peroxynitrite signaling through oxidation of NO and decomposition of peroxynitrite. The regulatory potential of catalase at the crosspoint of ROS and RNS chemical biology, as well as its high local concentration on the outside of the cell membrane of tumor cells, establish tight control of intercellular signaling and thus prevent tumor cell apoptosis. Therefore, inhibition of catalase or its inactivation by singlet oxygen reactivate intercellular apoptosis-inducing signaling. Nitric oxide and peroxynitrite are connected with catalase in multiple and meaningful ways, as (i) NO can be oxidated by compound I of catalase, (ii) NO can reversibly inhibit catalase, (iii) peroxynitrite can be decomposed by catalase and (iv) the interaction between peroxynitrite and H2O2 leads to the generation of singlet oxygen that inactivates catalase. Therefore, modulation of the concentration of free NO through addition of arginine, inhibition of arginase, induction of NOS expression or inhibition of NO dioxygenase triggers an autoamplificatory biochemical cascade that is based on initial formation of singlet oxygen, amplification of superoxide anion/H2O2 and NO generation through singlet oxygen dependent stimulation of the FAS receptor and caspase-8. Finally, singlet oxygen is generated at sufficiently high concentration to inactivate protective catalase and to reactivate intercellular apoptosis-inducing ROS signaling. This regulatory network allows to establish several pathways for synergistic interactions, like the combination of modulators of NO metabolism with enhancers of superoxide anion generation, modulators of NO metabolism that act at different targets and between modulators of NO metabolism and direct catalase inhibitors. The latter aspect is explicitely studied for the interaction between catalase inhibiting acetylsalicylic acid and an NO donor. It is also shown that hybrid molecules like NO-aspirin utilize this synergistic potential. Our data open novel approaches for rational tumor therapy based on specific ROS signaling and its control in tumor cells. PMID:26342455

Keratins Regulate p38MAPK-Dependent Desmoglein Binding Properties in Pemphigus

PubMed Central

Vielmuth, Franziska; Walter, Elias; Fuchs, Michael; Radeva, Mariya Y.; Buechau, Fanny; Magin, Thomas M.; Spindler, Volker; Waschke, Jens

2018-01-01

Keratins are crucial for the anchorage of desmosomes. Severe alterations of keratin organization and detachment of filaments from the desmosomal plaque occur in the autoimmune dermatoses pemphigus vulgaris and pemphigus foliaceus (PF), which are mainly caused by autoantibodies against desmoglein (Dsg) 1 and 3. Keratin alterations are a structural hallmark in pemphigus pathogenesis and correlate with loss of intercellular adhesion. However, the significance for autoantibody-induced loss of intercellular adhesion is largely unknown. In wild-type (wt) murine keratinocytes, pemphigus autoantibodies induced keratin filament retraction. Under the same conditions, we used murine keratinocytes lacking all keratin filaments (KtyII k.o.) as a model system to dissect the role of keratins in pemphigus. KtyII k.o. cells show compromised intercellular adhesion without antibody (Ab) treatment, which was not impaired further by pathogenic pemphigus autoantibodies. Nevertheless, direct activation of p38MAPK via anisomycin further decreased intercellular adhesion indicating that cell cohesion was not completely abrogated in the absence of keratins. Direct inhibition of Dsg3, but not of Dsg1, interaction via pathogenic autoantibodies as revealed by atomic force microscopy was detectable in both cell lines demonstrating that keratins are not required for this phenomenon. However, PF-IgG shifted Dsg1-binding events from cell borders toward the free cell surface in wt cells. This led to a distribution pattern of Dsg1-binding events similar to KtyII k.o. cells under resting conditions. In keratin-deficient keratinocytes, PF-IgG impaired Dsg1-binding strength, which was not different from wt cells under resting conditions. In addition, pathogenic autoantibodies were capable of activating p38MAPK in both KtyII wt and k.o. cells, the latter of which already displayed robust p38MAPK activation under resting conditions. Since inhibition of p38MAPK blocked autoantibody-induced loss of intercellular adhesion in wt cells and restored baseline cell cohesion in keratin-deficient cells, we conclude that p38MAPK signaling is (i) critical for regulation of cell adhesion, (ii) regulated by keratins, and (iii) targets both keratin-dependent and -independent mechanisms. PMID:29616033

Destruction of the hepatocyte junction by intercellular invasion of Leptospira causes jaundice in a hamster model of Weil's disease.

PubMed

Miyahara, Satoshi; Saito, Mitsumasa; Kanemaru, Takaaki; Villanueva, Sharon Y A M; Gloriani, Nina G; Yoshida, Shin-ichi

2014-08-01

Weil's disease, the most severe form of leptospirosis, is characterized by jaundice, haemorrhage and renal failure. The mechanisms of jaundice caused by pathogenic Leptospira remain unclear. We therefore aimed to elucidate the mechanisms by integrating histopathological changes with serum biochemical abnormalities during the development of jaundice in a hamster model of Weil's disease. In this work, we obtained three-dimensional images of infected hamster livers using scanning electron microscope together with freeze-cracking and cross-cutting methods for sample preparation. The images displayed the corkscrew-shaped bacteria, which infiltrated the Disse's space, migrated between hepatocytes, detached the intercellular junctions and disrupted the bile canaliculi. Destruction of bile canaliculi coincided with the elevation of conjugated bilirubin, aspartate transaminase and alkaline phosphatase levels in serum, whereas serum alanine transaminase and γ-glutamyl transpeptidase levels increased slightly, but not significantly. We also found in ex vivo experiments that pathogenic, but not non-pathogenic leptospires, tend to adhere to the perijunctional region of hepatocyte couplets isolated from hamsters and initiate invasion of the intercellular junction within 1 h after co-incubation. Our results suggest that pathogenic leptospires invade the intercellular junctions of host hepatocytes, and this invasion contributes in the disruption of the junction. Subsequently, bile leaks from bile canaliculi and jaundice occurs immediately. Our findings revealed not only a novel pathogenicity of leptospires, but also a novel mechanism of jaundice induced by bacterial infection. © 2014 The Authors. International Journal of Experimental Pathology © 2014 International Journal of Experimental Pathology.

Trpm7 Protein Contributes to Intercellular Junction Formation in Mouse Urothelium*

PubMed Central

Watanabe, Masaki; Suzuki, Yoshiro; Uchida, Kunitoshi; Miyazaki, Naoyuki; Murata, Kazuyoshi; Matsumoto, Seiji; Kakizaki, Hidehiro; Tominaga, Makoto

2015-01-01

Trpm7 is a divalent cation-permeable channel that has been reported to be involved in magnesium homeostasis as well as cellular adhesion and migration. We generated urothelium-specific Trpm7 knock-out (KO) mice to reveal the function of Trpm7 in vivo. A Trpm7 KO was induced by tamoxifen and was confirmed by genomic PCR and immunohistochemistry. By using patch clamp recordings in primary urothelial cells, we observed that Mg2+-inhibitable cation currents as well as acid-inducible currents were significantly smaller in Trpm7 KO urothelial cells than in cells from control mice. Assessment of voiding behavior indicated a significantly smaller voided volume in Trpm7 KO mice (mean voided volume 0.28 ± 0.08 g in KO mice and 0.36 ± 0.04 g in control mice, p < 0.05, n = 6–8). Histological analysis showed partial but substantial edema in the submucosal layer of Trpm7 KO mice, most likely due to inflammation. The expression of proinflammatory cytokines TNF-α and IL-1β was significantly higher in Trpm7 KO bladders than in controls. In transmission electron microscopic analysis, immature intercellular junctions were observed in Trpm7 KO urothelium but not in control mice. These results suggest that Trpm7 is involved in the formation of intercellular junctions in mouse urothelium. Immature intercellular junctions in Trpm7 knock-out mice might lead to a disruption of barrier function resulting in inflammation and hypersensitive bladder afferent nerves that may affect voiding behavior in vivo. PMID:26504086

Trpm7 Protein Contributes to Intercellular Junction Formation in Mouse Urothelium.

PubMed

Watanabe, Masaki; Suzuki, Yoshiro; Uchida, Kunitoshi; Miyazaki, Naoyuki; Murata, Kazuyoshi; Matsumoto, Seiji; Kakizaki, Hidehiro; Tominaga, Makoto

2015-12-11

Trpm7 is a divalent cation-permeable channel that has been reported to be involved in magnesium homeostasis as well as cellular adhesion and migration. We generated urothelium-specific Trpm7 knock-out (KO) mice to reveal the function of Trpm7 in vivo. A Trpm7 KO was induced by tamoxifen and was confirmed by genomic PCR and immunohistochemistry. By using patch clamp recordings in primary urothelial cells, we observed that Mg(2+)-inhibitable cation currents as well as acid-inducible currents were significantly smaller in Trpm7 KO urothelial cells than in cells from control mice. Assessment of voiding behavior indicated a significantly smaller voided volume in Trpm7 KO mice (mean voided volume 0.28 ± 0.08 g in KO mice and 0.36 ± 0.04 g in control mice, p < 0.05, n = 6-8). Histological analysis showed partial but substantial edema in the submucosal layer of Trpm7 KO mice, most likely due to inflammation. The expression of proinflammatory cytokines TNF-α and IL-1β was significantly higher in Trpm7 KO bladders than in controls. In transmission electron microscopic analysis, immature intercellular junctions were observed in Trpm7 KO urothelium but not in control mice. These results suggest that Trpm7 is involved in the formation of intercellular junctions in mouse urothelium. Immature intercellular junctions in Trpm7 knock-out mice might lead to a disruption of barrier function resulting in inflammation and hypersensitive bladder afferent nerves that may affect voiding behavior in vivo. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

Active properties of living tissues lead to size-dependent dewetting

NASA Astrophysics Data System (ADS)

Perez-Gonzalez, Carlos; Alert, Ricard; Blanch-Mercader, Carles; Gomez-Gonzalez, Manuel; Casademunt, Jaume; Trepat, Xavier

Key biological processes such as cancer and development are characterized by drastic transitions from 2D to a 3D geometry. These rearrangements have been classically studied as a wetting problem. According to this theory, wettability of a substrate by an epithelium is determined by the competition between cell-cell and cell-substrate adhesion energies. In contrast, we found that, far from a passive process, tissue dewetting is an active process driven by tissue internal forces. Experimentally, we reproduced epithelial dewetting by promoting a progressive formation of intercellular junctions in a monolayer of epithelial cells. Interestingly, the formation of intercellular junctions produces an increase in cell contractility, with the subsequent increase in traction and intercellular stress. At a certain time, tissue tension overcomes cell-substrate maximum adhesion and the monolayer spontaneously dewets the substrate. We developed an active polar fluid model, finding both theoretically and experimentally that critical contractility to promote wetting-dewetting transition depends on cell-substrate adhesion and, unexpectedly, on tissue size. As a whole, this work generalizes wetting theory to living tissues, unveiling unprecedented properties due to their unique active nature.

Twist1-positive epithelial cells retain adhesive and proliferative capacity throughout dissemination

PubMed Central

Shamir, Eliah R.; Coutinho, Kester; Georgess, Dan; Auer, Manfred

2016-01-01

ABSTRACT Dissemination is the process by which cells detach and migrate away from a multicellular tissue. The epithelial-to-mesenchymal transition (EMT) conceptualizes dissemination in a stepwise fashion, with downregulation of E-cadherin leading to loss of intercellular junctions, induction of motility, and then escape from the epithelium. This gain of migratory activity is proposed to be mutually exclusive with proliferation. We previously developed a dissemination assay based on inducible expression of the transcription factor Twist1 and here utilize it to characterize the timing and dynamics of intercellular adhesion, proliferation and migration during dissemination. Surprisingly, Twist1+ epithelium displayed extensive intercellular junctions, and Twist1– luminal epithelial cells could still adhere to disseminating Twist1+ cells. Although proteolysis and proliferation were both observed throughout dissemination, neither was absolutely required. Finally, Twist1+ cells exhibited a hybrid migration mode; their morphology and nuclear deformation were characteristic of amoeboid cells, whereas their dynamic protrusive activity, pericellular proteolysis and migration speeds were more typical of mesenchymal cells. Our data reveal that epithelial cells can disseminate while retaining competence to adhere and proliferate. PMID:27402962

Activation of L-type calcium channels is required for gap junction-mediated intercellular calcium signaling in osteoblastic cells

NASA Technical Reports Server (NTRS)

Jorgensen, Niklas Rye; Teilmann, Stefan Cuoni; Henriksen, Zanne; Civitelli, Roberto; Sorensen, Ole Helmer; Steinberg, Thomas H.

2003-01-01

The propagation of mechanically induced intercellular calcium waves (ICW) among osteoblastic cells occurs both by activation of P2Y (purinergic) receptors by extracellular nucleotides, resulting in "fast" ICW, and by gap junctional communication in cells that express connexin43 (Cx43), resulting in "slow" ICW. Human osteoblastic cells transmit intercellular calcium signals by both of these mechanisms. In the current studies we have examined the mechanism of slow gap junction-dependent ICW in osteoblastic cells. In ROS rat osteoblastic cells, gap junction-dependent ICW were inhibited by removal of extracellular calcium, plasma membrane depolarization by high extracellular potassium, and the L-type voltage-operated calcium channel inhibitor, nifedipine. In contrast, all these treatments enhanced the spread of P2 receptor-mediated ICW in UMR rat osteoblastic cells. Using UMR cells transfected to express Cx43 (UMR/Cx43) we confirmed that nifedipine sensitivity of ICW required Cx43 expression. In human osteoblastic cells, gap junction-dependent ICW also required activation of L-type calcium channels and influx of extracellular calcium.

The multicellular nature of filamentous heterocyst-forming cyanobacteria.

PubMed

Herrero, Antonia; Stavans, Joel; Flores, Enrique

2016-11-01

Cyanobacteria carry out oxygenic photosynthesis, play a key role in the cycling of carbon and nitrogen in the biosphere, and have had a large impact on the evolution of life and the Earth itself. Many cyanobacterial strains exhibit a multicellular lifestyle, growing as filaments that can be hundreds of cells long and endowed with intercellular communication. Furthermore, under depletion of combined nitrogen, filament growth requires the activity of two interdependent cell types: vegetative cells that fix CO2 and heterocysts that fix N2. Intercellular molecular transfer is essential for signaling involved in the regulation of heterocyst differentiation and for reciprocal nutrition of heterocysts and vegetative cells. Here we review various aspects of multicellularity in cyanobacterial filaments and their differentiation, including filament architecture with emphasis on the structures used for intercellular communication; we survey theoretical models that have been put forward to understand heterocyst patterning and discuss the factors that need to be considered for these models to reflect the biological entity; and finally, since cell division in filamentous cyanobacteria has the peculiarity of producing linked instead of independent cells, we review distinct aspects of cell division in these organisms.

Mechanisms for Cell-to-Cell Transmission of HIV-1

PubMed Central

Bracq, Lucie; Xie, Maorong; Benichou, Serge; Bouchet, Jérôme

2018-01-01

While HIV-1 infection of target cells with cell-free viral particles has been largely documented, intercellular transmission through direct cell-to-cell contact may be a predominant mode of propagation in host. To spread, HIV-1 infects cells of the immune system and takes advantage of their specific particularities and functions. Subversion of intercellular communication allows to improve HIV-1 replication through a multiplicity of intercellular structures and membrane protrusions, like tunneling nanotubes, filopodia, or lamellipodia-like structures involved in the formation of the virological synapse. Other features of immune cells, like the immunological synapse or the phagocytosis of infected cells are hijacked by HIV-1 and used as gateways to infect target cells. Finally, HIV-1 reuses its fusogenic capacity to provoke fusion between infected donor cells and target cells, and to form infected syncytia with high capacity of viral production and improved capacities of motility or survival. All these modes of cell-to-cell transfer are now considered as viral mechanisms to escape immune system and antiretroviral therapies, and could be involved in the establishment of persistent virus reservoirs in different host tissues. PMID:29515578

Studies of intercellular invasion in vitro using rabbit peritoneal neutrophil granulocytes (PMNS). I. Role of contact inhibition of locomotion

PubMed Central

1975-01-01

Intercellular invasion is the active migration of cells on one type into the interiors of tissues composed of cells of dissimilar cell types. Contact paralysis of locomotion is the cessation of forward extension of the pseudopods of a cell as a result of its collision with another cell. One hypothesis to account for intercellular invasion proposes that a necessary condition for a cell type to be invasive to a given host tissue is that it lack contact paralysis of locomotion during collision with cells of that host tissue. The hypothesis has been tested using rabbit peritoneal neutrophil granulocytes (PMNs) as the invasive cell type and chick embryo fibroblasts as the host tissue. In organ culture, PMNs rapidly invade aggregates of fibroblasts. The behavior of the pseudopods of PMNs during collision with fibroblasts was analyzed for contact paralysis by a study of time-lapse films of cells in mixed monolayer culture. In monolayer culture, PMNs show little sign of paralysis of the pseudopods upon collision with fibroblasts and thus conform in their behavior to that predicted by the hypothesis. PMID:1092702

Dilated intercellular spaces and chronic cough as an extra-oesophageal manifestation of gastrooesophageal reflux disease.

PubMed

Orlando, Roy C

2011-06-01

Chronic cough is one of the extra-oesophageal manifestations of gastrooesophageal reflux disease (GORD). It is presumed to occur either directly by microaspiration of acidic gastric contents into the airway or indirectly by a reflex triggered by contact of acidic refluxates with the oesophageal epithelium in GORD. How contact of the oesophageal epithelium with acidic refluxates promotes sensitization for chronic cough is unknown, but like heartburn, which is a necessary accompaniment, it requires acid activation of nociceptors within the oesophageal mucosa. Dilated intercellular spaces within the oesophageal epithelium, a reflection of an increase in paracellular permeability, is a histopathologic feature of both erosive and non-erosive forms of GORD. Since it correlates with the symptom of heartburn, it is hypothesized herein that the increase in paracellular permeability to acid reflected by dilated intercellular spaces in oesophageal epithelium also serves as mediator of the signals that produce the reflex-induced sensitization for cough--a sensitization that can occur centrally within the medullary Nucleus Tractus Solitarius or peripherally within the tracheobronchial tree. Copyright © 2010 Elsevier Ltd. All rights reserved.

Protective Effects of LSGYGP from Fish Skin Gelatin Hydrolysates on UVB-Induced MEFs by Regulation of Oxidative Stress and Matrix Metalloproteinase Activity.

PubMed

Ma, Qingyu; Liu, Qiuming; Yuan, Ling; Zhuang, Yongliang

2018-03-28

A previous study has shown that tilapia fish skin gelatin hydrolysates inhibited photoaging in vivo, and that, Leu-Ser-Gly-Tyr-Gly-Pro (LSGYGP) identified in the hydrolysate had a high hydroxyl radical scavenging activity. In this study, activities of LSGYGP were further evaluated using ultraviolet B (UVB)-induced mouse embryonic fibroblasts (MEFs). UVB irradiation significantly increased the intercellular reactive oxygen species (ROS) production and matrix metalloproteinases (MMPs) activities and decreased the content of collagen in MEFs. LSGYGP reduced the intercellular ROS generation in UVB-induced MEFs. Meanwhile, the decrease of superoxide dismutase (SOD) activity and the increase of malondiaidehyde (MDA) content were inhibited by LSGYGP. LSGYGP reduced MMP-1 and MMP-9 activities in a dose-dependent manner. Molecular docking simulation indicated that LSGYGP inhibited MMPs activities by docking the active sites of MMP-1 and MMP-9. Furthermore, LSGYGP also affected the intercellular phosphorylation of UVB-induced the mitogen-activated protein kinase pathway. LSGYGP could protect collagen synthesis in MEFs under UVB irradiation by inhibiting oxidative stress and regulating MMPs activities.

Protective Effects of LSGYGP from Fish Skin Gelatin Hydrolysates on UVB-Induced MEFs by Regulation of Oxidative Stress and Matrix Metalloproteinase Activity

PubMed Central

Ma, Qingyu; Liu, Qiuming; Yuan, Ling; Zhuang, Yongliang

2018-01-01

A previous study has shown that tilapia fish skin gelatin hydrolysates inhibited photoaging in vivo, and that, Leu-Ser-Gly-Tyr-Gly-Pro (LSGYGP) identified in the hydrolysate had a high hydroxyl radical scavenging activity. In this study, activities of LSGYGP were further evaluated using ultraviolet B (UVB)-induced mouse embryonic fibroblasts (MEFs). UVB irradiation significantly increased the intercellular reactive oxygen species (ROS) production and matrix metalloproteinases (MMPs) activities and decreased the content of collagen in MEFs. LSGYGP reduced the intercellular ROS generation in UVB-induced MEFs. Meanwhile, the decrease of superoxide dismutase (SOD) activity and the increase of malondiaidehyde (MDA) content were inhibited by LSGYGP. LSGYGP reduced MMP-1 and MMP-9 activities in a dose-dependent manner. Molecular docking simulation indicated that LSGYGP inhibited MMPs activities by docking the active sites of MMP-1 and MMP-9. Furthermore, LSGYGP also affected the intercellular phosphorylation of UVB-induced the mitogen-activated protein kinase pathway. LSGYGP could protect collagen synthesis in MEFs under UVB irradiation by inhibiting oxidative stress and regulating MMPs activities. PMID:29597313

Terbinafine inhibits gap junctional intercellular communication

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lee, Ju Yeun, E-mail: whitewndus@naver.com

Terbinafine is an antifungal agent that selectively inhibits fungal sterol synthesis by blocking squalene epoxidase. We evaluated the effect of terbinafine on gap junctional intercellular communication (GJIC). Fluorescence recovery after photobleaching (FRAP) and I-YFP GJIC assays revealed that terbinafine inhibits GJIC in a reversible and dose-dependent manner in FRT-Cx43 and LN215 cells. Treatment with terbinafine did not affect Cx43 phosphorylation status or intracellular Ca{sup 2+} concentration, well-known action mechanisms of various GJIC blockers. While a structurally related chemical, naftifine, attenuated GJIC, epigallocatechin gallate, another potent squalene epoxidase inhibitor with a different structure, did not. These results suggest that terbinafine inhibitsmore » GJIC with a so far unknown mechanism of action. - Highlights: • In vitro pharmacological studies were performed on FRT-Cx43 and LN215 cells. • Terbinafine inhibits gap junctional intercellular communication in both cell lines. • The inhibitory effect of terbinafine is reversible and dose-dependent. • Treatment of terbinafine does not alter Cx43 phosphorylation or cytosolic Ca{sup 2+} concentration. • Inhibition of squalene epoxidase is not involved in this new effect of terbinafine.« less

Zn(II)-cyclam based chromogenic sensors for recognition of ATP in aqueous solution under physiological conditions and their application as viable staining agents for microorganism.

PubMed

Mahato, Prasenjit; Ghosh, Amrita; Mishra, Sanjiv K; Shrivastav, Anupama; Mishra, Sandhya; Das, Amitava

2011-05-02

Two chromogenic complexes, L.Zn (where L is (E)-4-((4-(1,4,8,11-tetraazacyclotetradecan-1-ylsulfonyl)phenyl)diazenyl)-N,N-dimethylaniline) and its [2]pseudorotaxane form (α-CD.L.Zn), were found to bind preferentially to adenosine triphosphate (ATP), among all other common anions and biologically important phosphate (AMP, ADP, pyrophosphate, and phosphate) ions in aqueous HEPES buffer medium of pH 7.2. Studies with live cell cultures of prokaryotic microbes revealed that binding of these two reagents to intercellular ATP, produced in situ, could be used in delineating the gram-positive and the gram-negative bacteria. More importantly, these dyes were found to be nontoxic to living microbes (eukaryotes and prokaryotes) and could be used for studying the cell growth dynamics. Binding to these two viable staining agents to intercellular ATP was also confirmed by spectroscopic studies on cell growth in the presence of different respiratory inhibitors that influence the intercellular ATP generation. © 2011 American Chemical Society

Purification and Characterization of Peptides Inhibiting MMP-1 Activity with C Terminate of Gly-Leu from Simulated Gastrointestinal Digestion Hydrolysates of Tilapia (Oreochromis niloticus) Skin Gelatin.

PubMed

Liping, Sun; Qiuming, Liu; Jian, Fan; Xiao, Li; Yongliang, Zhuang

2018-01-24

Tilapia skin gelatin hydrolysates (TSGHs) were prepared by simulated gastrointestinal digestion and separated by gel filtration and semi-preparative reversed-phase high-performance liquid chromatography. The anti-photoaging effects were evaluated using an ultraviolet radiation B (UVB)-induced mouse embryonic fibroblast (MEF) photoaging model in vitro. Three fractions from TSGHs with high inhibitory intercellular matrix metalloproteinase-1 (MMP-1) activities and reactive oxygen species (ROS) production were obtained. Three key peptides, GYTGL, LGATGL, and VLGL, were identified, and their C terminate was Gly-Leu. Three peptides were synthesized and exhibited a significant inhibition of intercellular MMP-1 activity and ROS production. Furthermore, three peptides inhibiting MMP-1 activities were evaluated through their docking of S 1 ' and S 3 ' active pockets of MMP-1. Hydrogen bonds and C terminate Gly-Leu played important roles. Finally, the protective effects of three peptides on intercellular collagen in UVB-induced MEFs were compared. Our results indicated that tilapia gelatin peptides exhibited potential activities to prevent and regulate photoaging.

A nonpolio enterovirus with respiratory tropism causes poliomyelitis in intercellular adhesion molecule 1 transgenic mice.

PubMed

Dufresne, Andrew T; Gromeier, Matthias

2004-09-14

Coxsackievirus A21 (CAV21) is classified within the species Human enterovirus C (HEV-C) of the Enterovirus genus of picornaviruses. HEV-C share striking homology with the polioviruses (PV), their closest kin among the enteroviruses. Despite a high level of sequence identity, CAV21 and PV cause distinct clinical disease typically attributed to their differential use of host receptors. PV cause poliomyelitis, whereas CAV21 shares a receptor and a propensity to cause upper respiratory tract infections with the major group rhinoviruses. As a model for CAV21 infection, we have developed transgenic mice that express human intercellular adhesion molecule 1, the cell-surface receptor for CAV21. Surprisingly, CAV21 administered to these mice via the intramuscular route causes a paralytic condition consistent with poliomyelitis. The virus appears to invade the CNS by retrograde axonal transport, as has been demonstrated to occur in analogous PV infections. We detected human intercellular adhesion molecule 1 expression on both transgenic mouse and human spinal cord anterior horn motor neurons, indicating that members of HEV-C may share PV's potential to elicit poliomyelitis in humans.

Bridge effects on light harvesting of a DBfA type polymer system

NASA Astrophysics Data System (ADS)

Sun, Sam-Shajing; Hasib, Muhammad; Gavrilenko, Alexander V.; Devan, Joshua; Gavrilenko, Vladimir

2016-09-01

Plastic optoelectronic materials and thin film devices are very attractive in future optical sensor and solar energy applications due to their lightweight, flexible shape, high photon absorption coefficients, low cost, and environmental benefits. In this study, optoelectronic properties of D, D/fA blend, DfA, and a series of DBfA type of conjugated block copolymers has been investigated, where D is a donor type PPV conjugated block, B is a non-conjugated and flexible aliphatic hydrocarbon bridge chain containing different number of aliphatic methylene units, and fA is a fluorinated acceptor type PPV conjugated block. The optical absorptions of the D/fA blend, DfA, and DBfAs are typical overlaps of individual absorptions of D and fA blocks, while the solution steady state photoluminescence (PL) emission of D were quenched to different levels in blends and block copolymers, with DBfAs containing one methylene unit bridge (DB1fA) quenched most. This could be attributed to an intra-molecular photo induced electron transfer or charge separation in DBfA systems. Theoretical first principles study of the equilibrium atomic configuration of DfA reveals the existence of twisting angles between the D and fA blocks in DfA stable states which may account for a less PL quenching of DfA as compared to DB1fA. These results are important for designing and developing high efficiency polymer based optoelectronic systems.

High-efficiency emitting materials based on phenylquinoline/carbazole-based compounds for organic light emitting diode applications

NASA Astrophysics Data System (ADS)

Jin, Sung-Ho

2009-08-01

Highly efficient light-emitting materials based on phenylquinoline-carbazole derivative has been synthesized for organic-light emitting diodes (OLEDs). The materials form high quality amorphous thin films by thermal evaporation and the energy levels can be easily adjusted by the introduction of different electron donating and electron withdrawing groups on carbazoylphenylquinoline. Non-doped deep-blue OLEDs using Et-CVz-PhQ as the emitter show bright emission (CIE coordinates, x=0.156, y=0.093) with an external quantum efficiency of 2.45 %. Furthermore, the material works as an excellent host material for BCzVBi to get high-performance OLEDs with excellent deep-blue CIE coordinates (x=0.155, y=0.157), high power efficiency (5.98 lm/W), and high external quantum efficiency (5.22 %). Cyclometalated Ir(III) μ-chloride bridged dimers were synthesized by iridium trichloride hydrate with an excess of our developed deep-blue emitter, Et-CVz-PhQ. The Ir(III) complexes were prepared by the dimers with the corresponding ancillary ligands. The chloride bridged diiridium complexes can be easily converted to mononuclear Ir(III) complexes by replacing the two bridging chlorides with bidentate monoanionic ancillary ligands. Among the various types of ancillary ligands, we firstly used picolinic acid N-oxide, including picolinic acid and acetylacetone as an ancillary ligands for Ir(III) complexes. The PhOLEDs also shows reasonably high brightness and good luminance efficiency of 20,000 cd/m2 and 12 cd/A, respectively.

Self-repair of cracks in brittle material systems

NASA Astrophysics Data System (ADS)

Dry, Carolyn M.

2016-04-01

One of the most effective uses for self repair is in material systems that crack because the cracks can allow the repair chemical to flow into the crack damage sites in all three dimensions. In order for the repair chemical to stay in the damage site and flow along to all the crack and repair there must be enough chemical to fill the entire crack. The repair chemical must be designed appropriately for the particular crack size and total volume of cracks. In each of the three examples of self repair in crackable brittle systems, the viscosity and chemical makeup and volume of the repair chemicals used is different for each system. Further the chemical delivery system has to be designed for each application also. Test results from self repair of three brittle systems are discussed. In "Self Repair of Concrete Bridges and Infrastructure" two chemicals were used due to different placements in bridges to repair different types of cracks- surface shrinkage and shear cracks, In "Airplane Wings and Fuselage, in Graphite" the composite has very different properties than the concrete bridges. In the graphite for airplane components the chemical also had to survive the high processing temperatures. In this composite the cracks were so definite and deep and thin that the repair chemical could flow easily and repair in all layers of the composite. In "Ceramic/Composite Demonstrating Self Repair" the self repair system not only repaired the broken ceramic but also rebounded the composite to the ceramic layer

The functional response of bioactive titania-modified three-dimensional Ti-6Al-4V mesh structure toward providing a favorable pathway for intercellular communication and osteoincorporation.

PubMed

Nune, K C; Misra, R D K; Li, S J; Hao, Y L; Zhang, W

2016-10-01

The objective of the study is to fundamentally elucidate the biological response of 3D printed mesh structures subjected to plasma electrolytic oxidation process through the study of osteoblast functions. The cellular activity of plasma electrolytic-oxidized mesh structure was explored in terms of cell-to-cell communication involving proliferation, synthesis of extracellular and intracellular proteins, and mineralization. Upon plasma electrolytic oxidation of the mesh structure, a thin layer of bioactive titania with pore size 1-3 µm was nucleated on the surface. The combination of microporous bioactive titania and interconnected porous architecture provided the desired pathway for supply of nutrients and oxygen to cells and tissue and a favorable osteogenic microenvironment for tissue on-growth and in-growth, in relation to the unmodified mesh structure. The formation of a confluent layer as envisaged via electron microscopy and quantitative assessment of the expression level of proteins (actin, vinculin, and fibronectin) point toward the determining role of surface-modified mesh structure in modulating osteoblasts functions. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 2488-2501, 2016. © 2016 Wiley Periodicals, Inc.

Laser direct writing of combinatorial libraries of idealized cellular constructs: Biomedical applications

NASA Astrophysics Data System (ADS)

Schiele, Nathan R.; Koppes, Ryan A.; Corr, David T.; Ellison, Karen S.; Thompson, Deanna M.; Ligon, Lee A.; Lippert, Thomas K. M.; Chrisey, Douglas B.

2009-03-01

The ability to control cell placement and to produce idealized cellular constructs is essential for understanding and controlling intercellular processes and ultimately for producing engineered tissue replacements. We have utilized a novel intra-cavity variable aperture excimer laser operated at 193 nm to reproducibly direct write mammalian cells with micrometer resolution to form a combinatorial array of idealized cellular constructs. We deposited patterns of human dermal fibroblasts, mouse myoblasts, rat neural stem cells, human breast cancer cells, and bovine pulmonary artery endothelial cells to study aspects of collagen network formation, breast cancer progression, and neural stem cell proliferation, respectively. Mammalian cells were deposited by matrix assisted pulsed laser evaporation direct write from ribbons comprised of a UV transparent quartz coated with either a thin layer of extracellular matrix or triazene as a dynamic release layer using CAD/CAM control. We demonstrate that through optical imaging and incorporation of a machine vision algorithm, specific cells on the ribbon can be laser deposited in spatial coherence with respect to geometrical arrays and existing cells on the receiving substrate. Having the ability to direct write cells into idealized cellular constructs can help to answer many biomedical questions and advance tissue engineering and cancer research.

Direct observation of current-induced conductive path in colossal-electroresistance manganite thin films

NASA Astrophysics Data System (ADS)

Wei, Wengang; Zhu, Yinyan; Bai, Yu; Liu, Hao; Du, Kai; Zhang, Kai; Kou, Yunfang; Shao, Jian; Wang, Wenbin; Hou, Denglu; Dong, Shuai; Yin, Lifeng; Shen, Jian

2016-01-01

Manganites are known to often show colossal electroresistance (CER) in addition to colossal magnetoresistance. The (La1-yP ry) 1 -xC axMn O3 (LPCMO) system has a peculiar CER behavior in that little change of magnetization occurs. We use a magnetic force microscope to uncover the CER mechanism in the LPCMO system. In contrast to the previous belief that current reshapes the ferromagnetic metallic (FMM) domains, we show that the shape of the FMM domains remain virtually unchanged after passing electric current. Instead, it is the appearance of a tiny fraction of FMM "bridges" that is responsible for the CER behavior.

Temperature Regulation of Shigella Virulence: Identification of Temperature-Regulated Shigella Invasion Genes by the Isolation of inv::lacZ Operon Fusions and the Characterization of the Virulence Gene Regulator virR

DTIC Science & Technology

1991-04-10

Partial nucleotide sequence of viri? clone pAEH122 102 14. Effects of VirR’ activity on Ipa expression 106 15. Sequencing strategy for the 2.3 kb EcoRl...Confluent monolayers of mammalian cells are challenged with virulent organisms and invasion and intercellular spread result in a cytopathic effect ...destruction of the mucosal surface and an inflammatory response ensues which mimics the effects of invasion and intercellular spread in the mucosa of the

Ischemic preconditioning protects against gap junctional uncoupling in cardiac myofibroblasts.

PubMed

Sundset, Rune; Cooper, Marie; Mikalsen, Svein-Ole; Ytrehus, Kirsti

2004-01-01

Ischemic preconditioning increases the heart's tolerance to a subsequent longer ischemic period. The purpose of this study was to investigate the role of gap junction communication in simulated preconditioning in cultured neonatal rat cardiac myofibroblasts. Gap junctional intercellular communication was assessed by Lucifer yellow dye transfer. Preconditioning preserved intercellular coupling after prolonged ischemia. An initial reduction in coupling in response to the preconditioning stimulus was also observed. This may protect neighboring cells from damaging substances produced during subsequent regional ischemia in vivo, and may preserve gap junctional communication required for enhanced functional recovery during subsequent reperfusion.

Ultrastructural changes of the capillaries of the cat iris in experimental neuroparalytic keratitis.

PubMed

Saari, M; Huhtala, A; Johansson, G

1975-01-01

In order to study the morphological basis of the increased permeability of the capillaries of the iris in neuroparalytic keratitis the ophthalmic division of the trigeminal nerve in the cat was denervated using a stereotactic method. The homolateral iris was studied by electron microscopy three days after denervation. Abnormally large pinocytotic vacuoles were observed in the endothelial cells of the iris capillaries and the intercellular junctions of the endothelial cells showed widened inter-cellular space and macula occludens. These ultrastructural changes may explain the protein leakage into the anterior chamber in neuroparalytic keratitis.

Functional Dependence between Septal Protein SepJ from Anabaena sp. Strain PCC 7120 and an Amino Acid ABC-Type Uptake Transporter.

PubMed

Escudero, Leticia; Mariscal, Vicente; Flores, Enrique

2015-08-01

In the diazotrophic filaments of heterocyst-forming cyanobacteria, two different cell types, the CO2-fixing vegetative cells and the N2-fixing heterocysts, exchange nutrients, including some amino acids. In the model organism Anabaena sp. strain PCC 7120, the SepJ protein, composed of periplasmic and integral membrane (permease) sections, is located at the intercellular septa joining adjacent cells in the filament. The unicellular cyanobacterium Synechococcus elongatus strain PCC 7942 bears a gene, Synpcc7942_1024 (here designated dmeA), encoding a permease homologous to the SepJ permease domain. Synechococcus strains lacking dmeA or lacking dmeA and expressing Anabaena sepJ were constructed. The Synechococcus dmeA mutant showed a significant 22 to 32% decrease in the uptake of aspartate, glutamate, and glutamine, a phenotype that could be partially complemented by Anabaena sepJ. Synechococcus mutants of an ATP-binding-cassette (ABC)-type transporter for polar amino acids showed >98% decreased uptake of glutamate irrespective of the presence of dmeA or Anabaena sepJ in the same strain. Thus, Synechococcus DmeA or Anabaena SepJ is needed to observe full (or close to full) activity of the ABC transporter. An Anabaena sepJ deletion mutant was significantly impaired in glutamate and aspartate uptake, which also in this cyanobacterium requires the activity of an ABC-type transporter for polar amino acids. SepJ appears therefore to generally stimulate the activity of cyanobacterial ABC-type transporters for polar amino acids. Conversely, an Anabaena mutant of three ABC-type transporters for amino acids was impaired in the intercellular transfer of 5-carboxyfluorescein, a SepJ-related property. Our results unravel possible functional interactions in transport elements important for diazotrophic growth. Membrane transporters are essential for many aspects of cellular life, from uptake and export of substances in unicellular organisms to intercellular molecular exchange in multicellular organisms. Heterocyst-forming cyanobacteria such as Anabaena represent a unique case of multicellularity, in which two cell types exchange nutrients and regulators. The SepJ protein located at the intercellular septa in the filaments of Anabaena contains a permease domain of the drug/metabolite transporter (DMT) superfamily that somehow contributes to intercellular molecular transfer. In this work, we have found that SepJ stimulates the activity of a polar amino acid uptake transporter of the ATP-binding-cassette (ABC) superfamily, which could itself affect an intercellular transfer activity related to SepJ, thus unraveling possible functional interactions between these different transporters. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Ductile-Phase-Toughened Tungsten for Plasma-Facing Materials

NASA Astrophysics Data System (ADS)

Cunningham, Kevin Hawkins

A variety of processing approaches were employed to fabricate ductile-phase-toughened (DPT) tungsten (W) composites. Mechanical testing and analytical modeling were used to guide composite development. This work provides a basis for further development of W composites to be used in structural divertor components of future fusion reactors. W wire was tested in tension, showing significant ductility and strength. Coatings of copper (Cu) or tungsten carbide (WC) were applied to the W wire via electrodeposition and carburization, respectively. Composites were fabricated using spark plasma sintering (SPS) to consolidate W powders together with each type of coated W wire. DPT behavior, e.g. crack arrest and crack bridging, was not observed in three-point bend testing of the sintered composites. A laminate was fabricated by hot pressing W and Cu foils together with W wires, and subsequently tested in tension. This laminate was bonded via hot pressing to thick W plate as a reinforcing layer, and the composite was tested in three-point bending. Crack arrest was observed along with some fiber pullout, but significant transverse cracking in the W plate confounded further fracture toughness analysis. The fracture toughness of thin W plate was measured in three-point bending. W plates were brazed with Cu foils to form a laminate. Crack arrest and crack bridging were observed in three-point bend tests of the laminate, and fracture resistance curves were successfully calculated for this DPT composite. An analytical model of crack bridging was developed using the basis described by Chao in previous work by the group. The model uses the specimen geometry, matrix properties, and the stress-displacement function of a ductile reinforcement ("bridging law") to calculate the fracture resistance curve (R-curve) and load-displacement curve (P-D curve) for any test specimen geometry. The code was also implemented to estimate the bridging law of an arbitrary composite using R-curve data. Finally, a parametric study was performed to quantitatively determine the necessary mechanical properties of useful toughening reinforcements for a DPT W composite. The analytical model has a broad applicability for any DPT material.

A pore-level scenario for the development of mixed-wettability in oil reservoirs

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kovscek, A.R.; Wong, H.; Radke, C.J.

Understanding the role of thin films in porous media is vital if wettability is to be elucidated at the pore level. The type and thickness of films coating pore walls determines reservoir wettability and whether or not reservoir rock can be altered from its initial state of wettability. Pore shape, especially pore wall curvature, is an important factor in determining wetting-film thicknesses. Yet, pore shape and the physics of thin wetting films are generally neglected in models of flow in porous rocks. This paper incorporates thin-film forces into a collection of star-shaped capillary tubes model to describe the geological developmentmore » of mixed-wettability in reservoir rock. Here, mixed-wettability refers to continuous and distinct oil and water-wetting surfaces coexisting in the porous medium. The proposed model emphasizes the remarkable role of thin films. New pore-level fluid configurations arise that are quite unexpected. For example, efficient water displacement of oil (i.e, low residual oil saturation) characteristic of mixed-wettability porous media is ascribed to interconnected oil lenses or rivulets which bridge the walls adjacent to a pore corner. Predicted residual oil saturations are approximately 35 % less in mixed-wet rock compared to completely water-wet rock. Calculated capillary pressure curves mimic those of mixed-wet porous media in the primary drainage of water, imbibition of water, and secondary drainage modes. Amott-Harvey indices range from {minus}0.18 to 0.36 also in good agreement with experimental values. (Morrow et al, 1986; Judhunandan and Morrow, 1991).« less

Failure of physiologic transformation of spiral arteries, endothelial and trophoblast cell activation, and acute atherosis in the basal plate of the placenta.

PubMed

Labarrere, Carlos A; DiCarlo, Hector L; Bammerlin, Elaine; Hardin, James W; Kim, Yeon M; Chaemsaithong, Piya; Haas, David M; Kassab, Ghassan S; Romero, Roberto

2017-03-01

Failure of physiologic transformation of spiral arteries has been reported in preeclampsia, fetal growth restriction, fetal death, and spontaneous preterm labor with intact or ruptured membranes. Spiral arteries with failure of physiologic transformation are prone to develop atherosclerotic-like lesions of atherosis. There are striking parallels between preeclampsia and atherosclerotic disease, and between lesions of atherosis and atherosclerosis. Endothelial activation, identified by intercellular adhesion molecule-1 expression, is present in atherosclerotic-like lesions of heart transplantation, and is considered a manifestation of rejection. Similarly, endothelial activation/dysfunction has been implicated in the pathophysiology of atherosclerosis and preeclampsia. Intercellular adhesion molecule-1-overexpressing-activated endothelial cells are more resistant to trophoblast displacement than nonactivated endothelium, and may contribute to shallow spiral artery trophoblastic invasion in obstetrical syndromes having failure of physiologic transformation. We sought to determine whether failure of spiral artery physiologic transformation was associated with activation of interstitial extravillous trophoblasts and/or spiral artery endothelium and presence of acute atherosis in the placental basal plate. A cross-sectional study of 123 placentas (19-42 weeks' gestation) obtained from normal pregnancies (n = 22), preterm prelabor rupture of membranes (n = 26), preterm labor (n = 23), preeclampsia (n = 27), intrauterine fetal death (n = 15), and small for gestational age (n = 10) was performed. Failure of spiral artery physiologic transformation and presence of cell activation was determined using immunohistochemistry of placental basal plates containing a median of 4 (minimum: 1; maximum: 9) vessels per placenta. Endothelial/trophoblast cell activation was defined by the expression of intercellular adhesion molecule-1. Investigators examining microscopic sections were blinded to clinical diagnosis. Pairwise comparisons among placenta groups were performed with Fisher exact test and Wilcoxon rank sum test using a Bonferroni-adjusted level of significance (.025). We found that 87% (94/108) of placentas having spiral arteries with failure of physiologic transformation (actin-positive and cytokeratin-negative) in the basal plate, and 0% (0/15) of placentas having only spiral arteries with complete physiologic transformation (cytokeratin-positive and actin-negative), had arterial endothelial and/or interstitial extravillous trophoblasts reactive with the intercellular adhesion molecule-1 activation marker (P < .001). A significant correlation (R 2  = 0.84) was found between expression of spiral artery endothelial and interstitial extravillous trophoblast intercellular adhesion molecule-1 (P < .001) in activated placentas. Lesions of atherosis were found in 31.9% (30/94) of placentas with complete and/or partial failure of physiologic transformation of spiral arteries that were intercellular adhesion molecule-1-positive, in none of the 14 placentas with failure of physiologic transformation that were intercellular adhesion molecule-1-negative, and in none of the 15 placentas with complete spiral artery physiologic transformation without failure (P = .001). All placentas (30/30, 100%) with atherosis were identified in placentas having concomitant spiral artery endothelial and interstitial extravillous trophoblast activation. Failure of spiral artery physiologic transformation in the placental basal plate is associated with interstitial extravillous trophoblast and arterial endothelial activation along with increased frequency of spiral artery atherosis. These findings may be used to improve the characterization of different disorders of the placental bed such as in refining the existing tools for the early prediction of risk for preterm, preeclamptic, and other abnormal pregnancies. Copyright © 2016 Elsevier Inc. All rights reserved.

Effective growth-suppressive activity of maternal embryonic leucine-zipper kinase (MELK) inhibitor against small cell lung cancer.

PubMed

Inoue, Hiroyuki; Kato, Taigo; Olugbile, Sope; Tamura, Kenji; Chung, Suyoun; Miyamoto, Takashi; Matsuo, Yo; Salgia, Ravi; Nakamura, Yusuke; Park, Jae-Hyun

2016-03-22

Maternal embryonic leucine zipper kinase (MELK), that plays a critical role in maintenance of cancer stem cells (CSCs), is predominantly expressed in various types of human cancer including small cell lung cancer (SCLC). SCLC usually acquires resistance to anti-cancer drugs and portends dismal prognosis. We have delineated roles of MELK in development/progression of SCLC and examined anti-tumor efficacy of OTS167, a highly potent MELK inhibitor, against SCLC. MELK expression was highly upregulated in both SCLC cell lines and primary tumors. siRNA-mediated MELK knockdown induced significant growth inhibition in SCLC cell lines. Concordantly, treatment with OTS167 exhibited strong cytotoxicity against eleven SCLC cell lines with IC50 of < 10 nM. As similar to siRNA knockdown, OTS167 treatment induced cytokinetic defects with intercellular bridges, and in some cell lines we observed formation of neuronal protrusions accompanied with increase of a neuronal differentiation marker (CD56), indicating that the compound induced differentiation of cancer cells to neuron-like cells. Furthermore, the MELK inhibition decreased its downstream FOXM1 activity and Akt expression in SCLC cells, and led to apoptotic cell death. OTS167 appeared to be more effective to CSCs as measured by the sphere formation assay, thus MELK inhibition might become a promising treatment modality for SCLC.

TOPK (T-LAK cell-originated protein kinase) inhibitor exhibits growth suppressive effect on small cell lung cancer.

PubMed

Park, Jae-Hyun; Inoue, Hiroyuki; Kato, Taigo; Zewde, Makda; Miyamoto, Takashi; Matsuo, Yo; Salgia, Ravi; Nakamura, Yusuke

2017-03-01

T-lymphokine-activated killer cell-originated protein kinase (TOPK) plays critical roles in cancer cell proliferation as well as maintenance of cancer stem cells (CSC). Small cell lung cancer (SCLC) has highly aggressive phenotype, reveals early spread to distant sites, and results in dismal prognosis with little effective treatment. In this study, we demonstrate that TOPK expression was highly upregulated in both SCLC cell lines and primary tumors. Similar to siRNA-mediated TOPK knockdown effects, treatment with a potent TOPK inhibitor, OTS514, effectively suppressed growth of SCLC cell lines (IC 50 ; 0.4-42.6 nM) and led to their apoptotic cell death. TOPK inhibition caused cell morphologic changes in SCLC cells, elongation of intercellular bridges caused by cytokinesis defects or neuronal protrusions induced by neuronal differentiation in a subset of CSC-like SCLC cells. Treatment with OTS514 suppressed forkhead box protein M1 (FOXM1) activity, which was involved in stemness of CSC. Furthermore, OTS514 treatment reduced CD90-positive SCLC cells and showed higher cytotoxic effect against lung sphere-derived CSC-like SCLC cells. Collectively, our results suggest that targeting TOPK is a promising approach for SCLC therapy. © 2017 The Authors. Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.

The structure of plasmodesmata as revealed by plasmolysis, detergent extraction, and protease digestion

PubMed Central

1991-01-01

Plasmodesmata or intercellular bridges that connect plant cells are cylindrical channels approximately 40 nm in diameter. Running through the center of each is a dense rod, the desmotubule, that is connected to the endoplasmic reticulum of adjacent cells. Fern, Onoclea sensibilis, gametophytes were cut in half and the cut surfaces exposed to the detergent, Triton X 100, then fixed. Although the plasma membrane limiting the plasmodesma is solubilized partially or completely, the desmotubule remains intact. Alternatively, if the cut surface is exposed to papain, then fixed, the desmotubule disappears, but the plasma membrane limiting the plasmodesmata remains intact albeit swollen and irregular in profile. Gametophytes were plasmolyzed, and then fixed. As the cells retract from their cell walls they leave behind the plasmodesmata still inserted in the cell wall. They can break cleanly when the cell proper retracts or can pull away portions of the plasma membrane of the cell with them. Where the desmotubule remains intact, the plasmodesma retains its shape. These images and the results with detergents and proteases indicate that the desmotubule provides a cytoskeletal element for each plasmodesma, an element that not only stabilizes the whole structure, but also limits its size and porosity. It is likely to be composed in large part of protein. Suggestions are made as to why this structure has been selected for in evolution. PMID:1993740

Effective growth-suppressive activity of maternal embryonic leucine-zipper kinase (MELK) inhibitor against small cell lung cancer

PubMed Central

Inoue, Hiroyuki; Kato, Taigo; Olugbile, Sope; Tamura, Kenji; Chung, Suyoun; Miyamoto, Takashi; Matsuo, Yo; Salgia, Ravi; Nakamura, Yusuke; Park, Jae-Hyun

2016-01-01

Maternal embryonic leucine zipper kinase (MELK), that plays a critical role in maintenance of cancer stem cells (CSCs), is predominantly expressed in various types of human cancer including small cell lung cancer (SCLC). SCLC usually acquires resistance to anti-cancer drugs and portends dismal prognosis. We have delineated roles of MELK in development/progression of SCLC and examined anti-tumor efficacy of OTS167, a highly potent MELK inhibitor, against SCLC. MELK expression was highly upregulated in both SCLC cell lines and primary tumors. siRNA-mediated MELK knockdown induced significant growth inhibition in SCLC cell lines. Concordantly, treatment with OTS167 exhibited strong cytotoxicity against eleven SCLC cell lines with IC50 of < 10 nM. As similar to siRNA knockdown, OTS167 treatment induced cytokinetic defects with intercellular bridges, and in some cell lines we observed formation of neuronal protrusions accompanied with increase of a neuronal differentiation marker (CD56), indicating that the compound induced differentiation of cancer cells to neuron-like cells. Furthermore, the MELK inhibition decreased its downstream FOXM1 activity and Akt expression in SCLC cells, and led to apoptotic cell death. OTS167 appeared to be more effective to CSCs as measured by the sphere formation assay, thus MELK inhibition might become a promising treatment modality for SCLC. PMID:26871945

Chemotherapeutic Drug Based Metal-Organic Particles for Microvesicle-Mediated Deep Penetration and Programmable pH/NIR/Hypoxia Activated Cancer Photochemotherapy.

PubMed

Zhang, Da; Wu, Ming; Cai, Zhixiong; Liao, Naishun; Ke, Kun; Liu, Hongzhi; Li, Ming; Liu, Gang; Yang, Huanghao; Liu, Xiaolong; Liu, Jingfeng

2018-02-01

A novel metal-organic particle (MOP) based nanodrug formed by mild self-assembly of chemotherapeutic drugs, including banoxantrone and doxorubicin, through Cu(II)-mediated coordination effects, is reported. In this nanodrug, Cu(II) acts as a bridge to join AQ4N and DOX, and then, self-assembly of [-AQ4N-Cu(II)-(DOX) 2 -Cu(II)-] n complexes forms nanosized MOPs (referred to as ADMOPs) through multiple interactions including host-metal-guest coordination, hydrophobic interactions, π-stacking, and van der Waals force. The ADMOPs reported here have several important features over conventional drugs, including tumor microenvironment pH-sensitive drug release that can be tracked by "turning on" the fluorescence of AQ4N or DOX through proton competition with Cu(II) to break the coordination bonds and much deeper penetration into solid tumors via microvesicle-mediated intercellular transfer. Most strikingly, the ADMOPs can serve as stimuli-responsive nanocarriers to efficiently load the photosensitizer phthalocyanine due to their inherent highly porous characteristics. Thus, the ADMOPs significantly enhance the chemotherapeutic efficacy by "on-demand" photodynamic therapy, which further induces a hypoxic environment that enhances the reduction of AQ4N to systematically increase the therapeutic efficiency. Taken together, the designed ADMOPs composed of chemotherapeutic drugs may serve as a potential programmable controlled synergistic agent for cancer therapy.

Protein Phosphatase 1ß Limits Ring Canal Constriction during Drosophila Germline Cyst Formation

PubMed Central

Yamamoto, Shinya; Bayat, Vafa; Bellen, Hugo J.; Tan, Change

2013-01-01

Germline cyst formation is essential for the propagation of many organisms including humans and flies. The cytoplasm of germline cyst cells communicate with each other directly via large intercellular bridges called ring canals. Ring canals are often derived from arrested contractile rings during incomplete cytokinesis. However how ring canal formation, maintenance and growth are regulated remains unclear. To better understand this process, we carried out an unbiased genetic screen in Drosophila melanogaster germ cells and identified multiple alleles of flapwing (flw), a conserved serine/threonine-specific protein phosphatase. Flw had previously been reported to be unnecessary for early D. melanogaster oogenesis using a hypomorphic allele. We found that loss of Flw leads to over-constricted nascent ring canals and subsequently tiny mature ring canals, through which cytoplasmic transfer from nurse cells to the oocyte is impaired, resulting in small, non-functional eggs. Flw is expressed in germ cells undergoing incomplete cytokinesis, completely colocalized with the Drosophila myosin binding subunit of myosin phosphatase (DMYPT). This colocalization, together with genetic interaction studies, suggests that Flw functions together with DMYPT to negatively regulate myosin activity during ring canal formation. The identification of two subunits of the tripartite myosin phosphatase as the first two main players required for ring canal constriction indicates that tight regulation of myosin activity is essential for germline cyst formation and reproduction in D. melanogaster and probably other species as well. PMID:23936219

Myosin IXa Regulates Epithelial Differentiation and Its Deficiency Results in Hydrocephalus

PubMed Central

Abouhamed, Marouan; Grobe, Kay; Leefa Chong San, Isabelle V.; Thelen, Sabine; Honnert, Ulrike; Balda, Maria S.; Matter, Karl

2009-01-01

The ependymal multiciliated epithelium in the brain restricts the cerebrospinal fluid to the cerebral ventricles and regulates its flow. We report here that mice deficient for myosin IXa (Myo9a), an actin-dependent motor molecule with a Rho GTPase–activating (GAP) domain, develop severe hydrocephalus with stenosis and closure of the ventral caudal 3rd ventricle and the aqueduct. Myo9a is expressed in maturing ependymal epithelial cells, and its absence leads to impaired maturation of ependymal cells. The Myo9a deficiency further resulted in a distorted ependyma due to irregular epithelial cell morphology and altered organization of intercellular junctions. Ependymal cells occasionally delaminated, forming multilayered structures that bridged the CSF-filled ventricular space. Hydrocephalus formation could be significantly attenuated by the inhibition of the Rho-effector Rho-kinase (ROCK). Administration of ROCK-inhibitor restored maturation of ependymal cells, but not the morphological distortions of the ependyma. Similarly, down-regulation of Myo9a by siRNA in Caco-2 adenocarcinoma cells increased Rho-signaling and induced alterations in differentiation, cell morphology, junction assembly, junctional signaling, and gene expression. Our results demonstrate that Myo9a is a critical regulator of Rho-dependent and -independent signaling mechanisms that guide epithelial differentiation. Moreover, Rho-kinases may represent a new target for therapeutic intervention in some forms of hydrocephalus. PMID:19828736

Focus on the emerging new fields of network physiology and network medicine

NASA Astrophysics Data System (ADS)

Ivanov, Plamen Ch; Liu, Kang K. L.; Bartsch, Ronny P.

2016-10-01

Despite the vast progress and achievements in systems biology and integrative physiology in the last decades, there is still a significant gap in understanding the mechanisms through which (i) genomic, proteomic and metabolic factors and signaling pathways impact vertical processes across cells, tissues and organs leading to the expression of different disease phenotypes and influence the functional and clinical associations between diseases, and (ii) how diverse physiological systems and organs coordinate their functions over a broad range of space and time scales and horizontally integrate to generate distinct physiologic states at the organism level. Two emerging fields, network medicine and network physiology, aim to address these fundamental questions. Novel concepts and approaches derived from recent advances in network theory, coupled dynamical systems, statistical and computational physics show promise to provide new insights into the complexity of physiological structure and function in health and disease, bridging the genetic and sub-cellular level with inter-cellular interactions and communications among integrated organ systems and sub-systems. These advances form first building blocks in the methodological formalism and theoretical framework necessary to address fundamental problems and challenges in physiology and medicine. This ‘focus on’ issue contains 26 articles representing state-of-the-art contributions covering diverse systems from the sub-cellular to the organism level where physicists have key role in laying the foundations of these new fields.

Increased soluble vascular cell adhesion molecule-1 plasma levels and soluble intercellular adhesion molecule-1 during antiretroviral therapy interruption and retention of elevated soluble vascular cellular adhesion molecule-1 levels following resumption of antiretroviral therapy.

PubMed

Papasavvas, Emmanouil; Azzoni, Livio; Pistilli, Maxwell; Hancock, Aidan; Reynolds, Griffin; Gallo, Cecile; Ondercin, Joe; Kostman, Jay R; Mounzer, Karam; Shull, Jane; Montaner, Luis J

2008-06-19

We investigated the effect of short viremic episodes on soluble markers associated with endothelial stress and cardiovascular disease risk in chronically HIV-1-infected patients followed during continuous antiretroviral therapy, antiretroviral therapy interruption and antiretroviral therapy resumption. We assessed changes in plasma levels of von Willebrand factor, soluble vascular cell adhesion molecule-1 and intercellular adhesion molecule-1 by enzyme-linked immunosorbent assay, as well as T-cell activation (CD8+/CD38+, CD8+/HLA-DR+ and CD3+/CD95+) by flow cytometry, in 36 chronically HIV-1-infected patients participating in a randomized study. Patients were divided into the following three groups: a, on continuous antiretroviral therapy; b, on a 6-week antiretroviral therapy interruption; or c, on antiretroviral therapy interruption extended to the achievement of viral set point. Although all measurements remained stable over a 40-week follow-up on antiretroviral therapy, plasma levels of soluble vascular cell adhesion molecule-1 (P < 0.0001) and soluble intercellular adhesion molecule-1 (P = 0.003) increased during treatment interruption in correlation with viral rebound and T-cell activation. No significant changes in von Willebrand factor were observed in any of the groups. After resuming antiretroviral therapy, soluble vascular cell adhesion molecule-1 levels remained elevated even after achievement of viral suppression to less than 50 copies/ml. The prompt rise in plasma soluble vascular cell adhesion molecule-1 and soluble intercellular adhesion molecule-1 upon viral rebound suggests an acute increase in endothelial stress upon treatment interruption, which may persists after viral resuppression of virus. Thus, viral replication during short-term treatment interruption may increase the overall cardiovascular risk during and beyond treatment interruption.

Microtubules and epithem-cell morphogenesis in hydathodes of Pilea cadierei.

PubMed

Galatis, B

1988-12-01

When cell divisions have ceased, the epithem of the hydathodes of Pilea cadierei Gagnep. et Guill. consists of small polyhedral cells exhibiting a meristematic appearance, and completely lacks intercellular spaces. The cortical microtubules in epithem cells exhibit a unique organization: they are not scattered along the whole wall surface but form groups lying at some distance from each other. In sections, from two to eight groups of microtubules can be observed, each lining a wall region averaging between 0.5 and 1.5 μm in length. These groups represent sections of microtubule bundles girdling a major part or the whole of the cell periphery. They are connected to one another by anastomoses, forming a microtubular reticulum. The assembly of microtubule bundles is followed by the appearance of distinct local thickenings in the adjacent wall areas. The cellulose microfibrils in the thickenings are deposited in parallel to the underlying microtubules. Gradually, the vacuolating epithem cells undergo swelling, except for the areas bounded by the wall thickenings. Since the latter, and actually their constituent bundles of cellulose microfibrils, cannot extend in length the differential cell growth results in schizogenous formation of intercellular spaces between contiguous cell walls at their thickened regions. The spaces then broaden and merge to become an extensive intercellular space system. As a result of the above processes, the epithem cells become constricted and finally deeply lobed. The observations show that (i) the cortical microtubules are intimately involved in the morphogenesis of the epithem cells and (ii) the initiation and development of the epithem intercellular spaces is a phenomenon directly related to cell morphogenesis and therefore to the cortical microtubule cytoskeleton. The sites of initiation of these spaces are highly predictable.

Applying 3D-FRAP microscopy to analyse gap junction-dependent shuttling of small antisense RNAs between cardiomyocytes.

PubMed

Lemcke, Heiko; Peukert, Janine; Voronina, Natalia; Skorska, Anna; Steinhoff, Gustav; David, Robert

2016-09-01

Small antisense RNAs like miRNA and siRNA are of crucial importance in cardiac physiology, pathology and, moreover, can be applied as therapeutic agents for the treatment of cardiovascular diseases. Identification of novel strategies for miRNA/siRNA therapy requires a comprehensive understanding of the underlying mechanisms. Emerging data suggest that small RNAs are transferred between cells via gap junctions and provoke gene regulatory effects in the recipient cell. To elucidate the role of miRNA/siRNA as signalling molecules, suitable tools are required that will allow the analysis of these small RNAs at the cellular level. In the present study, we applied 3 dimensional fluorescence recovery after photo bleaching microscopy (3D-FRAP) to visualise and quantify the gap junctional exchange of small RNAs between neonatal cardiomyocytes in real time. Cardiomyocytes were transfected with labelled miRNA and subjected to FRAP microscopy. Interestingly, we observed recovery rates of 21% already after 13min, indicating strong intercellular shuttling of miRNA, which was significantly reduced when connexin43 was knocked down. Flow cytometry analysis confirmed our FRAP results. Furthermore, using an EGFP/siRNA reporter construct we demonstrated that the intercellular transfer does not affect proper functioning of small RNAs, leading to marker gene silencing in the recipient cell. Our results show that 3D-FRAP microscopy is a straightforward, non-invasive live cell imaging technique to evaluate the GJ-dependent shuttling of small RNAs with high spatio-temporal resolution. Moreover, the data obtained by 3D-FRAP confirm a novel pathway of intercellular gene regulation where small RNAs act as signalling molecules within the intercellular network. Copyright © 2016 Elsevier Ltd. All rights reserved.

Regulation Involved in Colonization of Intercellular Spaces of Host Plants in Ralstonia solanacearum

PubMed Central

Hikichi, Yasufumi; Mori, Yuka; Ishikawa, Shiho; Hayashi, Kazusa; Ohnishi, Kouhei; Kiba, Akinori; Kai, Kenji

2017-01-01

A soil-borne bacterium Ralstonia solanacearum invading plant roots first colonizes the intercellular spaces of the root, and eventually enters xylem vessels, where it replicates at high levels leading to wilting symptoms. After invasion into intercellular spaces, R. solanacearum strain OE1-1 attaches to host cells and expression of the hrp genes encoding components of the type III secretion system (T3SS). OE1-1 then constructs T3SS and secrets effectors into host cells, inducing expression of the host gene encoding phosphatidic acid phosphatase. This leads to suppressing plant innate immunity. Then, OE1-1 grows on host cells, inducing quorum sensing (QS). The QS contributes to regulation of OE1-1 colonization of intercellular spaces including mushroom-type biofilm formation on host cells, leading to its virulence. R. solanacearum strains AW1 and K60 produce methyl 3-hydroxypalmitate (3-OH PAME) as a QS signal. The methyltransferase PhcB synthesizes 3-OH PAME. When 3-OH PAME reaches a threshold level, it increases the ability of the histidine kinase PhcS to phosphorylate the response regulator PhcR. This results in elevated levels of functional PhcA, the global virulence regulator. On the other hand, strains OE1-1 and GMI1000 produce methyl 3-hydroxymyristate (3-OH MAME) as a QS signal. Among R. solanacearum strains, the deduced PhcB and PhcS amino acid sequences are related to the production of QS signals. R. solanacearum produces aryl-furanone secondary metabolites, ralfuranones, which are extracellularly secreted and required for its virulence, dependent on the QS. Interestingly, ralfuranones affect the QS feedback loop. Taken together, integrated signaling via ralfuranones influences the QS, contributing to pathogen virulence. PMID:28642776

Quantitative measurements of intercellular adhesion between a macrophage and cancer cells using a cup-attached AFM chip.

PubMed

Kim, Hyonchol; Yamagishi, Ayana; Imaizumi, Miku; Onomura, Yui; Nagasaki, Akira; Miyagi, Yohei; Okada, Tomoko; Nakamura, Chikashi

2017-07-01

Intercellular adhesion between a macrophage and cancer cells was quantitatively measured using atomic force microscopy (AFM). Cup-shaped metal hemispheres were fabricated using polystyrene particles as a template, and a cup was attached to the apex of the AFM cantilever. The cup-attached AFM chip (cup-chip) approached a murine macrophage cell (J774.2), the cell was captured on the inner concave of the cup, and picked up by withdrawing the cup-chip from the substrate. The cell-attached chip was advanced towards a murine breast cancer cell (FP10SC2), and intercellular adhesion between the two cells was quantitatively measured. To compare cell adhesion strength, the work required to separate two adhered cells (separation work) was used as a parameter. Separation work was almost 2-fold larger between a J774.2 cell and FP10SC2 cell than between J774.2 cell and three additional different cancer cells (4T1E, MAT-LyLu, and U-2OS), two FP10SC2 cells, or two J774.2 cells. FP10SC2 was established from 4T1E as a highly metastatic cell line, indicates separation work increased as the malignancy of cancer cells became higher. One possible explanation of the strong adhesion of macrophages to cancer cells observed in this study is that the measurement condition mimicked the microenvironment of tumor-associated macrophages (TAMs) in vivo, and J774.2 cells strongly expressed CD204, which is a marker of TAMs. The results of the present study, which were obtained by measuring cell adhesion strength quantitatively, indicate that the fabricated cup-chip is a useful tool for measuring intercellular adhesion easily and quantitatively. Copyright © 2017 Elsevier B.V. All rights reserved.

Critical role of ATP-induced ATP release for Ca2+ signaling in nonsensory cell networks of the developing cochlea

PubMed Central

Ceriani, Federico; Pozzan, Tullio; Mammano, Fabio

2016-01-01

Spatially and temporally coordinated variations of the cytosolic free calcium concentration ([Ca2+]c) play a crucial role in a variety of tissues. In the developing sensory epithelium of the mammalian cochlea, elevation of extracellular adenosine trisphosphate concentration ([ATP]e) triggers [Ca2+]c oscillations and propagation of intercellular inositol 1,4,5-trisphosphate (IP3)-dependent Ca2+ waves. What remains uncertain is the relative contribution of gap junction channels and connexin hemichannels to these fundamental mechanisms, defects in which impair hearing acquisition. Another related open question is whether [Ca2+]c oscillations require oscillations of the cytosolic IP3 concentration ([IP3]c) in this system. To address these issues, we performed Ca2+ imaging experiments in the lesser epithelial ridge of the mouse cochlea around postnatal day 5 and constructed a computational model in quantitative adherence to experimental data. Our results indicate that [Ca2+]c oscillations are governed by Hopf-type bifurcations within the experimental range of [ATP]e and do not require [IP3]c oscillations. The model replicates accurately the spatial extent and propagation speed of intercellular Ca2+ waves and predicts that ATP-induced ATP release is the primary mechanism underlying intercellular propagation of Ca2+ signals. The model also uncovers a discontinuous transition from propagating regimes (intercellular Ca2+ wave speed > 11 μm⋅s−1) to propagation failure (speed = 0), which occurs upon lowering the maximal ATP release rate below a minimal threshold value. The approach presented here overcomes major limitations due to lack of specific connexin channel inhibitors and can be extended to other coupled cellular systems. PMID:27807138

Inspirations on Virus Replication and Cell-to-Cell Movement from Studies Examining the Cytopathology Induced by Lettuce infectious yellows virus in Plant Cells

PubMed Central

Qiao, Wenjie; Medina, Vicente; Falk, Bryce W.

2017-01-01

Lettuce infectious yellows virus (LIYV) is the type member of the genus Crinivirus in the family Closteroviridae. Like many other positive-strand RNA viruses, LIYV infections induce a number of cytopathic changes in plant cells, of which the two most characteristic are: Beet yellows virus-type inclusion bodies composed of vesicles derived from cytoplasmic membranes; and conical plasmalemma deposits (PLDs) located at the plasmalemma over plasmodesmata pit fields. The former are not only found in various closterovirus infections, but similar structures are known as ‘viral factories’ or viroplasms in cells infected with diverse types of animal and plant viruses. These are generally sites of virus replication, virion assembly and in some cases are involved in cell-to-cell transport. By contrast, PLDs induced by the LIYV-encoded P26 non-virion protein are not involved in replication but are speculated to have roles in virus intercellular movement. These deposits often harbor LIYV virions arranged to be perpendicular to the plasma membrane over plasmodesmata, and our recent studies show that P26 is required for LIYV systemic plant infection. The functional mechanism of how LIYV P26 facilitates intercellular movement remains unclear, however, research on other plant viruses provides some insights on the possible ways of viral intercellular movement through targeting and modifying plasmodesmata via interactions between plant cellular components and viral-encoded factors. In summary, beginning with LIYV, we review the studies that have uncovered the biological determinants giving rise to these cytopathological effects and their importance in viral replication, virion assembly and intercellular movement during the plant infection by closteroviruses, and compare these findings with those for other positive-strand RNA viruses. PMID:29021801

Band-gap narrowing and magnetic behavior of Ni-doped Ba(Ti0.875Ce0.125)O3 thin films

NASA Astrophysics Data System (ADS)

Zhou, Wenliang; Deng, Hongmei; Yu, Lu; Yang, Pingxiong; Chu, Junhao

2015-11-01

Band-gap narrowing and magnetic effects have been observed in a Ni-doped Ba(Ti0.875Ce0.125)O3 (BTC) thin film. Structural characterizations and microstructural analysis show that the as-prepared Ba(Ti0.75Ce0.125Ni0.125)O3-δ (BTCN) thin film exhibits a cubic perovskite structure with an average grain size of 25 nm. The Ce doping at the Ti-site results in an increasing perovskite volume to favour an O-vacancy-stabilized Ni2+ substitution. Raman spectroscopy, however, shows the cubic symmetry of crystalline structures is locally lowered by the presence of dopants, significantly deviating from the ideal Pm3m space group. Moreover, BTCN presents a narrowed band-gap, much smaller than that of BaTiO3 and BTC, due to new states of both the highest occupied molecular orbital and the lowest unoccupied molecular orbital in an electronic structure with the presence of Ni. Also, magnetic enhancement driven by co-doping has been confirmed in the films, which mainly stems from the exchange interaction of Ni2+ ions via an electron trapped in a bridging oxygen vacancy. These findings may open an avenue to discover and design optimal perovskite compounds for solar-energy devices and information storage.

Growing Oxide Nanowires and Nanowire Networks by Solid State Contact Diffusion into Solution-Processed Thin Films.

PubMed

Glynn, Colm; McNulty, David; Geaney, Hugh; O'Dwyer, Colm

2016-11-01

New techniques to directly grow metal oxide nanowire networks without the need for initial nanoparticle seed deposition or postsynthesis nanowire casting will bridge the gap between bottom-up formation and top-down processing for many electronic, photonic, energy storage, and conversion technologies. Whether etched top-down, or grown from catalyst nanoparticles bottom-up, nanowire growth relies on heterogeneous material seeds. Converting surface oxide films, ubiquitous in the microelectronics industry, to nanowires and nanowire networks by the incorporation of extra species through interdiffusion can provide an alternative deposition method. It is shown that solution-processed thin films of oxides can be converted and recrystallized into nanowires and networks of nanowires by solid-state interdiffusion of ionic species from a mechanically contacted donor substrate. NaVO 3 nanowire networks on smooth Si/SiO 2 and granular fluorine-doped tin oxide surfaces can be formed by low-temperature annealing of a Na diffusion species-containing donor glass to a solution-processed V 2 O 5 thin film, where recrystallization drives nanowire growth according to the crystal habit of the new oxide phase. This technique illustrates a new method for the direct formation of complex metal oxide nanowires on technologically relevant substrates, from smooth semiconductors, to transparent conducting materials and interdigitated device structures. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Characterization of fully functional spray-on antibody thin films

NASA Astrophysics Data System (ADS)

Figueroa, Jhon; Magaña, Sonia; Lim, Daniel V.; Schlaf, Rudy

2014-02-01

The authors recently demonstrated that fully functional Escherichia coli O157:H7 antibody thin films can be prepared using a simple pneumatic nebulizer on glass surface [1]. This paper focuses on the investigation of the morphology and physical properties of these films with the aim to better understand their performance. A series of E. coli O157:H7 antibody spray-on thin films were investigated by ellipsometry, X-ray photoelectron spectroscopy (XPS), immunoassays, attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), fluorescence microscopy, atomic force microscope (AFM) and contact angle analysis. These data were compared to measurements on films prepared with the biotin-avidin covalent bonding scheme. The investigation showed that films created by a 2 min pneumatic spray deposition time can capture antigens similar as the avidin-biotin wet-chemical method. The results also suggests that an influential factor for the comparable capture cell ability between sprayed and covalent films is an increased antibody surface coverage for the sprayed films (non-equilibrium technique), which compensates for the lack of its antibody orientation. There was no significant antibody denaturation detected on any of the sprayed films. Both techniques led to the formation of cluster-aggregates, a factor that seems unavoidable due to the natural tendency of protein to cluster. The avidin-biotin bridge films generally had a higher roughness, which manifested itself in a higher wettability compared to the sprayed films.

The effects of gamma-ray irradiation on organic materials of different conjugation lengths

NASA Astrophysics Data System (ADS)

Zhang, Cheng; Taylor, Edward W.

2009-08-01

The radiation resistance of organic electro-optic and optoelectronic materials of different conjugation lengths for space applications is receiving increased attention. Earlier investigation reported that guest-host (G-H) poled polymer EO modulator devices composed of a phenyltetraene bridge-type chromophore in amorphous polycarbonate (CLD/APC) did not exhibit a decrease in EO response (i.e., an increase in modulation-switching voltage- Vπ) following irradiation by low dose [10-160 krad(Si)] 60Co gamma-rays. In this work, the post-irradiation responses of 60Co gamma-rays on CLD1/APC thin films are examined by various chemical and spectroscopic methods including: a solubility test, thin-layer chromatography, proton nuclear magnetic resonance spectroscopy, UV-vis absorption, and infra-red absorption. The results indicate that CLD1 and APC did not decompose under gamma-ray irradiation at dose levels ranging from 66-274 krad(Si) and from 61-154 krad(Si), respectively which support the previously reported data. A comparison with an in situ proton irradiated DRI/PMMA material is also presented.

Thin film strain transducer. [in-flight measurement of stress or strain in walls of high altitude balloons

NASA Technical Reports Server (NTRS)

Rand, J. L.

1981-01-01

Previous attempts to develop an appropriate sensor for measuring the stress or strain of high altitude balloons during flight are reviewed as well as the various conditions that must be met by such a device. The design, development and calibration of a transducer which promises to satisfy the necessary design constraints are described. The thin film strain transducer has a low effective modulus so as not to interfere with the strain that would naturally occur in the balloon. In addition, the transducer has a high sensitivity to longitudinal strain (7.216 mV/V/unit strain) which is constant for all temperature from room temperature to -80 C and all strains from 5 percent compression to 10 percent tensile strain. At the same time, the sensor is relatively insensitive (0.27 percent) to transverse forces. The device has a standard 350 ohm impedance which is compatible with available bridge balance, amplification and telemetry instrumentation now available for balloon flight. Recommendations are included for improved coatings to provide passive thermal control as well as model, tethered and full scale flight testing.

Novel fabrication method for 3D microstructures using surface-activated bonding and its application to micro-mechanical parts

NASA Astrophysics Data System (ADS)

Yamada, Takayuki; Takahashi, Mutsuya; Ozawa, Takashi; Tawara, Satoshi; Goto, Takayuki

2002-11-01

The purpose of this work is to demonstrate that a novel fabrication method for 3-D microstructures (FORMULA) is applicable to fabrication of micro mechanical parts with a large flexibility. This method is a kind of layer manufacturing method of thin films for metallic or dielectric microstructures using surface-activated bonding (SAB). The bonding interfaces of thin films are investigated by transmission electron microscope (TEM). Voids were observed at the interfaces of both pure aluminum films and Al-Cu alloy films. The ratio of void on the Al-Cu/Al-Cu interface is much larger than that of Al/Al interface, although the films have the same surface roughness of 3nm in Ra (average roughness). And approximately 10nm-thick amorphous intermediate layers were found at the interfaces. Furthermore, we have fabricated a micro gear of 900μm in diameter and 200μm in height, which is about ten times as large as our previous test pieces. Overhung structures such as a bridge structure and a cantilever were also fabricated without supporting layers beneath them.

Butt-coupled interface between stoichiometric Si3N4 and thin-film plasmonic waveguides

NASA Astrophysics Data System (ADS)

Dabos, G.; Ketzaki, D.; Tsiokos, D.; Pleros, N.

2017-02-01

Plasmonic technology has emerged as the most promising candidate to revolutionize future photonic-integrated-circuits (PICs) and deliver performance breakthroughs in diverse application areas by providing increased light-matter interaction at the nanometer scale, overcoming the diffraction limit. However, high insertion losses of plasmonic devices impede their practical deployment in PICs. To overcome this hurdle, selective integration of individual plasmonic devices on low-loss photonic platforms is considered, allowing for enhanced chip-scale functionalities with realistic power budgets. In this context, highly-efficient and fabrication-tolerant optical interfaces for co-planar plasmonic and photonic waveguides become essential, bridging these two "worlds" and ease combined high-volume manufacturing. Herein, a TM-mode butt-coupled interface for stoichiometric Si3N4 and Au-based thin-film plasmonic waveguides is proposed aiming to be utilized for bio-sensing applications. Following a systematic design process, this new configuration has been analyzed through 3D FDTD numerical simulations demonstrating coupling efficiencies up to 64% at the wavelength of 1.55 μm, with increased fabrication tolerance compared to silicon based waveguide alternatives.

Magnetic phase change in Mn-doped ZnSnAs2 thin films depending on Mn concentration

NASA Astrophysics Data System (ADS)

Uchitomi, Naotaka; Hidaka, Shiro; Saito, Shin; Asubar, Joel T.; Toyota, Hideyuki

2018-04-01

The relationship between Mn concentration and Curie temperature (TC) is studied for Mn-doped ZnSnAs2 ferromagnetic semiconductors, epitaxially grown on InP substrates by molecular beam epitaxy. In the ferromagnetic phase, Mn distributions in a (Zn,Mn,Sn)As2 thin film with 7.2 cation percent (cat. %) Mn are investigated using three-dimensional atom probe tomography. The results indicate an inhomogeneous distribution which spreads to a relatively high Mn concentration of 9.0 at. % (at. %). In the paramagnetic phase, it is found that the paramagnetic to ferromagnetic transition takes place sharply with a TC of 334 K when the Mn doping concentration increases to about 4 cat. % Mn, which corresponds to a magnetic percolation threshold for ferromagnetism in (Zn,Mn,Sn)As2. An effective Curie temperature ⟨TC⟩ is considered to bridge the Curie temperatures obtained experimentally to those calculated theoretically in inhomogeneous magnetic semiconductors. The behavior of magnetism in Mn-doped ZnSnAs2 can be explained by three different phases within the present framework.

A zinc phthalocyanine based periodic mesoporous organosilica exhibiting charge transfer to fullerenes.

PubMed

Auras, Florian; Li, Yan; Löbermann, Florian; Döblinger, Markus; Schuster, Jörg; Peter, Laurence M; Trauner, Dirk; Bein, Thomas

2014-11-10

Periodic mesoporous organosilica (PMO) materials offer a strategy to position molecular semiconductors within a highly defined, porous network. We developed thin films of a new semiconducting zinc phthalocyanine-bridged PMO exhibiting a face-centered orthorhombic pore structure with an average pore diameter of 11 nm. The exceptional degree of order achieved with this PMO enabled us to create thin films consisting of a single porous domain throughout their entire thickness, thus providing maximal accessibility for subsequent incorporation of a complementary phase. The phthalocyanine building blocks inside the pore walls were found to be well-aggregated, enabling electronic conductivity and extending the light-harvesting capabilities to the near IR region. Ordered 3D heterojunctions capable of promoting photo-induced charge transfer were constructed by impregnation of the PMO with a fullerene derivative. When integrated into a photovoltaic device, the infiltrated PMO is capable of producing a high open-circuit voltage and a considerable photocurrent, which represents a significant step towards potential applications of PMOs in optoelectronics. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Transition from reconstruction toward thin film on the (110) surface of strontium titanate

DOE PAGES

Wang, Z.; Loon, A.; Subramanian, A.; ...

2016-03-08

The surfaces of metal oxides often are reconstructed with a geometry and composition that is considerably different from a simple termination of the bulk. Such structures can also be viewed as ultrathin films, epitaxed on a substrate. Here, the reconstructions of the SrTiO 3 (110) surface are studied combining scanning tunneling microscopy (STM), transmission electron diffraction, and X-ray absorption spectroscopy (XAS), and analyzed with density functional theory calculations. Whereas SrTiO 3 (110) invariably terminates with an overlayer of titania, with increasing density its structure switches from n × 1 to 2 × n. At the same time the coordination ofmore » the Ti atoms changes from a network of corner-sharing tetrahedra to a double layer of edge-shared octahedra with bridging units of octahedrally coordinated strontium. Furthermore, this transition from the n × 1 to 2 × n reconstructions is a transition from a pseudomorphically stabilized tetrahedral network toward an octahedral titania thin film with stress-relief from octahedral strontia units at the surface.« less

Nonthermal response of YBa2Cu3O7-δ thin films to picosecond THz pulses

NASA Astrophysics Data System (ADS)

Probst, P.; Semenov, A.; Ries, M.; Hoehl, A.; Rieger, P.; Scheuring, A.; Judin, V.; Wünsch, S.; Il'in, K.; Smale, N.; Mathis, Y.-L.; Müller, R.; Ulm, G.; Wüstefeld, G.; Hübers, H.-W.; Hänisch, J.; Holzapfel, B.; Siegel, M.; Müller, A.-S.

2012-05-01

The photoresponse of YBa2Cu3O7-δ thin film microbridges with thicknesses between 15 and 50 nm was studied in the optical and terahertz frequency range. The voltage transients in response to short radiation pulses were recorded in real time with a resolution of a few tens of picoseconds. The bridges were excited by either femtosecond pulses at a wavelength of 0.8 μm or broadband (0.1-1.5 THz) picosecond pulses of coherent synchrotron radiation. The transients in response to optical radiation are qualitatively well explained in the framework of the two-temperature model with a fast component in the picosecond range and a bolometric nanosecond component whose decay time depends on the film thickness. The transients in the THz regime showed no bolometric component and had amplitudes up to three orders of magnitude larger than the two-temperature model predicts. Additionally THz field-dependent transients in the absence of DC bias were observed. We attribute the response in the THz regime to a rearrangement of vortices caused by high-frequency currents.

Drop formation, pinch-off dynamics and liquid transfer of simple and complex fluids

NASA Astrophysics Data System (ADS)

Dinic, Jelena; Sharma, Vivek

Liquid transfer and drop formation processes underlying jetting, spraying, coating, and printing - inkjet, screen, roller-coating, gravure, nanoimprint hot embossing, 3D - often involve formation of unstable columnar necks. Capillary-driven thinning of such necks and their pinchoff dynamics are determined by a complex interplay of inertial, viscous and capillary stresses for simple, Newtonian fluids. Micro-structural changes in response to extensional flow field that arises within the thinning neck give rise to additional viscoelastic stresses in complex, non- Newtonian fluids. Using FLOW-3D, we simulate flows realized in prototypical geometries (dripping and liquid bridge stretched between two parallel plates) used for studying pinch-off dynamics and influence of microstructure and viscoelasticity. In contrast with often-used 1D or 2D models, FLOW-3D allows a robust evaluation of the magnitude of the underlying stresses and extensional flow field (both uniformity and magnitude). We find that the simulated radius evolution profiles match the pinch-off dynamics that are experimentally-observed and theoretically-predicted for model Newtonian fluids and complex fluids.

Cell Alignment Required in Differentiation of Myxococcus xanthus

NASA Astrophysics Data System (ADS)

Kim, Seung K.; Kaiser, Dale

1990-08-01

During fruiting body morphogenesis of Myxococcus xanthus, cell movement is required for transmission of C-factor, a short range intercellular signaling protein necessary for sporulation and developmental gene expression. Nonmotile cells fail to sporulate and to express C-factor-dependent genes, but both defects were rescued by a simple manipulation of cell position that oriented the cells in aligned, parallel groups. A similar pattern of aligned cells normally results from coordinated recruitment of wild-type cells into multicellular aggregates, which later form mature fruiting bodies. It is proposed that directed cell movement establishes critical contacts between adjacent cells, which are required for efficient intercellular C-factor transmission.

[The biological role of exosomes in bone remodeling and bone diseases.

PubMed

Urabe, Fumihiko; Yoshioka, Yusuke; Ochiya, Takahiro

Exosomes are about 100nm membrane vesicles, and released from almost all cell types. They carry and transfer a wide variety of molecules, such as mRNAs, microRNAs, proteins, and lipids, as modulators of intercellular communication. Various studies have shown that this exosome-mediated intercellular communication lead to proliferation, invasion and metastasis of cancer cells. In addition to that, emerging data suggest that exosomes are also involved in physiological processes of bone remodeling and bone diseases. Increasing understanding of the working mechanism of exosomes will provide us with new therapeutic and diagnostic opportunities. Here we summarize the current research on exosomes in bone remodeling and bone diseases.

Teaching resources. Movement of macromolecules in plant cells through plasmodesmata.

PubMed

Jorgensen, Richard A; Lucas, William J

2006-02-21

Plasmodesmata are intercellular organelles in plants that allow the passage of molecules between plant cells. Movement through plasmodesmata may allow transcription factors expressed in one cell to move into adjacent cells, thereby regulating gene expression non-cell autonomously. The two animations illustrate (i) movement of a protein through an individual plasmodesma and (ii) an experiment to detect the movement of the transcription factor through plasmodesmata from the L1 layer of a plant meristem into the L2 and L3 layers. These two animations would be useful in teaching plant biology or plant development or a cell biology class discussing mechanisms of intercellular transport.

Temperature-dependence of isometric tension and cross-bridge kinetics of cardiac muscle fibers reconstituted with a tropomyosin internal deletion mutant.

PubMed

Lu, Xiaoying; Tobacman, Larry S; Kawai, Masataka

2006-12-01

The effect of temperature on isometric tension and cross-bridge kinetics was studied with a tropomyosin (Tm) internal deletion mutant AS-Delta23Tm (Ala-Ser-Tm Delta(47-123)) in bovine cardiac muscle fibers by using the thin filament extraction and reconstitution technique. The results are compared with those from actin reconstituted alone, cardiac muscle-derived control acetyl-Tm, and recombinant control AS-Tm. In all four reconstituted muscle groups, isometric tension and stiffness increased linearly with temperature in the range 5-40 degrees C for fibers activated in the presence of saturating ATP and Ca(2+). The slopes of the temperature-tension plots of the two controls were very similar, whereas the slope derived from fibers with actin alone had approximately 40% the control value, and the slope from mutant Tm had approximately 36% the control value. Sinusoidal analysis was performed to study the temperature dependence of cross-bridge kinetics. All three exponential processes A, B, and C were identified in the high temperature range (30-40 degrees C); only processes B and C were identified in the mid-temperature range (15-25 degrees C), and only process C was identified in the low temperature range (5-10 degrees C). At a given temperature, similar apparent rate constants (2pia, 2pib, 2pic) were observed in all four muscle groups, whereas their magnitudes were markedly less in the order of AS-Delta23Tm < Actin < AS-Tm approximately Acetyl-Tm groups. Our observations are consistent with the hypothesis that Tm enhances hydrophobic and stereospecific interactions (positive allosteric effect) between actin and myosin, but Delta23Tm decreases these interactions (negative allosteric effect). Our observations further indicate that tension/cross-bridge is increased by Tm, but is diminished by Delta23Tm. We conclude that Tm affects the conformation of actin so as to increase the area of hydrophobic interaction between actin and myosin molecules.

Microdosimetric Modeling of Biological Effectiveness for Boron Neutron Capture Therapy Considering Intra- and Intercellular Heterogeneity in 10B Distribution.

PubMed

Sato, Tatsuhiko; Masunaga, Shin-Ichiro; Kumada, Hiroaki; Hamada, Nobuyuki

2018-01-17

We here propose a new model for estimating the biological effectiveness for boron neutron capture therapy (BNCT) considering intra- and intercellular heterogeneity in 10 B distribution. The new model was developed from our previously established stochastic microdosimetric kinetic model that determines the surviving fraction of cells irradiated with any radiations. In the model, the probability density of the absorbed doses in microscopic scales is the fundamental physical index for characterizing the radiation fields. A new computational method was established to determine the probability density for application to BNCT using the Particle and Heavy Ion Transport code System PHITS. The parameters used in the model were determined from the measured surviving fraction of tumor cells administrated with two kinds of 10 B compounds. The model quantitatively highlighted the indispensable need to consider the synergetic effect and the dose dependence of the biological effectiveness in the estimate of the therapeutic effect of BNCT. The model can predict the biological effectiveness of newly developed 10 B compounds based on their intra- and intercellular distributions, and thus, it can play important roles not only in treatment planning but also in drug discovery research for future BNCT.

Treponema pallidum Invades Intercellular Junctions of Endothelial Cell Monolayers

NASA Astrophysics Data System (ADS)

Thomas, D. Denee; Navab, Mahamad; Haake, David A.; Fogelman, Alan M.; Miller, James N.; Lovett, Michael A.

1988-05-01

The pathogenesis of syphilis reflects invasive properties of Treponema pallidum, but the actual mode of tissue invasion is unknown. We have found two in vitro parallels of treponemal invasiveness. We tested whether motile T. pallidum could invade host cells by determining the fate of radiolabeled motile organisms added to a HeLa cell monolayer; 26% of treponemes associated with the monolayer in a trypsin-resistant niche, presumably between the monolayer and the surface to which it adhered, but did not attain intracellularity. Attachment of T. pallidum to cultured human and rabbit aortic and human umbilical vein endothelial cells was 2-fold greater than to HeLa cells. We added T. pallidum to aortic endothelial cells grown on membrane filters under conditions in which tight intercellular junctions had formed. T. pallidum was able to pass through the endothelial cell monolayers without altering tight junctions, as measured by electrical resistance. In contrast, heat-killed T. pallidum and the nonpathogen Treponema phagedenis biotype Reiter failed to penetrate the monolayer. Transmission electron micrographs of sections of the monolayer showed T. pallidum in intercellular junctions. Our in vitro observations suggest that these highly motile spirochetes may leave the circulation by invading the junctions between endothelial cells.

14-3-3 proteins regulate desmosomal adhesion via plakophilins.

PubMed

Rietscher, Katrin; Keil, René; Jordan, Annemarie; Hatzfeld, Mechthild

2018-05-22

Desmosomes are essential for strong intercellular adhesion and are abundant in tissues exposed to mechanical strain. At the same time, desmosomes need to be dynamic to allow for remodeling of epithelia during differentiation or wound healing. Phosphorylation of desmosomal plaque proteins appears to be essential for desmosome dynamics. However, the mechanisms of how context-dependent post-translational modifications regulate desmosome formation, dynamics or stability are incompletely understood. Here, we show that growth factor signaling regulates the phosphorylation-dependent association of plakophilins 1 and 3 (PKP1 and PKP3) with 14-3-3 protein isoforms, and uncover unique and partially antagonistic functions of members of the 14-3-3 family in the regulation of desmosomes. 14-3-3γ associated primarily with cytoplasmic PKP1 phosphorylated at S155 and destabilized intercellular cohesion of keratinocytes by reducing its incorporation into desmosomes. In contrast, 14-3-3σ (also known as stratifin, encoded by SFN ) interacted preferentially with S285-phosphorylated PKP3 to promote its accumulation at tricellular contact sites, leading to stable desmosomes. Taken together, our study identifies a new layer of regulation of intercellular adhesion by 14-3-3 proteins. © 2018. Published by The Company of Biologists Ltd.

Crystal structure of the Haemophilus influenzae Hap adhesin reveals an intercellular oligomerization mechanism for bacterial aggregation

PubMed Central

Meng, Guoyu; Spahich, Nicole; Kenjale, Roma; Waksman, Gabriel; St Geme, Joseph W

2011-01-01

Bacterial biofilms are complex microbial communities that are common in nature and are being recognized increasingly as an important determinant of bacterial virulence. However, the structural determinants of bacterial aggregation and eventual biofilm formation have been poorly defined. In Gram-negative bacteria, a major subgroup of extracellular proteins called self-associating autotransporters (SAATs) can mediate cell–cell adhesion and facilitate biofilm formation. In this study, we used the Haemophilus influenzae Hap autotransporter as a prototype SAAT to understand how bacteria associate with each other. The crystal structure of the H. influenzae HapS passenger domain (harbouring the SAAT domain) was determined to 2.2 Å by X-ray crystallography, revealing an unprecedented intercellular oligomerization mechanism for cell–cell interaction. The C-terminal SAAT domain folds into a triangular-prism-like structure that can mediate Hap–Hap dimerization and higher degrees of multimerization through its F1–F2 edge and F2 face. The intercellular multimerization can give rise to massive buried surfaces that are required for overcoming the repulsive force between cells, leading to bacterial cell–cell interaction and formation of complex microcolonies. PMID:21841773

Morphological examination of the effects of defibrotide on experimentally induced bladder injury and its relation to interstitial cystitis.

PubMed

Aydin, H; Ercan, F; Cetinel, S; San, T

2001-08-01

This morphological study aims to investigate the effects of defibrotide, a deoxyribonucleic acid derivative drug with cytoprotective, immunosuppressive and vasorelaxant effects, on protamine sulfate induced bladder injury. Wistar albino female rats were catheterized and intravesically infused with phosphate buffered solution (control group) or, either protamine sulfate (bladder injury group) or protamine sulfate+defibrotide (bladder injury+defibrotide group) dissolved in phosphate buffered solution. The morphology of the urinary bladder was investigated using light and electron microscopy. The number of mast cells in the mucosa, mucosal alterations, intercellular junctions, surface topography and the glycosaminoglycan (GAG) layer as well as microvillus formation on the luminal surface were evaluated. In the bladder injury group, ulcerated areas, irregularity of the GAG layer, increased number of mast cells, vacuole formation, dilated perinuclear cistern, formation of pleomorphic and uniform microvilli and dilatations in the intercellular spaces in the urothelium were observed. In the bladder injury+defibrotide group a relatively normal urothelial topography, GAG layer and a few mast cells in the mucosa, some dilatations between the intercellular areas, less uniform microvilli, regular perinuclear cistern and tight junctions were observed. These results show that defibrotide can inhibit PS induced bladder damage.

Multimodal quantitative phase and fluorescence imaging of cell apoptosis

NASA Astrophysics Data System (ADS)

Fu, Xinye; Zuo, Chao; Yan, Hao

2017-06-01

Fluorescence microscopy, utilizing fluorescence labeling, has the capability to observe intercellular changes which transmitted and reflected light microscopy techniques cannot resolve. However, the parts without fluorescence labeling are not imaged. Hence, the processes simultaneously happen in these parts cannot be revealed. Meanwhile, fluorescence imaging is 2D imaging where information in the depth is missing. Therefore the information in labeling parts is also not complete. On the other hand, quantitative phase imaging is capable to image cells in 3D in real time through phase calculation. However, its resolution is limited by the optical diffraction and cannot observe intercellular changes below 200 nanometers. In this work, fluorescence imaging and quantitative phase imaging are combined to build a multimodal imaging system. Such system has the capability to simultaneously observe the detailed intercellular phenomenon and 3D cell morphology. In this study the proposed multimodal imaging system is used to observe the cell behavior in the cell apoptosis. The aim is to highlight the limitations of fluorescence microscopy and to point out the advantages of multimodal quantitative phase and fluorescence imaging. The proposed multimodal quantitative phase imaging could be further applied in cell related biomedical research, such as tumor.

Electrotonic potentials in Aloe vera L.: Effects of intercellular and external electrodes arrangement.

PubMed

Volkov, Alexander G; Nyasani, Eunice K; Tuckett, Clayton; Scott, Jessenia M; Jackson, Mariah M Z; Greeman, Esther A; Greenidge, Ariane S; Cohen, Devin O; Volkova, Maia I; Shtessel, Yuri B

2017-02-01

Electrostimulation of plants can induce plant movements, activation of ion channels, ion transport, gene expression, enzymatic systems activation, electrical signaling, plant-cell damage, enhanced wound healing, and influence plant growth. Here we found that electrical networks in plant tissues have electrical differentiators. The amplitude of electrical responses decreases along a leaf and increases by decreasing the distance between polarizing Pt-electrodes. Intercellular Ag/AgCl electrodes inserted in a leaf and extracellular Ag/AgCl electrodes attached to the leaf surface were used to detect the electrotonic potential propagation along a leaf of Aloe vera. There is a difference in duration and amplitude of electrical potentials measured by electrodes inserted in a leaf and those attached to a leaf's surface. If the external reference electrode is located in the soil near the root, it changes the amplitude and duration of electrotonic potentials due to existence of additional resistance, capacitance, ion channels and ion pumps in the root. The information gained from this study can be used to elucidate extracellular and intercellular communication in the form of electrical signals within plants. Copyright © 2016 Elsevier B.V. All rights reserved.

Intercellular production of tamavidin 1, a biotin-binding protein from Tamogitake mushroom, confers resistance to the blast fungus Magnaporthe oryzae in transgenic rice.

PubMed

Takakura, Yoshimitsu; Oka, Naomi; Suzuki, Junko; Tsukamoto, Hiroshi; Ishida, Yuji

2012-05-01

The blast fungus Magnaporthe oryzae, one of the most devastating rice pathogens in the world, shows biotin-dependent growth. We have developed a strategy for creating disease resistance to M. oryzae whereby intercellular production of tamavidin 1, a biotin-binding protein from Pleurotus cornucopiae occurs in transgenic rice plants. The gene that encodes tamavidin 1, fused to the sequence for a secretion signal peptide derived from rice chitinase gene, was connected to the Cauliflower mosaic virus 35S promoter, and the resultant construct was introduced into rice. The tamavidin 1 was accumulated at levels of 0.1-0.2% of total soluble leaf proteins in the transgenic rice and it was localized in the intercellular space of rice leaves. The tamavidin 1 purified from the transgenic rice was active, it bound to biotin and inhibited in vitro growth of M. oryzae by causing biotin deficiency. The transgenic rice plants showed a significant resistance to M. oryzae. This study shows the possibility of a new strategy to engineer disease resistance in higher plants by taking advantage of a pathogen's auxotrophy.

Exocytosis of vacuolar apical compartment (VAC): a cell-cell contact controlled mechanism for the establishment of the apical plasma membrane domain in epithelial cells

PubMed Central

1988-01-01

The vacuolar apical compartment (VAC) is an organelle found in Madin- Darby canine kidney (MDCK) cells with incomplete intercellular contacts by incubation in 5 microM Ca++ and in cells without contacts (single cells in subconfluent culture); characteristically, it displays apical biochemical markers and microvilli and excludes basolateral markers (Vega-Salas, D. E., P. J. I. Salas, and E. Rodriguez-Boulan. 1987. J. Cell Biol. 104:1249-1259). The apical surface of cells kept under these culture conditions is immature, with reduced numbers of microvilli and decreased levels of an apical biochemical marker (184 kD), which is, however, still highly polarized (Vega-Salas, D. E., P. J. I. Salas, D. Gundersen, and E. Rodriguez-Boulan. 1987. J. Cell Biol. 104:905-916). We describe here the morphological stages of VAC exocytosis which ultimately lead to the establishment of a differentiated apical domain. Addition of 1.8 mM Ca++ to monolayers developed in 5 microM Ca++ causes the rapid (20-40 min) fusion of VACs with the plasma membrane and their accessibility to external antibodies, as demonstrated by immunofluorescence, immunoperoxidase EM, and RIA with antibodies against the 184-kD apical plasma membrane marker. Exocytosis occurs towards areas of cell-cell contact in the developing lateral surface where they form intercellular pockets; fusion images are always observed immediately adjacent to the incomplete junctional bands detected by the ZO-1 antibody (Stevenson, B. R., J. D. Siliciano, M. S. Mooseker, and D. A. Goodenough. 1986. J. Cell Biol. 103:755-766). Blocks of newly incorporated VAC microvilli and 184-kD protein progressively move from intercellular ("primitive" lateral) spaces towards the microvilli-poor free cell surface. The definitive lateral domain is sealed behind these blocks by the growing tight junctional fence. These results demonstrate a fundamental role of cell-cell contact-mediated VAC exocytosis in the establishment of epithelial surface polarity. Because isolated stages (intercellular pockets) of the stereotyped sequence of events triggered by the establishment of intercellular contacts in MDCK cells have been reported during normal differentiation of intestine epithelium (Colony, P. C., and M. R. Neutra. 1983. Dev. Biol. 97:349-363), we speculate that the mechanism we describe here plays an important role in the establishment of epithelial cell polarity in vivo. PMID:3053735

Increasing the endogenous NO level causes catalase inactivation and reactivation of intercellular apoptosis signaling specifically in tumor cells.

PubMed

Bauer, Georg

2015-12-01

Tumor cells generate extracellular superoxide anions and are protected against intercellular apoptosis-inducing HOCl- and NO/peroxynitrite signaling through the expression of membrane-associated catalase. This enzyme decomposes H2O2 and thus prevents HOCl synthesis. It efficiently interferes with NO/peroxynitrite signaling through oxidation of NO and decomposition of peroxynitrite. The regulatory potential of catalase at the crosspoint of ROS and RNS chemical biology, as well as its high local concentration on the outside of the cell membrane of tumor cells, establish tight control of intercellular signaling and thus prevent tumor cell apoptosis. Therefore, inhibition of catalase or its inactivation by singlet oxygen reactivate intercellular apoptosis-inducing signaling. Nitric oxide and peroxynitrite are connected with catalase in multiple and meaningful ways, as (i) NO can be oxidated by compound I of catalase, (ii) NO can reversibly inhibit catalase, (iii) peroxynitrite can be decomposed by catalase and (iv) the interaction between peroxynitrite and H2O2 leads to the generation of singlet oxygen that inactivates catalase. Therefore, modulation of the concentration of free NO through addition of arginine, inhibition of arginase, induction of NOS expression or inhibition of NO dioxygenase triggers an autoamplificatory biochemical cascade that is based on initial formation of singlet oxygen, amplification of superoxide anion/H2O2 and NO generation through singlet oxygen dependent stimulation of the FAS receptor and caspase-8. Finally, singlet oxygen is generated at sufficiently high concentration to inactivate protective catalase and to reactivate intercellular apoptosis-inducing ROS signaling. This regulatory network allows to establish several pathways for synergistic interactions, like the combination of modulators of NO metabolism with enhancers of superoxide anion generation, modulators of NO metabolism that act at different targets and between modulators of NO metabolism and direct catalase inhibitors. The latter aspect is explicitely studied for the interaction between catalase inhibiting acetylsalicylic acid and an NO donor. It is also shown that hybrid molecules like NO-aspirin utilize this synergistic potential. Our data open novel approaches for rational tumor therapy based on specific ROS signaling and its control in tumor cells. Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.

TopoIIα prevents telomere fragility and formation of ultra thin DNA bridges during mitosis through TRF1-dependent binding to telomeres.

PubMed

d'Alcontres, Martina Stagno; Palacios, Jose Alejandro; Mejias, Diego; Blasco, Maria A

2014-01-01

Telomeres are repetitive nucleoprotein structures at the ends of chromosomes. Like most genomic regions consisting of repetitive DNA, telomeres are fragile sites prone to replication fork stalling and generation of chromosomal instability. In particular, abrogation of the TRF1 telomere binding protein leads to stalled replication forks and aberrant telomere structures known as "multitelomeric signals". Here, we report that TRF1 deficiency also leads to the formation of "ultra-fine bridges" (UFB) during mitosis, and to an increased time to complete mitosis mediated by the spindle assembly checkpoint proteins (SAC). We find that topoisomerase IIα (TopoIIα), an enzyme essential for resolution of DNA replication intermediates, binds telomeres in a TRF1-mediated manner. Indeed, similar to TRF1 abrogation, TopoIIα downregulation leads to telomere fragility and UFB, suggesting that these phenotypes are due to decreased TopoIIα at telomeres. We find that SAC proteins bind telomeres in vivo, and that this is disrupted upon TRF1 deletion. These findings suggest that TRF1 links TopoIIα and SAC proteins in a pathway that ensures correct telomere replication and mitotic segregation, unveiling how TRF1 protects from telomere fragility and mitotic defects.

Is Pulp Inflammation a Prerequisite for Pulp Healing and Regeneration?

PubMed

Goldberg, Michel; Njeh, Akram; Uzunoglu, Emel

2015-01-01

The importance of inflammation has been underestimated in pulpal healing, and in the past, it has been considered only as an undesirable effect. Associated with moderate inflammation, necrosis includes pyroptosis, apoptosis, and nemosis. There are now evidences that inflammation is a prerequisite for pulp healing, with series of events ahead of regeneration. Immunocompetent cells are recruited in the apical part. They slide along the root and migrate toward the crown. Due to the high alkalinity of the capping agent, pulp cells display mild inflammation, proliferate, and increase in number and size and initiate mineralization. Pulp fibroblasts become odontoblast-like cells producing type I collagen, alkaline phosphatase, and SPARC/osteonectin. Molecules of the SIBLING family, matrix metalloproteinases, and vascular and nerve mediators are also implicated in the formation of a reparative dentinal bridge, osteo/orthodentin closing the pulp exposure. Beneath a calciotraumatic line, a thin layer identified as reactionary dentin underlines the periphery of the pulp chamber. Inflammatory and/or noninflammatory processes contribute to produce a reparative dentinal bridge closing the pulp exposure, with minute canaliculi and large tunnel defects. Depending on the form and severity of the inflammatory and noninflammatory processes, and according to the capping agent, pulp reactions are induced specifically.

Is Pulp Inflammation a Prerequisite for Pulp Healing and Regeneration?

PubMed Central

Goldberg, Michel; Njeh, Akram; Uzunoglu, Emel

2015-01-01

The importance of inflammation has been underestimated in pulpal healing, and in the past, it has been considered only as an undesirable effect. Associated with moderate inflammation, necrosis includes pyroptosis, apoptosis, and nemosis. There are now evidences that inflammation is a prerequisite for pulp healing, with series of events ahead of regeneration. Immunocompetent cells are recruited in the apical part. They slide along the root and migrate toward the crown. Due to the high alkalinity of the capping agent, pulp cells display mild inflammation, proliferate, and increase in number and size and initiate mineralization. Pulp fibroblasts become odontoblast-like cells producing type I collagen, alkaline phosphatase, and SPARC/osteonectin. Molecules of the SIBLING family, matrix metalloproteinases, and vascular and nerve mediators are also implicated in the formation of a reparative dentinal bridge, osteo/orthodentin closing the pulp exposure. Beneath a calciotraumatic line, a thin layer identified as reactionary dentin underlines the periphery of the pulp chamber. Inflammatory and/or noninflammatory processes contribute to produce a reparative dentinal bridge closing the pulp exposure, with minute canaliculi and large tunnel defects. Depending on the form and severity of the inflammatory and noninflammatory processes, and according to the capping agent, pulp reactions are induced specifically. PMID:26538825

Strain and Temperature Sensing Properties of Multiwalled Carbon Nanotube Yarn Composites

NASA Technical Reports Server (NTRS)

Kahng, Seun K.; Gates, Thomas S.; Jefferson, Gail D.

2008-01-01

Strain and temperature response of Multiwalled Carbon Nanotube (MWCNT/CNT) yarns on a stainless steel test beam has been studied. The carbon nanotube yarns are spun from a multiwalled carbon nanotube forest grown on a silicon substrate to a 4-ply yarn with a diameter of about 15-20 microns. Four of the 4-ply CNT yarns are arranged in a Wheatstone bridge configuration on the stainless steel test beam using a thin layer of polyurethane resin that insulates and protects the yarns from the test beam. Strain sensitivities of the CNT yarn sensors range from 1.39 to 1.75 mV/V/1000 microstrain at room temperature, and temperature sensitivity of the CNT yarn bridge is 91 microA/degC. Resistance of the yarns range from 215 to 270 ohms for CNT yarn length of approximately 5 mm. Processes used in attaching the CNT yarns on the test beam and experimental procedures used for the measurements are described. Conventional metallic foil strain gages are attached to the test beam to compare with the CNT sensors. The study demonstrates multifunctional capability of the sensor for strain and temperature measurements and shows its applicability where engineering strain is less than 3%.

Microscale Investigation of Thermo-Fluid Transport in the Transition FIL, Region of an Evaporating Capillary Meniscus Using a Microgravity Environment

NASA Technical Reports Server (NTRS)

Kihm, K. D.; Allen, J. S.; Hallinan, K. P.; Pratt, D. M.

2004-01-01

In order to enhance the fundamental understanding of thin film evaporation and thereby improve the critical design concept for two-phase heat transfer devices, microscale heat and mass transport is to be investigated for the transition film region using state-of-the-art optical diagnostic techniques. By utilizing a microgravity environment, the length scales of the transition film region can be extended sufficiently, from submicron to micron, to probe and measure the microscale transport fields which are affected by intermolecular forces. Extension of the thin film dimensions under microgravity will be achieved by using a conical evaporator made of a thin silicon substrate under which concentric and individually controlled micro-heaters are vapor-deposited to maintain either a constant surface temperature or a controlled temperature variation. Local heat transfer rates, required to maintain the desired wall temperature boundary condition, will be measured and recorded by the concentric thermoresistance heaters controlled by a Wheatstone bridge circuit, The proposed experiment employs a novel technique to maintain a constant liquid volume and liquid pressure in the capillary region of the evaporating meniscus so as to maintain quasi-stationary conditions during measurements on the transition film region. Alternating use of Fizeau interferometry via white and monochromatic light sources will measure the thin film slope and thickness variation, respectively. Molecular Fluorescence Tracking Velocimetry (MFTV), utilizing caged fluorophores of approximately 10-nm in size as seeding particles, will be used to measure the velocity profiles in the thin film region. An optical sectioning technique using confocal microscopy will allow submicron depthwise resolution for the velocity measurements within the film for thicknesses on the order of a few microns. Digital analysis of the fluorescence image-displacement PDFs, as described in the main proposal, can further enhance the depthwise resolution.

Radial stiffness characteristics of the overlap regions of sarcomeres in isolated skeletal myofibrils in pre-force generating state.

PubMed

Miyashiro, Daisuke; Ohtsuki, Misato; Shimamoto, Yuta; Wakayama, Jun'ichi; Kunioka, Yuki; Kobayashi, Takakazu; Ishiwata, Shin'ichi; Yamada, Takenori

2017-01-01

We have studied the stiffness of myofilament lattice in sarcomeres in the pre-force generating state, which was realized by a relaxing reagent, BDM (butane dione monoxime). First, the radial stiffness for the overlap regions of sarcomeres of isolated single myofibrils was estimated from the resulting decreases in diameter by osmotic pressure applied with the addition of Dextran. Then, the radial stiffness was also estimated from force-distance curve measurements with AFM technology. The radial stiffness for the overlap regions thus obtained was composed of a soft and a rigid component. The soft component visco-elastically changed in a characteristic fashion depending on the physiological conditions of myofibrils, suggesting that it comes from cross-bridge structures. BDM treatments significantly affected the soft radial component of contracting myofibrils depending on the approach velocity of cantilever: It was nearly equal to that in the contracting state at high approach velocity, whereas as low as that in the relaxing state at low approach velocity. However, comparable BDM treatments greatly suppressed the force production and the axial stiffness in contracting glycerinated muscle fibers and also the sliding velocity of actin filaments in the in vitro motility assay. Considering that BDM shifts the cross-bridge population from force generating to pre-force generating states in contracting muscle, the obtained results strongly suggest that cross-bridges in the pre-force generating state are visco-elastically attached to the thin filaments in such a binding manner that the axial stiffness is low but the radial stiffness significantly high similar to that in force generating state.

Thin film strain transducer. [suitable for in-flight measurement of scientific balloon strain

NASA Technical Reports Server (NTRS)

Rand, J. L. (Inventor)

1985-01-01

A strain transducer system and process for making same is disclosed wherein a beryllium-copper ring having four strain gages disposed thereon is electrically connected in Wheatstone bridge fashion to output instrumentation. Tabs are bonded to a balloon or like surface with strain on the surface causing bending of the ring and providing an electrical signal through the gages proportional to the surface strain. A figure is provided which illustrates a pattern of a one-half ring segment as placed on a sheet of beryllium-copper for chem-mill etch formation, prior to bending and welding of a pair of the segments to form a ring structure.

Growth of carbon nanotubes by Fe-catalyzed chemical vapor processes on silicon-based substrates

NASA Astrophysics Data System (ADS)

Angelucci, Renato; Rizzoli, Rita; Vinciguerra, Vincenzo; Fortuna Bevilacqua, Maria; Guerri, Sergio; Corticelli, Franco; Passini, Mara

2007-03-01

In this paper, a site-selective catalytic chemical vapor deposition synthesis of carbon nanotubes on silicon-based substrates has been developed in order to get horizontally oriented nanotubes for field effect transistors and other electronic devices. Properly micro-fabricated silicon oxide and polysilicon structures have been used as substrates. Iron nanoparticles have been obtained both from a thin Fe film evaporated by e-gun and from iron nitrate solutions accurately dispersed on the substrates. Single-walled nanotubes with diameters as small as 1 nm, bridging polysilicon and silicon dioxide “pillars”, have been grown. The morphology and structure of CNTs have been characterized by SEM, AFM and Raman spectroscopy.

Femtosecond ultrasonic spectroscopy using a piezoelectric nanolayer: Hypersound attenuation in vitreous silica films

NASA Astrophysics Data System (ADS)

Wen, Yu-Chieh; Guol, Shi-Hao; Chen, Hung-Pin; Sheu, Jinn-Kong; Sun, Chi-Kuang

2011-08-01

We report ultra-broadband ultrasonic spectroscopy with an impedance-matched piezoelectric nanolayer, which enables optical generation and detection of a 730-fs acoustic pulse (the width of ten lattice constants). The bandwidth improvement facilitates THz laser ultrasonics to bridge the spectral gap between inelastic light and x-ray scatterings (0.1-1 THz) in the studies of lattice dynamics. As a demonstration, this method is applied to measure sound attenuation α in a vitreous SiO2 thin film. Our results extend the existing low-frequency data obtained by ultrasonic-based and light scattering methods and also show a α∝ f2 behavior for frequencies f up to 650 GHz.

FAST OPENING SWITCH

DOEpatents

Bender, M.; Bennett, F.K.; Kuckes, A.F.

1963-09-17

A fast-acting electric switch is described for rapidly opening a circuit carrying large amounts of electrical power. A thin, conducting foil bridges a gap in this circuit and means are provided for producing a magnetic field and eddy currents in the foil, whereby the foil is rapidly broken to open the circuit across the gap. Advantageously the foil has a hole forming two narrow portions in the foil and the means producing the magnetic field and eddy currents comprises an annular coil having its annulus coaxial with the hole in the foil and turns adjacent the narrow portions of the foil. An electrical current flows through the coil to produce the magnetic field and eddy currents in the foil. (AEC)

Opening and retraction of particulate soap films

NASA Astrophysics Data System (ADS)

Timounay, Yousra; Lorenceau, Elise; Rouyer, Florence

2015-07-01

We study for the first time the bursting dynamics of thin liquid films laden with hydrophobic micronic particles either with free or constrained edges. We highlight that the particles can arrange in bilayer or monolayer configurations and explore a range of particles coverage from zero to random close packing. When the particles bridge the two interfaces (monolayer configuration) of free-edge films, the hole opens intermittently. For the other cases, we observe constant retraction velocities, modeled by balancing liquid and particles inertia against surface tension as in Taylor-Culick theory. But, this approach is only valid up to a critical value of particles coverage due to the interplay between the interfaces and the friction between particles.

A numerical study of mobility in thin films of fullerene derivatives.

PubMed

Mackenzie, Roderick C I; Frost, Jarvist M; Nelson, Jenny

2010-02-14

The effect of functional group size on the electron mobility in films of fullerene derivatives is investigated numerically. A series of four C(60) derivatives are formed by attaching saturated hydrocarbon chains to the C(60) cage via a methano bridge. For each of the derivatives investigated, molecular dynamics is used to generate a realistic material morphology. Quantum chemical methods are then used to calculate intermolecular charge transfer rates. Finally, Monte Carlo methods are used to simulate time-of-flight experiments and thus calculate the electron mobility. It is found that as the length of the aliphatic side chain increases, the configurational disorder increases and thus the mobility decreases.

Oocyte Polarization Is Coupled to the Chromosomal Bouquet, a Conserved Polarized Nuclear Configuration in Meiosis

PubMed Central

Elkouby, Yaniv M.; Jamieson-Lucy, Allison; Mullins, Mary C.

2016-01-01

The source of symmetry breaking in vertebrate oocytes is unknown. Animal—vegetal oocyte polarity is established by the Balbiani body (Bb), a conserved structure found in all animals examined that contains an aggregate of specific mRNAs, proteins, and organelles. The Bb specifies the oocyte vegetal pole, which is key to forming the embryonic body axes as well as the germline in most vertebrates. How Bb formation is regulated and how its asymmetric position is established are unknown. Using quantitative image analysis, we trace oocyte symmetry breaking in zebrafish to a nuclear asymmetry at the onset of meiosis called the chromosomal bouquet. The bouquet is a universal feature of meiosis where all telomeres cluster to one pole on the nuclear envelope, facilitating chromosomal pairing and meiotic recombination. We show that Bb precursor components first localize with the centrosome to the cytoplasm adjacent to the telomere cluster of the bouquet. They then aggregate around the centrosome in a specialized nuclear cleft that we identified, assembling the early Bb. We show that the bouquet nuclear events and the cytoplasmic Bb precursor localization are mechanistically coordinated by microtubules. Thus the animal—vegetal axis of the oocyte is aligned to the nuclear axis of the bouquet. We further show that the symmetry breaking events lay upstream to the only known regulator of Bb formation, the Bucky ball protein. Our findings link two universal features of oogenesis, the Bb and the chromosomal bouquet, to oocyte polarization. We propose that a meiotic—vegetal center couples meiosis and oocyte patterning. Our findings reveal a novel mode of cellular polarization in meiotic cells whereby cellular and nuclear polarity are aligned. We further reveal that in zygotene nests, intercellular cytoplasmic bridges remain between oocytes and that the position of the cytoplasmic bridge coincides with the location of the centrosome meiotic—vegetal organizing center. These results suggest that centrosome positioning is set by the last mitotic oogonial division plane. Thus, oocytes are polarized in two steps: first, mitotic divisions preset the centrosome with no obvious polarization yet, then the meiotic—vegetal center forms at zygotene bouquet stages, when symmetry is, in effect, broken. PMID:26741740

Active cell-matrix coupling regulates cellular force landscapes of cohesive epithelial monolayers

NASA Astrophysics Data System (ADS)

Zhao, Tiankai; Zhang, Yao; Wei, Qiong; Shi, Xuechen; Zhao, Peng; Chen, Long-Qing; Zhang, Sulin

2018-03-01

Epithelial cells can assemble into cohesive monolayers with rich morphologies on substrates due to competition between elastic, edge, and interfacial effects. Here we present a molecularly based thermodynamic model, integrating monolayer and substrate elasticity, and force-mediated focal adhesion formation, to elucidate the active biochemical regulation over the cellular force landscapes in cohesive epithelial monolayers, corroborated by microscopy and immunofluorescence studies. The predicted extracellular traction and intercellular tension are both monolayer size and substrate stiffness dependent, suggestive of cross-talks between intercellular and extracellular activities. Our model sets a firm ground toward a versatile computational framework to uncover the molecular origins of morphogenesis and disease in multicellular epithelia.

Low-temperature and conventional scanning electron microscopy of human urothelial neoplasms.

PubMed

Hopkins, D M; Morris, J A; Oates, K; Huddart, H; Staff, W G

1989-05-01

The appearance of neoplastic human urothelium viewed by low-temperature scanning electron microscopy (LTSEM) and conventional scanning electron microscopy (CSEM) was compared. Fixed, dehydrated neoplastic cells viewed by CSEM had well-defined, often raised cell junctions; no intercellular gaps; and varying degrees of pleomorphic surface microvilli. The frozen hydrated material viewed by LTSEM, however, was quite different. The cells had a flat or dimpled surface, but no microvilli. There were labyrinthine lateral processes which interdigitated with those of adjacent cells and outlined large intercellular gaps. The process of fixation and dehydration will inevitably distort cell contours and on theoretical grounds, the images of frozen hydrated material should more closely resemble the in vivo appearance.

Flavonoids (apigenin, tangeretin) counteract tumor promoter-induced inhibition of intercellular communication of rat liver epithelial cells.

PubMed

Chaumontet, C; Droumaguet, C; Bex, V; Heberden, C; Gaillard-Sanchez, I; Martel, P

1997-03-19

We have shown previously that two flavonoids, apigenin and tangeretin, enhance gap junctional intercellular communication (GJIC) in rat liver epithelial cells, named REL cells. Here, we show that these two flavones also antagonize the inhibition of GJIC induced by tumor promoters like 12-O-tetradecanoyl-phorbol-acetate (TPA) and 3,5,di-tertio-butyl-4-hydroxytoluene (BHT). Their preventive effect is rapid. It does not seem to involve any change of the amount of the connexin expressed in REL cells, connexin 43 (Cx 43), and in its phosphorylation state. Other flavonoids tested including naringenin, myricetin, catechin and chrysin did not enhance GJIC nor counteract TPA-induced inhibition of GJIC.

Cleavage of transmembrane junction proteins and their role in regulating epithelial homeostasis

PubMed Central

Nava, Porfirio; Kamekura, Ryuta; Nusrat, Asma

2013-01-01

Epithelial tissues form a selective barrier that separates the external environment from the internal tissue milieu. Single epithelial cells are densely packed and associate via distinct intercellular junctions. Intercellular junction proteins not only control barrier properties of the epithelium but also play an important role in regulating epithelial homeostasis that encompasses cell proliferation, migration, differentiation and regulated shedding. Recent studies have revealed that several proteases target epithelial junction proteins during physiological maturation as well as in pathologic states such as inflammation and cancer. This review discusses mechanisms and biological consequences of transmembrane junction protein cleavage. The influence of junction protein cleavage products on pathogenesis of inflammation and cancer is discussed. PMID:24665393

Astroglial Metabolic Networks Sustain Hippocampal Synaptic Transmission

NASA Astrophysics Data System (ADS)

Rouach, Nathalie; Koulakoff, Annette; Abudara, Veronica; Willecke, Klaus; Giaume, Christian

2008-12-01

Astrocytes provide metabolic substrates to neurons in an activity-dependent manner. However, the molecular mechanisms involved in this function, as well as its role in synaptic transmission, remain unclear. Here, we show that the gap-junction subunit proteins connexin 43 and 30 allow intercellular trafficking of glucose and its metabolites through astroglial networks. This trafficking is regulated by glutamatergic synaptic activity mediated by AMPA receptors. In the absence of extracellular glucose, the delivery of glucose or lactate to astrocytes sustains glutamatergic synaptic transmission and epileptiform activity only when they are connected by gap junctions. These results indicate that astroglial gap junctions provide an activity-dependent intercellular pathway for the delivery of energetic metabolites from blood vessels to distal neurons.

Evolution of altruism in spatial prisoner's dilemma: Intra- and inter-cellular interactions

NASA Astrophysics Data System (ADS)

Yokoi, Hiroki; Uehara, Takashi; Sakata, Tomoyuki; Naito, Hiromi; Morita, Satoru; Tainaka, Kei-ichi

2014-12-01

Iterated prisoner's dilemma game is carried out on lattice with “colony” structure. Each cell is regarded as a colony which contains plural players with an identical strategy. Both intra- and inter-cellular interactions are assumed. In the former a player plays with all other players in the same colony, while in the latter he plays with one player each from adjacent colonies. Spatial patterns among four typical strategies exhibit various dynamics and winners. Both theory and simulation reveal that All Cooperation (AC) wins, when the members of colony or the intensity of noise increases. This result explains the evolution of altruism in animal societies, even though errors easily occur in animal communications.

Pemphigus vulgaris associated with autoimmune hemolytic anemia and elevated TNF alpha.

PubMed

Ujihara, M; Hamanaka, S; Matsuda, S; Numa, F; Kato, H

1994-01-01

A 76-year-old female was admitted with many bullae and erythema on her trunk and extremities. A biopsy specimen showed significant intercellular edema in the lower epidermis and eosinophilic infiltration into the dermis and the epidermis. Immunofluorescent staining revealed the deposition of IgG in the intercellular area of her prickle cells. From these histologic findings and the typical clinical features, we diagnosed her as having pemphigus vulgaris. Examination of her blood revealed that she also suffered from autoimmune hemolytic anemia. Despite intensive treatment with prednisolone, she finally died. This case is of interest because of its rarity and the TNF alpha detected significantly in the blister fluid of this patient.

Astroglial metabolic networks sustain hippocampal synaptic transmission.

PubMed

Rouach, Nathalie; Koulakoff, Annette; Abudara, Veronica; Willecke, Klaus; Giaume, Christian

2008-12-05

Astrocytes provide metabolic substrates to neurons in an activity-dependent manner. However, the molecular mechanisms involved in this function, as well as its role in synaptic transmission, remain unclear. Here, we show that the gap-junction subunit proteins connexin 43 and 30 allow intercellular trafficking of glucose and its metabolites through astroglial networks. This trafficking is regulated by glutamatergic synaptic activity mediated by AMPA receptors. In the absence of extracellular glucose, the delivery of glucose or lactate to astrocytes sustains glutamatergic synaptic transmission and epileptiform activity only when they are connected by gap junctions. These results indicate that astroglial gap junctions provide an activity-dependent intercellular pathway for the delivery of energetic metabolites from blood vessels to distal neurons.

Ice-Ocean Interactions to the North-West of Greenland: Glaciers, Straits, Ice Bridges, and the Rossby Radius (Invited)

NASA Astrophysics Data System (ADS)

Muenchow, A.; Falkner, K. K.; Melling, H.; Johnson, H. L.; Huntley, H. S.; Ryan, P.; Friends Of Petermann

2010-12-01

Petermann Glacier at 81 N latitude is a major outlet glacier adjacent to Nares Strait. It terminates in a long (70 km), narrow (16 km) and thin (50 m) floating tongue and has a grounding line more than 500 m below sea level. A calving event in 2010 reduced the floating area by 25% and produced a single 240 km2 ice island currently moving south in Nares Strait where it will likely interact with island to potentially create a temporary polynya in Nares Strait. The 2010 calving from Petermann Glacier contributes <10% to its mass balance as more than 80% is lost due to basal melting by the ocean. Hence the largely unexplored physics at the ice-ocean interface determine how a changing climate impacts this outlet glacier. Conducting exploratory surveys inside Petermann Fjord in 2003, 2007, and 2009, we find a 1100 m deep fjord connected to Nares Strait via a sill at 350-450 m depth. The fjord receives about 3 times the amount of heat required for the basal melt rates. Furthermore, limited data and analytical modeling suggests a 3-dimensional circulation over the upper 300-m of the water column with a coastally trapped buoyant outflow. We integrate these findings with more complete oceanic time series data from an array moored in Nares Strait from 2003 through 2009 near 80.5 N. In the past Nares Strait and Petermann Fjord were covered by land fast sea ice during the 9-10 month long winter season. Archeological and remotely sensed records indicate that an ice bridge formed regularly at the southern end of Nares Strait creating the North-Water polynya near 79 N latitude. Since 2006 this ice bridge has largely failed to form, leading, perhaps, to the occasional formation of a secondary ice bridge 300 km to the north where Nares Strait connects to the Arctic Ocean. However, this ice bridge appears to form for shorter periods only. Consequently Arctic sea ice can now exit the Arctic in winter via pathways to the west of Greenland all year. We speculate that this changed ocean and sea ice regime in Nares Strait and the Arctic Ocean may contribute to the recently observed calving events in Petermann Fjord.

Construction of the bridge in the cavern in the Vrata tunnel (Croatia)

NASA Astrophysics Data System (ADS)

Garasic, Mladen; Sasa Kovacevic, Meho; Juric-Kacunic, Danijela

2010-05-01

In the Dinaric karst system in Croatia some 11500 speleological objects have been explored so far, more than 1000 of which were discovered during construction works. Such speleological objects without natural entrance on the terrain surface (which are called "caverns") have been discovered on the construction sites of the highways. Over the past twenty years they have been systematically investigated and treated. A special kind of remediation was conducted in the cavern's large hall of the "Vrata" tunnel on the Zagreb - Rijeka highway. Due to size, shape, cavern's position and hydrogeological parameters (fissured and karstified aquifers) within the karst system it was necessary to design and construct a 58 m bridge over the cavern. In addition, the cavern's vault had to be reinforced and stabilized, as the overburden was very thin. The beam-and -stringer grid with special anchors was used. The cavern's rehabilitation in the "Vrata" tunnel was a unique undertaking, and the bridge (without piers) is the cavern's longest bridge in the world. A speleological object of large dimensions was discovered in the "Vrata"tunnel's right tube on the Rijeka-Zagreb highway. Speleological, geotechnical, engineering geological and hydrogeological investigation works were carried out for the purpose of preservation the speleological object (cavern). On the basis of classification results of rock masses and conducted numerical analyses the support system for the cavern's vault stabilization was selected. The support system's elements include the beam-and-stringer grid constructed on the terrain's surface above the cavern, tendons and geotechnical anchors. To ensure stability of the speleological object, and to conduct the backward numerical analyses the measurement of vertical deformations from the terrain's surface along the rock's mass by means of sliding micrometers was undertaken. Backward numerical analyses combined with geotechnical measurements enable safer and more rational approach to design and construction of underground structures. They contribute to the knowledge on rock mass performance and to determination of its physical and mechanical parameters connecting them with rock classification results. The analyses are a great help in verification or modification of elements' features of primary support system. Tunnel and bridge in tunnel "Vrata" were opened for traffic in November 2008. Keywords: speleology, cave, Dinaric karst, Croatia, tunnel, karst phenomena, geotechnical engineering.

FIB-SEM tomography of human skin telocytes and their extracellular vesicles

PubMed Central

Cretoiu, Dragos; Gherghiceanu, Mihaela; Hummel, Eric; Zimmermann, Hans; Simionescu, Olga; Popescu, Laurentiu M

2015-01-01

We have shown in 2012 the existence of telocytes (TCs) in human dermis. TCs were described by transmission electron microscopy (TEM) as interstitial cells located in non-epithelial spaces (stroma) of many organs (see www.telocytes.com). TCs have very long prolongations (tens to hundreds micrometers) named Telopodes (Tps). These Tps have a special conformation with dilated portions named podoms (containing mitochondria, endoplasmic reticulum and caveolae) and very thin segments (below resolving power of light microscopy), called podomers. To show the real 3D architecture of TC network, we used the most advanced available electron microscope technology: focused ion beam scanning electron microscopy (FIB-SEM) tomography. Generally, 3D reconstruction of dermal TCs by FIB-SEM tomography revealed the existence of Tps with various conformations: (i) long, flattened irregular veils (ribbon-like segments) with knobs, corresponding to podoms, and (ii) tubular structures (podomers) with uneven calibre because of irregular dilations (knobs) – the podoms. FIB-SEM tomography also showed numerous extracellular vesicles (diameter 438.6 ± 149.1 nm, n = 30) released by a human dermal TC. Our data might be useful for understanding the role(s) of TCs in intercellular signalling and communication, as well as for comprehension of pathologies like scleroderma, multiple sclerosis, psoriasis, etc. PMID:25823591

Eavesdropping on altered cell-to-cell signaling in cancer by secretome profiling.

PubMed

Klinke, David J

2016-01-01

In the past decade, cumulative clinical experiences with molecular targeted therapies and immunotherapies for cancer have promoted a shift in our conceptual understanding of cancer. This view shifted from viewing solid tumors as a homogeneous mass of malignant cells to viewing tumors as heterogeneous structures that are dynamically shaped by intercellular interactions among the variety of stromal, immune, and malignant cells present within the tumor microenvironment. As in any dynamic system, identifying how cells communicate to maintain homeostasis and how this communication is altered during oncogenesis are key hurdles for developing therapies to restore normal tissue homeostasis. Here, I discuss tissues as dynamic systems, using the mammary gland as an example, and the evolutionary concepts applied to oncogenesis. Drawing from these concepts, I present 2 competing hypotheses for how intercellular communication might be altered during oncogenesis. As an initial test of these competing hypotheses, a recent secretome comparison between normal human mammary and HER2+ breast cancer cell lines suggested that the particular proteins secreted by the malignant cells reflect a convergent evolutionary path associated with oncogenesis in a specific anatomical niche, despite arising in different individuals. Overall, this study illustrates the emerging power of secretome proteomics to probe, in an unbiased way, how intercellular communication changes during oncogenesis.

β-Catenin Serves as a Clutch between Low and High Intercellular E-Cadherin Bond Strengths

PubMed Central

Bajpai, Saumendra; Feng, Yunfeng; Wirtz, Denis; Longmore, Gregory D.

2013-01-01

A wide range of invasive pathological outcomes originate from the loss of epithelial phenotype and involve either loss of function or downregulation of transmembrane adhesive receptor complexes, including Ecadherin (Ecad) and binding partners β-catenin and α-catenin at adherens junctions. Cellular pathways regulating wild-type β-catenin level, or direct mutations in β-catenin that affect the turnover of the protein have been shown to contribute to cancer development, through induction of uncontrolled proliferation of transformed tumor cells, particularly in colon cancer. Using single-molecule force spectroscopy, we show that depletion of β-catenin or the prominent cancer-related S45 deletion mutation in β-catenin present in human colon cancers both weaken tumor intercellular Ecad/Ecad bond strength and diminishes the capacity of specific extracellular matrix proteins—including collagen I, collagen IV, and laminin V—to modulate intercellular Ecad/Ecad bond strength through α-catenin and the kinase activity of glycogen synthase kinase 3 (GSK-3β). Thus, in addition to regulating tumor cell proliferation, cancer-related mutations in β-catenin can influence tumor progression by weakening the adhesion of tumor cells to one another through reduced individual Ecad/Ecad bond strength and cellular adhesion to specific components of the extracellular matrix and the basement membrane. PMID:24268141

Phloroglucinol functions as an intracellular and intercellular chemical messenger influencing gene expression in Pseudomonas protegens.

PubMed

Clifford, Jennifer C; Buchanan, Alex; Vining, Oliver; Kidarsa, Teresa A; Chang, Jeff H; McPhail, Kerry L; Loper, Joyce E

2016-10-01

Bacteria can be both highly communicative and highly competitive in natural habitats and antibiotics are thought to play a role in both of these processes. The soil bacterium Pseudomonas protegens Pf-5 produces a spectrum of antibiotics, two of which, pyoluteorin and 2,4-diacetylphloroglucinol (DAPG), function in intracellular and intercellular communication, both as autoinducers of their own production. Here, we demonstrate that phloroglucinol, an intermediate in DAPG biosynthesis, can serve as an intercellular signal influencing the expression of pyoluteorin biosynthesis genes, the production of pyoluteorin, and inhibition of Pythium ultimum, a phytopathogenic oomycete sensitive to pyoluteorin. Through analysis of RNAseq data sets, we show that phloroglucinol had broad effects on the transcriptome of Pf-5, significantly altering the transcription of more than two hundred genes. The effects of nanomolar versus micromolar concentrations of phloroglucinol differed both quantitatively and qualitatively, influencing the expression of distinct sets of genes or having opposite effects on transcript abundance of certain genes. Therefore, our results support the concept of hormesis, a phenomenon associated with signalling molecules that elicit distinct responses at different concentrations. Phloroglucinol is the first example of an intermediate of antibiotic biosynthesis that functions as a chemical messenger influencing gene expression in P. protegens. © 2015 Society for Applied Microbiology and John Wiley & Sons Ltd.

Eavesdropping on altered cell-to-cell signaling in cancer by secretome profiling

PubMed Central

Klinke, David J

2016-01-01

In the past decade, cumulative clinical experiences with molecular targeted therapies and immunotherapies for cancer have promoted a shift in our conceptual understanding of cancer. This view shifted from viewing solid tumors as a homogeneous mass of malignant cells to viewing tumors as heterogeneous structures that are dynamically shaped by intercellular interactions among the variety of stromal, immune, and malignant cells present within the tumor microenvironment. As in any dynamic system, identifying how cells communicate to maintain homeostasis and how this communication is altered during oncogenesis are key hurdles for developing therapies to restore normal tissue homeostasis. Here, I discuss tissues as dynamic systems, using the mammary gland as an example, and the evolutionary concepts applied to oncogenesis. Drawing from these concepts, I present 2 competing hypotheses for how intercellular communication might be altered during oncogenesis. As an initial test of these competing hypotheses, a recent secretome comparison between normal human mammary and HER2+ breast cancer cell lines suggested that the particular proteins secreted by the malignant cells reflect a convergent evolutionary path associated with oncogenesis in a specific anatomical niche, despite arising in different individuals. Overall, this study illustrates the emerging power of secretome proteomics to probe, in an unbiased way, how intercellular communication changes during oncogenesis. PMID:27308541

Development of microarray device for functional evaluation of PC12D cell axonal extension ability

NASA Astrophysics Data System (ADS)

Nakamachi, Eiji; Yanagimoto, Junpei; Murakami, Shinya; Morita, Yusuke

2014-04-01

In this study, we developed a microarray bio-MEMS device that could trap PC12D (rat pheochromocytoma cells) cells to examine the intercellular interaction effect on the cell activation and the axonal extension ability. This is needed to assign particular patterns of PC12D cells to establish a cell functional evaluation technique. This experimental observation-based technique can be used for design of the cell sheet and scaffold for peripheral and central nerve regeneration. We have fabricated a micropillar-array bio-MEMS device, whose diameter was approximately 10 μm, by using thick photoresist SU-8 on the glass slide substrate. A maximum trapped PC12D cell ratio, 48.5%, was achieved. Through experimental observation of patterned PC12D "bi-cells" activation, we obtained the following results. Most of the PC12D "bi-cells" which had distances between 40 and 100 μm were connected after 24 h with a high probability. On the other hand, "bi-cells" which had distances between 110 and 200 μm were not connected. In addition, we measured axonal extension velocities in cases where the intercellular distance was between 40 and 100 μm. A maximum axonal extension velocity, 86.4 μm/h, was obtained at the intercellular distance of 40 μm.

[Influence of Cx26/Cx32 gap junction channel on antineoplastic effect of etoposide in Hela cells].

PubMed

Tong, Xu-Hui; Dong, Shu-Ying; Jiang, Guo-Jun; Fan, Gao-Fu

2012-03-01

To observe the influence of Cx26/Cx32 gap junction channel on the antineoplastic effect of etoposide in Hela cervical cancer cells. Fluorescence trace was used to assay the gap junction intercellular communication mediated by Cx26/Cx32 in Hela cells and its functional modulation by the pharmacological agents (oleamide, retinoid acid). A standard colony-forming assay was applied to determine the cell growth-inhibiting effect of etoposide in Hela cells with functional modulation of the gap junction. Hoechst 33258 staining was used to assess the changes in etoposide-induced apoptosis of Hela cells with altered gap junction functions. Oleamide markedly decreased while retinoid acid obviously increased the gap junction function in Hela cells. Standard colony-forming assay showed that etoposide produced a lowered antiproliferative effect in Hela cells with reduced gap junction and an increased antiproliferative effect in cells with enhanced gap junction function. In cells with a reduced gap junction function, etoposide induced a lowered apoptosis rate, which increased obviously in cells with an enhanced gap junction function. The antineoplastic effect of etoposide is reduced in Hela cells with a decreased gap junction intercellular communication mediated by Cx26/Cx32 and is enhanced in cells with an increased gap junction intercellular communication.

Symplastic Transport of Carboxyfluorescein in Staminal Hairs of Setcreasea purpurea Is Diffusive and Includes Loss to the Vacuole.

PubMed

Tucker, J E; Mauzerall, D; Tucker, E B

1989-07-01

The kinetics of symplastic transport in staminal hairs of Setcreasea purpurea was studied. The tip cell of a staminal hair was microinjected with carboxyfluorescein (CF) and the symplastic transport of this CF was videotaped and the digital data analyzed to produce kinetic curves. Using a finite difference equation for diffusion between cells and for loss of dye into the vacuole, kinetic curves were calculated and fitted to the observed data. These curves were matched with data from actual microinjection experiments by adjusting K (the coefficient of intercellular junction diffusion) and L (the coefficient of intracellular loss) until a minimum in the least squares difference between the curves was obtained. (a) Symplastic transport of CF was governed by diffusion through intercellular pores (plasmodesmata) and intracellular loss. Diffusion within the cell cytoplasm was never limiting. (b) Each cell and its plasmodesmata must be considered as its own diffusion system. Therefore, a diffusion coefficient cannot be calculated for an entire chain of cells. (c) The movement through plasmodesmata in either direction was the same since the data are fit by a diffusion equation. (d) Diffusion through the intercellular pores was estimated to be slower than diffusion through similar pores filled with water.

Symplastic Transport of Carboxyfluorescein in Staminal Hairs of Setcreasea purpurea Is Diffusive and Includes Loss to the Vacuole 1

PubMed Central

Tucker, Joseph E.; Mauzerall, David; Tucker, Edward B.

1989-01-01

The kinetics of symplastic transport in staminal hairs of Setcreasea purpurea was studied. The tip cell of a staminal hair was microinjected with carboxyfluorescein (CF) and the symplastic transport of this CF was videotaped and the digital data analyzed to produce kinetic curves. Using a finite difference equation for diffusion between cells and for loss of dye into the vacuole, kinetic curves were calculated and fitted to the observed data. These curves were matched with data from actual microinjection experiments by adjusting K (the coefficient of intercellular junction diffusion) and L (the coefficient of intracellular loss) until a minimum in the least squares difference between the curves was obtained. (a) Symplastic transport of CF was governed by diffusion through intercellular pores (plasmodesmata) and intracellular loss. Diffusion within the cell cytoplasm was never limiting. (b) Each cell and its plasmodesmata must be considered as its own diffusion system. Therefore, a diffusion coefficient cannot be calculated for an entire chain of cells. (c) The movement through plasmodesmata in either direction was the same since the data are fit by a diffusion equation. (d) Diffusion through the intercellular pores was estimated to be slower than diffusion through similar pores filled with water. PMID:16666864

Gap-junction-mediated communication in human periodontal ligament cells.

PubMed

Kato, R; Ishihara, Y; Kawanabe, N; Sumiyoshi, K; Yoshikawa, Y; Nakamura, M; Imai, Y; Yanagita, T; Fukushima, H; Kamioka, H; Takano-Yamamoto, T; Yamashiro, T

2013-07-01

Periodontal tissue homeostasis depends on a complex cellular network that conveys cell-cell communication. Gap junctions (GJs), one of the intercellular communication systems, are found between adjacent human periodontal ligament (hPDL) cells; however, the functional GJ coupling between hPDL cells has not yet been elucidated. In this study, we investigated functional gap-junction-mediated intercellular communication in isolated primary hPDL cells. SEM images indicated that the cells were in contact with each other via dendritic processes, and also showed high anti-connexin43 (Cx43) immunoreactivity on these processes. Gap-junctional intercellular communication (GJIC) among hPDL cells was assessed by fluorescence recovery after a photobleaching (FRAP) analysis, which exhibited dye coupling between hPDL cells, and was remarkably down-regulated when the cells were treated with a GJ blocker. Additionally, we examined GJs under hypoxic stress. The fluorescence recovery and expression levels of Cx43 decreased time-dependently under the hypoxic condition. Exposure to GJ inhibitor or hypoxia increased RANKL expression, and decreased OPG expression. This study shows that GJIC is responsible for hPDL cells and that its activity is reduced under hypoxia. This is consistent with the possible role of hPDL cells in regulating the biochemical reactions in response to changes in the hypoxic environment.

Structure and function of gap junction proteins: role of gap junction proteins in embryonic heart development.

PubMed

Ahir, Bhavesh K; Pratten, Margaret K

2014-01-01

Intercellular (cell-to-cell) communication is a crucial and complex mechanism during embryonic heart development. In the cardiovascular system, the beating of the heart is a dynamic and key regulatory process, which is functionally regulated by the coordinated spread of electrical activity through heart muscle cells. Heart tissues are composed of individual cells, each bearing specialized cell surface membrane structures called gap junctions that permit the intercellular exchange of ions and low molecular weight molecules. Gap junction channels are essential in normal heart function and they assist in the mediated spread of electrical impulses that stimulate synchronized contraction (via an electrical syncytium) of cardiac tissues. This present review describes the current knowledge of gap junction biology. In the first part, we summarise some relevant biochemical and physiological properties of gap junction proteins, including their structure and function. In the second part, we review the current evidence demonstrating the role of gap junction proteins in embryonic development with particular reference to those involved in embryonic heart development. Genetics and transgenic animal studies of gap junction protein function in embryonic heart development are considered and the alteration/disruption of gap junction intercellular communication which may lead to abnormal heart development is also discussed.

Gap junctional intercellular communication is required to maintain embryonic stem cells in a non-differentiated and proliferative state.

PubMed

Todorova, Mariana G; Soria, Bernat; Quesada, Ivan

2008-02-01

Pluripotent embryonic stem (ES) cells are capable of maintaining a self-renewal state and have the potential to differentiate into derivatives of all three embryonic germ layers. Despite their importance in cell therapy and developmental biology, the mechanisms whereby ES cells remain in a proliferative and pluripotent state are still not fully understood. Here we establish a critical role of gap junctional intercellular communication (GJIC) and connexin43 (Cx43) in both processes. Pharmacological blockers of GJIC and Cx43 down-regulation by small interfering RNA (siRNA) caused a profound inhibitory effect on GJIC, as evidenced by experiments of fluorescence recovery after photobleaching. This deficient intercellular communication in ES cells induced a loss of their pluripotent state, which was manifested in morphological changes, a decrease in alkaline phosphatase activity, Oct-3/4 and Nanog expression, as well as an up-regulation of several differentiation markers. A decrease in the proliferation rate was also detected. Under these conditions, the formation of embryoid bodies from mouse ES cells was impaired, although this inhibition was reversible upon restoration of GJIC. Our findings define a major function of GJIC in the regulation of self-renewal and maintenance of pluripotency in ES cells. (c) 2007 Wiley-Liss, Inc.

[Intercellular relationship of notochord determination of Xenopus laevis].

PubMed

Zeng, M B; Zhou, M Y; Wang, Y

1995-09-01

During the process of determination, the presumptive notochord is situated beneath neuroepithelium, flanked at two sides by presumptive somites and underlain with archenteron roof ventrally. Among these neighbouring embryonic tissues, presumptive somites were found to exert the main influence on notochord determination. By electron microscopic observations, the presumptive notochord and somite cells were seen to situate either close to each other (plate I, Fig. 1) or connected by cytoplasmic processes forming intercellular lumen (plate I, Fig. 5). Coated pits and coated vesicles appeared at the outer surface of both types of cells (plate I, Figs. 1-4). For the presumptive somite cells, spherical bodies of different sizes and variable contents were observed either near or protruding from the outer surface (plate II, Figs. 6-10). The spherical bodies were also found in the intercellular lumen (plate III, Fig. 11). These spherical bodies were mainly composed of granules, loosely scattered or densely packed. The granules were of similar size and similar shade of electron staining as those of ribosomes of the presumptive somite cells. For the presumptive notochord cells, no spherical bodies of the above mentioned type were found, but phenomenon of engulfing luminal material was observed (plate III, Fig. 12). The significance of the appearance of these spherical bodies in the determination of notochord cells has been discussed.

A systems biology approach to study systemic inflammation.

PubMed

Chen, Bor-Sen; Wu, Chia-Chou

2014-01-01

Systemic inflammation needs a precise control on the sequence and magnitude of occurring events. The high throughput data on the host-pathogen interactions gives us an opportunity to have a glimpse on the systemic inflammation. In this article, a dynamic Candida albicans-zebrafish interactive infectious network is built as an example to demonstrate how systems biology approach can be used to study systematic inflammation. In particular, based on microarray data of C. albicans and zebrafish during infection, the hyphal growth, zebrafish, and host-pathogen intercellular PPI networks were combined to form an integrated infectious PPI network that helps us understand the systematic mechanisms underlying the pathogenicity of C. albicans and the immune response of the host. The signaling pathways for morphogenesis and hyphal growth of C. albicans were 2 significant interactions found in the intercellular PPI network. Two cellular networks were also developed corresponding to the different infection stages (adhesion and invasion), and then compared with each other to identify proteins to gain more insight into the pathogenic role of hyphal development in the C. albicans infection process. Important defense-related proteins in zebrafish were predicted using the same approach. This integrated network consisting of intercellular invasion and cellular defense processes during infection can improve medical therapies and facilitate development of new antifungal drugs.

Long-range intercellular Ca2+ wave patterns

NASA Astrophysics Data System (ADS)

Tabi, C. B.; Maïna, I.; Mohamadou, A.; Ekobena, H. P. F.; Kofané, T. C.

2015-10-01

Modulational instability is utilized to investigate intercellular Ca2+ wave propagation in an array of diffusively coupled cells. Cells are supposed to be connected via paracrine signaling, where long-range effects, due to the presence of extracellular messengers, are included. The multiple-scale expansion is used to show that the whole dynamics of Ca2+ waves, from the endoplasmic reticulum to the cytosol, can be reduced to a single differential-difference nonlinear equation whose solutions are assumed to be plane waves. Their linear stability analysis is studied, with emphasis on the impact of long-range coupling, via the range parameter s. It is shown that s, as well as the number of interacting cells, importantly modifies the features of modulational instability, as small values of s imply a strong coupling, and increasing its value rather reduces the problem to a first-neighbor one. Our theoretical findings are numerically tested, as the generic equations are fully integrated, leading to the emergence of nonlinear patterns of Ca2+ waves. Strong long-range coupling is pictured by extended trains of breather-like structures whose frequency decreases with increasing s. We also show numerically that the number of interacting cells plays on the spatio-temporal formation of Ca2+ patterns, whilst the quasi-perfect intercellular communication depends on the paracrine coupling parameter.

Special sensors for deformation measurements of different construction materials and structures

NASA Astrophysics Data System (ADS)

Glisic, Branko; Inaudi, Daniele; Kronenberg, Pascal; LLoret, Sandra; Vurpillot, Samuel

1999-05-01

SOFO is a fiber optic sensor system that allows the monitoring of micrometer deformations over measurement bases up to a few meters. It is particularly adapted to measure civil structures built with conventional civil engineering materials (concrete, steel and timber). It has been successfully tested in different types of structures such as bridges, tunnels and piles. The application of the system is however limited in some case when unusual materials are used in the construction and in other cases by the dimensions of standard SOFO sensors. To extend the domain of application of the current system, special sensors have been developed. In this paper we present four special SOFO sensors: long, membrane, thin and stiff sensors. The long sensor has a measurement basis of several tenths of meters and its purpose is the measurement of deformations in massive and large structures (dames, tunnels). The membrane sensor is for use on laminated materials (e.g. membrane roofing) and it is easy to install by simply gluing it to the structure to be monitored. Since standard sensors can not be used for thin mortar layers because of their cross- section, a thin sensor has been developed, too. Finally, the aim of the stiff sensor is to determine the hardening (solidification) time of concrete. This time is determined by comparing the deformations of a stiff and a standard sensor, closely placed in the concrete at the very early age. The design of these sensors is presented along with significant application examples.

Intercellular communication in the immune system: differential expression of connexin40 and 43, and perturbation of gap junction channel functions in peripheral blood and tonsil human lymphocyte subpopulations

PubMed Central

Oviedo‐orta, E; Hoy, T; Evans, W H

2000-01-01

The distribution and function of connexins (integral membrane proteins assembled into gap junction intercellular communication channels) were studied in human lymphocyte subpopulations. The expression of mRNA encoding connexins in peripheral blood and tonsil‐derived T, B and natural killer (NK) lymphocytes was examined. Connexin43 (Cx43) mRNA was expressed in peripheral blood and tonsil lymphocytes, but Cx40 mRNA expression was confined to tonsil‐derived T and B lymphocytes; Cx26, Cx32, Cx37 and Cx45 were not detected by reverse transcription–polymerase chain reaction (RT–PCR). Western blot analysis also demonstrated the presence of Cx40 and Cx43 proteins in T and B lymphocytes in a manner coincidental to the mRNA detection. Stimulation in vitro of T and B lymphocytes with phytohaemagglutinin (PHA) and lipopolysaccharide (LPS), respectively, increased Cx40 and Cx43 protein expression. Flow cytometric analysis, using antibodies to extracellular loop amino acid sequences of connexins, confirmed the surface expression of connexins in all lymphocyte subpopulations. Assembly of connexins into gap junctions providing direct intercellular channels linking attached lymphocytes was demonstrated by using a dye transfer technique. The exchange of dye between lymphocytes was inhibited by a connexin extracellular loop mimetic peptide and α‐glycyrrhetinic acid, two reagents that restrict intercellular communication across gap junctions. Dye coupling occurred between homologous and heterologous co‐cultures of T and B lymphocytes, and was not influenced by their stimulation with PHA and LPS. The connexin mimetic peptide caused a significant decrease in the in vitro synthesis of immunoglobulin M (IgM) by T‐ and B‐lymphocyte co‐cultured populations in the presence or absence of stimulation by PHA. The results identify connexins as important cell surface components that modulate immune processes. PMID:10792506

The tight junction protein ZO-2 and Janus kinase 1 mediate intercellular communications in vascular smooth muscle cells

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tkachuk, Natalia; Tkachuk, Sergey; Patecki, Margret

2011-07-08

Highlights: {yields} The tight junction protein ZO-2 associates with Jak1 in vascular smooth muscle cells via ZO-2 N-terminal fragment. {yields} Jak1 mediates ZO-2 tyrosine phosphorylation and ZO-2 localization to the sites of homotypic intercellular contacts. {yields} The urokinase receptor uPAR regulates ZO-2/Jak1 functional association. {yields} The ZO-2/Jak1/uPAR signaling complex is required for vascular smooth muscle cells functional network formation. -- Abstract: Recent evidence points to a multifunctional role of ZO-2, the tight junction protein of the MAGUK (membrane-associated guanylate kinase-like) family. Though ZO-2 has been found in cell types lacking tight junction structures, such as vascular smooth muscle cells (VSMC),more » little is known about ZO-2 function in these cells. We provide evidence that ZO-2 mediates specific homotypic cell-to-cell contacts between VSMC. Using mass spectrometry we found that ZO-2 is associated with the non-receptor tyrosine kinase Jak1. By generating specific ZO-2 constructs we further found that the N-terminal fragment of ZO-2 molecule is responsible for this interaction. Adenovirus-based expression of Jak1 inactive mutant demonstrated that Jak1 mediates ZO-2 tyrosine phosphorylation. By means of RNA silencing, expression of Jak1 mutant form and fluorescently labeled ZO-2 fusion protein we further specified that active Jak1, but not Jak1 inactive mutant, mediates ZO-2 localization to the sites of intercellular contacts. We identified the urokinase receptor uPAR as a pre-requisite for these cellular events. Functional requirement of the revealed signaling complex for VSMC network formation was confirmed in experiments using Matrigel and in contraction assay. Our findings imply involvement of the ZO-2 tight junction independent signaling complex containing Jak1 and uPAR in VSMC intercellular communications. This mechanism may contribute to vascular remodeling in occlusive cardiovascular diseases and in arteriogenesis.« less

A distinct profile of serum levels of soluble intercellular adhesion molecule-1 and intercellular adhesion molecule-3 in mycosis fungoides and Sézary syndrome.

PubMed

López-Lerma, Ingrid; Estrach, Maria Teresa

2009-08-01

Cell adhesion molecules (CAMs) play a pivotal role in cutaneous localization of T cells. Tissue-selective localization of T lymphocytes to the skin is crucial for immune surveillance and in the pathogenesis of skin disorders. To detect the profile of soluble CAMs in patients with cutaneous T-cell lymphoma (CTCL), we investigated the levels of intercellular adhesion molecule-1 (ICAM-1, soluble ICAM-1 [sICAM-1]); intercellular adhesion molecule-3 (sICAM-3); vascular cell adhesion molecule-1 (sVCAM-1); and E-selectin (sE-selectin) in sera from patients with T-cell-mediated skin diseases. Serum levels of the 4 CAMs were measured by enzyme-linked immunosorbent assay in 42 participants including 11 patients with early stages of CTCL; 7 with advanced stages of CTCL including Sézary syndrome; 12 with inflammatory skin diseases (psoriasis and atopic dermatitis); 8 with skin diseases that may evolve into CTCL; and healthy individuals. Levels were correlated with biological parameters known as prognostic factors in non-Hodgkin lymphomas. In patients with CTCL, significantly increased levels of sICAM-1 and sICAM-3 were found when compared with healthy individuals and patients with inflammatory dermatosis. Soluble E-selectin and sVCAM-1 levels were not increased. There were significant positive correlations between sICAM-1 and sICAM-3 levels and each of them with beta2-microglobulin levels. Limited number of patients was a limitation. There is a distinct profile of soluble CAMs in patients with CTCL. However, future studies with a larger group of patients are needed to confirm these findings. We propose that high sICAM-1 and sICAM-3 levels have important implications in the context of immune response and immune surveillance in these patients.

Re-establishment of gap junctional intercellular communication (GJIC) between human endometrial carcinomas by prostaglandin E(2).

PubMed

Schlemmer, Scott R; Kaufman, David G

2012-12-01

Reduced intercellular communication via gap junctions is correlated with carcinogenesis. Gap junctional intercellular communication (GJIC), between normal human endometrial epithelial cells is enhanced when endometrial stromal cells were present in culture. This enhancement of GJIC between normal epithelial cells also occurs when they are cultured in medium conditioned by stromal cells. This observation indicated that a soluble compound (or compounds) produced and secreted by stromal cells mediates GJIC in epithelial cells. Previous studies have shown that endometrial stromal cells release prostaglandin E(2) (PGE(2)) and prostaglandin F(2α) (PGF(2α)) under physiological conditions. When we evaluated the response of normal endometrial epithelial cells to various concentrations of PGE(2,) we found enhanced GJIC with 1nM PGE(2). This is a smaller increase in GJIC than that induced by medium conditioned by stromal cells. When the extracellular concentration of PGE(2) was measured after incubation with stromal cells, it was found to be similar to the concentrations showing maximal GJIC between the normal epithelial cells. When indomethacin was used to inhibit prostaglandin synthesis by stromal cells, GJIC was reduced but not eliminated between normal endometrial epithelial cells. These observations suggest that although PGE(2) secreted by stromal cells is an important mediator of GJIC between the epithelial cells, it is not the sole mediator. Transformed endometrial epithelial cells did not demonstrate GJIC even in the presence of stromal cells. However, we were able to re-establish GJIC in transformed epithelial cells when we added PGE(2) to the cells. Our findings show that PGE(2) may serve as an intercellular mediator between stromal and epithelial cells that regulates GJIC in normal and malignant epithelial cells. This suggests that maintenance of GJIC by preserving or replacing PGE(2) secretion by endometrial stromal cells may have the potential to suppress carcinogenesis in endometrial epithelial cells. Copyright © 2012 Elsevier Inc. All rights reserved.

Improving the Post-Stroke Therapeutic Potency of Mesenchymal Multipotent Stromal Cells by Cocultivation With Cortical Neurons: The Role of Crosstalk Between Cells.

PubMed

Babenko, Valentina A; Silachev, Denis N; Zorova, Ljubava D; Pevzner, Irina B; Khutornenko, Anastasia A; Plotnikov, Egor Y; Sukhikh, Gennady T; Zorov, Dmitry B

2015-09-01

The goal of the present study was to maximally alleviate the negative impact of stroke by increasing the therapeutic potency of injected mesenchymal multipotent stromal cells (MMSCs). To pursue this goal, the intercellular communications of MMSCs and neuronal cells were studied in vitro. As a result of cocultivation of MMSCs and rat cortical neurons, we proved the existence of intercellular contacts providing transfer of cellular contents from one cell to another. We present evidence of intercellular exchange with fluorescent probes specifically occupied by cytosol with preferential transfer from neurons toward MMSCs. In contrast, we observed a reversed transfer of mitochondria (from MMSCs to neural cells). Intravenous injection of MMSCs in a postischemic period alleviated the pathological indexes of a stroke, expressed as a lower infarct volume in the brain and partial restoration of neurological status. Also, MMSCs after cocultivation with neurons demonstrated more profound neuroprotective effects than did unprimed MMSCs. The production of the brain-derived neurotrophic factor was slightly increased in MMSCs, and the factor itself was redistributed in these cells after cocultivation. The level of Miro1 responsible for intercellular traffic of mitochondria was increased in MMSCs after cocultivation. We conclude that the exchange by cellular compartments between neural and stem cells improves MMSCs' protective abilities for better rehabilitation after stroke. This could be used as an approach to enhance the therapeutic benefits of stem cell therapy to the damaged brain. The idea of priming stem cells before practical use for clinical purposes was applied. Thus, cells were preconditioned by coculturing them with the targeted cells (i.e., neurons for the treatment of brain pathological features) before the transfusion of stem cells to the organism. Such priming improved the capacity of stem cells to treat stroke. Some additional minimal study will be required to develop a detailed protocol for coculturing followed by cell separation. ©AlphaMed Press.

Intercellular Calcium Waves in HeLa Cells Expressing GFP-labeled Connexin 43, 32, or 26

PubMed Central

Paemeleire, Koen; Martin, Patricia E. M.; Coleman, Sharon L.; Fogarty, Kevin E.; Carrington, Walter A.; Leybaert, Luc; Tuft, Richard A.; Evans, W. Howard; Sanderson, Michael J.

2000-01-01

This study was undertaken to obtain direct evidence for the involvement of gap junctions in the propagation of intercellular Ca2+ waves. Gap junction-deficient HeLa cells were transfected with plasmids encoding for green fluorescent protein (GFP) fused to the cytoplasmic carboxyl termini of connexin 43 (Cx43), 32 (Cx32), or 26 (Cx26). The subsequently expressed GFP-labeled gap junctions rendered the cells dye- and electrically coupled and were detected at the plasma membranes at points of contact between adjacent cells. To correlate the distribution of gap junctions with the changes in [Ca2+]i associated with Ca2+ waves and the distribution of the endoplasmic reticulum (ER), cells were loaded with fluorescent Ca2+-sensitive (fluo-3 and fura-2) and ER membrane (ER-Tracker) dyes. Digital high-speed microscopy was used to collect a series of image slices from which the three-dimensional distribution of the gap junctions and ER were reconstructed. Subsequently, intercellular Ca2+ waves were induced in these cells by mechanical stimulation with or without extracellular apyrase, an ATP-degrading enzyme. In untransfected HeLa cells and in the absence of apyrase, cell-to-cell propagating [Ca2+]i changes were characterized by initiating Ca2+ puffs associated with the perinuclear ER. By contrast, in Cx–GFP-transfected cells and in the presence of apyrase, [Ca2+]i changes were propagated without initiating perinuclear Ca2+ puffs and were communicated between cells at the sites of the Cx–GFP gap junctions. The efficiency of Cx expression determined the extent of Ca2+ wave propagation. These results demonstrate that intercellular Ca2+ waves may be propagated simultaneously via an extracellular pathway and an intracellular pathway through gap junctions and that one form of communication may mask the other. PMID:10793154

Transfected connexin45 alters gap junction permeability in cells expressing endogenous connexin43

PubMed Central

1995-01-01

Many cells express multiple connexins, the gap junction proteins that interconnect the cytosol of adjacent cells. Connexin43 (Cx43) channels allow intercellular transfer of Lucifer Yellow (LY, MW = 443 D), while connexin45 (Cx45) channels do not. We transfected full-length or truncated chicken Cx45 into a rat osteosarcoma cell line ROS-17/2.8, which expresses endogenous Cx43. Both forms of Cx45 were expressed at high levels and colocalized with Cx43 at plasma membrane junctions. Cells transfected with full-length Cx45 (ROS/Cx45) and cells transfected with Cx45 missing the 37 carboxyl-terminal amino acids (ROS/Cx45tr) showed 30-60% of the gap junctional conductance exhibited by ROS cells. Intercellular transfer of three negatively charged fluorescent reporter molecules was examined. In ROS cells, microinjected LY was transferred to an average of 11.2 cells/injected cell, while dye transfer between ROS/Cx45 cells was reduced to 3.9 transfer between ROS/Cx45 cells was reduced to 3.9 cells. In contrast, ROS/Cx45tr cells transferred LY to > 20 cells. Transfer of calcein (MW = 623 D) was also reduced by approximately 50% in ROS/Cx45 cells, but passage of hydroxycoumarin carboxylic acid (HCCA; MW = 206 D) was only reduced by 35% as compared to ROS cells. Thus, introduction of Cx45 altered intercellular coupling between cells expressing Cx43, most likely the result of direct interaction between Cx43 and Cx45. Transfection of Cx45tr and Cx45 had different effects in ROS cells, consistent with a role of the carboxyl-terminal domain of Cx45 in determining gap junction permeability or interactions between connexins. These data suggest that coexpression of multiple connexins may enable cells to achieve forms of intercellular communication that cannot be attained by expression of a single connexin. PMID:7642714

The Peptidoglycan-Binding Protein SjcF1 Influences Septal Junction Function and Channel Formation in the Filamentous Cyanobacterium Anabaena.

PubMed

Rudolf, Mareike; Tetik, Nalan; Ramos-León, Félix; Flinner, Nadine; Ngo, Giang; Stevanovic, Mara; Burnat, Mireia; Pernil, Rafael; Flores, Enrique; Schleiff, Enrico

2015-06-30

Filamentous, heterocyst-forming cyanobacteria exchange nutrients and regulators between cells for diazotrophic growth. Two alternative modes of exchange have been discussed involving transport either through the periplasm or through septal junctions linking adjacent cells. Septal junctions and channels in the septal peptidoglycan are likely filled with septal junction complexes. While possible proteinaceous factors involved in septal junction formation, SepJ (FraG), FraC, and FraD, have been identified, little is known about peptidoglycan channel formation and septal junction complex anchoring to the peptidoglycan. We describe a factor, SjcF1, involved in regulation of septal junction channel formation in the heterocyst-forming cyanobacterium Anabaena sp. strain PCC 7120. SjcF1 interacts with the peptidoglycan layer through two peptidoglycan-binding domains and is localized throughout the cell periphery but at higher levels in the intercellular septa. A strain with an insertion in sjcF1 was not affected in peptidoglycan synthesis but showed an altered morphology of the septal peptidoglycan channels, which were significantly wider in the mutant than in the wild type. The mutant was impaired in intercellular exchange of a fluorescent probe to a similar extent as a sepJ deletion mutant. SjcF1 additionally bears an SH3 domain for protein-protein interactions. SH3 binding domains were identified in SepJ and FraC, and evidence for interaction of SjcF1 with both SepJ and FraC was obtained. SjcF1 represents a novel protein involved in structuring the peptidoglycan layer, which links peptidoglycan channel formation to septal junction complex function in multicellular cyanobacteria. Nonetheless, based on its subcellular distribution, this might not be the only function of SjcF1. Cell-cell communication is central not only for eukaryotic but also for multicellular prokaryotic systems. Principles of intercellular communication are well established for eukaryotes, but the mechanisms and components involved in bacteria are just emerging. Filamentous heterocyst-forming cyanobacteria behave as multicellular organisms and represent an excellent model to study prokaryotic cell-cell communication. A path for intercellular metabolite exchange appears to involve transfer through molecular structures termed septal junctions. They are reminiscent of metazoan gap junctions that directly link adjacent cells. In cyanobacteria, such structures need to traverse the peptidoglycan layers in the intercellular septa of the filament. Here we describe a factor involved in the formation of channels across the septal peptidoglycan layers, thus contributing to the multicellular behavior of these organisms. Copyright © 2015 Rudolf et al.

Separation and Measurement of Direct and Indirect Effects of Light on Stomata 1

PubMed Central

Sharkey, Thomas D.; Raschke, Klaus

1981-01-01

Conductance for water vapor, assimilation of CO2, and intercellular CO2 concentration of leaves of five species were determined at various irradiances and ambient CO2 concentrations. Conductance and assimilation were then plotted as functions of irradiance and intercellular CO2 concentration. The slopes of these curves allowed us to estimate infinitesimal changes in conductance (and assimilation) that occurred when irradiance changed and intercellular CO2 concentration was constant, and when CO2 concentration changed and irradiance was constant. On leaves of Xanthium strumarium L., Gossypium hirsutum L., Phaseolus vulgaris L., and Perilla frutescens (L.), Britt., the stomatal response to light was determined to be mainly a direct response to light and to a small extent only a response to changes in intercellular CO2 concentration. This was also true for stomata of Zea mays L., except at irradiances < 150 watts per square meter, when stomata responded primarily to the depletion of the intercellular spaces of CO2 which in turn was caused by changes in the assimilation of CO2. Stomata responded to light even in leaves whose net exchange of CO2 was reduced to zero through application of the inhibitor of photosynthetic electron transport, cyanazine (2-chloro-4[1-cyano-1-methylethylamino]-6-ethylamino-S-triazine). When leaves were inverted and irradiated on the abaxial surface, conductance decreased in the shaded and increased in the illuminated epidermis, indicating that the photoreceptor pigment(s) involved are located in the epidermis (presumably in the guard cells). In leaves of X. strumarium, the direct effect of light on conductance is primarily a response to blue light. Stomatal responses to CO2 and to light opposed each other. In X. strumarium, stomatal opening in response to light was strongest in CO2 free air and saturated at lower irradiances than in CO2 containing air. Conversely, stomatal closure in response to CO2 was strongest in darkness and it decreased as irradiance increased. In X. strumarium, P. vulgaris, and P. frutescens, an irradiance of 300 watts per square meter was sufficient to eliminate the stomatal response to CO2 altogether. Application of abscisic acid, or an increase in vapor pressure deficit, or a decrease in leaf temperature reduced the stomatal conductance at light saturation, but when the data were normalized with respect to the conductance at the highest irradiance, the various curves were congruent. PMID:16661884

The study of the intercellular trafficking of the fusion proteins of herpes simplex virus protein VP22.

PubMed

Xue, Xiaodong; Huang, Jianhua; Wang, Huishan

2014-01-01

Genetic modifications can improve the therapeutic efficacy of mesenchymal stem cell (MSC) transplantation in myocardial infarction. However, so far, the efficiency of MSC modification is very low. Seeking for a more efficient way of MSC modification, we investigated the possibility of employing the intercellular trafficking capacity of the herpes simplex virus type-1 tegument protein VP22 on the enhancement of MSC modification. Plasmids pVP22-myc, pVP22-EGFP, pEGFP-VP22, pVP22-hBcl-xL and phBcl-xL-VP22 were constructed for the expressions of the myc-tagged VP22 and the fusion proteins VP22-EGFP, EGFP-VP22, VP22-hBcl-xL and hBcl-xL-VP22. MSCs were isolated from rat bone marrow and the surface markers were identified by Flowcytometry. COS-1 cells were transfected with the above plasmids and co-cultured with untransfected MSCs, the intercellular transportations of the constructed proteins were studied by immunofluorescence. The solubility of VP22-hBcl-xL and hBcl-xL-VP22 was analyzed by Western blot. VP22-myc could be expressed in and spread between COS-1 cells, which indicates the validity of our VP22 expression construct. Flowcytometry analysis revealed that the isolated MSCs were CD29, CD44, and CD90 positive and were negative for the hematopoietic markers, CD34 and CD45. The co-culturing and immunofluorescence assay showed that VP22-myc, VP22-EGFP and EGFP-VP22 could traffic between COS-1 cells and MSCs, while the evidence of intercellular transportation of VP22-hBcl-xL and hBcl-xL-VP22 was not detected. Western blot analysis showed that VP22-hBcl-xL and hBcl-xL-VP22 were both insoluble in the cell lysate suggesting interactions of the fusion proteins with other cellular components. The intercellular trafficking of VP22-myc, VP22-EGFP and EGFP-VP22 between COS-1 cells and MSCs presents an intriguing prospect in the therapeutic application of VP22 as a delivery vehicle which enhances genetic modifications of MSCs. However, VP22-hBcl-xL and hBcl-xL-VP22 failed to spread between cells, which are due to the insolubility of the fusion protein incurred by interactions with other cellular components.

The Study of the Intercellular Trafficking of the Fusion Proteins of Herpes Simplex Virus Protein VP22

PubMed Central

Xue, Xiaodong; Huang, Jianhua; Wang, Huishan

2014-01-01

Background Genetic modifications can improve the therapeutic efficacy of mesenchymal stem cell (MSC) transplantation in myocardial infarction. However, so far, the efficiency of MSC modification is very low. Seeking for a more efficient way of MSC modification, we investigated the possibility of employing the intercellular trafficking capacity of the herpes simplex virus type-1 tegument protein VP22 on the enhancement of MSC modification. Methods Plasmids pVP22-myc, pVP22-EGFP, pEGFP-VP22, pVP22-hBcl-xL and phBcl-xL-VP22 were constructed for the expressions of the myc-tagged VP22 and the fusion proteins VP22-EGFP, EGFP-VP22, VP22-hBcl-xL and hBcl-xL-VP22. MSCs were isolated from rat bone marrow and the surface markers were identified by Flowcytometry. COS-1 cells were transfected with the above plasmids and co-cultured with untransfected MSCs, the intercellular transportations of the constructed proteins were studied by immunofluorescence. The solubility of VP22-hBcl-xL and hBcl-xL-VP22 was analyzed by Western blot. Results VP22-myc could be expressed in and spread between COS-1 cells, which indicates the validity of our VP22 expression construct. Flowcytometry analysis revealed that the isolated MSCs were CD29, CD44, and CD90 positive and were negative for the hematopoietic markers, CD34 and CD45. The co-culturing and immunofluorescence assay showed that VP22-myc, VP22-EGFP and EGFP-VP22 could traffic between COS-1 cells and MSCs, while the evidence of intercellular transportation of VP22-hBcl-xL and hBcl-xL-VP22 was not detected. Western blot analysis showed that VP22-hBcl-xL and hBcl-xL-VP22 were both insoluble in the cell lysate suggesting interactions of the fusion proteins with other cellular components. Conclusions The intercellular trafficking of VP22-myc, VP22-EGFP and EGFP-VP22 between COS-1 cells and MSCs presents an intriguing prospect in the therapeutic application of VP22 as a delivery vehicle which enhances genetic modifications of MSCs. However, VP22-hBcl-xL and hBcl-xL-VP22 failed to spread between cells, which are due to the insolubility of the fusion protein incurred by interactions with other cellular components. PMID:24955582

Structure of a novel antibacterial toxin that exploits elongation factor Tu to cleave specific transfer RNAs

DOE PAGES

Michalska, Karolina; Gucinski, Grant C.; Garza-Sanchez, Fernando; ...

2017-08-11

Contact-dependent growth inhibition (CDI) is a mechanism of inter-cellular competition in which Gram-negative bacteria exchange polymorphic toxins using type V secretion systems. Here, we present structures of the CDI toxin from Escherichia coli NC101 in ternary complex with its cognate immunity protein and elongation factor Tu (EF-Tu). The toxin binds exclusively to domain 2 of EF-Tu, partially overlapping the site that interacts with the 3'-end of aminoacyl-tRNA (aa-tRNA). The toxin exerts a unique ribonuclease activity that cleaves the single-stranded 3'-end from tRNAs that contain guanine discriminator nucleotides. EF-Tu is required to support this tRNase activity in vitro, suggesting the toxinmore » specifically cleaves substrate in the context of GTP·EF-Tu·aa-tRNA complexes. However, superimposition of the toxin domain onto previously solved GTP·EF-Tu·aa-tRNA structures reveals potential steric clashes with both aa-tRNA and the switch I region of EF-Tu. Further, the toxin induces conformational changes in EF-Tu, displacing a β-hairpin loop that forms a critical salt-bridge contact with the 3'-terminal adenylate of aa-tRNA. Altogether, these observations suggest that the toxin remodels GTP·EF-Tu·aa-tRNA complexes to free the 3'-end of aa-tRNA for entry into the nuclease active site.« less

Structure of a novel antibacterial toxin that exploits elongation factor Tu to cleave specific transfer RNAs

DOE Office of Scientific and Technical Information (OSTI.GOV)

Michalska, Karolina; Gucinski, Grant C.; Garza-Sanchez, Fernando

Contact-dependent growth inhibition (CDI) is a mechanism of inter-cellular competition in which Gram-negative bacteria exchange polymorphic toxins using type V secretion systems. Here, we present structures of the CDI toxin from Escherichia coli NC101 in ternary complex with its cognate immunity protein and elongation factor Tu (EF-Tu). The toxin binds exclusively to domain 2 of EF-Tu, partially overlapping the site that interacts with the 3'-end of aminoacyl-tRNA (aa-tRNA). The toxin exerts a unique ribonuclease activity that cleaves the single-stranded 3'-end from tRNAs that contain guanine discriminator nucleotides. EF-Tu is required to support this tRNase activity in vitro, suggesting the toxinmore » specifically cleaves substrate in the context of GTP·EF-Tu·aa-tRNA complexes. However, superimposition of the toxin domain onto previously solved GTP·EF-Tu·aa-tRNA structures reveals potential steric clashes with both aa-tRNA and the switch I region of EF-Tu. Further, the toxin induces conformational changes in EF-Tu, displacing a β-hairpin loop that forms a critical salt-bridge contact with the 3'-terminal adenylate of aa-tRNA. Altogether, these observations suggest that the toxin remodels GTP·EF-Tu·aa-tRNA complexes to free the 3'-end of aa-tRNA for entry into the nuclease active site.« less

Understanding the Low Photosynthetic Rates of Sun and Shade Coffee Leaves: Bridging the Gap on the Relative Roles of Hydraulic, Diffusive and Biochemical Constraints to Photosynthesis

PubMed Central

Martins, Samuel C. V.; Galmés, Jeroni; Cavatte, Paulo C.; Pereira, Lucas F.; Ventrella, Marília C.; DaMatta, Fábio M.

2014-01-01

It has long been held that the low photosynthetic rates (A) of coffee leaves are largely associated with diffusive constraints to photosynthesis. However, the relative limitations of the stomata and mesophyll to the overall diffusional constraints to photosynthesis, as well as the coordination of leaf hydraulics with photosynthetic limitations, remain to be fully elucidated in coffee. Whether the low actual A under ambient CO2 concentrations is associated with the kinetic properties of Rubisco and high (photo)respiration rates also remains elusive. Here, we provide a holistic analysis to understand the causes associated with low A by measuring a variety of key anatomical/hydraulic and photosynthetic traits in sun- and shade-grown coffee plants. We demonstrate that leaf hydraulic architecture imposes a major constraint on the maximisation of the photosynthetic gas exchange of coffee leaves. Regardless of the light treatments, A was mainly limited by stomatal factors followed by similar limitations associated with the mesophyll and biochemical constraints. No evidence of an inefficient Rubisco was found; rather, we propose that coffee Rubisco is well tuned for operating at low chloroplastic CO2 concentrations. Finally, we contend that large diffusive resistance should lead to large CO2 drawdown from the intercellular airspaces to the sites of carboxylation, thus favouring the occurrence of relatively high photorespiration rates, which ultimately leads to further limitations to A. PMID:24743509

Initial characterization of a low-molecular-weight factor enhancing the checkpoint response.

PubMed

Fan, Xiaoxiang; Cheong, Nge; Iliakis, George

2010-10-01

In higher eukaryotes, DNA double-strand breaks (DSBs) induced by ionizing radiation activate checkpoints that delay progression through the cell cycle. Compared to delays in other phases of the cell cycle, delays induced in G(2) are longer and frequently correlate with resistance to killing by radiation. Therefore, modulation of the G(2) checkpoint offers a means to modulate cellular radiosensitivity. Although compounds are known that reduce the G(2) checkpoint and act as radiosensitizers, compounds enhancing this checkpoint have not been reported. Here we summarize evidence for a factor with such properties. We show that a highly radioresistant rat embryo fibroblast (REF) cell line displays a strong G(2) checkpoint partly as a result of a factor excreted into the growth medium by nonirradiated cells. Various tests indicate that this G(2)-arrest modulating activity (GAMA) is a small molecule showing detectable retention only after passing through filters with a molecular weight cutoff limit of less than 1,000 Da. GAMA is heat stable and resistant to treatment with proteases or nucleases. Electroelution tests show that GAMA is uncharged at neutral pH, a result that is in agreement with the observed failure to bind S- or Q-Sepharose. Investigations on the mechanism of GAMA function indicate ligand-receptor interactions and allow the classification of cells as producers, responders or both. Compounds with properties such as those of GAMA bridge intercellular communication with the DNA damage response and may function as radioprotectors.

Gifford-McMahon refrigerator with split cold head

NASA Technical Reports Server (NTRS)

Forth, H. J.; Heisig, R.; Klein, H. H.

1983-01-01

Leybold-Heraeus Co. have developed, built and successfully tested a Gifford-McMahon cryocooler with splitted cold head for cooling a cryopump. The refrigerating part of the cold head and the gas flow control device have been separated (splitted cold head) and the distance between them is bridged by only two thin lines for carrying the working gas. Due to this separation the size of the refrigerating part is virtually defined only by the size of the displacers whilst the gas flow control device can be of any desired design. It has been shown that dimensioning of the connecting lines and the corresponding losses became less critical with increasing size of the expander, but additional cooling in proportion to the refrigerating capacity is required.

MEMS switches having non-metallic crossbeams

NASA Technical Reports Server (NTRS)

Scardelletti, Maximillian C (Inventor)

2009-01-01

A RF MEMS switch comprising a crossbeam of SiC, supported by at least one leg above a substrate and above a plurality of transmission lines forming a CPW. Bias is provided by at least one layer of metal disposed on a top surface of the SiC crossbeam, such as a layer of chromium followed by a layer of gold, and extending beyond the switch to a biasing pad on the substrate. The switch utilizes stress and conductivity-controlled non-metallic thin cantilevers or bridges, thereby improving the RF characteristics and operational reliability of the switch. The switch can be fabricated with conventional silicon integrated circuit (IC) processing techniques. The design of the switch is very versatile and can be implemented in many transmission line mediums.

Thickness effect of nickel oxide thin films on associated solution-processed write-once-read-many-times memory devices

NASA Astrophysics Data System (ADS)

Wang, Xiao Lin; Liu, Zhen; Wen, Chao; Liu, Yang; Wang, Hong Zhe; Chen, T. P.; Zhang, Hai Yan

2018-06-01

With self-prepared nickel acetate based solution, NiO thin films with different thicknesses have been fabricated by spin coating followed by thermal annealing. By forming a two-terminal Ag/NiO/ITO structure on glass, write-once-read-many-times (WORM) memory devices are realized. The WORM memory behavior is based on a permanent switching from an initial high-resistance state (HRS) to an irreversible low-resistance state (LRS) under the application of a writing voltage, due to the formation of a solid bridge across Ag and ITO electrodes by conductive filaments (CFs). The memory performance is investigated as a function of the NiO film thickness, which is determined by the number of spin-coated NiO layers. For devices with 4 and 6 NiO layers, data retention up to 104 s and endurance of 103 reading operations in the measurement range have been obtained with memory window maintained above four orders for both HRS and LRS. Before and after writing, the devices show the hopping and ohmic conduction behaviors, respectively, confirming that the CF formation could be the mechanism responsible for writing in the WORM memory devices.

Evaluation of Human Corneal Lenticule Quality After SMILE With Different Cap Thicknesses Using Scanning Electron Microscopy.

PubMed

Weng, Shengbei; Liu, Manli; Yang, Xiaonan; Liu, Fang; Zhou, Yugui; Lin, Haiqin; Liu, Quan

2018-01-01

To evaluate the surface characteristics of lenticules created by small-incision lenticule extraction (SMILE) with different cap thicknesses. This prospective study included 20 consecutive patients who underwent bilateral SMILE. Surface regularity of the extracted corneal lenticule was analyzed using scanning electron microscopy (SEM) combined with 2 methods: qualitative and quantitative regularity. Qualitative regularity of SEM images was graded by masked observers using an established scoring system. Quantitative regularity of SEM images was assessed by counting the total number and areas of tissue bridges using Image-Pro Plus software. Four different cap thickness of 120, 130, 140, and 150 μm were compared. Refractive outcomes of patients were measured at baseline and 1 month after surgery. As 10 specimens were not analyzable, only 30 eyes were included. Postoperatively, all eyes had postoperative uncorrected distance visual acuity of 20/20 or better; 43% had an unchanged corrected distance visual acuity; 43% gained 1 line; 10% lost 1 line. Ultrastructurally, surface irregularity was primarily caused by tissue bridges. The average surface regularity score obtained was 10.87 ± 2.40 for 120 μm, 10.78 ± 2.60 for 130 μm, 8.76 ± 2.16 for 140 μm, and 8.70 ± 2.66 for 150 μm (P < 0.001). The total number and areas of tissue bridges of 120 to 130 μm were significantly less than 140 to 150 μm (P < 0.05). Surface regularity decreased as cap thickness increased (P < 0.05). There is smoother appearance of the lenticular surface as seen through SEM when a thin cap is created compared with a thick cap qualitatively and quantitatively.

Ultrastructural study of the human neurohypophysis. III. Vascular and perivascular structures.

PubMed

Seyama, S; Pearl, G S; Takei, Y

1980-01-01

The vascular and perivascular regions of the human neurohypophysis were studied electron microscopically. The abluminal basement membrane, perivascular space, luminal basement membrane and endothelium are interposed between the neural parenchyma and the blood stream. The capillaries are fenestrated, with pores measuring 30 to 50 nm in diameter. The perivascular and intercellular spaces form prominent networks that penetrate between rows of neurohypophysial parenchymal cells. The perivascular space contains pericytes, histiocytes, fibroblasts and mast cells, with ultrastructural features typical of each cell type. No transitional forms between histiocytes and pericytes were observed. A schema for the extracellular flow of neurohypophysial hormones through the sinusoidal and perivascular spaces is proposed, suggesting an important role for the pituicytes and their intercellular junctions in the control of hormone release.

Receptor Complex Mediated Regulation of Symplastic Traffic.

PubMed

Stahl, Yvonne; Faulkner, Christine

2016-05-01

Plant receptor kinases (RKs) and receptor proteins (RPs) are involved in a plethora of cellular processes, including developmental decisions and immune responses. There is increasing evidence that plasmodesmata (PD)-localized RKs and RPs act as nexuses that perceive extracellular signals and convey them into intra- and intercellular responses by regulating the exchange of molecules through PD. How RK/RP complexes regulate the specific and nonspecific traffic of molecules through PD, and how these receptors are specifically targeted to PD, have been elusive but underpin comprehensive understanding of the function and regulation of the symplast. In this review we gather the current knowledge of RK/RP complex function at PD and how they might regulate intercellular traffic. Copyright © 2015 Elsevier Ltd. All rights reserved.

Terbinafine inhibits gap junctional intercellular communication.

PubMed

Lee, Ju Yeun; Yoon, Sei Mee; Choi, Eun Ju; Lee, Jinu

2016-09-15

Terbinafine is an antifungal agent that selectively inhibits fungal sterol synthesis by blocking squalene epoxidase. We evaluated the effect of terbinafine on gap junctional intercellular communication (GJIC). Fluorescence recovery after photobleaching (FRAP) and I-YFP GJIC assays revealed that terbinafine inhibits GJIC in a reversible and dose-dependent manner in FRT-Cx43 and LN215 cells. Treatment with terbinafine did not affect Cx43 phosphorylation status or intracellular Ca(2+) concentration, well-known action mechanisms of various GJIC blockers. While a structurally related chemical, naftifine, attenuated GJIC, epigallocatechin gallate, another potent squalene epoxidase inhibitor with a different structure, did not. These results suggest that terbinafine inhibits GJIC with a so far unknown mechanism of action. Copyright © 2016 Elsevier Inc. All rights reserved.

Enzyme-activated intracellular drug delivery with tubule clay nanoformulation

PubMed Central

Dzamukova, Maria R.; Naumenko, Ekaterina A.; Lvov, Yuri M.; Fakhrullin, Rawil F.

2015-01-01

Fabrication of stimuli-triggered drug delivery vehicle s is an important milestone in treating cancer. Here we demonstrate the selective anticancer drug delivery into human cells with biocompatible 50-nm diameter halloysite nanotube carriers. Physically-adsorbed dextrin end stoppers secure the intercellular release of brilliant green. Drug-loaded nanotubes penetrate through the cellular membranes and their uptake efficiency depends on the cells growth rate. Intercellular glycosyl hydrolases-mediated decomposition of the dextrin tube-end stoppers triggers the release of the lumen-loaded brilliant green, which allowed for preferable elimination of human lung carcinoma cells (А549) as compared with hepatoma cells (Hep3b). The enzyme-activated intracellular delivery of brilliant green using dextrin-coated halloysite nanotubes is a promising platform for anticancer treatment. PMID:25976444

Metal-electrode-free Window-like Organic Solar Cells with p-Doped Carbon Nanotube Thin-film Electrodes

PubMed Central

Jeon, Il; Delacou, Clement; Kaskela, Antti; Kauppinen, Esko I.; Maruyama, Shigeo; Matsuo, Yutaka

2016-01-01

Organic solar cells are flexible and inexpensive, and expected to have a wide range of applications. Many transparent organic solar cells have been reported and their success hinges on full transparency and high power conversion efficiency. Recently, carbon nanotubes and graphene, which meet these criteria, have been used in transparent conductive electrodes. However, their use in top electrodes has been limited by mechanical difficulties in fabrication and doping. Here, expensive metal top electrodes were replaced with high-performance, easy-to-transfer, aerosol-synthesized carbon nanotubes to produce transparent organic solar cells. The carbon nanotubes were p-doped by two new methods: HNO3 doping via ‘sandwich transfer’, and MoOx thermal doping via ‘bridge transfer’. Although both of the doping methods improved the performance of the carbon nanotubes and the photovoltaic performance of devices, sandwich transfer, which gave a 4.1% power conversion efficiency, was slightly more effective than bridge transfer, which produced a power conversion efficiency of 3.4%. Applying a thinner carbon nanotube film with 90% transparency decreased the efficiency to 3.7%, which was still high. Overall, the transparent solar cells had an efficiency of around 50% that of non-transparent metal-based solar cells (7.8%). PMID:27527565

Metal-electrode-free Window-like Organic Solar Cells with p-Doped Carbon Nanotube Thin-film Electrodes.

PubMed

Jeon, Il; Delacou, Clement; Kaskela, Antti; Kauppinen, Esko I; Maruyama, Shigeo; Matsuo, Yutaka

2016-08-16

Organic solar cells are flexible and inexpensive, and expected to have a wide range of applications. Many transparent organic solar cells have been reported and their success hinges on full transparency and high power conversion efficiency. Recently, carbon nanotubes and graphene, which meet these criteria, have been used in transparent conductive electrodes. However, their use in top electrodes has been limited by mechanical difficulties in fabrication and doping. Here, expensive metal top electrodes were replaced with high-performance, easy-to-transfer, aerosol-synthesized carbon nanotubes to produce transparent organic solar cells. The carbon nanotubes were p-doped by two new methods: HNO3 doping via 'sandwich transfer', and MoOx thermal doping via 'bridge transfer'. Although both of the doping methods improved the performance of the carbon nanotubes and the photovoltaic performance of devices, sandwich transfer, which gave a 4.1% power conversion efficiency, was slightly more effective than bridge transfer, which produced a power conversion efficiency of 3.4%. Applying a thinner carbon nanotube film with 90% transparency decreased the efficiency to 3.7%, which was still high. Overall, the transparent solar cells had an efficiency of around 50% that of non-transparent metal-based solar cells (7.8%).

Trinuclear ruthenium dioxolene complexes based on the bridging ligand hexahydroxytriphenylene: electrochemistry, spectroscopy, and near-infrared electrochromic behaviour associated with a reversible seven-membered redox chain.

PubMed

Grange, Christopher S; Meijer, Anthony J H M; Ward, Michael D

2010-01-07

The trinuclear complexes [{(R2bipy)2Ru}3(mu3-HHTP)](PF6)3 [1(PF6)3, R = H; 2(PF6)3, R = 4-tBu] contain three {Ru(R2bipy)2}2+ fragments connected to the triangular tris-chelating ligand hexahydroxytriphenylene (H6HHTP). This bridging ligand contains three dioxolene-type binding sites, each of which can reversibly convert between dianionic catecholate (cat), monoanionic semiquinone (sq) or neutral quinone (q) redox states. The bridging ligand as a whole can therefore exist in seven different redox states from fully reduced [cat,cat,cat]6- through to fully oxidised, neutral [q,q,q]. Cyclic voltammetry of 1(PF6)3 in MeCN reveals six redox processes of which the three at more positive potentials (the sq/q couples) are reversible but the three at more negative potentials (the sq/cat couples) are irreversible with distorted wave shapes due to the insolubility of the reduced forms of the complex. In contrast, the more soluble complex 2(PF6)3 displays six reversible one-electron redox processes making all components of a seven-membered redox chain accessible. UV/Vis/NIR spectro-electrochemical studies reveal rich spectroscopic behaviour, with--in particular--very intense transitions in the near-IR region in many of the oxidation states associated with Ru(II)-->(dioxolene) MLCT and bridging ligand centred pi-pi* transitions. TDDFT calculations were used to analyse the electronic spectra in all seven oxidation states; the calculated spectra generally show very good agreement with experiment, which has allowed a fairly complete assignment of the low-energy transitions. The strong electrochromism of the complexes in the near-IR region has formed the basis of an optical window in which a thin film of 1(PF6)3 or 2(PF6)3 on a conductive glass surface can be reversibly and rapidly switched between redox states that alternate between strongly absorbing or near-transparent at 1100 nm, with--for 2(PF6)3--the switching being stable and reversible in water over thousands of cycles.

Effect of apical hyperosmotic sodium challenge and amiloride on sodium transport in human bronchial epithelial cells from cystic fibrosis donors.

PubMed

Rasgado-Flores, Hector; Krishna Mandava, Vamsi; Siman, Homayoun; Van Driessche, Willy; Pilewski, Joseph M; Randell, Scott H; Bridges, Robert J

2013-12-01

Hypertonic saline (HS) inhalation therapy benefits cystic fibrosis (CF) patients [Donaldson SH, Bennet WD, Zeman KL, Knowles MR, Tarran R, Boucher RC. N Engl J Med 354: 241-250, 2006; Elkins MR, Robinson M, Rose BR, Harbour C, Moriarty CP, Marks GB, Belousova EG, Xuan W, Bye PT; the National Hypertonic Saline in Cystic Fibrosis (NHSCF) Study Group. N Engl J Med 354: 229-240, 2006]. Surprisingly, these benefits are long-lasting and are diminished by the epithelial Na(+) channel blocker amiloride (Donaldson SH, Bennet WD, Zeman KL, Knowles MR, Tarran R, Boucher RC. N Engl J Med 354: 241-250, 2006). Our aim was to explain these effects. Human bronchial epithelial (hBE) cells from CF lungs were grown in inserts and were used in three experimental approaches: 1) Ussing chambers to measure amiloride-sensitive short-circuit currents (INa); 2) continuous perfusion Ussing chambers; and 3) near "thin-film" conditions in which the airway surface of the inserts was exposed to a small volume (30 μl) of isosmotic or HS solution as the inserts were kept in their incubation tray and were subsequently used to measure INa under isosmotic conditions (near thin-film experiments; Tarran R, Boucher RC. Methods Mol Med 70: 479-492, 2002). HS solutions (660 mosmol/kgH2O) were prepared by adding additional NaCl to the isosmotic buffer. The transepithelial short-circuit current (ISC), conductance (GT), and capacitance (CT) were measured by transepithelial impedance analysis (Danahay H, Atherton HC, Jackson AD, Kreindler JL, Poll CT, Bridges RJ. Am J Physiol Lung Cell Mol Physiol 290: L558-L569, 2006; Singh AK, Singh S, Devor DC, Frizzell RA, van Driessche W, Bridges RJ. Methods Mol Med 70: 129-142, 2002). Exposure to apical HS inhibited INa, GT, and CT. The INa inhibition required 60 min of reexposure to the isosmotic solution to recover 75%. The time of exposure to HS required to inhibit INa was <2.5 min. Under near thin-film conditions, apical exposure to HS inhibited INa, but as osmotically driven water moved to the apical surface, the aqueous apical volume increased, leading to an amiloride-insensitive decrease in its osmolality and to recovery of INa that lagged behind the osmotic recovery. Amiloride significantly accelerated the recovery of INa following exposure to HS. Our conclusions are that exposure to HS inhibits hBE INa and that amiloride diminishes this effect.

Intercellular Communication by Exosome-Derived microRNAs in Cancer

PubMed Central

Hannafon, Bethany N.; Ding, Wei-Qun

2013-01-01

The development of human cancers is a multistep process in which normal cells acquire characteristics that ultimately lead to their conversion into cancer cells. Many obstacles must be overcome for this process to occur; of these obstacles, is the ability to survive an inhospitable microenvironment. It is recognized that the intercommunication between tumor cells and their surrounding microenvironment is essential to overcoming this obstacle and for the tumor to progress, metastasize and establish itself at distant sites. Exosomes are membrane-derived vesicles that have recently been recognized as important mediators of intercellular communication, as they carry lipids, proteins, mRNAs and microRNAs that can be transferred to a recipient cell via fusion of the exosome with the target cell membrane. In the context of cancer cells, this process entails the transfer of cancer-promoting cellular contents to surrounding cells within the tumor microenvironment or into the circulation to act at distant sites, thereby enabling cancer progression. In this process, the transfer of exosomal microRNAs to a recipient cell where they can regulate target gene expression is of particular interest, both in understanding the basic biology of cancer progression and for the development of therapeutic approaches. This review discusses the exosome-mediated intercellular communication via microRNAs within the tumor microenvironment in human cancers, with a particular focus on breast cancer exosomes. PMID:23839094

β-Catenin serves as a clutch between low and high intercellular E-cadherin bond strengths.

PubMed

Bajpai, Saumendra; Feng, Yunfeng; Wirtz, Denis; Longmore, Gregory D

2013-11-19

A wide range of invasive pathological outcomes originate from the loss of epithelial phenotype and involve either loss of function or downregulation of transmembrane adhesive receptor complexes, including Ecadherin (Ecad) and binding partners β-catenin and α-catenin at adherens junctions. Cellular pathways regulating wild-type β-catenin level, or direct mutations in β-catenin that affect the turnover of the protein have been shown to contribute to cancer development, through induction of uncontrolled proliferation of transformed tumor cells, particularly in colon cancer. Using single-molecule force spectroscopy, we show that depletion of β-catenin or the prominent cancer-related S45 deletion mutation in β-catenin present in human colon cancers both weaken tumor intercellular Ecad/Ecad bond strength and diminishes the capacity of specific extracellular matrix proteins-including collagen I, collagen IV, and laminin V-to modulate intercellular Ecad/Ecad bond strength through α-catenin and the kinase activity of glycogen synthase kinase 3 (GSK-3β). Thus, in addition to regulating tumor cell proliferation, cancer-related mutations in β-catenin can influence tumor progression by weakening the adhesion of tumor cells to one another through reduced individual Ecad/Ecad bond strength and cellular adhesion to specific components of the extracellular matrix and the basement membrane. Copyright © 2013 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Pathogenetic role of the deafness-related M34T mutation of Cx26

PubMed Central

Bicego, Massimiliano; Beltramello, Martina; Melchionda, Salvatore; Carella, Massimo; Piazza, Valeria; Zelante, Leopoldo; Bukauskas, Feliksas F.; Arslan, Edoardo; Cama, Elona; Pantano, Sergio; Bruzzone, Roberto; D’Andrea, Paola; Mammano, Fabio

2010-01-01

Mutations in the GJB2 gene, which encodes the gap junction protein connexin26 (Cx26), are the major cause of genetic non-syndromic hearing loss. The role of the allelic variant M34T in causing hereditary deafness remains controversial. By combining genetic, clinical, biochemical, electrophysiological and structural modeling studies, we have re-assessed the pathogenetic role of the M34T mutation. Genetic and audiological data indicate that the majority of heterozygous carriers and all five compound heterozygotes exhibited an impaired auditory function. Functional expression in transiently transfected HeLa cells showed that, although M34T was correctly synthesized and targeted to the plasma membrane, it inefficiently formed intercellular channels that displayed an abnormal electrical behavior and retained only 11% of the unitary conductance of the wild-type protein (HCx26wt). Moreover, M34T channels failed to support the intercellular diffusion of Lucifer Yellow and the spreading of mechanically induced intercellular Ca2+ waves. When co-expressed together with HCx26wt, M34T exerted dominant-negative effects on cell–cell coupling. Our findings are consistent with a structural model, predicting that the mutation leads to a constriction of the channel pore. These data support the view that M34T is a pathological variant of Cx26 associated with hearing impairment. PMID:16849369

Autoamplificatory singlet oxygen generation sensitizes tumor cells for intercellular apoptosis-inducing signaling.

PubMed

Bauer, Georg

2018-06-01

Tumor cells express NADPH oxidase-1 (NOX1) in their membrane and control NOX1-based intercellular reactive oxygen and nitrogen species (ROS/RNS)-dependent apoptosis-inducing signaling through membrane-associated catalase and superoxide dismutase. of tumor cells with high concentrations of H 2 O 2 , peroxnitrite, HOCl, or increasing the concentration of cell-derived NO causes initial generation of singlet oxygen and local inactivation of membrane-associated catalase. As a result, free peroxynitrite and H 2 O 2 interact and generate secondary singlet oxygen. Inactivation of further catalase molecules by secondary singlet oxygen leads to auto-amplification of singlet oxygen generation and catalase inactivation. This allows reactivation of intercellular ROS/RNS-signaling and selective apoptosis induction in tumor cells. The initial singlet oxygen generation seems to be the critical point in this complex biochemical multistep mechanism. Initial singlet oxygen generation requires the interaction between distinct tumor cell-derived ROS and RNS and may also depend on either the induction of NO synthase expression or NOX1 activation through the FAS receptor. FAS receptor activation can be achieved by singlet oxygen. Autoamplificatory generation of singlet oxygen through the interaction between peroxynitrite and hydrogen peroxide inherits a rich potential for the establishment of synergistic effects that may be instrumental for novel approaches of tumor therapy with high selectivity towards malignant cells. Copyright © 2017 Elsevier B.V. All rights reserved.

Two dimensional finite element modelling for dynamic water diffusion through stratum corneum.

PubMed

Xiao, Perry; Imhof, Robert E

2012-10-01

Solvents penetration through in vivo human stratum corneum (SC) has always been an interesting research area for trans-dermal drug delivery studies, and the importance of intercellular routes (diffuse in between corneocytes) and transcellular routes (diffuse through corneocytes) during diffusion is often debatable. In this paper, we have developed a two dimensional finite element model to simulate the dynamic water diffusion through the SC. It is based on the brick-and-mortar model, with brick represents corneocytes and mortar represents lipids, respectively. It simulates the dynamic water diffusion process through the SC from pre-defined initial conditions and boundary conditions. Although the simulation is based on water diffusions, the principles can also be applied to the diffusions of other topical applied substances. The simulation results show that both intercellular routes and transcellular routes are important for water diffusion. Although intercellular routes have higher flux rates, most of the water still diffuse through transcellular routes because of the high cross area ratio of corneocytes and lipids. The diffusion water flux, or trans-epidermal water loss (TEWL), is reversely proportional to corneocyte size, i.e. the larger the corneocyte size, the lower the TEWL, and vice versa. There is also an effect of the SC thickness, external air conditions and diffusion coefficients on the water diffusion through SC on the resulting TEWL. Copyright © 2012 Elsevier B.V. All rights reserved.

Investigation of the mechanism for penetration of low density lipoprotein into the arterial wall

NASA Astrophysics Data System (ADS)

Glukhova, O. E.; Zyktin, A. A.; Slepchenkov, M. M.

2018-02-01

Currently, the pathology of the cardiovascular system is an extremely urgent problem of fundamental and clinical medicine. These diseases are caused, mainly, by atherosclerotic changes in the wall of blood vessels. The predominant role in the development of atherosclerosis is attributed to the penetration of various kinds of lipoproteins into the arterial intima. In this paper, we in silico investigated the dynamics of the penetration of low density lipoprotein (LDL) through the intercellular gap using molecular modeling methods. The simulation was carried out in the GROMACS software package using a coarse-grained MARTINI model. During investigation we carried out the LDL self-assembly for the first time. The coarse-grained model of LDL was collected from the following molecules: POPC (phosphatidylcholine) - 630 molecules, LPC (lysophosphatidylcholine) - 80 molecules CHOL (cholesterol) - 600 molecules CHYO (cholesteryl oleate) - 1600 molecules TOG (glycerol trioleate) 180 Molecules. The coarse-grained model of the intercellular endothelial gap was based on a model of lipid bilayer consisting of DPPC phospholipids and cholesterol in a percentage ratio of 70% and 30%, respectively. Based on the obtained results, we can predict the mechanism of LDL diffusion. Lipoproteins can be deformed so as to pass through narrow gaps. Our investigations open the way for the research of the behavior dynamics of LDL moving with the blood flow rate when interacting with the intercellular gaps of the endothelial layer of the vessel inner wall.

Overexpression of SepJ alters septal morphology and heterocyst pattern regulated by diffusible signals in Anabaena.

PubMed

Mariscal, Vicente; Nürnberg, Dennis J; Herrero, Antonia; Mullineaux, Conrad W; Flores, Enrique

2016-09-01

Filamentous, N2 -fixing, heterocyst-forming cyanobacteria grow as chains of cells that are connected by septal junctions. In the model organism Anabaena sp. strain PCC 7120, the septal protein SepJ is required for filament integrity, normal intercellular molecular exchange, heterocyst differentiation, and diazotrophic growth. An Anabaena strain overexpressing SepJ made wider septa between vegetative cells than the wild type, which correlated with a more spread location of SepJ in the septa as observed with a SepJ-GFP fusion, and contained an increased number of nanopores, the septal peptidoglycan perforations that likely accommodate septal junctions. The septa between heterocysts and vegetative cells, which are narrow in wild-type Anabaena, were notably enlarged in the SepJ-overexpressing mutant. Intercellular molecular exchange tested with fluorescent tracers was increased for the SepJ-overexpressing strain specifically in the case of calcein transfer between vegetative cells and heterocysts. These results support an association between calcein transfer, SepJ-related septal junctions, and septal peptidoglycan nanopores. Under nitrogen deprivation, the SepJ-overexpressing strain produced an increased number of contiguous heterocysts but a decreased percentage of total heterocysts. These effects were lost or altered in patS and hetN mutant backgrounds, supporting a role of SepJ in the intercellular transfer of regulatory signals for heterocyst differentiation. © 2016 John Wiley & Sons Ltd.

Intercellular transfer along the trichomes of the invasive terminal heterocyst forming cyanobacterium Cylindrospermopsis raciborskii CS-505.

PubMed

Plominsky, Álvaro M; Delherbe, Nathalie; Mandakovic, Dinka; Riquelme, Brenda; González, Karen; Bergman, Birgitta; Mariscal, Vicente; Vásquez, Mónica

2015-03-01

Cylindrospermopsis raciborskii CS-505 is an invasive freshwater filamentous cyanobacterium that when grown diazotrophically may develop trichomes of up to 100 vegetative cells while differentiating only two end heterocysts, the sole sites for their N2-fixation process. We examined the diazotrophic growth and intercellular transfer mechanisms in C. raciborskii CS-505. Subjecting cultures to a combined-nitrogen-free medium to elicit N2 fixation, the trichome length remained unaffected while growth rates decreased. The structures and proteins for intercellular communication showed that while a continuous periplasmic space was apparent along the trichomes, the putative septal junction sepJ gene is divided into two open reading frames and lacks several transmembrane domains unlike the situation in Anabaena, differentiating a 5-fold higher frequency of heterocysts. FRAP analyses also showed that the dyes calcein and 5-CFDA were taken up by heterocysts and vegetative cells, and that the transfer from heterocysts and 'terminal' vegetative cells showed considerably higher transfer rates than that from vegetative cells located in the middle of the trichomes. The data suggest that C. raciborskii CS-505 compensates its low-frequency heterocyst phenotype by a highly efficient transfer of the fixed nitrogen towards cells in distal parts of the trichomes (growing rapidly) while cells in central parts suffers (slow growth). © FEMS 2014. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Extracellular localization of catalase is associated with the transformed state of malignant cells.

PubMed

Böhm, Britta; Heinzelmann, Sonja; Motz, Manfred; Bauer, Georg

2015-12-01

Oncogenic transformation is dependent on activated membrane-associated NADPH oxidase (NOX). However, the resultant extracellular superoxide anions are also driving the NO/peroxynitrite and the HOCl pathway, which eliminates NOX-expressing transformed cells through selective apoptosis induction. Tumor progression is dependent on dominant interference with intercellular apoptosis-inducing ROS signaling through membrane-associated catalase, which decomposes H2O2 and peroxynitrite and oxidizes NO. Particularly, the decomposition of extracellular peroxynitrite strictly requires membrane-associated catalase. We utilized small interfering RNA (siRNA)-mediated knockdown of catalase and neutralizing antibodies directed against the enzyme in combination with challenging H2O2 or peroxynitrite to determine activity and localization of catalase in cells from three distinct steps of multistage oncogenesis. Nontransformed cells did not generate extracellular superoxide anions and only showed intracellular catalase activity. Transformed cells showed superoxide anion-dependent intercellular apoptosis-inducing ROS signaling in the presence of suboptimal catalase activity in their membrane. Tumor cells exhibited tight control of intercellular apoptosis-inducing ROS signaling through a high local concentration of membrane-associated catalase. These data demonstrate that translocation of catalase to the outside of the cell membrane is already associated with the transformation step. A strong local increase in the concentration of membrane-associated catalase is achieved during tumor progression and is controlled by tumor cell-derived H2O2 and by transglutaminase.

Dissipative particle dynamics simulations of deformation and aggregation of healthy and diseased red blood cells in a tube flow

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ye, Ting; Phan-Thien, Nhan, E-mail: Nhan@nus.edu.sg; Khoo, Boo Cheong

In this paper, we report simulation results assessing the deformation and aggregation of mixed healthy and malaria-infected red blood cells (RBCs) in a tube flow. A three dimensional particle model based on Dissipative Particle Dynamics (DPD) is developed to predict the tube flow containing interacting cells. The cells are also modelled by DPD, with a Morse potential to characterize the cell-cell interaction. As validation tests, a single RBC in a tube flow and two RBCs in a static flow are simulated to examine the cell deformation and intercellular interaction, respectively. The study of two cells, one healthy and the othermore » malaria-infected RBCs in a tube flow demonstrates that the malaria-infected RBC (in the leading position along flow direction) has different effects on the healthy RBC (in the trailing position) at the different stage of parasite development or at the different capillary number. With parasitic development, the malaria-infected RBC gradually loses its deformability, and in turn the corresponding trailing healthy RBC also deforms less due to the intercellular interaction. With increasing capillary number, both the healthy and malaria-infected RBCs are likely to undergo an axisymmetric motion. The minimum intercellular distance becomes small enough so that rouleaux is easily formed, i.e., the healthy and malaria-infected RBCs are difficultly disaggregated.« less

Electric Stimulus Opens Intercellular Spaces in Skin*

PubMed Central

Hama, Susumu; Kimura, Yuki; Mikami, Aya; Shiota, Kanako; Toyoda, Mao; Tamura, Atsushi; Nagasaki, Yukio; Kanamura, Kiyoshi; Kajimoto, Kazuaki; Kogure, Kentaro

2014-01-01

Iontophoresis is a technology for transdermal delivery of ionic small medicines by faint electricity. Since iontophoresis can noninvasively deliver charged molecules into the skin, this technology could be a useful administration method that may enhance patient comfort. Previously, we succeeded in the transdermal penetration of positively charged liposomes (diameters: 200–400 nm) encapsulating insulin by iontophoresis (Kajimoto, K., Yamamoto, M., Watanabe, M., Kigasawa, K., Kanamura, K., Harashima, H., and Kogure, K. (2011) Int. J. Pharm. 403, 57–65). However, the mechanism by which these liposomes penetrated the skin was difficult to define based on general knowledge of principles such as electro-repulsion and electro-osmosis. In the present study, we confirmed that rigid nanoparticles could penetrate into the epidermis by iontophoresis. We further found that levels of the gap junction protein connexin 43 protein significantly decreased after faint electric stimulus (ES) treatment, although occludin, CLD-4, and ZO-1 levels were unchanged. Moreover, connexin 43 phosphorylation and filamentous actin depolymerization in vivo and in vitro were observed when permeation of charged liposomes through intercellular spaces was induced by ES. Ca2+ inflow into cells was promoted by ES with charged liposomes, while a protein kinase C inhibitor prevented ES-induced permeation of macromolecules. Consequently, we demonstrate that ES treatment with charged liposomes induced dissociation of intercellular junctions via cell signaling pathways. These findings suggest that ES could be used to regulate skin physiology. PMID:24318878

Biomarker-guided screening of Juzen-taiho-to, an oriental herbal formulation for immunostimulation.

PubMed

Takaoka, Anna; Iacovidou, Maria; Hasson, Tal H; Montenegro, Diego; Li, Xiangming; Tsuji, Moriya; Kawamura, Akira

2014-03-01

Juzen-taiho-to is an immunostimulatory herbal formulation that is clinically used in East Asia for cancer patients undergoing chemotherapy and radiation. The formulation stimulates various leukocytes, including T, B, and NK cells and macrophages. Although Juzen-taiho-to is known to contain numerous compounds with various pharmacological activities, it is not clear which compounds are responsible for the stimulation of individual cell types. Here, we conducted what we call "biomarker-guided screening" to purify compounds responsible for the macrophages stimulatory activity. To this end, gene expression was analyzed by a DNA array for macrophages treated with Juzen-taiho-to and DMSO (vehicle control), which identified intercellular adhesion molecule 1 as a biomarker of macrophage stimulation by Juzen-taiho-to. A quantitative reverse transcription polymerase chain reaction assay of intercellular adhesion molecule 1 was then used to guide the purification of active compounds. The screening resulted in the purification of a glycolipid mixture, containing β-glucosylceramides. The glycolipid mixture potently stimulated intercellular adhesion molecule 1 expression in primary dendritic cells as well as in primary CD14+ (macrophages) cells. The identification of this glycolipid mixture opens up an opportunity for further studies to understand how plant-derived glycolipids stimulate macrophages and dendritic cells in a safe and effective manner as demonstrated by Juzen-taiho-to. Georg Thieme Verlag KG Stuttgart · New York.

Functional assessment of gap junctions in monolayer and three-dimensional cultures of human tendon cells using fluorescence recovery after photobleaching

PubMed Central

Kuzma-Kuzniarska, Maria; Yapp, Clarence; Pearson-Jones, Thomas W.; Jones, Andrew K.; Hulley, Philippa A.

2014-01-01

Abstract. Gap junction-mediated intercellular communication influences a variety of cellular activities. In tendons, gap junctions modulate collagen production, are involved in strain-induced cell death, and are involved in the response to mechanical stimulation. The aim of the present study was to investigate gap junction-mediated intercellular communication in healthy human tendon-derived cells using fluorescence recovery after photobleaching (FRAP). The FRAP is a noninvasive technique that allows quantitative measurement of gap junction function in living cells. It is based on diffusion-dependent redistribution of a gap junction-permeable fluorescent dye. Using FRAP, we showed that human tenocytes form functional gap junctions in monolayer and three-dimensional (3-D) collagen I culture. Fluorescently labeled tenocytes following photobleaching rapidly reacquired the fluorescent dye from neighboring cells, while HeLa cells, which do not communicate by gap junctions, remained bleached. Furthermore, both 18 β-glycyrrhetinic acid and carbenoxolone, standard inhibitors of gap junction activity, impaired fluorescence recovery in tendon cells. In both monolayer and 3-D cultures, intercellular communication in isolated cells was significantly decreased when compared with cells forming many cell-to-cell contacts. In this study, we used FRAP as a tool to quantify and experimentally manipulate the function of gap junctions in human tenocytes in both two-dimensional (2-D) and 3-D cultures. PMID:24390370

Contractility in type III cochlear fibrocytes is dependent on non-muscle myosin II and intercellular gap junctional coupling.

PubMed

Kelly, John J; Forge, Andrew; Jagger, Daniel J

2012-08-01

The cochlear spiral ligament is a connective tissue that plays diverse roles in normal hearing. Spiral ligament fibrocytes are classified into functional sub-types that are proposed to carry out specialized roles in fluid homeostasis, the mediation of inflammatory responses to trauma, and the fine tuning of cochlear mechanics. We derived a secondary sub-culture from guinea pig spiral ligament, in which the cells expressed protein markers of type III or "tension" fibrocytes, including non-muscle myosin II (nmII), α-smooth muscle actin (αsma), vimentin, connexin43 (cx43), and aquaporin-1. The cells formed extensive stress fibers containing αsma, which were also associated intimately with nmII expression, and the cells displayed the mechanically contractile phenotype predicted by earlier modeling studies. cx43 immunofluorescence was evident within intercellular plaques, and the cells were coupled via dye-permeable gap junctions. Coupling was blocked by meclofenamic acid (MFA), an inhibitor of cx43-containing channels. The contraction of collagen lattice gels mediated by the cells could be prevented reversibly by blebbistatin, an inhibitor of nmII function. MFA also reduced the gel contraction, suggesting that intercellular coupling modulates contractility. The results demonstrate that these cells can impart nmII-dependent contractile force on a collagenous substrate, and support the hypothesis that type III fibrocytes regulate tension in the spiral ligament-basilar membrane complex, thereby determining auditory sensitivity.

Smad ubiquitination regulatory factor-2 controls gap junction intercellular communication by modulating endocytosis and degradation of connexin43.

PubMed

Fykerud, Tone Aase; Kjenseth, Ane; Schink, Kay Oliver; Sirnes, Solveig; Bruun, Jarle; Omori, Yasufumi; Brech, Andreas; Rivedal, Edgar; Leithe, Edward

2012-09-01

Gap junctions consist of arrays of intercellular channels that enable adjacent cells to communicate both electrically and metabolically. Gap junction channels are made of a family of integral membrane proteins called connexins, of which the best-studied member is connexin43. Gap junctions are dynamic plasma membrane domains, and connexin43 has a high turnover rate in most tissue types. However, the mechanisms involved in the regulation of connexin43 endocytosis and transport to lysosomes are still poorly understood. Here, we demonstrate by live-cell imaging analysis that treatment of cells with 12-O-tetradecanoylphorbol 13-acetate (TPA) induces endocytosis of subdomains of connexin43 gap junctions. The internalized, connexin43-enriched vesicles were found to fuse with early endosomes, which was followed by transport of connexin43 to the lumen of early endosomes. The HECT E3 ubiquitin ligase smad ubiquitination regulatory factor-2 (Smurf2) was found to be recruited to connexin43 gap junctions in response to TPA treatment. Depletion of Smurf2 by small interfering RNA resulted in enhanced levels of connexin43 gap junctions between adjacent cells and increased gap junction intercellular communication. Smurf2 depletion also counteracted the TPA-induced endocytosis and degradation of connexin43. Collectively, these data identify Smurf2 as a novel regulator of connexin43 gap junctions.

The Sleep-inducing Lipid Oleamide Deconvolutes Gap Junction Communication and Calcium Wave Transmission in Glial Cells

PubMed Central

Guan, Xiaojun; Cravatt, Benjamin F.; Ehring, George R.; Hall, James E.; Boger, Dale L.; Lerner, Richard A.; Gilula, Norton B.

1997-01-01

Oleamide is a sleep-inducing lipid originally isolated from the cerebrospinal fluid of sleep-deprived cats. Oleamide was found to potently and selectively inactivate gap junction–mediated communication between rat glial cells. In contrast, oleamide had no effect on mechanically stimulated calcium wave transmission in this same cell type. Other chemical compounds traditionally used as inhibitors of gap junctional communication, like heptanol and 18β-glycyrrhetinic acid, blocked not only gap junctional communication but also intercellular calcium signaling. Given the central role for intercellular small molecule and electrical signaling in central nervous system function, oleamide- induced inactivation of glial cell gap junction channels may serve to regulate communication between brain cells, and in doing so, may influence higher order neuronal events like sleep induction. PMID:9412472

Neisseria meningitidis colonization of the brain endothelium and cerebrospinal fluid invasion.

PubMed

Miller, Florence; Lécuyer, Hervé; Join-Lambert, Olivier; Bourdoulous, Sandrine; Marullo, Stefano; Nassif, Xavier; Coureuil, Mathieu

2013-04-01

The brain and meningeal spaces are protected from bacterial invasion by the blood-brain barrier, formed by specialized endothelial cells and tight intercellular junctional complexes. However, once in the bloodstream, Neisseria meningitidis crosses this barrier in about 60% of the cases. This highlights the particular efficacy with which N. meningitidis targets the brain vascular cell wall. The first step of central nervous system invasion is the direct interaction between bacteria and endothelial cells. This step is mediated by the type IV pili, which induce a remodelling of the endothelial monolayer, leading to the opening of the intercellular space. In this review, strategies used by the bacteria to survive in the bloodstream, to colonize the brain vasculature and to cross the blood-brain barrier will be discussed. © 2012 Blackwell Publishing Ltd.

Intercellular Adhesion Molecule-5 Induces Dendritic Outgrowth by Homophilic Adhesion

PubMed Central

Tian, Li; Nyman, Henrietta; Kilgannon, Patrick; Yoshihara, Yoshihiro; Mori, Kensaku; Andersson, Leif C.; Kaukinen, Sami; Rauvala, Heikki; Gallatin, W. Michael; Gahmberg, Carl G.

2000-01-01

Intercellular adhesion molecule-5 (ICAM-5) is a dendritically polarized membrane glycoprotein in telencephalic neurons, which shows heterophilic binding to leukocyte β2-integrins. Here, we show that the human ICAM-5 protein interacts in a homophilic manner through the binding of the immunoglobulin domain 1 to domains 4–5. Surface coated ICAM-5-Fc promoted dendritic outgrowth and arborization of ICAM- 5–expressing hippocampal neurons. During dendritogenesis in developing rat brain, ICAM-5 was in monomer form, whereas in mature neurons it migrated as a high molecular weight complex. The findings indicate that its homophilic binding activity was regulated by nonmonomer/monomer transition. Thus, ICAM-5 displays two types of adhesion activity, homophilic binding between neurons and heterophilic binding between neurons and leukocytes. PMID:10893271

Enzyme-activated intracellular drug delivery with tubule clay nanoformulation

DOE PAGES

Dzamukova, Maria R.; Naumenko, Ekaterina A.; Lvov, Yuri M.; ...

2015-05-15

Fabrication of stimuli-triggered drug delivery vehicle is is an important milestone in treating cancer. Here we demonstrate the selective anticancer drug delivery into human cells with biocompatible 50-nm diameter halloysite nanotube carriers. Physically-adsorbed dextrin end stoppers secure the intercellular release of brilliant green. Drug-loaded nanotubes penetrate through the cellular membranes and their uptake efficiency depends on the cells growth rate. Intercellular glycosyl hydrolases-mediated decomposition of the dextrin tube-end stoppers triggers the release of the lumen-loaded brilliant green, which allowed for preferable elimination of human lung carcinoma cells (А549) as compared with hepatoma cells (Hep3b). In conclusion, the enzyme-activated intracellular deliverymore » of brilliant green using dextrin-coated halloysite nanotubes is a promising platform for anticancer treatment.« less

Growth directions of C8-BTBT thin films during drop-casting

NASA Astrophysics Data System (ADS)

Iizuka, Naoki; Zanka, Tomohiko; Onishi, Yosuke; Fujieda, Ichiro

2016-02-01

Because charge transport in a single crystal is anisotropic, control of its orientation is important for enhancing electrical characteristics and reducing variations among devices. For growing an organic thin film, a solution process such as inkjet printing offers advantages in throughput. We have proposed to apply an external temperature gradient during drop-casting and to control the direction of solvent evaporation. In experiment, a temperature gradient was generated in a bare Si substrate by placing it on a Si plate bridging two heat stages. When a solution containing 2,7-dioctyl [1]benzothieno[3,2-b]benzothiophene (C8-BTBT) was dropped on the substrate, evaporation started at the hotter side of the droplet and proceeded toward the colder side. The front line of the liquid was not pinned and the solution extended toward the colder region. As a result, a thin film was formed in a 7mm-long region. The peripheral region of the film was significantly thicker due to the coffee ring effect. The surface of the rest of the film was mostly smooth and terrace structures with 2.6nm steps were observed. The step roughly corresponds to the length of the C8-BTBT molecule. The film thickness varied from 20nm to 50nm over the distance of 3mm. Another film was grown on a glass substrate under a similar condition. Observation of the film with a polarizing microscope revealed that fan-shaped domains were formed in the film and that their optical axes were mostly along the directions of the solvent evaporation.

Titin strain contributes to the Frank–Starling law of the heart by structural rearrangements of both thin- and thick-filament proteins

PubMed Central

Ait-Mou, Younss; Hsu, Karen; Farman, Gerrie P.; Kumar, Mohit; Greaser, Marion L.; Irving, Thomas C.; de Tombe, Pieter P.

2016-01-01

The Frank–Starling mechanism of the heart is due, in part, to modulation of myofilament Ca2+ sensitivity by sarcomere length (SL) [length-dependent activation (LDA)]. The molecular mechanism(s) that underlie LDA are unknown. Recent evidence has implicated the giant protein titin in this cellular process, possibly by positioning the myosin head closer to actin. To clarify the role of titin strain in LDA, we isolated myocardium from either WT or homozygous mutant (HM) rats that express a giant splice isoform of titin, and subjected the muscles to stretch from 2.0 to 2.4 μm of SL. Upon stretch, HM compared with WT muscles displayed reduced passive force, twitch force, and myofilament LDA. Time-resolved small-angle X-ray diffraction measurements of WT twitching muscles during diastole revealed stretch-induced increases in the intensity of myosin (M2 and M6) and troponin (Tn3) reflections, as well as a reduction in cross-bridge radial spacing. Independent fluorescent probe analyses in relaxed permeabilized myocytes corroborated these findings. X-ray electron density reconstruction revealed increased mass/ordering in both thick and thin filaments. The SL-dependent changes in structure observed in WT myocardium were absent in HM myocardium. Overall, our results reveal a correlation between titin strain and the Frank–Starling mechanism. The molecular basis underlying this phenomenon appears not to involve interfilament spacing or movement of myosin toward actin but, rather, sarcomere stretch-induced simultaneous structural rearrangements within both thin and thick filaments that correlate with titin strain and myofilament LDA. PMID:26858417

KSC-98pc1086

NASA Image and Video Library

1998-09-14

KENNEDY SPACE CENTER, FLA. The International Extreme Ultraviolet Hitchhiker-3 (IEH-3), one of the payloads for the STS-95 mission, is prepared for launch in the Multi-Payload Processing Facility. IEH-3 is comprised of seven experiments, including one that will be deployed on Flight Day 3. It is the small, non-recoverable Petite Amateur Navy Satellite (PANSAT) which will store and transmit digital communications. Other IEH investigations are the Solar Constant Experiment (SOLCON), Solar Extreme Ultraviolet Hitchhiker (SEH), Spectrograph/Telescope for Astronomical Research (STAR-LITE), Ultraviolet Spectrograph Telescope for Astronomical Research (UVSTAR), Consortium for Materials Development in Space Complex Autonomous Payloads (CONCAP-IV) for growing thin films via physical vapor transport, and two Get-Away Special (GAS) canister experiments. The experiments will be mounted on a hitchhiker bridge in Discovery's payload bay

Annular array and method of manufacturing same

DOEpatents

Day, Robert A.

1989-01-01

A method for manufacturing an annular acoustic transducer array from a plate of transducer material, which enables production of precision aligned arrays at low cost. The circular plate is sawed along at least two lines that are radial to the axis of the plate. At steps along each radial cut, the plate is rotated first in one direction and then in an opposite direction by a predetermined angle such as slightly less than 90.degree.. The cuts result in the forming of several largely ring-shaped lands, each largely ring-shaped land being joined to the other rings of different radii by thin portions of the plate, and each ring being cut into segments. The bridges that join different rings, hold the transducer together until it can be mounted on a lens.

Means of manufacturing annular arrays

DOEpatents

Day, R.A.

1985-10-10

A method is described for manufacturing an annular acoustic transducer array from a plate of transducer material, which enables production of precision aligned arrays at low cost. The circular plate is sawed along at least two lines that are radial to the axis of the plate. At steps along each radial cut, the plate is rotated first in one direction and then in an opposite direction by a predetermined angle such as slightly less than 90/sup 0/. The cuts result in the forming of several largely ring-shaped lands, each largely ring-shaped land being joined to the other rings of different radii by thin portions of the plate, and each ring being cut into segments. The bridges that join different rings hold the transducer together until it can be mounted on a lens.

Magnetic current sensor

NASA Technical Reports Server (NTRS)

Black, Jr., William C. (Inventor); Hermann, Theodore M. (Inventor)

1998-01-01

A current determiner having an output at which representations of input currents are provided having an input conductor for the input current and a current sensor supported on a substrate electrically isolated from one another but with the sensor positioned in the magnetic fields arising about the input conductor due to any input currents. The sensor extends along the substrate in a direction primarily perpendicular to the extent of the input conductor and is formed of at least a pair of thin-film ferromagnetic layers separated by a non-magnetic conductive layer. The sensor can be electrically connected to a electronic circuitry formed in the substrate including a nonlinearity adaptation circuit to provide representations of the input currents of increased accuracy despite nonlinearities in the current sensor, and can include further current sensors in bridge circuits.

Hypoparathyroidism-retardation-Dysmorphism (HRD) syndrome--a review.

PubMed

Hershkovitz, Eli; Parvari, Ruti; Diaz, George A; Gorodischer, Rafael

2004-12-01

Hypoparathyroidism, retardation, and dysmorphism (HRD) is a newly recognized genetic syndrome, described in patients of Arab origin. The syndrome consists of permanent congenital hypoparathyroidism, severe prenatal and postnatal growth retardation, and profound global developmental delay. The patients are susceptible to severe infections including life-threatening pneumococcal infections especially during infancy. The main dysmorphic features are microcephaly, deep-set eyes or microphthalmia, ear abnormalities, depressed nasal bridge, thin upper lip, hooked small nose, micrognathia, and small hands and feet. A single 12-bp deletion (del52-55) in the second coding exon of the tubulin cofactor E (TCFE) gene, located on the long arm of chromosome 1, is the cause of HRD among Arab patients. Early recognition and therapy of hypocalcemia is important as is daily antibiotic prophylaxis against pneumococcal infections.

Cohesive zone model for direct silicon wafer bonding

NASA Astrophysics Data System (ADS)

Kubair, D. V.; Spearing, S. M.

2007-05-01

Direct silicon wafer bonding and decohesion are simulated using a spectral scheme in conjunction with a rate-dependent cohesive model. The cohesive model is derived assuming the presence of a thin continuum liquid layer at the interface. Cohesive tractions due to the presence of a liquid meniscus always tend to reduce the separation distance between the wafers, thereby opposing debonding, while assisting the bonding process. In the absence of the rate-dependence effects the energy needed to bond a pair of wafers is equal to that needed to separate them. When rate-dependence is considered in the cohesive law, the experimentally observed asymmetry in the energetics can be explained. The derived cohesive model has the potential to form a bridge between experiments and a multiscale-modelling approach to understand the mechanics of wafer bonding.

Bernoulli Suction Effect on Soap Bubble Blowing?

NASA Astrophysics Data System (ADS)

Davidson, John; Ryu, Sangjin

2015-11-01

As a model system for thin-film bubble with two gas-liquid interfaces, we experimentally investigated the pinch-off of soap bubble blowing. Using the lab-built bubble blower and high-speed videography, we have found that the scaling law exponent of soap bubble pinch-off is 2/3, which is similar to that of soap film bridge. Because air flowed through the decreasing neck of soap film tube, we studied possible Bernoulli suction effect on soap bubble pinch-off by evaluating the Reynolds number of airflow. Image processing was utilized to calculate approximate volume of growing soap film tube and the volume flow rate of the airflow, and the Reynolds number was estimated to be 800-3200. This result suggests that soap bubbling may involve the Bernoulli suction effect.

Low temperature InP /Si wafer bonding using boride treated surface

NASA Astrophysics Data System (ADS)

Huang, Hui; Ren, Xiaomin; Wang, Wenjuan; Song, Hailan; Wang, Qi; Cai, Shiwei; Huang, Yongqing

2007-04-01

An approach for InP /Si wafer bonding based on boride-solution treatment was presented. The bonding energy is higher than the InP fracture energy by annealing at 280°C. An In0.53Ga0.47As/InP multiple-quantum-well (MQW) structure grown on InP was transferred onto Si substrate via the bonding process. X-ray diffraction and photoluminescence reveal that crystal quality of the bonded MQW was preserved. A thin B2O3-POx-SiO2 oxide layer of about 28nm thick at the bonding interface was detected. X-ray photoelectron spectroscopy and Raman analyses indicate that the formation of oxygen bridging bonds by boride treatment is responsible for the strong fusion obtained at such low temperature.

Enhancement of Resonant Energy Transfer Due to an Evanescent Wave from the Metal.

PubMed

Poudel, Amrit; Chen, Xin; Ratner, Mark A

2016-03-17

The high density of evanescent modes in the vicinity of a metal leads to enhancement of the near-field Förster resonant energy transfer (FRET) rate. We present a classical approach to calculate the FRET rate based on the dyadic Green's function of an arbitrary dielectric environment and consider the nonlocal limit of material permittivity in the case of the metallic half-space and thin film. In a dimer system, we find that the FRET rate is enhanced due to shared evanescent photon modes bridging a donor and an acceptor. Furthermore, a general expression for the FRET rate for multimer systems is derived. The presence of a dielectric environment and the path interference effect enhance the transfer rate, depending on the combination of distance and geometry.

Comparison of ultrastructure, tight junction-related protein expression and barrier function of human corneal epithelial cells cultivated on amniotic membrane with and without air-lifting.

PubMed

Ban, Yuriko; Cooper, Leanne J; Fullwood, Nigel J; Nakamura, Takahiro; Tsuzuki, Masakatsu; Koizumi, Noriko; Dota, Atsuyoshi; Mochida, Chikako; Kinoshita, Shigeru

2003-06-01

To evaluate the usefulness of the air-lifting technique for culturing corneal limbal epithelial cells on amniotic membrane (AM) for use in ocular surface reconstruction. A cultured sheet that has a good barrier function should be better for this purpose. In corneal epithelium, tight junctions (TJ) play a vital role in the barrier function. The TJ complex includes the integral transmembrane proteins occludin and the claudins, and some membrane-associated proteins such as ZO-1. In this paper, we investigated the barrier function and the expression of TJ related proteins. Corneal limbal epithelium obtained from donor corneas and cultivated on acellular AM was divided into two groups. These were the non-air-lifting (Non-AL) group, which was continuously submerged in medium, and the air-lifting (AL) group, which was submerged in medium for 3 weeks, then exposed to air by lowering the medium level. Morphology and the permeability to horseradish peroxidase (HRP) were determined by electron microscopy. Tight junction (TJ)-related protein and mRNA expression changes were assessed by immunoblotting and reverse transcription-polymerase chain reaction. The cultures of both groups formed 4-5-layer-thick, well-stratified epithelium. The AL cultures had tightly packed epithelial cells with all the HRP/diaminobenzidine (DAB) reaction product accumulated on the apical surface of the superficial cells. The Non-AL culture, by contrast, had more loosely packed epithelial cells with larger intercellular spaces. The HRP/DAB reaction product penetrated the intercellular space to a depth of 3-4 cell layers. Statistically, there was a significant difference in intercellular spaces and desmosome count in the superficial cells between the groups. With AL, TJ-related proteins localized at the apical portion of the lateral membrane. TJ-related protein and mRNA amounts were not changed by AL while claudin subtype expression became more consistent and closer to that of in vivo corneal epithelium. The AL technique reduces intercellular spaces in the superficial cells and promotes the formation of the barrier function. It is useful in culturing corneal epithelial cells for use in ocular surface reconstruction.

Deformation and breakup of a stretching liquid bridge covered with an insoluble surfactant monolayer

NASA Astrophysics Data System (ADS)

Liao, Ying-Chih; Franses, Elias I.; Basaran, Osman A.

2006-02-01

The breakup of surfactant-laden drops and jets is of technological interest and fundamental scientific importance. Surfactants are routinely used to control the breakup of drops and jets in applications ranging from inkjet printing to crop spraying. Accurate computation of breakup of surfactant-laden drops and jets is often the key to the development of new applications and to providing a rational fundamental understanding of both existing and emerging applications. While highly accurate algorithms for studying the breakup of surfactant-free drops and jets are well documented and much is now known about the dynamics in such situations, little is known by contrast about the closely related problem of interface rupture when surfactant effects cannot be neglected. The deformation and breakup of a stretching liquid bridge of an incompressible Newtonian fluid whose surface is covered with an insoluble surfactant monolayer are analyzed here experimentally and computationally. In the experiments, high-speed visualization is used to capture the transient deformation of a bridge. The dynamic shapes of bridges (captive between two rods of 3.15 mm diameter) are captured and analyzed with a time resolution of 1 ms. The bridge lengths are 3.15 mm initially and about 4-7 mm at breakup, which occurs after stretching for about 0.1-0.2 s, depending on the volume and viscosity of the liquid and the surface density of spread monolayers. The dynamics of a surfactant-covered bridge is governed by the Navier-Stokes and convection-diffusion equations. First, these equations are solved with a three-dimensional, but axisymmetric, or two-dimensional (2D), finite element algorithm using elliptic mesh generation. Second, the governing set of 2D equations is reduced to a set of one-dimensional (1D) equations by means of the slender-jet approximation and the resulting set of 1D equations is solved with a 1D finite element algorithm. The presence of surfactant results not only in the lowering of surface tension and the capillary pressure, but also in surface tension gradients and Marangoni stresses, both of which affect the transient dynamics leading to breakup. In particular, the role of Marangoni stresses in delaying bridge breakup and on formation of satellite droplets is investigated as a function of the initial surface density and surface activity of the surfactant, and surface Peclet number that measures the importance of convection relative to diffusion. The predictions of the 2D algorithm are confirmed to be faithful to the physics by demonstrating that the computed results accord well with the experiments and existing scaling theories. In the pinch-off region, the surfactant is swept out of a thinning neck by strong convection. The calculations thus reveal that the scaling behavior in the presence of surfactant parallels that observed in the absence of surfactant, in accordance with recent reports by others. The 2D computations and the experiments are used in tandem to identify regions in the space of governing parameters where the 1D equations can be used with confidence.

Structure of the stigma and style of Callaeum psilophyllum (Malpighiaceae) and its relation with potential pollinators.

PubMed

Aliscioni, Sandra Silvina; Gotelli, Marina; Torretta, Juan Pablo

2018-03-29

The family Malpighiaceae, particularly in the Neotropic, shows a similar floral morphology. Although floral attraction and rewards to pollinators are alike, stigmas and styles show more diversity. The stigmas were described covered with a thin and impermeable cuticle that needs to be ruptured by the mechanical action of the pollinators. However, this characteristic was only mentioned for a few species and the anatomy and ultrastructure of the stigmas were not explored. In this work, we analyze the morphology, anatomy, and ultrastructure of the stigma and style of Callaeum psilophyllum. Moreover, we identify the potential pollinators in order to evaluate how the disposition of the stigmas is related with their size and its role in the exposure of the receptive stigmatic surface. Our observations indicate that Centris flavifrons, C. fuscata, C. tarsata, and C. trigonoides are probably efficient pollinators of C. psilophyllum. The three stigmas are covered by a cuticle that remained intact in bagged flowers. The flowers exposed to visitors show the cuticle broken, more secretion in the intercellular spaces between sub-stigmatic cells and abundant electron-dense components inside vacuoles in stigmatic papillae. This indicates that the stigmas prepares in similar ways to receive pollen grains, but the pollinator action is required to break the cuticle, and once pollen tubes start growing, stigmatic and sub-stigmatic cells release more secretion by a granulocrine process.

A distinct type of cell in myocardium: interstitial Cajal-like cells (ICLCs)

PubMed Central

Kostin, S; Popescu, L M

2009-01-01

Abstract The existence of a novel type of interstitial cells in the heart, interstitial Cajal-like cells (ICLCs), had been described for the first time in 2005. Their identification was mainly based on ultrastructural criteria: very long (tens up to hundreds of micrometres) and moniliform prolongations, which are extremely thin (less than 0.2 μm), below the resolving power of light microscopy. Myocardial ICLCs were also identified by methylene-blue vital staining, silver impregnation, and immunoreactivity for CD 34, vimentin, CD117/c-kit, etc. Although a series of studies provided evidence for the existence of ICLCs in human atria and rat ventricles, further investigations in other laboratories, using additional techniques, are required to substantiate the consistency of these findings. Here we provide further evidence for the existence of ICLCs in human and mammalian hearts (by transmission and scanning electron microscopy, as well as confocal laser scanning microscopy). Noteworthy, we confirm that ICLCs communicate with neighbouring cells via shedding (micro)vesicles. Although these so-called ICLCs represent a distinct type of cells, different from classical interstitial cells of Cajal, or fibroblasts, their role(s) in myocardium remain(s) to be established. Several hypotheses are proposed: (i) adult stromal (mesenchymal) stem cells, which might participate in cardiac repair/remodelling; (ii) intercellular signalling (e.g. via shedding microvesicles); (iii) chemo-mechanical transducers and (iv) players in pacemaking and/or arrhytmogenesis, and so on. PMID:19183408

FIB-SEM tomography of human skin telocytes and their extracellular vesicles.

PubMed

Cretoiu, Dragos; Gherghiceanu, Mihaela; Hummel, Eric; Zimmermann, Hans; Simionescu, Olga; Popescu, Laurentiu M

2015-04-01

We have shown in 2012 the existence of telocytes (TCs) in human dermis. TCs were described by transmission electron microscopy (TEM) as interstitial cells located in non-epithelial spaces (stroma) of many organs (see www.telocytes.com). TCs have very long prolongations (tens to hundreds micrometers) named Telopodes (Tps). These Tps have a special conformation with dilated portions named podoms (containing mitochondria, endoplasmic reticulum and caveolae) and very thin segments (below resolving power of light microscopy), called podomers. To show the real 3D architecture of TC network, we used the most advanced available electron microscope technology: focused ion beam scanning electron microscopy (FIB-SEM) tomography. Generally, 3D reconstruction of dermal TCs by FIB-SEM tomography revealed the existence of Tps with various conformations: (i) long, flattened irregular veils (ribbon-like segments) with knobs, corresponding to podoms, and (ii) tubular structures (podomers) with uneven calibre because of irregular dilations (knobs) - the podoms. FIB-SEM tomography also showed numerous extracellular vesicles (diameter 438.6 ± 149.1 nm, n = 30) released by a human dermal TC. Our data might be useful for understanding the role(s) of TCs in intercellular signalling and communication, as well as for comprehension of pathologies like scleroderma, multiple sclerosis, psoriasis, etc. © 2015 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Emperipolesis of erythroblasts within Kupffer cells during hepatic hemopoiesis in human fetus.

PubMed

Lee, W B; Erm, S K; Kim, K Y; Becker, R P

1999-10-01

The state in which cells can inhabit other cells without damage is known as emperipolesis. Emperipolesis has been found in various physiological and pathological conditions. We performed a study of emperipolesis of erythroblasts within Kupffer cells in the human fetal liver. We found that Kupffer cells, identified by CD68 immunolabeling, contained 4-8 erythroblasts in a hypertrophic cytoplasm on light microscopy. Emperipoletic erythroblasts were present in various maturation stages from proerythroblast to reticulocyte. By electron microscopy, we found that erythroblasts occupied membrane-bound vacuoles that were separated from each other by thin partitions of Kupffer cell cytoplasm. Neither emperipoletic erythroblasts nor their Kupffer cell hosts showed evidence of damage. Emperipoletic cells in mitosis were found, which suggests the capacity for the proliferation of erythroblasts within Kupffer cells. Some Kupffer cells were seen to contain both emperipoletic cells and phagosomes, without evidence of interaction. Erythroblasts and other hemopoietic cells were also found to be closely associated with the sinusoidal surface of Kupffer cells. However, intercellular junctions, if present, were inconspicuous. On occasion, Kupffer cells engorged with erythroblasts nearly occluded the sinusoidal lumen. Our results demonstrate that emperipolesis of erythroblasts within Kupffer cells occurs in human fetal hepatic hemopoiesis. We suggest that emperipolesis may be one of the mechanisms that support the maturation of erythroblasts in the fetal liver. Copyright 1999 Wiley-Liss, Inc.

Neuroblastoma SH-SY5Y cell-derived exosomes stimulate dendrite-like outgrowths and modify the differentiation of A375 melanoma cells.

PubMed

Park, Seyeon; Ahn, Eun Sook; Kim, Yunjoo

2015-04-01

The identification of small vesicles released by many cell types as tools of intercellular communication is proposed. Here, we identify SH-SY5Y neuroblastoma-derived exosomes comprised of major histocompatibility complex II (MHC II), Hsp90 and flotillin-1. Our data also suggest that, when applied extracellularly, exosomes released from neuronal cells stimulate dendrite-like outgrowth and melanogenesis of A375 melanoma cells through the mitogen-activated protein kinase (MAP kinase), extracellular signal-regulated kinase 1 (ERK1) activation. These results suggest a modification of differentiation of melanocyte by the treatment of neuronal cell exosomes. Since exosomes from neuronal cells have the capacity to affect melanoma cells, they could be generally implicated in intercellular communication between different types of cells. © 2014 International Federation for Cell Biology.

The spread of prion-like proteins by lysosomes and tunneling nanotubes: Implications for neurodegenerative diseases.

PubMed

Victoria, Guiliana Soraya; Zurzolo, Chiara

2017-09-04

Progression of pathology in neurodegenerative diseases is hypothesized to be a non-cell-autonomous process that may be mediated by the productive spreading of prion-like protein aggregates from a "donor cell" that is the source of misfolded aggregates to an "acceptor cell" in which misfolding is propagated by conversion of the normal protein. Although the proteins involved in the various diseases are unrelated, common pathways appear to be used for their intercellular propagation and spreading. Here, we summarize recent evidence of the molecular mechanisms relevant for the intercellular trafficking of protein aggregates involved in prion, Alzheimer's, Huntington's, and Parkinson's diseases. We focus in particular on the common roles that lysosomes and tunneling nanotubes play in the formation and spreading of prion-like assemblies. © 2017 Victoria and Zurzolo.

The spread of prion-like proteins by lysosomes and tunneling nanotubes: Implications for neurodegenerative diseases

PubMed Central

Victoria, Guiliana Soraya

2017-01-01

Progression of pathology in neurodegenerative diseases is hypothesized to be a non–cell-autonomous process that may be mediated by the productive spreading of prion-like protein aggregates from a “donor cell” that is the source of misfolded aggregates to an “acceptor cell” in which misfolding is propagated by conversion of the normal protein. Although the proteins involved in the various diseases are unrelated, common pathways appear to be used for their intercellular propagation and spreading. Here, we summarize recent evidence of the molecular mechanisms relevant for the intercellular trafficking of protein aggregates involved in prion, Alzheimer’s, Huntington’s, and Parkinson’s diseases. We focus in particular on the common roles that lysosomes and tunneling nanotubes play in the formation and spreading of prion-like assemblies. PMID:28724527

Amphiregulin enhances intercellular adhesion molecule-1 expression and promotes tumor metastasis in human osteosarcoma

PubMed Central

Liu, Ju-Fang; Tsao, Ya-Ting; Hou, Chun-Han

2015-01-01

Osteosarcoma is a common, high malignant, and metastatic bone cancer. Amphiregulin (AREG) has been associated with cancer cellular activities. However, the effect of AREG on metastasis activity in human osteosarcoma cells has yet to be determined. We determined that AREG increases the expression of intercellular adhesion molecule-1 (ICAM-1) through PI3K/Akt signaling pathway via its interaction with the epidermal growth factor receptor, thus resulting in the enhanced cell migration of osteosarcoma. Furthermore, AREG stimulation increased the association of NF-κB to ICAM-1 promoter which then up-regulated ICAM-1 expression. Finally, we observed that shRNA silencing of AREG decreased osteosarcoma metastasis in vivo. Our findings revealed a relationship between osteosarcoma metastatic potential and AREG expression and the modulating effect of AREG on ICAM-1 expression. PMID:26503469

Primordial oscillations in life: Direct observation of glycolytic oscillations in individual HeLa cervical cancer cells

NASA Astrophysics Data System (ADS)

Amemiya, Takashi; Shibata, Kenichi; Itoh, Yoshihiro; Itoh, Kiminori; Watanabe, Masatoshi; Yamaguchi, Tomohiko

2017-10-01

We report the first direct observation of glycolytic oscillations in HeLa cervical cancer cells, which we regard as primordial oscillations preserved in living cells. HeLa cells starved of glucose or both glucose and serum exhibited glycolytic oscillations in nicotinamide adenine dinucleotide (NADH), exhibiting asynchronous intercellular behaviors. Also found were spatially homogeneous and inhomogeneous intracellular NADH oscillations in the individual cells. Our results demonstrate that starved HeLa cells may be induced to exhibit glycolytic oscillations by either high-uptake of glucose or the enhancement of a glycolytic pathway (Crabtree effect or the Warburg effect), or both. Their asynchronous collective behaviors in the oscillations were probably due to a weak intercellular coupling. Elucidation of the relationship between the mechanism of glycolytic dynamics in cancer cells and their pathophysiological characteristics remains a challenge in future.

Structure of Peptide Sex Pheromone Receptor PrgX and PrgX/Pheromone Complexes and Regulation of Conjugation in Enterococcus faecalis

DOE Office of Scientific and Technical Information (OSTI.GOV)

Shi,K.; Brown, C.; Gu, Z.

2005-01-01

Many bacterial activities, including expression of virulence factors, horizontal genetic transfer, and production of antibiotics, are controlled by intercellular signaling using small molecules. To date, understanding of the molecular mechanisms of peptide-mediated cell-cell signaling has been limited by a dearth of published information about the molecular structures of the signaling components. Here, we present the molecular structure of PrgX, a DNA- and peptide-binding protein that regulates expression of the conjugative transfer genes of the Enterococcus faecalis plasmid pCF10 in response to an intercellular peptide pheromone signal. Comparison of the structures of PrgX and the PrgX/pheromone complex suggests that pheromone bindingmore » destabilizes PrgX tetramers, opening a 70-bp pCF10 DNA loop required for conjugation repression.« less

Endocytosis and Signaling during Development

PubMed Central

Bökel, Christian

2014-01-01

The development of multicellular organisms relies on an intricate choreography of intercellular communication events that pattern the embryo and coordinate the formation of tissues and organs. It is therefore not surprising that developmental biology, especially using genetic model organisms, has contributed significantly to the discovery and functional dissection of the associated signal-transduction cascades. At the same time, biophysical, biochemical, and cell biological approaches have provided us with insights into the underlying cell biological machinery. Here we focus on how endocytic trafficking of signaling components (e.g., ligands or receptors) controls the generation, propagation, modulation, reception, and interpretation of developmental signals. A comprehensive enumeration of the links between endocytosis and signal transduction would exceed the limits of this review. We will instead use examples from different developmental pathways to conceptually illustrate the various functions provided by endocytic processes during key steps of intercellular signaling. PMID:24591521

Glial response to polyglutamine-mediated stress

PubMed Central

Vig, Parminder J.S.; Shao, Qingmei; Lopez, Maripar E

2009-01-01

Neurodegenerative trinucleotide (CAG) repeat disorders are caused by the expansion of polyglutamine tracts within the disease proteins. Some of these proteins have an unknown function. How does expanded polyglutamine cause target neurons to degenerate, is not clear. Recent evidence suggests that intercellular miscommunication may contribute to polyglutamine pathogenesis in CAG repeat disorders. Polyglutamine induced degeneration of the target neuron can be mediated via glia-neuron interactions. Here we hypothesize during neurodegenerative process the failure of cell: cell interactions have more severe consequences than alterations in intracellular neuron biology. We further believe that bidirectional communication between neurons and glia are prerequisite for the normal development and function of either cell-type. Understanding intercellular signaling mechanisms such as glial trophic factors and their receptors, cell adhesion or other well-defined signaling molecules provide opportunities for developing potential therapies. PMID:20046986

Effect of Cr Contents and Heat Treating on Reverted Austenite in Maraging Steel Weldments

NASA Astrophysics Data System (ADS)

Kim, S. W.; Lee, H. W.

2018-05-01

By conducting flux cored arc welding (FCAW) on maraging steels with Cr contents of 1.4 and 5.2 wt%, this study observed the effects of Cr content and heat treating on reverted austenite formation in welded maraging steel. Aging treatment was carried out at the temperatures of 450, 480 and 530 °C for 3 h in each condition. As the aging temperature increased, reverted austenite was formed along the interdendritic and intercellular grain boundaries, and the proportion of reverted austenite increased with increasing Cr addition. The aging process led to the segregation of Ti and Mo along the interdendritic and intercellular grain boundaries. Some of the welded specimens were subjected to solution heat treatment at 820 and 1250 °C for 1 h after welding, resulting in a decrease in reverted austenite fraction.

Faster embryonic segmentation through elevated Delta-Notch signalling

PubMed Central

Liao, Bo-Kai; Jörg, David J.; Oates, Andrew C.

2016-01-01

An important step in understanding biological rhythms is the control of period. A multicellular, rhythmic patterning system termed the segmentation clock is thought to govern the sequential production of the vertebrate embryo's body segments, the somites. Several genetic loss-of-function conditions, including the Delta-Notch intercellular signalling mutants, result in slower segmentation. Here, we generate DeltaD transgenic zebrafish lines with a range of copy numbers and correspondingly increased signalling levels, and observe faster segmentation. The highest-expressing line shows an altered oscillating gene expression wave pattern and shortened segmentation period, producing embryos with more, shorter body segments. Our results reveal surprising differences in how Notch signalling strength is quantitatively interpreted in different organ systems, and suggest a role for intercellular communication in regulating the output period of the segmentation clock by altering its spatial pattern. PMID:27302627

Effect of Cr Contents and Heat Treating on Reverted Austenite in Maraging Steel Weldments

NASA Astrophysics Data System (ADS)

Kim, S. W.; Lee, H. W.

2018-03-01

By conducting flux cored arc welding (FCAW) on maraging steels with Cr contents of 1.4 and 5.2 wt%, this study observed the effects of Cr content and heat treating on reverted austenite formation in welded maraging steel. Aging treatment was carried out at the temperatures of 450, 480 and 530 °C for 3 h in each condition. As the aging temperature increased, reverted austenite was formed along the interdendritic and intercellular grain boundaries, and the proportion of reverted austenite increased with increasing Cr addition. The aging process led to the segregation of Ti and Mo along the interdendritic and intercellular grain boundaries. Some of the welded specimens were subjected to solution heat treatment at 820 and 1250 °C for 1 h after welding, resulting in a decrease in reverted austenite fraction.

Does rat granulation tissue maturation involve gap junction communications?

PubMed

Au, Katherine; Ehrlich, H Paul

2007-07-01

Wound healing, a coordinated process, proceeds by sequential changes in cell differentiation and terminates with the deposition of a new connective tissue matrix, a scar. Initially, there is the migratory fibroblast, followed by the proliferative fibroblast, then the synthetic fibroblast, which transforms into the myofibroblast, and finally the apoptotic fibroblast. Gap junction intercellular communications are proposed to coordinate the stringent control of fibroblast phenotypic changes. Does added oleamide, a natural fatty acid that blocks gap junction intercellular communications, alter the phenotypic progression of wound fibroblasts? Pairs of polyvinyl alcohol sponges attached to Alzet pumps, which constantly pumped either oleamide or vehicle solvent, were implanted subcutaneously into three rats. On day 8, implants were harvested and evaluated histologically and biochemically. The capsule of oleamide-treated sponge contained closely packed fibroblasts with little connective tissue between them. The birefringence intensity of that connective tissue was reduced, indicating a reduced density of collagen fiber bundles. Myofibroblasts, identified immunohistologically by alpha-smooth muscle actin-stained stress fibers, were reduced in oleamide-treated implants. Western blot analysis showing less alpha-smooth muscle actin confirmed the reduced density of myofibroblasts. It appears that oleamide retards the progression of wound repair, where less connective tissue is deposited, the collagen is less organized, and the appearance of myofibroblasts is impaired. These findings support the hypothesis that gap junction intercellular communications between wound fibroblasts in granulation tissue play a role in the progression of repair and the maturation of granulation tissue into scar.

Metabolite pools and carbon flow during C4 photosynthesis in maize: 13CO2 labeling kinetics and cell type fractionation.

PubMed

Arrivault, Stéphanie; Obata, Toshihiro; Szecówka, Marek; Mengin, Virginie; Guenther, Manuela; Hoehne, Melanie; Fernie, Alisdair R; Stitt, Mark

2017-01-01

Worldwide efforts to engineer C 4 photosynthesis into C 3 crops require a deep understanding of how this complex pathway operates. CO 2 is incorporated into four-carbon metabolites in the mesophyll, which move to the bundle sheath where they are decarboxylated to concentrate CO 2 around RuBisCO. We performed dynamic 13 CO 2 labeling in maize to analyze C flow in C 4 photosynthesis. The overall labeling kinetics reflected the topology of C 4 photosynthesis. Analyses of cell-specific labeling patterns after fractionation to enrich bundle sheath and mesophyll cells revealed concentration gradients to drive intercellular diffusion of malate, but not pyruvate, in the major CO 2 -concentrating shuttle. They also revealed intercellular concentration gradients of aspartate, alanine, and phosphenolpyruvate to drive a second phosphoenolpyruvate carboxykinase (PEPCK)-type shuttle, which carries 10-14% of the carbon into the bundle sheath. Gradients also exist to drive intercellular exchange of 3-phosphoglycerate and triose-phosphate. There is rapid carbon exchange between the Calvin-Benson cycle and the CO 2 -concentrating shuttle, equivalent to ~10% of carbon gain. In contrast, very little C leaks from the large pools of metabolites in the C concentration shuttle into respiratory metabolism. We postulate that the presence of multiple shuttles, alongside carbon transfer between them and the Calvin-Benson cycle, confers great flexibility in C 4 photosynthesis. © The Author 2016. Published by Oxford University Press on behalf of the Society for Experimental Biology.

KinG Is a Plant-Specific Kinesin That Regulates Both Intra- and Intercellular Movement of SHORT-ROOT.

PubMed

Spiegelman, Ziv; Lee, Chin-Mei; Gallagher, Kimberly L

2018-01-01

Both endogenous plant proteins and viral movement proteins associate with microtubules to promote their movement through plasmodesmata. The association of viral movement proteins with microtubules facilitates the formation of virus-associated replication complexes, which are required for the amplification and subsequent spread of the virus. However, the role of microtubules in the intercellular movement of plant proteins is less clear. Here we show that the SHORT-ROOT (SHR) protein, which moves between cells in the root to regulate root radial patterning, interacts with a type-14 kinesin, KINESIN G (KinG). KinG is a calponin homology domain kinesin that directly interacts with the SHR-binding protein SIEL (SHR-INTERACING EMBRYONIC LETHAL) and localizes to both microtubules and actin. Since SIEL and SHR associate with endosomes, we suggest that KinG serves as a linker between SIEL, SHR, and the plant cytoskeleton. Loss of KinG function results in a decrease in the intercellular movement of SHR and an increase in the sensitivity of SHR movement to treatment with oryzalin. Examination of SHR and KinG localization and dynamics in live cells suggests that KinG is a nonmotile kinesin that promotes the pausing of SHR-associated endosomes. We suggest a model in which interaction of KinG with SHR allows for the formation of stable movement complexes that facilitate the cell-to-cell transport of SHR. © 2018 American Society of Plant Biologists. All Rights Reserved.

Effect of detergents on the physicochemical properties of skin stratum corneum: a two-photon excitation fluorescence microscopy study.

PubMed

Bloksgaard, M; Brewer, J R; Pashkovski, E; Ananthapadmanabhan, K P; Sørensen, J A; Bagatolli, L A

2014-02-01

Understanding the structural and dynamical features of skin is critical for advancing innovation in personal care and drug discovery. Synthetic detergent mixtures used in commercially available body wash products are thought to be less aggressive towards the skin barrier when compared to conventional detergents. The aim of this work is to comparatively characterize the effect of a mild synthetic cleanser mixture (SCM) and sodium dodecyl sulphate (SDS) on the hydration state of the intercellular lipid matrix and on proton activity of excised skin stratum corneum (SC). Experiments were performed using two-photon excitation fluorescence microscopy. Fluorescent images of fluorescence reporters sensitive to proton activity and hydration of SC were obtained in excised skin and examined in the presence and absence of SCM and SDS detergents. Hydration of the intercellular lipid matrix to a depth of 10 μm into the SC was increased upon treatment with SCM, whereas SDS shows this effect only at the very surface of SC. The proton activity of SC remained unaffected by treatment with either detergent. While our study indicates that the SC is very resistant to external stimuli, it also shows that, in contrast to the response to SDS, SCM to some extent modulates the in-depth hydration properties of the intercellular lipid matrix within excised skin SC. © 2013 Society of Cosmetic Scientists and the Société Française de Cosmétologie.

Celecoxib increases lung cancer cell lysis by lymphokine-activated killer cells via upregulation of ICAM-1.

PubMed

Schellhorn, Melina; Haustein, Maria; Frank, Marcus; Linnebacher, Michael; Hinz, Burkhard

2015-11-17

The antitumorigenic mechanism of the selective cyclooxygenase-2 (COX-2) inhibitor celecoxib is still a matter of debate. Using lung cancer cell lines (A549, H460) and metastatic cells derived from a lung cancer patient, the present study investigates the impact of celecoxib on the expression of intercellular adhesion molecule 1 (ICAM-1) and cancer cell lysis by lymphokine-activated killer (LAK) cells. Celecoxib, but not other structurally related selective COX-2 inhibitors (i.e., etoricoxib, rofecoxib, valdecoxib), was found to cause a substantial upregulation of ICAM-1 protein levels. Likewise, ICAM-1 mRNA expression was increased by celecoxib. Celecoxib enhanced the susceptibility of cancer cells to be lysed by LAK cells with the respective effect being reversed by a neutralizing ICAM-1 antibody. In addition, enhanced killing of celecoxib-treated cancer cells was reversed by preincubation of LAK cells with an antibody to lymphocyte function associated antigen 1 (LFA-1), suggesting intercellular ICAM-1/LFA-1 crosslink as crucial event within this process. Finally, celecoxib elicited no significant increase of LAK cell-mediated lysis of non-tumor bronchial epithelial cells, BEAS-2B, associated with a far less ICAM-1 induction as compared to cancer cells. Altogether, our data demonstrate celecoxib-induced upregulation of ICAM-1 on lung cancer cells to be responsible for intercellular ICAM-1/LFA-1 crosslink that confers increased cancer cell lysis by LAK cells. These findings provide proof for a novel antitumorigenic mechanism of celecoxib.

Celecoxib increases lung cancer cell lysis by lymphokine-activated killer cells via upregulation of ICAM-1

PubMed Central

Frank, Marcus; Linnebacher, Michael; Hinz, Burkhard

2015-01-01

The antitumorigenic mechanism of the selective cyclooxygenase-2 (COX-2) inhibitor celecoxib is still a matter of debate. Using lung cancer cell lines (A549, H460) and metastatic cells derived from a lung cancer patient, the present study investigates the impact of celecoxib on the expression of intercellular adhesion molecule 1 (ICAM-1) and cancer cell lysis by lymphokine-activated killer (LAK) cells. Celecoxib, but not other structurally related selective COX-2 inhibitors (i.e., etoricoxib, rofecoxib, valdecoxib), was found to cause a substantial upregulation of ICAM-1 protein levels. Likewise, ICAM-1 mRNA expression was increased by celecoxib. Celecoxib enhanced the susceptibility of cancer cells to be lysed by LAK cells with the respective effect being reversed by a neutralizing ICAM-1 antibody. In addition, enhanced killing of celecoxib-treated cancer cells was reversed by preincubation of LAK cells with an antibody to lymphocyte function associated antigen 1 (LFA-1), suggesting intercellular ICAM-1/LFA-1 crosslink as crucial event within this process. Finally, celecoxib elicited no significant increase of LAK cell-mediated lysis of non-tumor bronchial epithelial cells, BEAS-2B, associated with a far less ICAM-1 induction as compared to cancer cells. Altogether, our data demonstrate celecoxib-induced upregulation of ICAM-1 on lung cancer cells to be responsible for intercellular ICAM-1/LFA-1 crosslink that confers increased cancer cell lysis by LAK cells. These findings provide proof for a novel antitumorigenic mechanism of celecoxib. PMID:26513172

Determining the impact of cell mixing on signaling during development.

PubMed

Uriu, Koichiro; Morelli, Luis G

2017-06-01

Cell movement and intercellular signaling occur simultaneously to organize morphogenesis during embryonic development. Cell movement can cause relative positional changes between neighboring cells. When intercellular signals are local such cell mixing may affect signaling, changing the flow of information in developing tissues. Little is known about the effect of cell mixing on intercellular signaling in collective cellular behaviors and methods to quantify its impact are lacking. Here we discuss how to determine the impact of cell mixing on cell signaling drawing an example from vertebrate embryogenesis: the segmentation clock, a collective rhythm of interacting genetic oscillators. We argue that comparing cell mixing and signaling timescales is key to determining the influence of mixing. A signaling timescale can be estimated by combining theoretical models with cell signaling perturbation experiments. A mixing timescale can be obtained by analysis of cell trajectories from live imaging. After comparing cell movement analyses in different experimental settings, we highlight challenges in quantifying cell mixing from embryonic timelapse experiments, especially a reference frame problem due to embryonic motions and shape changes. We propose statistical observables characterizing cell mixing that do not depend on the choice of reference frames. Finally, we consider situations in which both cell mixing and signaling involve multiple timescales, precluding a direct comparison between single characteristic timescales. In such situations, physical models based on observables of cell mixing and signaling can simulate the flow of information in tissues and reveal the impact of observed cell mixing on signaling. © 2017 Japanese Society of Developmental Biologists.

An EGFR autocrine loop encodes a slow-reacting but dominant mode of mechanotransduction in a polarized epithelium

PubMed Central

Kojic, Nikola; Chung, Euiheon; Kho, Alvin T.; Park, Jin-Ah; Huang, Austin; So, Peter T. C.; Tschumperlin, Daniel J.

2010-01-01

The mechanical landscape in biological systems can be complex and dynamic, with contrasting sustained and fluctuating loads regularly superposed within the same tissue. How resident cells discriminate between these scenarios to respond accordingly remains largely unknown. Here, we show that a step increase in compressive stress of physiological magnitude shrinks the lateral intercellular space between bronchial epithelial cells, but does so with strikingly slow exponential kinetics (time constant ∼110 s). We confirm that epidermal growth factor (EGF)-family ligands are constitutively shed into the intercellular space and demonstrate that a step increase in compressive stress enhances EGF receptor (EGFR) phosphorylation with magnitude and onset kinetics closely matching those predicted by constant-rate ligand shedding in a slowly shrinking intercellular geometry. Despite the modest degree and slow nature of EGFR activation evoked by compressive stress, we find that the majority of transcriptomic responses to sustained mechanical loading require ongoing activity of this autocrine loop, indicating a dominant role for mechanotransduction through autocrine EGFR signaling in this context. A slow deformation response to a step increase in loading, accompanied by synchronous increases in ligand concentration and EGFR activation, provides one means for cells to mount a selective and context-appropriate response to a sustained change in mechanical environment.—Kojic, N., Chung, E., Kho, A. T., Park, J.-A., Huang, A., So, P. T. C., Tschumperlin, D. J. An EGFR autocrine loop encodes a slow-reacting but dominant mode of mechanotransduction in a polarized epithelium. PMID:20056713

Intercellular adhesion molecule-1 blockade attenuates inflammatory response and improves microvascular perfusion in rat pancreas grafts.

PubMed

Preissler, Gerhard; Eichhorn, Martin; Waldner, Helmut; Winter, Hauke; Kleespies, Axel; Massberg, Steffen

2012-10-01

After pancreas transplantation (PTx), early capillary malperfusion and leukocyte recruitment indicate the manifestation of severe ischemia/reperfusion injury (IRI). Oscillatory blood-flow redistribution (intermittent capillary perfusion, IP), leading to an overall decrease in erythrocyte flux, precedes complete microvascular perfusion failure with persistent blood flow cessation. We addressed the role of intercellular adhesion molecule-1 (ICAM-1) for leukocyte-endothelial interactions (LEIs) after PTx and evaluated the contribution of IP and malperfusion. Pancreas transplantation was performed in rats after 18-hour preservation, receiving either isotype-matched IgG or monoclonal anti-ICAM-1 antibodies (10 mg/kg intravenously) once before reperfusion. Leukocyte-endothelial interaction, IP, erythrocyte flux, and functional capillary density, respectively, were examined in vivo during 2-hour reperfusion. Nontransplanted animals served as controls. Tissue samples were analyzed by histomorphometry. In grafts of IgG-treated animals, IP was encountered already at an early stage after reperfusion and steadily increased over 2 hours, whereas erythrocyte flux declined continuously. In contrast, inhibition of ICAM-1 significantly improved erythrocyte flux and delayed IP appearance by 2 hours. Further, anti-ICAM-1 significantly reduced LEI and leukocyte tissue infiltration when compared to IgG; edema development was less pronounced in response to anti-ICAM-1 monoclonal antibody. Intercellular adhesion molecule-1 blockade significantly attenuates IRI via immediate reduction of LEI and concomitant improvement of capillary perfusion patterns, emphasizing its central role during IRI in PTx.

Identification of heparin-binding EGF-like growth factor as a target in intercellular regulation of epidermal basal cell growth by suprabasal retinoic acid receptors.

PubMed Central

Xiao, J H; Feng, X; Di, W; Peng, Z H; Li, L A; Chambon, P; Voorhees, J J

1999-01-01

The role of retinoic acid receptors (RARs) in intercellular regulation of cell growth was assessed by targeting a dominant-negative RARalpha mutant (dnRARalpha) to differentiated suprabasal cells of mouse epidermis. dnRARalpha lacks transcriptional activation but not DNA-binding and receptor dimerization functions. Analysis of transgenic mice revealed that dnRARalpha dose-dependently impaired induction of basal cell proliferation and epidermal hyperplasia by all-trans RA (tRA). dnRARalpha formed heterodimers with endogenous retinoid X receptor-alpha (RXRalpha) over RA response elements in competition with remaining endogenous RARgamma-RXRalpha heterodimers, and dose-dependently impaired retinoid-dependent gene transcription. To identify genes regulated by retinoid receptors and involved in cell growth control, we analyzed the retinoid effects on expression of the epidermal growth factor (EGF) receptor, EGF, transforming growth factor-alpha, heparin-binding EGF-like growth factor (HB-EGF) and amphiregulin genes. In normal epidermis, tRA rapidly and selectively induced expression of HB-EGF but not the others. This induction occurred exclusively in suprabasal cells. In transgenic epidermis, dnRARalpha dose-dependently inhibited tRA induction of suprabasal HB-EGF and subsequent basal cell hyperproliferation. Together, our observations suggest that retinoid receptor heterodimers located in differentiated suprabasal cells mediate retinoid induction of HB-EGF, which in turn stimulates basal cell growth via intercellular signaling. These events may underlie retinoid action in epidermal regeneration during wound healing. PMID:10075925

Intercellular signaling pathways active during and after growth and differentiation of the lumbar vertebral growth plate.

PubMed

Dahia, Chitra Lekha; Mahoney, Eric J; Durrani, Atiq A; Wylie, Christopher

2011-06-15

Vertebral growth plates at different postnatal ages were assessed for active intercellular signaling pathways. To generate a spatial and temporal map of the major signaling pathways active in the postnatal mouse lumbar vertebral growth plate. The growth of all long bones is known to occur by cartilaginous growth plates. The growth plate is composed of layers of chondrocyets that actively proliferate, differentiate, die and, are replaced by bone. The role of major cell signaling pathways has been suggested for regulation of the fetal long bones. But not much is known about the molecular or cellular signals that control the postnatal vertebral growth plate and hence postnatal vertebral bone growth. Understanding such molecular mechanisms will help design therapeutic treatments for vertebral growth disorders such as scoliosis. Antibodies against activated downstream intermediates were used to identify cells in the growth plate responding to BMP, TGFβ, and FGF in cryosections of lumbar vertebrae from different postnatal age mice to identify the zones that were responding to these signals. Reporter mice were used to identify the chondrocytes responding to hedgehog (Ihh), and Wnt signaling. We present a spatial/temporal map of these signaling pathways during growth, and differentiation of the mouse lumbar vertebral growth plate. During growth and differentiation of the vertebral growth plate, its different components respond at different times to different intercellular signaling ligands. Response to most of these signals is dramatically downregulated at the end of vertebral growth.

Visualization of glucagon secretion from pancreatic α cells by bioluminescence video microscopy: Identification of secretion sites in the intercellular contact regions

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yokawa, Satoru; School of Pharmacy, Aichi Gakuin University, Nagoya 464-8650; Suzuki, Takahiro

We have firstly visualized glucagon secretion using a method of video-rate bioluminescence imaging. The fusion protein of proglucagon and Gaussia luciferase (PGCG-GLase) was used as a reporter to detect glucagon secretion and was efficiently expressed in mouse pancreatic α cells (αTC1.6) using a preferred human codon-optimized gene. In the culture medium of the cells expressing PGCG-GLase, luminescence activity determined with a luminometer was increased with low glucose stimulation and KCl-induced depolarization, as observed for glucagon secretion. From immunochemical analyses, PGCG-GLase stably expressed in clonal αTC1.6 cells was correctly processed and released by secretory granules. Luminescence signals of the secreted PGCG-GLase frommore » the stable cells were visualized by video-rate bioluminescence microscopy. The video images showed an increase in glucagon secretion from clustered cells in response to stimulation by KCl. The secretory events were observed frequently at the intercellular contact regions. Thus, the localization and frequency of glucagon secretion might be regulated by cell-cell adhesion. - Highlights: • The fused protein of proglucagon to Gaussia luciferase was used as a reporter. • The fusion protein was highly expressed using a preferred human-codon optimized gene. • Glucagon secretion stimulated by depolarization was determined by luminescence. • Glucagon secretion in α cells was visualized by bioluminescence imaging. • Glucagon secretion sites were localized in the intercellular contact regions.« less