Identification and functional analysis of endothelial tip cell-enriched genes.

PubMed

del Toro, Raquel; Prahst, Claudia; Mathivet, Thomas; Siegfried, Geraldine; Kaminker, Joshua S; Larrivee, Bruno; Breant, Christiane; Duarte, Antonio; Takakura, Nobuyuki; Fukamizu, Akiyoshi; Penninger, Josef; Eichmann, Anne

2010-11-11

Sprouting of developing blood vessels is mediated by specialized motile endothelial cells localized at the tips of growing capillaries. Following behind the tip cells, endothelial stalk cells form the capillary lumen and proliferate. Expression of the Notch ligand Delta-like-4 (Dll4) in tip cells suppresses tip cell fate in neighboring stalk cells via Notch signaling. In DLL4(+/-) mouse mutants, most retinal endothelial cells display morphologic features of tip cells. We hypothesized that these mouse mutants could be used to isolate tip cells and so to determine their genetic repertoire. Using transcriptome analysis of retinal endothelial cells isolated from DLL4(+/-) and wild-type mice, we identified 3 clusters of tip cell-enriched genes, encoding extracellular matrix degrading enzymes, basement membrane components, and secreted molecules. Secreted molecules endothelial-specific molecule 1, angiopoietin 2, and apelin bind to cognate receptors on endothelial stalk cells. Knockout mice and zebrafish morpholino knockdown of apelin showed delayed angiogenesis and reduced proliferation of stalk cells expressing the apelin receptor APJ. Thus, tip cells may regulate angiogenesis via matrix remodeling, production of basement membrane, and release of secreted molecules, some of which regulate stalk cell behavior.

Electromagnetic needles with submicron pole tip radii for nanomanipulation of biomolecules and living cells

NASA Astrophysics Data System (ADS)

Matthews, Benjamin D.; LaVan, David A.; Overby, Darryl R.; Karavitis, John; Ingber, Donald E.

2004-10-01

We describe the design and fabrication of a temperature-controlled electromagnetic microneedle (EMN) to generate custom magnetic field gradients for biomedical and biophysical applications. An electropolishing technique was developed to sharpen the EMN pole tip to any desired radius between 100 nm and 20 μm. The EMN can be used to apply strong static or dynamic forces (>50nN) to micrometer- or nanometer-sized magnetic beads without producing significant heating or needle movement. Large tip radii (20 μm) allow magnetic force application to multiple magnetic beads over a large area, while small radii (0.1-6 μm) can be used to selectively pull or capture single magnetic beads from within a large population of similar particles. The customizable EMN is thus well suited for micro- and nanomanipulation of magnetic particles linked to biomolecules or living cells.

Chromosomal Fragmentation: A Possible Marker for the Selection of High Gymnemic Acid Yielding Accessions of Gymnema sylvestre R. Br.

PubMed

Verma, Ashutosh Kumar; Dhawan, Sunita Singh

2017-10-01

Gymnema sylvestre R. Br. a member of family Asclepiadaceae as mentioned in Indian Pharmacopoeia popular among the researchers because of stimulatory effect of its phytoconstituent on pancreatic cells and potential to treat Type I and II type of diabetes. Development of cost-effective marker system for the selection of high gymnemic acid yielding accessions of G. sylvestre . Presoaked seeds of Brassica campestris treated with different dilutions of gymnemagenin and 10% leaf extract of twenty different accessions of G. sylvestre . Root tips of germinated seeds were fixed, and chromosomal studies were made by root tip bioassay method. Exposure of seeds to treatment solutions promotes various types of chromosomal anomalies in root meristem, and surprisingly, direct correlation between the percentage of chromosomal fragmentation and the percentage of gymnemic acid shared by treatment solution were observed. Later finding may be explored for the development of a novel methodology or marker system for the selection of high active principle yielding accessions of G. sylvestre . An experiment was carried out using root tip bioassay method for the study of effect of different dilutions of standard gymnemic acid and 10% leaf extract of twenty different accessions of Gymnema sylvestre on root tip meristem of Brassica campestris . Various types of chromosomal anomalies were observed. Of which, percentage of chromosomal fragmentation was showed a direct (∞) relationship with the percentage of gymnemic acid shared by treatment solution. This interesting result after more and more exploration and revalidation could be utilized for the development of a novel methodology for the selection of high active principle yielding accessions of G. sylvestre . Abbreviations used: MI: Mitotic index; CP: Condensed prophase; CM: Clumped metaphase; MC: Metaphase cleft; FR: Fragmentation; AP: Anaphase with persistent nucleolous; LA: Laggard, BR: Bridge; BI: Bi-nucleated cell; DA: Disturbed anaphasic polarity.

Chromosomal Fragmentation: A Possible Marker for the Selection of High Gymnemic Acid Yielding Accessions of Gymnema sylvestre R. Br

PubMed Central

Verma, Ashutosh Kumar; Dhawan, Sunita Singh

2017-01-01

Background: Gymnema sylvestre R. Br. a member of family Asclepiadaceae as mentioned in Indian Pharmacopoeia popular among the researchers because of stimulatory effect of its phytoconstituent on pancreatic cells and potential to treat Type I and II type of diabetes. Objectives: Development of cost-effective marker system for the selection of high gymnemic acid yielding accessions of G. sylvestre. Materials and Methods: Presoaked seeds of Brassica campestris treated with different dilutions of gymnemagenin and 10% leaf extract of twenty different accessions of G. sylvestre. Root tips of germinated seeds were fixed, and chromosomal studies were made by root tip bioassay method. Results: Exposure of seeds to treatment solutions promotes various types of chromosomal anomalies in root meristem, and surprisingly, direct correlation between the percentage of chromosomal fragmentation and the percentage of gymnemic acid shared by treatment solution were observed. Conclusion: Later finding may be explored for the development of a novel methodology or marker system for the selection of high active principle yielding accessions of G. sylvestre. SUMMARY An experiment was carried out using root tip bioassay method for the study of effect of different dilutions of standard gymnemic acid and 10% leaf extract of twenty different accessions of Gymnema sylvestre on root tip meristem of Brassica campestris. Various types of chromosomal anomalies were observed. Of which, percentage of chromosomal fragmentation was showed a direct (∞) relationship with the percentage of gymnemic acid shared by treatment solution. This interesting result after more and more exploration and revalidation could be utilized for the development of a novel methodology for the selection of high active principle yielding accessions of G. sylvestre. Abbreviations used: MI: Mitotic index; CP: Condensed prophase; CM: Clumped metaphase; MC: Metaphase cleft; FR: Fragmentation; AP: Anaphase with persistent nucleolous; LA: Laggard, BR: Bridge; BI: Bi-nucleated cell; DA: Disturbed anaphasic polarity. PMID:29142402

Modulating p56Lck in T-Cells by a Chimeric Peptide Comprising Two Functionally Different Motifs of Tip from Herpesvirus saimiri

PubMed Central

Vernot, Jean-Paul; Perdomo-Arciniegas, Ana María; Pérez-Quintero, Luis Alberto; Martínez, Diego Fernando

2015-01-01

The Lck interacting protein Tip of Herpesvirus saimiri is responsible for T-cell transformation both in vitro and in vivo. Here we designed the chimeric peptide hTip-CSKH, comprising the Lck specific interacting motif CSKH of Tip and its hydrophobic transmembrane sequence (hTip), the latter as a vector targeting lipid rafts. We found that hTip-CSKH can induce a fivefold increase in proliferation of human and Aotus sp. T-cells. Costimulation with PMA did not enhance this proliferation rate, suggesting that hTip-CSKH is sufficient and independent of further PKC stimulation. We also found that human Lck phosphorylation was increased earlier after stimulation when T-cells were incubated previously with hTip-CSKH, supporting a strong signalling and proliferative effect of the chimeric peptide. Additionally, Lck downstream signalling was evident with hTip-CSKH but not with control peptides. Importantly, hTip-CSKH could be identified in heavy lipid rafts membrane fractions, a compartment where important T-cell signalling molecules (LAT, Ras, and Lck) are present during T-cell activation. Interestingly, hTip-CSKH was inhibitory to Jurkat cells, in total agreement with the different signalling pathways and activation requirements of this leukemic cell line. These results provide the basis for the development of new compounds capable of modulating therapeutic targets present in lipid rafts. PMID:26539553

Modulating p56Lck in T-Cells by a Chimeric Peptide Comprising Two Functionally Different Motifs of Tip from Herpesvirus saimiri.

PubMed

Vernot, Jean-Paul; Perdomo-Arciniegas, Ana María; Pérez-Quintero, Luis Alberto; Martínez, Diego Fernando

2015-01-01

The Lck interacting protein Tip of Herpesvirus saimiri is responsible for T-cell transformation both in vitro and in vivo. Here we designed the chimeric peptide hTip-CSKH, comprising the Lck specific interacting motif CSKH of Tip and its hydrophobic transmembrane sequence (hTip), the latter as a vector targeting lipid rafts. We found that hTip-CSKH can induce a fivefold increase in proliferation of human and Aotus sp. T-cells. Costimulation with PMA did not enhance this proliferation rate, suggesting that hTip-CSKH is sufficient and independent of further PKC stimulation. We also found that human Lck phosphorylation was increased earlier after stimulation when T-cells were incubated previously with hTip-CSKH, supporting a strong signalling and proliferative effect of the chimeric peptide. Additionally, Lck downstream signalling was evident with hTip-CSKH but not with control peptides. Importantly, hTip-CSKH could be identified in heavy lipid rafts membrane fractions, a compartment where important T-cell signalling molecules (LAT, Ras, and Lck) are present during T-cell activation. Interestingly, hTip-CSKH was inhibitory to Jurkat cells, in total agreement with the different signalling pathways and activation requirements of this leukemic cell line. These results provide the basis for the development of new compounds capable of modulating therapeutic targets present in lipid rafts.

Fiber-optic Singlet Oxygen [1O2 (1Δg)] Generator Device Serving as a Point Selective Sterilizer

PubMed Central

Aebisher, David; Zamadar, Matibur; Mahendran, Adaickapillai; Ghosh, Goutam; McEntee, Catherine; Greer, Alexander

2016-01-01

Traditionally, Type II heterogeneous photo-oxidations produce singlet oxygen via external irradiation of a sensitizer and external supply of ground-state oxygen. A potential improvement is reported here. A hollow-core fiber-optic device was developed with an “internal” supply of light and flowing oxygen, and a porous photosensitizer-end capped configuration. Singlet oxygen was delivered through the fiber tip. The singlet oxygen steady-state concentration in the immediate vicinity of the probe tip was ca 20 fM by N-benzoyl-DL-methionine trapping. The device is portable and the singlet oxygen-generating tip is maneuverable, which opened the door to simple disinfectant studies. Complete Escherichia coli inactivation was observed in 2 h when the singlet oxygen sensitizing probe tip was immersed in 0.1 mL aqueous samples of 0.1–4.4 × 107 cells. Photobleaching of the probe tip occurred after ca 12 h of use, requiring baking and sensitizer reloading steps for reuse. PMID:20497367

Cotton fiber tips have diverse morphologies and show evidence of apical cell wall synthesis

PubMed Central

Stiff , Michael R.; Haigler, Candace H.

2016-01-01

Cotton fibers arise through highly anisotropic expansion of a single seed epidermal cell. We obtained evidence that apical cell wall synthesis occurs through examining the tips of young elongating Gossypium hirsutum (Gh) and G. barbadense (Gb) fibers. We characterized two tip types in Gh fiber (hemisphere and tapered), each with distinct apical diameter, central vacuole location, and distribution of cell wall components. The apex of Gh hemisphere tips was enriched in homogalacturonan epitopes, including a relatively high methyl-esterified form associated with cell wall pliability. Other wall components increased behind the apex including cellulose and the α-Fuc-(1,2)-β-Gal epitope predominantly found in xyloglucan. Gb fibers had only one narrow tip type featuring characters found in each Gh tip type. Pulse-labeling of cell wall glucans indicated wall synthesis at the apex of both Gh tip types and in distal zones. Living Gh hemisphere and Gb tips ruptured preferentially at the apex upon treatment with wall degrading enzymes, consistent with newly synthesized wall at the apex. Gh tapered tips ruptured either at the apex or distantly. Overall, the results reveal diverse cotton fiber tip morphologies and support primary wall synthesis occurring at the apex and discrete distal regions of the tip. PMID:27301434

Substitution of a single amino acid residue in the aromatic/arginine selectivity filter alters the transport profiles of tonoplast aquaporin homologs.

PubMed

Azad, Abul Kalam; Yoshikawa, Naoki; Ishikawa, Takahiro; Sawa, Yoshihiro; Shibata, Hitoshi

2012-01-01

Aquaporins are integral membrane proteins that facilitate the transport of water and some small solutes across cellular membranes. X-ray crystallography of aquaporins indicates that four amino acids constitute an aromatic/arginine (ar/R) pore constriction known as the selectivity filter. On the basis of these four amino acids, tonoplast aquaporins called tonoplast intrinsic proteins (TIPs) are divided into three groups in Arabidopsis. Herein, we describe the characterization of two group I TIP1s (TgTIP1;1 and TgTIP1;2) from tulip (Tulipa gesneriana). TgTIP1;1 and TgTIP1;2 have a novel isoleucine in loop E (LE2 position) of the ar/R filter; the residue at LE2 is a valine in all group I TIPs from model plants. The homologs showed mercury-sensitive water channel activity in a fast kinetics swelling assay upon heterologous expression in Pichia pastoris. Heterologous expression of both homologs promoted the growth of P. pastoris on ammonium or urea as sole sources of nitrogen and decreased growth and survival in the presence of H(2)O(2). TgTIP1;1- and TgTIP1;2-mediated H(2)O(2) conductance was demonstrated further by a fluorescence assay. Substitutions in the ar/R selectivity filter of TgTIP1;1 showed that mutants that mimicked the ar/R constriction of group I TIPs could conduct the same substrates that were transported by wild-type TgTIP1;1. In contrast, mutants that mimicked group II TIPs showed no evidence of urea or H(2)O(2) conductance. These results suggest that the amino acid residue at LE2 position is critical for the transport selectivity of the TIP homologs and group I TIPs might have a broader spectrum of substrate selectivity than group II TIPs. Copyright © 2011 Elsevier B.V. All rights reserved.

Automatic Test, Configuration, and Repair of Cellular Arrays

DTIC Science & Technology

1975-06-01

one cell to loed any good cell that is not walled off by flawed cells. The loading Information which the computer sends to the array may select one...to loed that cell’s loader and function-specificatioi state bits. After the cell is loaded» it« loader state bits may specify which of the cell’s...Mechanism With Options FUNCTION-SPEC, STATE BITS D.OUT INO INI OUTO 0UT1 LOO LO PAGE 99 LOADER TIP BITS CHANGER ill LSTA S.U.IN S.R.IN

The effect of the serum corona on interactions between a single nano-object and a living cell

NASA Astrophysics Data System (ADS)

Dror, Yael; Sorkin, Raya; Brand, Guy; Boubriak, Olga; Urban, Jill; Klein, Jacob

2017-04-01

Nanoparticles (NPs) which enter physiological fluids are rapidly coated by proteins, forming a so-called corona which may strongly modify their interaction with tissues and cells relative to the bare NPs. In this work the interactions between a living cell and a nano-object, and in particular the effect on this of the adsorption of serum proteins, are directly examined by measuring the forces arising as an Atomic Force Microscope tip (diameter 20 nm) - simulating a nano-object - approaches and contacts a cell. We find that the presence of a serum protein corona on the tip strongly modifies the interaction as indicated by pronounced increase in the indentation, hysteresis and work of adhesion compared to a bare tip. Classically one expects an AFM tip interacting with a cell surface to be repelled due to cell elastic distortion, offset by tip-cell adhesion, and indeed such a model fits the bare-tip/cell interaction, in agreement with earlier work. However, the force plots obtained with serum-modified tips are very different, indicating that the cell is much more compliant to the approaching tip. The insights obtained in this work may promote better design of NPs for drug delivery and other nano-medical applications.

The effect of the serum corona on interactions between a single nano-object and a living cell

PubMed Central

Dror, Yael; Sorkin, Raya; Brand, Guy; Boubriak, Olga; Urban, Jill; Klein, Jacob

2017-01-01

Nanoparticles (NPs) which enter physiological fluids are rapidly coated by proteins, forming a so-called corona which may strongly modify their interaction with tissues and cells relative to the bare NPs. In this work the interactions between a living cell and a nano-object, and in particular the effect on this of the adsorption of serum proteins, are directly examined by measuring the forces arising as an Atomic Force Microscope tip (diameter 20 nm) - simulating a nano-object - approaches and contacts a cell. We find that the presence of a serum protein corona on the tip strongly modifies the interaction as indicated by pronounced increase in the indentation, hysteresis and work of adhesion compared to a bare tip. Classically one expects an AFM tip interacting with a cell surface to be repelled due to cell elastic distortion, offset by tip-cell adhesion, and indeed such a model fits the bare-tip/cell interaction, in agreement with earlier work. However, the force plots obtained with serum-modified tips are very different, indicating that the cell is much more compliant to the approaching tip. The insights obtained in this work may promote better design of NPs for drug delivery and other nano-medical applications. PMID:28383528

Vegetative and sperm cell-specific aquaporins of Arabidopsis highlight the vacuolar equipment of pollen and contribute to plant reproduction.

PubMed

Wudick, Michael M; Luu, Doan-Trung; Tournaire-Roux, Colette; Sakamoto, Wataru; Maurel, Christophe

2014-04-01

The water and nutrient status of pollen is crucial to plant reproduction. Pollen grains of Arabidopsis (Arabidopsis thaliana) contain a large vegetative cell and two smaller sperm cells. Pollen grains express AtTIP1;3 and AtTIP5;1, two members of the Tonoplast Intrinsic Protein subfamily of aquaporins. To address the spatial and temporal expression pattern of the two homologs, C-terminal fusions of AtTIP1;3 and AtTIP5;1 with green fluorescent protein and mCherry, respectively, were expressed in transgenic Arabidopsis under the control of their native promoter. Confocal laser scanning microscopy revealed that AtTIP1;3 and AtTIP5;1 are specific for the vacuoles of the vegetative and sperm cells, respectively. The tonoplast localization of AtTIP5;1 was established by reference to fluorescent protein markers for the mitochondria and vacuoles of sperm and vegetative cells and is at variance with the claim that AtTIP5;1 is localized in vegetative cell mitochondria. AtTIP1;3-green fluorescent protein and AtTIP5;1-mCherry showed concomitant expression, from first pollen mitosis up to pollen tube penetration in the ovule, thereby revealing the dynamics of vacuole morphology in maturating and germinating pollen. Transfer DNA insertion mutants for either AtTIP1;3 or AtTIP5;1 showed no apparent growth phenotype and had no significant defect in male transmission of the mutated alleles. By contrast, a double knockout displayed an abnormal rate of barren siliques, this phenotype being more pronounced under limited water or nutrient supply. The overall data indicate that vacuoles of vegetative and sperm cells functionally interact and contribute to male fertility in adverse environmental conditions.

Applications of fiber-optics-based nanosensors to drug discovery.

PubMed

Vo-Dinh, Tuan; Scaffidi, Jonathan; Gregas, Molly; Zhang, Yan; Seewaldt, Victoria

2009-08-01

Fiber-optic nanosensors are fabricated by heating and pulling optical fibers to yield sub-micron diameter tips and have been used for in vitro analysis of individual living mammalian cells. Immobilization of bioreceptors (e.g., antibodies, peptides, DNA) selective to targeting analyte molecules of interest provides molecular specificity. Excitation light can be launched into the fiber, and the resulting evanescent field at the tip of the nanofiber can be used to excite target molecules bound to the bioreceptor molecules. The fluorescence or surface-enhanced Raman scattering produced by the analyte molecules is detected using an ultra-sensitive photodetector. This article provides an overview of the development and application of fiber-optic nanosensors for drug discovery. The nanosensors provide minimally invasive tools to probe subcellular compartments inside single living cells for health effect studies (e.g., detection of benzopyrene adducts) and medical applications (e.g., monitoring of apoptosis in cells treated with anticancer drugs).

Jagged gives endothelial tip cells an edge.

PubMed

Suchting, Steven; Eichmann, Anne

2009-06-12

Sprouting blood vessels have tip cells that lead and stalk cells that follow. Benedito et al. (2009) now show that competition between endothelial cells for the tip position is regulated by glycosylation of Notch receptors and by the opposing actions of the Notch ligands Jagged1 and Delta-like 4.

TNF-α-inducing protein of Helicobacter pylori induces epithelial-mesenchymal transition (EMT) in gastric cancer cells through activation of IL-6/STAT3 signaling pathway

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

Chen, Guodong; Tang, Na; Wang, Chao

Tumor necrosis factor (TNF)-α-inducing protein (Tipα) is a newly identified carcinogenic factor secreted by Helicobacter pylori (H. pylori). Although it has been proved that Tipα is a strong inducer of epithelial-mesenchymal transition (EMT), a crucial process of migration, the exact molecular mechanism is unknown. Current evidence indicates that the oncogenic transcription factor signal transducers and activators of transcription 3 (STAT3) is inappropriately activated in multiple malignancies, including gastric cancer. In this study, we showed that Tipα significantly down-regulated the expression of EMT-related markers E-cadherin as well as up-regulated N-cadherin and vimentin in SGC7901 cells, with typical morphological changes of EMT. Tipα alsomore » promoted proliferation and migration of SGC7901 cells. Furthermore, Tipα activated interleukin-6 (IL-6)/STAT3 signaling pathway in SGC7901 cells. The effects of Tipα treatment observed was abolished when we block IL-6/STAT3 signaling pathway. Altogether, our data demonstrated that Tipα may accelerate tumor aggressiveness in gastric cancer by promoting EMT through activation of IL-6/STAT3 pathway. - Highlights: • Tipα induces EMT and activates IL-6/STAT3 pathway in gastric cancer cells. • IL-6/STAT3 pathway inhibition reverses Tipα-induced proliferation and migration in gastric cancer cells. • Tipα induces EMT in gastric cancer cells via IL-6/STAT3 pathway activation.« less

Relocalization of human chromatin remodeling cofactor TIP48 in mitosis

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

Sigala, Barbara; Edwards, Mina; Puri, Teena

2005-11-01

TIP48 is a highly conserved eukaryotic AAA{sup +} protein which is an essential cofactor for several complexes involved in chromatin acetylation and remodeling, transcriptional and developmental regulation and nucleolar organization and trafficking. We show that TIP48 abundance in HeLa cells did not change during the cell cycle, nor did its distribution in various biochemical fractions. However, we observed distinct changes in the subcellular localization of TIP48 during M phase using immunofluorescence microscopy. Our studies demonstrate that in interphase cells TIP48 was found mainly in the nucleus and exhibited a distinct localization in the nuclear periphery. As the cells entered mitosis,more » TIP48 was excluded from the condensing chromosomes but showed association with the mitotic apparatus. During anaphase, some TIP48 was detected in the centrosome colocalizing with tubulin but the strongest staining appeared in the mitotic equator associated with the midzone central spindle. Accumulation of TIP48 in the midzone and the midbody was observed in late telophase and cytokinesis. This redeployment of TIP48 during anaphase and cytokinesis was independent of microtubule assembly. The relocation of endogenous TIP48 to the midzone/midbody under physiological conditions suggests a novel and distinct function for TIP48 in mitosis and possible involvement in the exit of mitosis.« less

Comprehensive analysis of tobacco pollen transcriptome unveils common pathways in polar cell expansion and underlying heterochronic shift during spermatogenesis

PubMed Central

2012-01-01

Background Many flowering plants produce bicellular pollen. The two cells of the pollen grain are destined for separate fates in the male gametophyte, which provides a unique opportunity to study genetic interactions that govern guided single-cell polar expansion of the growing pollen tube and the coordinated control of germ cell division and sperm cell fate specification. We applied the Agilent 44 K tobacco gene chip to conduct the first transcriptomic analysis of the tobacco male gametophyte. In addition, we performed a comparative study of the Arabidopsis root-hair trichoblast transcriptome to evaluate genetic factors and common pathways involved in polarized cell-tip expansion. Results Progression of pollen grains from freshly dehisced anthers to pollen tubes 4 h after germination is accompanied with > 5,161 (14.9%) gametophyte-specific expressed probes active in at least one of the developmental stages. In contrast, > 18,821 (54.4%) probes were preferentially expressed in the sporophyte. Our comparative approach identified a subset of 104 pollen tube-expressed genes that overlap with root-hair trichoblasts. Reverse genetic analysis of selected candidates demonstrated that Cu/Zn superoxide dismutase 1 (CSD1), a WD-40 containing protein (BP130384), and Replication factor C1 (NtRFC1) are among the central regulators of pollen-tube tip growth. Extension of our analysis beyond the second haploid mitosis enabled identification of an opposing-dynamic accumulation of core regulators of cell proliferation and cell fate determinants in accordance with the progression of the germ cell cycle. Conclusions The current study provides a foundation to isolate conserved regulators of cell tip expansion and those that are unique for pollen tube growth to the female gametophyte. A transcriptomic data set is presented as a benchmark for future functional studies using developing pollen as a model. Our results demonstrated previously unknown functions of certain genes in pollen-tube tip growth. In addition, we highlighted the molecular dynamics of core cell-cycle regulators in the male gametophyte and postulated the first genetic model to account for the differential timing of spermatogenesis among angiosperms and its coordination with female gametogenesis. PMID:22340370

Role of Tat-interacting protein of 110 kDa and microRNAs in the regulation of hematopoiesis.

PubMed

Liu, Ying; He, Johnny J

2016-07-01

Hematopoiesis is regulated by cellular factors including transcription factors, microRNAs, and epigenetic modifiers. Understanding how these factors regulate hematopoiesis is pivotal for manipulating them to achieve their desired potential. In this review, we will focus on HIV-1 Tat-interacting protein of 110 kDa (Tip110) and its regulation of hematopoiesis. There are several pathways in hematopoiesis that involve Tip110 regulation. Tip110 is expressed in human cord blood CD34 cells; its expression decreases when CD34 cells begin to differentiate. Tip110 is also expressed in mouse marrow hematopoietic stem cells (HSC) and hematopoietic progenitor cells (HPC). Tip110 expression increases the number, survival, and cell cycling of HPC. Tip110-mediated regulation of hematopoiesis has been linked to its reciprocal control of proto-oncogene expression. Small noncoding microRNAs (miRs) have been shown to play important roles in regulation of hematopoiesis. miR-124 specifically targets 3'-untranslated region of Tip110 and subsequently regulates Tip110 expression in HSC. Our recent findings for manipulating expression levels of Tip110 in HSC and HPC could be useful for expanding HSC and HPC and for improving engraftment of cord blood HSC/HPC.

Endothelial Ca2+ oscillations reflect VEGFR signaling-regulated angiogenic capacity in vivo

PubMed Central

Yokota, Yasuhiro; Nakajima, Hiroyuki; Wakayama, Yuki; Muto, Akira; Kawakami, Koichi; Fukuhara, Shigetomo; Mochizuki, Naoki

2015-01-01

Sprouting angiogenesis is a well-coordinated process controlled by multiple extracellular inputs, including vascular endothelial growth factor (VEGF). However, little is known about when and how individual endothelial cell (EC) responds to angiogenic inputs in vivo. Here, we visualized endothelial Ca2+ dynamics in zebrafish and found that intracellular Ca2+ oscillations occurred in ECs exhibiting angiogenic behavior. Ca2+ oscillations depended upon VEGF receptor-2 (Vegfr2) and Vegfr3 in ECs budding from the dorsal aorta (DA) and posterior cardinal vein, respectively. Thus, visualizing Ca2+ oscillations allowed us to monitor EC responses to angiogenic cues. Vegfr-dependent Ca2+ oscillations occurred in migrating tip cells as well as stalk cells budding from the DA. We investigated how Dll4/Notch signaling regulates endothelial Ca2+ oscillations and found that it was required for the selection of single stalk cell as well as tip cell. Thus, we captured spatio-temporal Ca2+ dynamics during sprouting angiogenesis, as a result of cellular responses to angiogenic inputs. DOI: http://dx.doi.org/10.7554/eLife.08817.001 PMID:26588168

Direct mechanical stimulation of tip links in hair cells through DNA tethers

PubMed Central

Basu, Aakash; Lagier, Samuel; Vologodskaia, Maria; Fabella, Brian A; Hudspeth, AJ

2016-01-01

Mechanoelectrical transduction by hair cells commences with hair-bundle deflection, which is postulated to tense filamentous tip links connected to transduction channels. Because direct mechanical stimulation of tip links has not been experimentally possible, this hypothesis has not been tested. We have engineered DNA tethers that link superparamagnetic beads to tip links and exert mechanical forces on the links when exposed to a magnetic-field gradient. By pulling directly on tip links of the bullfrog's sacculus we have evoked transduction currents from hair cells, confirming the hypothesis that tension in the tip links opens transduction channels. This demonstration of direct mechanical access to tip links additionally lays a foundation for experiments probing the mechanics of individual channels. DOI: http://dx.doi.org/10.7554/eLife.16041.001 PMID:27331611

Integrin-specific mechanoresponses to compression and extension probed by cylindrical flat-ended AFM tips in lung cells.

PubMed

Acerbi, Irene; Luque, Tomás; Giménez, Alícia; Puig, Marta; Reguart, Noemi; Farré, Ramon; Navajas, Daniel; Alcaraz, Jordi

2012-01-01

Cells from lung and other tissues are subjected to forces of opposing directions that are largely transmitted through integrin-mediated adhesions. How cells respond to force bidirectionality remains ill defined. To address this question, we nanofabricated flat-ended cylindrical Atomic Force Microscopy (AFM) tips with ~1 µm(2) cross-section area. Tips were uncoated or coated with either integrin-specific (RGD) or non-specific (RGE/BSA) molecules, brought into contact with lung epithelial cells or fibroblasts for 30 s to form focal adhesion precursors, and used to probe cell resistance to deformation in compression and extension. We found that cell resistance to compression was globally higher than to extension regardless of the tip coating. In contrast, both tip-cell adhesion strength and resistance to compression and extension were the highest when probed at integrin-specific adhesions. These integrin-specific mechanoresponses required an intact actin cytoskeleton, and were dependent on tyrosine phosphatases and Ca(2+) signaling. Cell asymmetric mechanoresponse to compression and extension remained after 5 minutes of tip-cell adhesion, revealing that asymmetric resistance to force directionality is an intrinsic property of lung cells, as in most soft tissues. Our findings provide new insights on how lung cells probe the mechanochemical properties of the microenvironment, an important process for migration, repair and tissue homeostasis.

Dll4-containing exosomes induce capillary sprout retraction in a 3D microenvironment

PubMed Central

Sharghi-Namini, Soheila; Tan, Evan; Ong, Lee-Ling Sharon; Ge, Ruowen; Asada, H. Harry

2014-01-01

Delta-like 4 (Dll4), a membrane-bound Notch ligand, plays a fundamental role in vascular development and angiogenesis. Dll4 is highly expressed in capillary endothelial tip cells and is involved in suppressing neighboring stalk cells to become tip cells during angiogenesis. Dll4-Notch signaling is mediated either by direct cell-cell contact or by Dll4-containing exosomes from a distance. However, whether Dll4-containing exosomes influence tip cells of existing capillaries is unknown. Using a 3D microfluidic device and time-lapse confocal microscopy, we show here for the first time that Dll4-containing exosomes causes tip cells to lose their filopodia and trigger capillary sprout retraction in collagen matrix. We demonstrate that Dll4 exosomes can freely travel through 3D collagen matrix and transfer Dll4 protein to distant tip cells. Upon reaching endothelial sprout, it causes filopodia and tip cell retraction. Continuous application of Dll4 exosomes from a distance lead to significant reduction of sprout formation. This effect correlates with Notch signaling activation upon Dll4-containing exosome interaction with recipient endothelial cells. Furthermore, we show that Dll4-containing exosomes increase endothelial cell motility while suppressing their proliferation. These data revealed novel functions of Dll4 in angiogenesis through exosomes. PMID:24504253

Pattern selection and tip perturbations in the Saffman-Taylor problem

NASA Technical Reports Server (NTRS)

Hong, D. C.; Langer, J. S.

1987-01-01

An analytic approach to the Saffman-Taylor problem of predicting the width of a viscous finger in a Hele-Shaw cell is presented. The first purpose is to provide a systematic description of the way in which the singular perturbation introduced by capillary forces leads to a solvability mechanism for pattern selection. It is then shown how recent experimental observations by Couder et al. (1986) may be interpreted in terms suggested by this mechanism.

A role for CSLD3 during cell-wall synthesis in apical plasma membranes of tip-growing root-hair cells.

PubMed

Park, Sungjin; Szumlanski, Amy L; Gu, Fangwei; Guo, Feng; Nielsen, Erik

2011-07-17

In plants, cell shape is defined by the cell wall, and changes in cell shape and size are dictated by modification of existing cell walls and deposition of newly synthesized cell-wall material. In root hairs, expansion occurs by a process called tip growth, which is shared by root hairs, pollen tubes and fungal hyphae. We show that cellulose-like polysaccharides are present in root-hair tips, and de novo synthesis of these polysaccharides is required for tip growth. We also find that eYFP-CSLD3 proteins, but not CESA cellulose synthases, localize to a polarized plasma-membrane domain in root hairs. Using biochemical methods and genetic complementation of a csld3 mutant with a chimaeric CSLD3 protein containing a CESA6 catalytic domain, we provide evidence that CSLD3 represents a distinct (1→4)-β-glucan synthase activity in apical plasma membranes during tip growth in root-hair cells.

Angiogenesis in the reparatory mucosa of the mandibular edentulous ridge is driven by endothelial tip cells.

PubMed

Stănescu, Ruxandra; Didilescu, Andreea Cristiana; Jianu, Adelina Maria; Rusu, M C

2012-01-01

Sprouting angiogenesis is led by specialized cell--the endothelial tip cells (ETCs) which can be targeted by pro- or anti-angiogenic therapies. We aimed to perform a qualitative study in order to assess the guidance by tip cells of the endothelial sprouts in the repairing mucosa of the edentulous mandibular crest. Mucosa of the mandibular edentulous ridge was collected from six adult patients, prior to healing abutment placement (second surgery). Slides were prepared and immunostained with antibodies for CD34 and Ki67. The abundant vasculature of the lamina propria was observed on slides and the CD34 antibodies labeled endothelial tip cells in various stages of the endothelial sprouts. Ki67 identified positive endothelial cells, confirming the proliferative status of the microvascular bed. According to the results, the in situ sprouting angiogenesis is driven by tip cells in the oral mucosa of the edentulous ridge and these cells can be targeted by various therapies, as required by the local pathologic or therapeutic conditions.

[Study on teratogenic effect of potassium dichromate on Vicia faba root tip cells].

PubMed

Qian, Xiao-Wei

2004-05-01

We studied the aberrant effects of different concentrations of potassium dichromate on Vicia faba root tip cells. The micronucleus and chromosome aberration assay was conducted to determine the micronucleus rate and chromosome aberration rate of Vicia faba root tip cells induced by potassium dichromate. The result indicated that potassium dichromate could increase the micronucleus rate of Vicia faba root tip cells. Within certain range of concentration the rate of micronucleus was found to be increased with the increase of potassium dichromate concentration,but beyond this range the rate of micronucleus decreased with further increase of potassium dichromate concentration. The potassium dichromate at different concentrations could increase the cell mitosis index. Besides,it also caused various types of chromosome aberration,and the rates of chromosome aberration were always higher than that of the control group. The conclusion of this study was that potassium dichromate has obvious teratogenic effect on Vicia faba root tip cells.

Ultrastructural analysis of cell component distribution in the apical cell of Ceratodon protonemata

NASA Technical Reports Server (NTRS)

Walker, L. M.; Sack, F. D.

1995-01-01

A distinctive feature of tip-growing plant cells is that cell components are distributed differentially along the length of the cell, although most ultrastructural analyses have been qualitative. The longtitudinal distribution of cell components was studied both qualitatively and quantitatively in the apical cell of dark-grown protonemata of the moss Ceratodon. The first 35 micrometers of the apical cell was analyzed stereologically using transmission electron microscopy. There were four types of distributions along the cell's axis, three of them differential: (1) tubular endoplasmic reticulum was evenly distributed, (2) cisternal endoplasmic reticulum and Golgi vesicles were distributed in a tip-to-base gradient, (3) plastids, vacuoles, and Golgi stacks were enriched in specific areas, although the locations of the enrichments varied, and (4) mitochondria were excluded in the tip-most 5 micrometers and evenly distributed throughout the remaining 30 micrometers. This study provides one of the most comprehensive quantitative, ultrastructural analyses of the distribution of cell components in the apex of any tip-growing plant cell. The finding that almost every component had its own spatial arrangement demonstrates the complexity of the organization and regulation of the distribution of components in tip-growing cells.

Tip60 degradation by adenovirus relieves transcriptional repression of viral transcriptional activator EIA.

PubMed

Gupta, A; Jha, S; Engel, D A; Ornelles, D A; Dutta, A

2013-10-17

Adenoviruses are linear double-stranded DNA viruses that infect human and rodent cell lines, occasionally transform them and cause tumors in animal models. The host cell challenges the virus in multifaceted ways to restrain viral gene expression and DNA replication, and sometimes even eliminates the infected cells by programmed cell death. To combat these challenges, adenoviruses abrogate the cellular DNA damage response pathway. Tip60 is a lysine acetyltransferase that acetylates histones and other proteins to regulate gene expression, DNA damage response, apoptosis and cell cycle regulation. Tip60 is a bona fide tumor suppressor as mice that are haploid for Tip60 are predisposed to tumors. We have discovered that Tip60 is degraded by adenovirus oncoproteins EIB55K and E4orf6 by a proteasome-mediated pathway. Tip60 binds to the immediate early adenovirus promoter and suppresses adenovirus EIA gene expression, which is a master regulator of adenovirus transcription, at least partly through retention of the virally encoded repressor pVII on this promoter. Thus, degradation of Tip60 by the adenoviral early proteins is important for efficient viral early gene transcription and for changes in expression of cellular genes.

HIV-1 Tat targets Tip60 to impair the apoptotic cell response to genotoxic stresses

PubMed Central

Col, Edwige; Caron, Cécile; Chable-Bessia, Christine; Legube, Gaelle; Gazzeri, Sylvie; Komatsu, Yasuhiko; Yoshida, Minoru; Benkirane, Monsef; Trouche, Didier; Khochbin, Saadi

2005-01-01

HIV-1 transactivator Tat uses cellular acetylation signalling by targeting several cellular histone acetyltransferases (HAT) to optimize its various functions. Although Tip60 was the first HAT identified to interact with Tat, the biological significance of this interaction has remained obscure. We had previously shown that Tat represses Tip60 HAT activity. Here, a new mechanism of Tip60 neutralization by Tat is described, where Tip60 is identified as a substrate for the newly reported p300/CBP-associated E4-type ubiquitin-ligase activity, and Tat uses this mechanism to induce the polyubiquitination and degradation of Tip60. Tip60 targeting by Tat results in a dramatic impairment of the Tip60-dependent apoptotic cell response to DNA damage. These data reveal yet unknown strategies developed by HIV-1 to increase cell resistance to genotoxic stresses and show a role of Tat as a modulator of cellular protein ubiquitination. PMID:16001085

[Effects of chlorobenzene stress on seedling growth and cell division of Vicia faba].

PubMed

Liu, Wan; Zhou, Qixing; Li, Peijun; Sun, Tieheng; Tai, Peidong; Xu, Huaxia; Zhang, Chungui; Zhang, Hairong

2003-04-01

Effects of 1, 2, 4-trichlorobenzene (TCB) stress on seedling growth, cell division and chromosomal aberration frequency of root-tip cells of Vicia faba were studied. The results indicated that the growth of the root length and mitotic index of root tip cells were successively decreased and even stopped with the increase of TCB concentrations and treatment duration. Numerical and structural chromosomal aberrations at metaphase and anaphase of root-tip cells in Vicia faba seedlings were produced by 50-300 micrograms.g-1 TCB treatment for 12-96 h. The percentage of c-mitosis, chromosomal bridge and chromosomal asymmetry array in root tip cells exposed to 50-100 micrograms.g-1 TCB for 12-24 h was up to 1.0-10.3%. The percentage of chromosomal stickness (S), chromosomal stickiness + chromosomal breakage (S + B), chromosomal stickness + chromosomal ring (S + R), chromosomal stickiness + chromosomal asymmetry array (S + A) and chromosomal stickness + chromosomal bridge (S + Be) in root tip cells reached 47.9-88.9%, and 18.1-29.6% for different kinds of chromosomal breakage at 300 micrograms.g-1 TCB for 12-96 h. Thus, the chromosomal aberration of root tip cells in Vicia faba seedlings could be used as a sensitive biomarker of monitoring soil contaminated with TCB.

Adhesion between peptides/antibodies and breast cancer cells

NASA Astrophysics Data System (ADS)

Meng, J.; Paetzell, E.; Bogorad, A.; Soboyejo, W. O.

2010-06-01

Atomic force microscopy (AFM) techniques were used to measure the adhesion forces between the receptors on breast cancer cells specific to human luteinizing hormone-releasing hormone (LHRH) peptides and antibodies specific to the EphA2 receptor. The adhesion forces between LHRH-coated AFM tips and human MDA-MB-231 cells (breast cancer cells) were shown to be about five times greater than those between LHRH-coated AFM tips and normal Hs578Bst breast cells. Similarly, those between EphA2 antibody-coated AFM tips and breast cancer cells were over five times greater than those between EphA2 antibody-coated AFM tips and normal breast cells. The results suggest that AFM can be used for the detection of breast cancer cells in biopsies. The implications of the results are also discussed for the early detection and localized treatment of cancer.

DNA-binding activity of TNF-{alpha} inducing protein from Helicobacter pylori

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

Kuzuhara, T.; Suganuma, M.; Oka, K.

2007-11-03

Tumor necrosis factor-{alpha} (TNF-{alpha}) inducing protein (Tip{alpha}) is a carcinogenic factor secreted from Helicobacter pylori (H. pylori), mediated through both enhanced expression of TNF-{alpha} and chemokine genes and activation of nuclear factor-{kappa}B. Since Tip{alpha} enters gastric cancer cells, the Tip{alpha} binding molecules in the cells should be investigated. The direct DNA-binding activity of Tip{alpha} was observed by pull down assay using single- and double-stranded genomic DNA cellulose. The surface plasmon resonance assay, indicating an association between Tip{alpha} and DNA, revealed that the affinity of Tip{alpha} for (dGdC)10 is 2400 times stronger than that of del-Tip{alpha}, an inactive Tip{alpha}. This suggestsmore » a strong correlation between DNA-binding activity and carcinogenic activity of Tip{alpha}. And the DNA-binding activity of Tip{alpha} was first demonstrated with a molecule secreted from H. pylori.« less

Filter presence and tipping paper color influence consumer perceptions of cigarettes.

PubMed

O'Connor, Richard J; Bansal-Travers, Maansi; Cummings, K Michael; Hammond, David; Thrasher, James F; Tworek, Cindy

2015-12-22

Cigarettes are marketed in a wide array of packaging and product configurations, and these may impact consumers' perceptions of product health effects and attractiveness. Filtered cigarettes are typically perceived as less hazardous and white tipping paper (as opposed to cork) often conveys 'lightness'. This study examined cigarette-related perceptions among 1220 young adult (age 18-35) current, ever, and never smokers recruited from three eastern U.S. cities (Buffalo NY, Columbia SC, Morgantown WV). Participants rated three cigarette sticks: two filtered cigarettes 85 mm in length, differing only in tipping paper color (cork versus white), and an unfiltered 70 mm cigarette. Overall, the cork-tipped cigarette was most commonly selected on taste and attractiveness, the white-tipped on least dangerous, and the unfiltered on most dangerous. Current smokers were more likely to select white-tipped (OR = 1.98) and cork-tipped (OR = 3.42) cigarettes, while ever smokers more commonly selected the cork-tipped (OR = 1.96), as most willing to try over the other products. Those willing to try the filtered white-tipped cigarette were more likely to have rated that cigarette as best tasting (OR = 11.10), attracting attention (OR = 17.91), and lowest health risk (OR = 1.94). Similarly, those willing to try cork tipped or unfiltered cigarettes rated those as best testing, attracting attention, and lowest health risk, respectively. Findings from this study demonstrate that consumer product perceptions can be influenced by elements of cigarette design, such as the presence and color of the filter tip.

Reduction of TIP30 in esophageal squamous cell carcinoma cells involves promoter methylation and microRNA-10b

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

Dong, Wenjie, E-mail: dongwenjie200581@126.com; Shen, Ruizhe; Cheng, Shidan

2014-10-31

Highlights: • TIP30 expression is frequently suppressed in ESCC. • TIP30 was hypermethylated in ESCC. • Reduction of TIP30 was significantly correlated with LN metastasis. • miR-10b is a direct regulator of TIP30. - Abstract: TIP30 is a putative tumor suppressor that can promote apoptosis and inhibit angiogenesis. However, the role of TIP30 in esophageal squamous cell carcinoma (ESCC) biology has not been investigated. Immunohistochemistry was used to investigate the expression of TIP30 in 70 ESCC. Hypermethylation of TIP30 was evaluated by the methylation specific PCR (MSP) method in ESCC (tumor and paired adjacent non-tumor tissues). Lost expression of TIP30more » was observed in 50 of 70 (71.4%) ESCC. 61.4% (43 of 70) of primary tumors analyzed displayed TIP30 hypermethylation, indicating that this aberrant characteristic is common in ESCC. Moreover, a statistically significant inverse association was found between TIP30 methylation status and expression of the TIP30 protein in tumor tissues (p = 0.001). We also found that microRNA-10b (miR-10b) targets a homologous DNA region in the 3′untranslated region of the TIP30 gene and represses its expression at the transcriptional level. Reporter assay with 3′UTR of TIP30 cloned downstream of the luciferase gene showed reduced luciferase activity in the presence of miR-10b, providing strong evidence that miR-10b is a direct regulator of TIP30. These results suggest that TIP30 expression is regulated by promoter methylation and miR-10b in ESCC.« less

Imaging of soft material with carbon nanotube tip using near-field scanning microwave microscopy.

PubMed

Wu, Zhe; Sun, Wei-Qiang; Feng, Tao; Tang, Shawn Wenjie; Li, Gang; Jiang, Kai-Li; Xu, Sheng-Yong; Ong, Chong Kim

2015-01-01

In this manuscript, a near-field scanning microwave microscope (NSMM) of our own design is introduced while using a multi-walled carbon nanotube (MWCNT) bundle as the tip (referred to as 'CNT tip'). Clear images of gold-patterned numbers, photoresist stripes and corneal endothelial cells (cell line B4G12) were obtained by mapping the resonant frequency fr and S11 amplitude of a given area while the NSMM is operating in tapping mode. The CNT tip helps to improve image quality and reveals more information about the sample as compared to a traditional metallic tip. The CNT tip is flexible and does not scratch the surface of the sample during the scan, which is useful for imaging soft material in biological science. In the imaging of the B4G12 endothelial cells, the nuclei and cytoplasm can be clearly distinguished from the rest of the cell and its surrounding medium. Copyright © 2014 Elsevier B.V. All rights reserved.

Means and methods for cytometric therapies

DOEpatents

Gillies, George T.; Fillmore, Helen; Broaddus, William C.; Evans, III, Boyd M.; Allison, Stephen W.

2013-03-26

A functionalized tip is incorporated into catheters for the cytometric delivery of cells into the brain and other body parts. For use in the brain, the tip forms part of a neurosurgical probe having a proximal end and a distal end. In addition to the functionalized tip, the probe has at least one cell slurry delivery lumen and a plurality of optical fibers configured along the probe, terminating in the tip to provide the photo-optical capability needed to monitor the viability and physiological behavior of the grafted cells as well as certain characteristics of the cellular environment. Details are also presented of the use of a neurocatheter having a cytometric tip of the type disclosed in the invention, as employed within the context of a feedback and control system for regulating the number of cells delivered to the brain of a patient.

The type III secretion system needle tip complex mediates host cell sensing and translocon insertion.

PubMed

Veenendaal, Andreas K J; Hodgkinson, Julie L; Schwarzer, Lynn; Stabat, David; Zenk, Sebastian F; Blocker, Ariel J

2007-03-01

Type III secretion systems (T3SSs) are essential virulence determinants of many Gram-negative bacterial pathogens. The Shigella T3SS consists of a cytoplasmic bulb, a transmembrane region and a hollow 'needle' protruding from the bacterial surface. Physical contact with host cells initiates secretion and leads to assembly of a pore, formed by IpaB and IpaC, in the host cell membrane, through which proteins that facilitate host cell invasion are translocated. As the needle is implicated in host cell sensing and secretion regulation, its tip should contain components that initiate host cell contact. Through biochemical and immunological studies of wild-type and mutant Shigella T3SS needles, we reveal tip complexes of differing compositions and functional states, which appear to represent the molecular events surrounding host cell sensing and pore formation. Our studies indicate that the interaction between IpaB and IpaD at needle tips is key to host cell sensing, orchestration of IpaC secretion and its subsequent assembly at needle tips. This allows insertion into the host cell membrane of a translocation pore that is continuous with the needle.

Molecular tips for scanning tunneling microscopy: intermolecular electron tunneling for single-molecule recognition and electronics.

PubMed

Nishino, Tomoaki

2014-01-01

This paper reviews the development of molecular tips for scanning tunneling microscopy (STM). Molecular tips offer many advantages: first is their ability to perform chemically selective imaging because of chemical interactions between the sample and the molecular tip, thus improving a major drawback of conventional STM. Rational design of the molecular tip allows sophisticated chemical recognition; e.g., chiral recognition and selective visualization of atomic defects in carbon nanotubes. Another advantage is that they provide a unique method to quantify electron transfer between single molecules. Understanding such electron transfer is mandatory for the realization of molecular electronics.

23 CFR 450.330 - Project selection from the TIP.

Code of Federal Regulations, 2013 CFR

2013-04-01

... 23 Highways 1 2013-04-01 2013-04-01 false Project selection from the TIP. 450.330 Section 450.330 Highways FEDERAL HIGHWAY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION PLANNING AND RESEARCH PLANNING ASSISTANCE AND STANDARDS Metropolitan Transportation Planning and Programming § 450.330 Project selection...

23 CFR 450.330 - Project selection from the TIP.

Code of Federal Regulations, 2012 CFR

2012-04-01

... 23 Highways 1 2012-04-01 2012-04-01 false Project selection from the TIP. 450.330 Section 450.330 Highways FEDERAL HIGHWAY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION PLANNING AND RESEARCH PLANNING ASSISTANCE AND STANDARDS Metropolitan Transportation Planning and Programming § 450.330 Project selection...

23 CFR 450.330 - Project selection from the TIP.

Code of Federal Regulations, 2014 CFR

2014-04-01

... 23 Highways 1 2014-04-01 2014-04-01 false Project selection from the TIP. 450.330 Section 450.330 Highways FEDERAL HIGHWAY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION PLANNING AND RESEARCH PLANNING ASSISTANCE AND STANDARDS Metropolitan Transportation Planning and Programming § 450.330 Project selection...

23 CFR 450.330 - Project selection from the TIP.

Code of Federal Regulations, 2011 CFR

2011-04-01

... 23 Highways 1 2011-04-01 2011-04-01 false Project selection from the TIP. 450.330 Section 450.330 Highways FEDERAL HIGHWAY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION PLANNING AND RESEARCH PLANNING ASSISTANCE AND STANDARDS Metropolitan Transportation Planning and Programming § 450.330 Project selection...

Adenylyl cyclase A expression is tip-specific in Dictyostelium slugs and directs StatA nuclear translocation and CudA gene expression.

PubMed

Verkerke-van Wijk, I; Fukuzawa, M; Devreotes, P N; Schaap, P

2001-06-01

cAMP oscillations, generated by adenylyl cyclase A (ACA), coordinate cell aggregation in Dictyostelium and have also been implicated in organizer function during multicellular development. We used a gene fusion of the ACA promoter with a labile lacZ derivative to study the expression pattern of ACA. During aggregation, most cells expressed ACA, but thereafter expression was lost in all cells except those of the anterior tip. Before aggregation, ACA transcription was strongly upregulated by nanomolar cAMP pulses. Postaggregative transcription was sustained by nanomolar cAMP pulses, but downregulated by a continuous micromolar cAMP stimulus and by the stalk-cell-inducing factor DIF. Earlier work showed that the transcription factor StatA displays tip-specific nuclear translocation and directs tip-specific expression of the nuclear protein CudA, which is essential for culmination. Both StatA and CudA were present in nuclei throughout the entire slug in an aca null mutant that expresses ACA from the constitutive actin15 promoter. This suggests that the tip-specific expression of ACA directs tip-specific nuclear translocation of StatA and tip-specific expression of CudA. Copyright 2001 Academic Press.

Mechanotransduction current is essential for stability of the transducing stereocilia in mammalian auditory hair cells

PubMed Central

Vélez-Ortega, A Catalina; Freeman, Mary J; Indzhykulian, Artur A; Grossheim, Jonathan M; Frolenkov, Gregory I

2017-01-01

Mechanotransducer channels at the tips of sensory stereocilia of inner ear hair cells are gated by the tension of 'tip links' interconnecting stereocilia. To ensure maximal sensitivity, tip links are tensioned at rest, resulting in a continuous influx of Ca2+ into the cell. Here, we show that this constitutive Ca2+ influx, usually considered as potentially deleterious for hair cells, is in fact essential for stereocilia stability. In the auditory hair cells of young postnatal mice and rats, a reduction in mechanotransducer current, via pharmacological channel blockers or disruption of tip links, leads to stereocilia shape changes and shortening. These effects occur only in stereocilia that harbor mechanotransducer channels, recover upon blocker washout or tip link regeneration and can be replicated by manipulations of extracellular Ca2+ or intracellular Ca2+ buffering. Thus, our data provide the first experimental evidence for the dynamic control of stereocilia morphology by the mechanotransduction current. DOI: http://dx.doi.org/10.7554/eLife.24661.001 PMID:28350294

Rapid Polymer Sequencer

NASA Technical Reports Server (NTRS)

Stolc, Viktor (Inventor); Brock, Mathew W. (Inventor)

2011-01-01

Method and system for rapid and accurate determination of each of a sequence of unknown polymer components, such as nucleic acid components. A self-assembling monolayer of a selected substance is optionally provided on an interior surface of a pipette tip, and the interior surface is immersed in a selected liquid. A selected electrical field is impressed in a longitudinal or transverse direction at the tip, a polymer sequence is passed through the tip, and a change in an electrical current signal is measured as each polymer component passes through the tip. Each measured change in electrical current signals is compared with a database of reference signals, with each reference signal identified with a polymer component, to identify the unknown polymer component. The tip preferably has a pore inner diameter of no more than about 40 nm and is prepared by heating and pulling a very small section of a glass tubing.

Physcomitrella patens: a model for tip cell growth and differentiation.

PubMed

Vidali, Luis; Bezanilla, Magdalena

2012-12-01

The moss Physcomitrella patens has emerged as an excellent model system owing to its amenability to reverse genetics. The moss gametophyte has three filamentous tissues that grow by tip growth: chloronemata, caulonemata, and rhizoids. Because establishment of the moss plant relies on this form of growth, it is particularly suited for dissecting the molecular basis of tip growth. Recent studies demonstrate that a core set of actin cytoskeletal proteins is essential for tip growth. Additional actin cytoskeletal components are required for modulating growth to produce caulonemata and rhizoids. Differentiation into these cell types has previously been linked to auxin, light and nutrients. Recent studies have identified that core auxin signaling components as well as transcription factors that respond to auxin or nutrient levels are required for tip-growing cell differentiation. Future studies may establish a connection between the actin cytoskeleton and auxin or nutrient-induced cell differentiation. Copyright © 2012 Elsevier Ltd. All rights reserved.

Joint effects of microwave and chromium trioxide on root tip cells of Vicia faba *

PubMed Central

Qian, Xiao-Wei; Luo, Wei-Hua; Zheng, Ou-Xiang

2006-01-01

The mutagenic effects of microwave and chromium trioxide (CrO3) on Vicia faba root tip were studied. Micronucleus assay and chromosomal aberration assay were used to determine the mitotic index, the micronucleus frequency and chromosomal aberration frequency of Vicia faba root tip cells induced by microwave and CrO3. The results showed that the micronucleus frequency decreased, and that the mitotic index and chromosomal aberration frequency showed linear dose responses to CrO3, in treatment of microwave for 5 s. In microwave of 25 s, the mitotic index decreased, the micronucleus frequency and chromosomal aberration frequency increased with increase of CrO3 concentration. We concluded that microwave and CrO3 had antagonistic effect on the mitotic index of Vicia faba root tip cells, but had synergetic effect on micronucleus frequency and chromosomal aberration frequency of Vicia faba root tip cells. PMID:16502510

Joint effects of microwave and chromium trioxide on root tip cells of Vicia faba.

PubMed

Qian, Xiao-wei; Luo, Wei-hua; Zheng, Ou-xiang

2006-03-01

The mutagenic effects of microwave and chromium trioxide (CrO(3)) on Vicia faba root tip were studied. Micronucleus assay and chromosomal aberration assay were used to determine the mitotic index, the micronucleus frequency and chromosomal aberration frequency of Vicia faba root tip cells induced by microwave and CrO(3). The results showed that the micronucleus frequency decreased, and that the mitotic index and chromosomal aberration frequency showed linear dose responses to CrO(3), in treatment of microwave for 5 s. In microwave of 25 s, the mitotic index decreased, the micronucleus frequency and chromosomal aberration frequency increased with increase of CrO(3) concentration. We concluded that microwave and CrO(3) had antagonistic effect on the mitotic index of Vicia faba root tip cells, but had synergetic effect on micronucleus frequency and chromosomal aberration frequency of Vicia faba root tip cells.

Optically triggering spatiotemporally confined GPCR activity in a cell and programming neurite initiation and extension

PubMed Central

Karunarathne, W. K. Ajith; Giri, Lopamudra; Kalyanaraman, Vani; Gautam, N.

2013-01-01

G-protein–coupled receptor (GPCR) activity gradients evoke important cell behavior but there is a dearth of methods to induce such asymmetric signaling in a cell. Here we achieved reversible, rapidly switchable patterns of spatiotemporally restricted GPCR activity in a single cell. We recruited properties of nonrhodopsin opsins—rapid deactivation, distinct spectral tuning, and resistance to bleaching—to activate native Gi, Gq, or Gs signaling in selected regions of a cell. Optical inputs were designed to spatiotemporally control levels of second messengers, IP3, phosphatidylinositol (3,4,5)-triphosphate, and cAMP in a cell. Spectrally selective imaging was accomplished to simultaneously monitor optically evoked molecular and cellular response dynamics. We show that localized optical activation of an opsin-based trigger can induce neurite initiation, phosphatidylinositol (3,4,5)-triphosphate increase, and actin remodeling. Serial optical inputs to neurite tips can refashion early neuron differentiation. Methods here can be widely applied to program GPCR-mediated cell behaviors. PMID:23479634

The Rice Basic Helix-Loop-Helix Transcription Factor TDR INTERACTING PROTEIN2 Is a Central Switch in Early Anther Development[C][W

PubMed Central

Fu, Zhenzhen; Yu, Jing; Cheng, Xiaowei; Zong, Xu; Xu, Jie; Chen, Mingjiao; Li, Zongyun; Zhang, Dabing; Liang, Wanqi

2014-01-01

In male reproductive development in plants, meristemoid precursor cells possessing transient, stem cell–like features undergo cell divisions and differentiation to produce the anther, the male reproductive organ. The anther contains centrally positioned microsporocytes surrounded by four distinct layers of wall: the epidermis, endothecium, middle layer, and tapetum. Here, we report that the rice (Oryza sativa) basic helix-loop-helix (bHLH) protein TDR INTERACTING PROTEIN2 (TIP2) functions as a crucial switch in the meristemoid transition and differentiation during early anther development. The tip2 mutants display undifferentiated inner three anther wall layers and abort tapetal programmed cell death, causing complete male sterility. TIP2 has two paralogs in rice, TDR and EAT1, which are key regulators of tapetal programmed cell death. We revealed that TIP2 acts upstream of TDR and EAT1 and directly regulates the expression of TDR and EAT1. In addition, TIP2 can interact with TDR, indicating a role of TIP2 in later anther development. Our findings suggest that the bHLH proteins TIP2, TDR, and EAT1 play a central role in regulating differentiation, morphogenesis, and degradation of anther somatic cell layers, highlighting the role of paralogous bHLH proteins in regulating distinct steps of plant cell–type determination. PMID:24755456

Rapid change of chromomeric and pairing patterns of polytene chromosome tips in D. melanogaster: migration of polytene-nonpolytene transition zone?

PubMed

Roberts, P A

1979-07-01

The high variability of chromomeric patterns in near-terminal regions of polytene chromosome arms has been explored in a number of races, strains and hybrids of Drosophila melanogaster. Traditional explanations for tip differences between strains (differential compaction of chromatin, somatic or germinal deletion) are examined and, in the light of the reported observations, rejected. The range of polytene tip variability and rates of change in wild races are greater than has been supposed: strains formerly considered to be terminally deleted appear to gain terminal bands; others, formerly considered normal, appear to have lost them. Strains with high cell-to-cell tip variability are also described. Cell-to-cell variations, as well as much of the observed rapid changes in tip appearance, are probably due to heritable differences in the location of an abrupt transition zone between polytene and nonpolytene chromatin. A quantitative relationship between the amount of certain subterminal bands present and the frequency of tip association of nonhomologous chromosomes is shown and its possible significance for chromosome is shown and its possible for chromosome pairing discussed.

Mutations in cadherin 23 affect tip links in zebrafish sensory hair cells.

PubMed

Söllner, Christian; Rauch, Gerd-Jörg; Siemens, Jan; Geisler, Robert; Schuster, Stephan C; Müller, Ulrich; Nicolson, Teresa

2004-04-29

Hair cells have highly organized bundles of apical projections, or stereocilia, that are deflected by sound and movement. Displacement of stereocilia stretches linkages at the tips of stereocilia that are thought to gate mechanosensory channels. To identify the molecular machinery that mediates mechanotransduction in hair cells, zebrafish mutants were identified with defects in balance and hearing. In sputnik mutants, stereociliary bundles are splayed to various degrees, with individuals displaying reduced or absent mechanotransduction. Here we show that the defects in sputnik mutants are caused by mutations in cadherin 23 (cdh23). Mutations in Cdh23 also cause deafness and vestibular defects in mice and humans, and the protein is present in hair bundles. We show that zebrafish Cdh23 protein is concentrated near the tips of hair bundles, and that tip links are absent in homozygous sputnik(tc317e) larvae. Moreover, tip links are absent in larvae carrying weak alleles of cdh23 that affect mechanotransduction but not hair bundle integrity. We conclude that Cdh23 is an essential tip link component required for hair-cell mechanotransduction.

Raman Imaging in Cell Membranes, Lipid-Rich Organelles, and Lipid Bilayers.

PubMed

Syed, Aleem; Smith, Emily A

2017-06-12

Raman-based optical imaging is a promising analytical tool for noninvasive, label-free chemical imaging of lipid bilayers and cellular membranes. Imaging using spontaneous Raman scattering suffers from a low intensity that hinders its use in some cellular applications. However, developments in coherent Raman imaging, surface-enhanced Raman imaging, and tip-enhanced Raman imaging have enabled video-rate imaging, excellent detection limits, and nanometer spatial resolution, respectively. After a brief introduction to these commonly used Raman imaging techniques for cell membrane studies, this review discusses selected applications of these modalities for chemical imaging of membrane proteins and lipids. Finally, recent developments in chemical tags for Raman imaging and their applications in the analysis of selected cell membrane components are summarized. Ongoing developments toward improving the temporal and spatial resolution of Raman imaging and small-molecule tags with strong Raman scattering cross sections continue to expand the utility of Raman imaging for diverse cell membrane studies.

Genome-wide identification, characterization, and expression profile of aquaporin gene family in flax (Linum usitatissimum)

PubMed Central

Shivaraj, S. M.; Deshmukh, Rupesh K.; Rai, Rhitu; Bélanger, Richard; Agrawal, Pawan K.; Dash, Prasanta K.

2017-01-01

Membrane intrinsic proteins (MIPs) form transmembrane channels and facilitate transport of myriad substrates across the cell membrane in many organisms. Majority of plant MIPs have water transporting ability and are commonly referred as aquaporins (AQPs). In the present study, we identified aquaporin coding genes in flax by genome-wide analysis, their structure, function and expression pattern by pan-genome exploration. Cross-genera phylogenetic analysis with known aquaporins from rice, arabidopsis, and poplar showed five subgroups of flax aquaporins representing 16 plasma membrane intrinsic proteins (PIPs), 17 tonoplast intrinsic proteins (TIPs), 13 NOD26-like intrinsic proteins (NIPs), 2 small basic intrinsic proteins (SIPs), and 3 uncharacterized intrinsic proteins (XIPs). Amongst aquaporins, PIPs contained hydrophilic aromatic arginine (ar/R) selective filter but TIP, NIP, SIP and XIP subfamilies mostly contained hydrophobic ar/R selective filter. Analysis of RNA-seq and microarray data revealed high expression of PIPs in multiple tissues, low expression of NIPs, and seed specific expression of TIP3 in flax. Exploration of aquaporin homologs in three closely related Linum species bienne, grandiflorum and leonii revealed presence of 49, 39 and 19 AQPs, respectively. The genome-wide identification of aquaporins, first in flax, provides insight to elucidate their physiological and developmental roles in flax. PMID:28447607

Genome-wide identification, characterization, and expression profile of aquaporin gene family in flax (Linum usitatissimum).

PubMed

Shivaraj, S M; Deshmukh, Rupesh K; Rai, Rhitu; Bélanger, Richard; Agrawal, Pawan K; Dash, Prasanta K

2017-04-27

Membrane intrinsic proteins (MIPs) form transmembrane channels and facilitate transport of myriad substrates across the cell membrane in many organisms. Majority of plant MIPs have water transporting ability and are commonly referred as aquaporins (AQPs). In the present study, we identified aquaporin coding genes in flax by genome-wide analysis, their structure, function and expression pattern by pan-genome exploration. Cross-genera phylogenetic analysis with known aquaporins from rice, arabidopsis, and poplar showed five subgroups of flax aquaporins representing 16 plasma membrane intrinsic proteins (PIPs), 17 tonoplast intrinsic proteins (TIPs), 13 NOD26-like intrinsic proteins (NIPs), 2 small basic intrinsic proteins (SIPs), and 3 uncharacterized intrinsic proteins (XIPs). Amongst aquaporins, PIPs contained hydrophilic aromatic arginine (ar/R) selective filter but TIP, NIP, SIP and XIP subfamilies mostly contained hydrophobic ar/R selective filter. Analysis of RNA-seq and microarray data revealed high expression of PIPs in multiple tissues, low expression of NIPs, and seed specific expression of TIP3 in flax. Exploration of aquaporin homologs in three closely related Linum species bienne, grandiflorum and leonii revealed presence of 49, 39 and 19 AQPs, respectively. The genome-wide identification of aquaporins, first in flax, provides insight to elucidate their physiological and developmental roles in flax.

Root Border Cells and Their Role in Plant Defense.

PubMed

Hawes, Martha; Allen, Caitilyn; Turgeon, B Gillian; Curlango-Rivera, Gilberto; Minh Tran, Tuan; Huskey, David A; Xiong, Zhongguo

2016-08-04

Root border cells separate from plant root tips and disperse into the soil environment. In most species, each root tip can produce thousands of metabolically active cells daily, with specialized patterns of gene expression. Their function has been an enduring mystery. Recent studies suggest that border cells operate in a manner similar to mammalian neutrophils: Both cell types export a complex of extracellular DNA (exDNA) and antimicrobial proteins that neutralize threats by trapping pathogens and thereby preventing invasion of host tissues. Extracellular DNases (exDNases) of pathogens promote virulence and systemic spread of the microbes. In plants, adding DNase I to root tips eliminates border cell extracellular traps and abolishes root tip resistance to infection. Mutation of genes encoding exDNase activity in plant-pathogenic bacteria (Ralstonia solanacearum) and fungi (Cochliobolus heterostrophus) results in reduced virulence. The study of exDNase activities in plant pathogens may yield new targets for disease control.

Intracellular chloride ion concentration in differentiating neuronal cell and its role in growing neurite

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

Nakajima, Ken-ichi; Marunaka, Yoshinori; Department of Bio-Ionomics, Kyoto Prefectural University of Medicine, Graduate School of Medical Science, Kyoto 602-8566

Chloride ion (Cl{sup −}) is one of the most abundant anions in our body. Increasing evidence suggests that Cl{sup −} plays fundamental roles in various cellular functions. We have previously reported that electroneutral cation-chloride cotransporters, such as Na{sup +}-K{sup +}-2Cl{sup −} cotransporter 1 (NKCC1) and K{sup +}-Cl{sup −} cotransporter 1 (KCC1), are involved in neurite outgrowth during neuronal differentiation. In the present study, we studied if there is correlation between intracellular Cl{sup −} concentrations ([Cl{sup −}]{sub i}) and the length of growing neurites. We measured [Cl{sup −}]{sub i} in the cell body and growing neurite tips using halide-sensitive fluorescent dyemore » N-(ethoxycarbonylmethyl)-6-methoxyquinolinium bromide (MQAE), revealing that [Cl{sup −}]{sub i} in the tip of growing neurite was higher than that in cell body in a single cell. Importantly, there was a significant positive correlation between the length of growing neurite and [Cl{sup −}]{sub i} in neurite tip. Bumtanide (BMT), an inhibitor of NKCC1, significantly inhibited neurite outgrowth and decreased [Cl{sup −}]{sub i} in neurite tip. The results obtained in the present study and our previous studies together strongly suggest that high [Cl{sup −}]{sub i} in neurite tip region is crucial for efficient neurite outgrowth. - Highlights: • Intracellular Cl{sup −} concentrations ([Cl{sup −}]{sub i}) in the tip of growing neurite is higher than that in cell body in a single cell. • There is a significant positive correlation between the length of growing neurite and [Cl{sup −}]{sub i} in neurite tip. • Bumetanide significantly inhibits neurite outgrowth and decreased [Cl{sup −}]{sub i} in neurite tip. • High [Cl{sup −}]{sub i} in neurite tip region is crucial for efficient neurite outgrowth.« less

Electron Transfer from Triplet State of TIPS-Pentacene Generated by Singlet Fission Processes to CH3NH3PbI3 Perovskite.

PubMed

Lee, Sangsu; Hwang, Daesub; Jung, Seok Il; Kim, Dongho

2017-02-16

To reveal the applicability of singlet fission processes in perovskite solar cell, we investigated electron transfer from TIPS-pentacene to CH 3 NH 3 PbI 3 (MAPbI 3 ) perovskite in film phase. Through the observation of the shorter fluorescence lifetime in TIPS-pentacene/MAPbI 3 perovskite bilayer film (5 ns) compared with pristine MAPbI 3 perovskite film (20 ns), we verified electron-transfer processes between TIPS-pentacene and MAPbI 3 perovskite. Furthermore, the observation of singlet fission processes, a faster decay rate, TIPS-pentacene cations, and the analysis of kinetic profiles of the intensity ratio between 500 and 525 nm in the TA spectra of the TIPS-pentacene/MAPbI 3 perovskite bilayer film indicate that electron transfer occurs from triplet state of TIPS-pentacene generated by singlet fission processes to MAPbI 3 perovskite conduction band. We believe that our results can provide useful information on the design of solar cells sensitized by singlet fission processes and pave the way for new types of perovskite solar cells.

The Physiology of Mechanoelectrical Transduction Channels in Hearing

PubMed Central

Fettiplace, Robert; Kim, Kyunghee X.

2014-01-01

Much is known about the mechanotransducer (MT) channels mediating transduction in hair cells of the vertrbrate inner ear. With the use of isolated preparations, it is experimentally feasible to deliver precise mechanical stimuli to individual cells and record the ensuing transducer currents. This approach has shown that small (1–100 nm) deflections of the hair-cell stereociliary bundle are transmitted via interciliary tip links to open MT channels at the tops of the stereocilia. These channels are cation-permeable with a high selectivity for Ca2+; two channels are thought to be localized at the lower end of the tip link, each with a large single-channel conductance that increases from the low- to high-frequency end of the cochlea. Ca2+ influx through open channels regulates their resting open probability, which may contribute to setting the hair cell resting potential in vivo. Ca2+ also controls transducer fast adaptation and force generation by the hair bundle, the two coupled processes increasing in speed from cochlear apex to base. The molecular intricacy of the stereocilary bundle and the transduction apparatus is reflected by the large number of single-gene mutations that are linked to sensorineural deafness, especially those in Usher syndrome. Studies of such mutants have led to the discovery of many of the molecules of the transduction complex, including the tip link and its attachments to the stereociliary core. However, the MT channel protein is still not firmly identified, nor is it known whether the channel is activated by force delivered through accessory proteins or by deformation of the lipid bilayer. PMID:24987009

Customized atomic force microscopy probe by focused-ion-beam-assisted tip transfer

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

Wang, Andrew; Butte, Manish J., E-mail: manish.butte@stanford.edu

2014-08-04

We present a technique for transferring separately fabricated tips onto tipless atomic force microscopy (AFM) cantilevers, performed using focused ion beam-assisted nanomanipulation. This method addresses the need in scanning probe microscopy for certain tip geometries that cannot be achieved by conventional lithography. For example, in probing complex layered materials or tall biological cells using AFM, a tall tip with a high-aspect-ratio is required to avoid artifacts caused by collisions of the tip's sides with the material being probed. We show experimentally that tall (18 μm) cantilever tips fabricated by this approach reduce squeeze-film damping, which fits predictions from hydrodynamic theory, andmore » results in an increased quality factor (Q) of the fundamental flexural mode. We demonstrate that a customized tip's well-defined geometry, tall tip height, and aspect ratio enable improved measurement of elastic moduli by allowing access to low-laying portions of tall cells (T lymphocytes). This technique can be generally used to attach tips to any micromechanical device when conventional lithography of tips cannot be accomplished.« less

Regulation of Histone Acetyltransferase TIP60 Function by Histone Deacetylase 3

PubMed Central

Yi, Jingjie; Huang, Xiangyang; Yang, Yuxia; Zhu, Wei-Guo; Gu, Wei; Luo, Jianyuan

2014-01-01

The key member of the MOZ (monocyticleukaemia zinc finger protein), Ybf2/Sas3, Sas2, and TIP60 acetyltransferases family, Tat-interactive protein, 60 kD (TIP60), tightly modulates a wide array of cellular processes, including chromatin remodeling, gene transcription, apoptosis, DNA repair, and cell cycle arrest. The function of TIP60 can be regulated by SIRT1 through deacetylation. Here we found that TIP60 can also be functionally regulated by HDAC3. We identified six lysine residues as its autoacetylation sites. Mutagenesis of these lysines to arginines completely abolishes the autoacetylation of TIP60. Overexpression of HDAC3 increases TIP60 ubiquitination levels. However, unlike SIRT1, HDAC3 increased the half-life of TIP60. Further study found that HDAC3 colocalized with TIP60 both in the nucleus and the cytoplasm, which could be the reason why HDAC3 can stabilize TIP60. The deacetylation of TIP60 by both SIRT1 and HDAC3 reduces apoptosis induced by DNA damage. Knockdown of HDAC3 in cells increased TIP60 acetylation levels and increased apoptosis after DNA damage. Together, our findings provide a better understanding of TIP60 regulation mechanisms, which is a significant basis for further studies of its cellular functions. PMID:25301942

Similarity is not enough: Tipping points of Ebola Zaire mortalities

NASA Astrophysics Data System (ADS)

Phillips, J. C.

2015-06-01

In early 2014 an outbreak of a slightly mutated Zaire Ebola subtype appeared in West Africa which is less virulent than 1976 and 1994 strains. The numbers of cases per year appear to be ∼1000 times larger than the earlier strains, suggesting a greatly enhanced transmissibility. Although the fraction of the 2014 spike glycoprotein mutations is very small (∼3%), the mortality is significantly reduced, while the transmission appears to have increased strongly. Bioinformatic scaling had previously shown similar inversely correlated trends in virulence and transmission in N1 (H1N1) and N2 (H3N2) influenza spike glycoprotein mutations. These trends appear to be related to various external factors (migration, availability of pure water, and vaccination programs). The molecular mechanisms for Ebola's mutational response involve mainly changes in the disordered mucin-like domain (MLD) of its spike glycoprotein amino acids. The MLD has been observed to form the tip of an oligomeric amphiphilic wedge that selectively pries apart cell-cell interfaces via an oxidative mechanism.

The Steerable Microcatheter: A New Device for Selective Catheterisation

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

Soyama, Takeshi; Yoshida, Daisuke; Sakuhara, Yusuke, E-mail: yusaku@med.hokudai.ac.jp

The steerable microcatheter (SwiftNINJA, Sumitomo Bakelite, Tokyo, Japan), which has a remote-controlled flexible tip manipulated using a dial in the handgrip, was recently developed and delivered to the market. This device enables the user to change the angle of the microcatheter tip manually, and potentially makes selective catheterisation easier. We evaluated its unique characteristics and utility in selective catheterisation and coil embolization. This article describes: (1) the advantages of this device in catheterisations involving acute angle branches, and (2) a new technique of compact coil packing with the use of intentional folding by the bendable tip of the catheter.

Alleviation effect of alginate-derived oligosaccharides on Vicia faba root tip cells damaged by cadmium.

PubMed

Ma, L J; Zhang, Y; Bu, N; Wang, S H

2010-02-01

Cadmium has been shown to prevent Vicia faba growth by inhibiting cell mitosis. In this study we investigated the role of Alginate-derived Oligosaccharides (ADO) in alleviating Vicia faba root tip cells damaged by 6 and 8 mg L(-1) CdCl2. Micronucleus assay and chromosomal aberration assay were used to determine mitotic index, micronucleus frequency and chromosomal aberration frequency. The results showed that micronucleus frequency of Vicia faba root tip cells was inhibited under all the ADO concentrations. Especially, the inhibition ratio of 0.125% ADO highly reached 66.11 and 67.17% in 6 and 8 mg L(-1) CdCl2, respectively. Furthermore, the mitotic index increased (p < 0.05) and chromosomal aberration frequency decreased (p < 0.05) under all the ADO concentrations. This indicated that ADO had a significant alleviation effect on Vicia faba root tip cells damaged by cadmium.

Living cell manipulation, manageable sampling, and shotgun picoliter electrospray mass spectrometry for profiling metabolites.

PubMed

Gholipour, Yousef; Erra-Balsells, Rosa; Hiraoka, Kenzo; Nonami, Hiroshi

2013-02-01

A modified cell pressure probe and an online Orbitrap mass spectrometer were used to sample in situ plant single cells without any additional manipulation. The cell pressure probe, a quartz capillary tip filled with an oil mixture, was penetrated to various depths into parenchyma cells of tulip bulb scale, followed by a hydraulic continuity test to determine the exact location of the tip inside target cells. The operation was conducted under a digital microscope, and the capillary tip was photographed to calculate the volume of the cell sap sucked. The cell sap sample was then directly nebulized/ionized under high-voltage conditions at the entrance of the mass spectrometer. Several sugars, amino acids, organic acids, vitamins, fatty acids, and secondary metabolites were detected. Because picoliter solutions can be accurately handled and measured, known volumes of standard solutions can be added to cell sap samples inside the capillary tip to be used as references for metabolite characterization and relative quantitation. The high precision and sensitivity of the cell pressure probe and Orbitrap mass spectrometer allow for the manipulation and analysis of both femtoliter cell sap samples and standard solutions. Copyright © 2012 Elsevier Inc. All rights reserved.

Methods for Improving the Curvature of Steerable Needles in Biological Tissue

PubMed Central

Adebar, Troy K.; Greer, Joseph D.; Laeseke, Paul F.; Hwang, Gloria L.; Okamura, Allison M.

2016-01-01

Robotic needle steering systems have the potential to improve percutaneous interventions such as radiofrequency ablation of liver tumors, but steering techniques described to date have not achieved sufficiently small radius of curvature in biological tissue to be relevant to this application. In this work, the impact of tip geometry on steerable needle curvature is examined. Finite-element simulations and experiments with bent-tip needles in ex vivo liver tissue demonstrate that selection of tip length and angle can greatly improve curvature, with radius of curvature below 5 cm in liver tissue possible through judicious selection of these parameters. Motivated by the results of this analysis, a new articulated-tip steerable needle is described, in which a distal section is actively switched by a robotic system between a straight tip (resulting in a straight path) and a bent tip (resulting in a curved path). This approach allows the tip length and angle to be increased, while the straight configuration allows the needle tip to still pass through an introducer sheath and rotate inside the body. Validation testing in liver tissue shows that the new articulated-tip steerable needle achieves smaller radius of curvature compared to bent-tip needles described in previous work. Steerable needles with optimized tip parameters, which can generate tight curves in liver tissue, increase the clinical relevance of needle steering to percutaneous interventions. PMID:26441438

The coalescent of a sample from a binary branching process.

PubMed

Lambert, Amaury

2018-04-25

At time 0, start a time-continuous binary branching process, where particles give birth to a single particle independently (at a possibly time-dependent rate) and die independently (at a possibly time-dependent and age-dependent rate). A particular case is the classical birth-death process. Stop this process at time T>0. It is known that the tree spanned by the N tips alive at time T of the tree thus obtained (called a reduced tree or coalescent tree) is a coalescent point process (CPP), which basically means that the depths of interior nodes are independent and identically distributed (iid). Now select each of the N tips independently with probability y (Bernoulli sample). It is known that the tree generated by the selected tips, which we will call the Bernoulli sampled CPP, is again a CPP. Now instead, select exactly k tips uniformly at random among the N tips (a k-sample). We show that the tree generated by the selected tips is a mixture of Bernoulli sampled CPPs with the same parent CPP, over some explicit distribution of the sampling probability y. An immediate consequence is that the genealogy of a k-sample can be obtained by the realization of k random variables, first the random sampling probability Y and then the k-1 node depths which are iid conditional on Y=y. Copyright © 2018. Published by Elsevier Inc.

Towards force spectroscopy of single tip-link bonds

NASA Astrophysics Data System (ADS)

Koussa, Mounir A.; Sotomayor, Marcos; Wong, Wesley P.; Corey, David P.

2015-12-01

Inner-ear mechanotransduction relies on tip links, fine protein filaments made of cadherin-23 and protocadherin-15 that convey tension to mechanosensitive channels at the tips of hair-cell stereocilia. The tip-link cadherins are thought to form a heterotetrameric complex, with two cadherin-23 molecules forming the upper part of the filament and two protocadherin-15 molecules forming the lower end. The interaction between cadherin-23 and protocadherin-15 is mediated by their N-terminal tips. Missense mutations that modify the interaction interface impair binding and lead to deafness. Molecular dynamics simulations predict that the tip-link bond is mechanically strong enough to withstand forces in hair cells, but its experimentally determined strength is unknown. We have developed molecular tools to facilitate single-molecule force spectroscopy on the tip link bond. Self-assembling DNA nanoswitches are functionalized with the interacting tips of cadherin-23 and protocadherin-15 using the enzyme sortase under conditions that preserve protein function. These tip link nanoswitches are designed to provide a signature force-extension profile. This molecular signature should allow us to identify single-molecule rupture events in pulling experiments.

Calcium in Gravitropism of the Moss Pohlia nutans (Hedw.) Lindb. protonemata

NASA Astrophysics Data System (ADS)

Khorkavtsiv, O. Ya; Demkiv, O. T.; Khorkavtsiv, Ya. D.

Protonemata of mosses of Pohlia nutans grow both by extension and division of single apical cells which are negatively gravitropic in darkness. The fluorescence of Ca2+ increased in the tip of apical cells from the first hours of gravitropism stimulation. Cytochemical investigations confirmed the existence of a well pronounced tip-to-base Ca2+-gradient, its formation being favoured by localization of calcium influx in the tip of the apical cell. Measurement of the cytochemical reaction intensity showed that the level of Ca2+-ATFase activity is low in apex and increases towards the base of the apical cell. The gravitropic protonemata and filaments which grew on the clinostat showed similar distributions of the Ca2+ and Ca2+-ATFase activity along the apical cell axis. Thus, these data demonstrate that growing apical cells of gravitropic protonemata have a high tip-to-base Ca2+ gradient, the Ca2+ transport being afforded by Ca2+-ATFase.

Human Papillomavirus Type 18 E6 Protein Binds the Cellular PDZ Protein TIP-2/GIPC, Which Is Involved in Transforming Growth Factor β Signaling and Triggers Its Degradation by the Proteasome

PubMed Central

Favre-Bonvin, Arnaud; Reynaud, Caroline; Kretz-Remy, Carole; Jalinot, Pierre

2005-01-01

Several viral proteins expressed by DNA or RNA transforming viruses have the particular property of binding via their C-terminal end to various cellular proteins with PDZ domains. This study is focused on the PDZ protein TIP-2/GIPC, which was originally identified in two-hybrid screens performed with two different baits: the human T-cell leukemia virus type 1 Tax oncoprotein and the regulator of G signaling RGS-GAIP. Further studies have shown that TIP-2/GIPC is also able to associate with the cytoplasmic domains of various transmembrane proteins. In this report we show that TIP-2/GIPC interacts with the E6 protein of human papillomavirus type 18 (HPV-18). This event triggers polyubiquitination and proteasome-mediated degradation of the cellular protein. In agreement with this observation, silencing of E6 by RNA interference in HeLa cells causes an increase in the intracellular TIP-2/GIPC level. This PDZ protein has been previously found to be involved in transforming growth factor β (TGF-β) signaling by favoring expression of the TGF-β type III receptor at the cell membrane. In line with this activity of TIP-2/GIPC, we observed that depletion of this protein in HeLa cells hampers induction of the Id3 gene by TGF-β treatment and also diminishes the antiproliferative effect of this cytokine. Conversely, silencing of E6 increases the expression of Id3 and blocks proliferation of HeLa cells. These results support the notion that HPV-18 E6 renders cells less sensitive to the cytostatic effect of TGF-β by lowering the intracellular amount of TIP-2/GIPC. PMID:15767424

Single-cell isolation by a modular single-cell pipette for RNA-sequencing.

PubMed

Zhang, Kai; Gao, Min; Chong, Zechen; Li, Ying; Han, Xin; Chen, Rui; Qin, Lidong

2016-11-29

Single-cell transcriptome sequencing highly requires a convenient and reliable method to rapidly isolate a live cell into a specific container such as a PCR tube. Here, we report a modular single-cell pipette (mSCP) consisting of three modular components, a SCP-Tip, an air-displacement pipette (ADP), and ADP-Tips, that can be easily assembled, disassembled, and reassembled. By assembling the SCP-Tip containing a hydrodynamic trap, the mSCP can isolate single cells from 5-10 cells per μL of cell suspension. The mSCP is compatible with microscopic identification of captured single cells to finally achieve 100% single-cell isolation efficiency. The isolated live single cells are in submicroliter volumes and well suitable for single-cell PCR analysis and RNA-sequencing. The mSCP possesses merits of convenience, rapidness, and high efficiency, making it a powerful tool to isolate single cells for transcriptome analysis.

A tip-localized RhoGAP controls cell polarity by globally inhibiting Rho GTPase at the cell apex.

PubMed

Hwang, Jae-Ung; Vernoud, Vanessa; Szumlanski, Amy; Nielsen, Erik; Yang, Zhenbiao

2008-12-23

Highly elongated eukaryotic cells (e.g., neuronal axons, fungal hyphae, and pollen tubes) are generated through continuous apically restricted growth (tip growth), which universally requires tip-localized Rho GTPases. We used the oscillating pollen tube as a model system to determine the function and regulation of Rho GTPases in tip growth. Our previous work showed that the spatiotemporal dynamics of the apical cap of the activated Rho-like GTPase from Plant 1 (ROP1) are critical for tip growth in pollen tubes. However, the underlying mechanism for the generation and maintenance of this dynamic apical cap is poorly understood. A screen for mutations that enhance ROP1-overexpression-induced depolarization of pollen-tube growth identified REN1 (ROP1 enhancer 1) in Arabidopsis, whose null mutations turn elongated pollen tubes into bulbous cells. REN1 encodes a novel Rho GTPase-activating protein (RhoGAP) required for restricting the ROP1 activity to the pollen-tube tip. REN1 was localized to exocytic vesicles accumulated in the pollen-tube apex, as well as to the apical plasma membrane at the site of ROP1 activation. The apical localization of REN1 and its function in controlling growth polarity was compromised by disruption of ROP1-dependent F-actin and vesicular trafficking, which indicates that REN1 targeting and function is regulated by ROP1 downstream signaling. Our findings suggest that the REN1 RhoGAP controls a negative-feedback-based global inhibition of ROP1. This function provides a critical self-organizing mechanism, by which ROP signaling is spatially limited to the growth site and temporally oscillates during continuous tip growth. Similar spatiotemporal control of Rho GTPase signaling may also play an important role in cell-polarity control in other systems, including tip growth in fungi and cell movement in animals.

Negative regulators of vessel patterning.

PubMed

Suchting, Steven; Freitas, Catarina; le Noble, Ferdinand; Benedito, Rui; Bréant, Christiane; Duarte, Antonio; Eichmann, Anne

2007-01-01

Blood vessels and nerves are structurally similar, complex branched networks that require guidance to ensure their proper positioning in the body. Recent studies have demonstrated that specialized endothelial cells, resembling axonal growth cones, are located at the tips of growing capillaries. These endothelial tip cells guide outgrowing capillaries in response to gradients of extracellular matrix-bound vascular endothelial growth factor (VEGF). Here we show that endothelial tip cell formation and vessel branching are negatively regulated by the Notch ligand Delta-like 4 (Dll4). Heterozygous deletion of Dll4 or pharmacological inhibition of Notch signalling using gamma-secretase inhibitor revealed a striking vascular phenotype, with greatly increased numbers of filopodia-extending endothelial tip cells and increased expression of tip cell marker genes compared to controls. Filopodia extension in Dll4+/- retinal vessels required VEGF and was inhibited when VEGF signalling was blocked. While VEGF expression was not significantly altered in Dll4+- retinas, Dll4+/- vessels showed increased expression of VEGF Receptor 2 and decreased expression of VEGF Receptor 1 compared to wildtype, suggesting that they could be more responsive to VEGF stimulation. In addition, expression of Dll4 in wildtype tip cells was itself decreased when VEGF signalling was blocked, indicating that Dll4 may act downstream of VEGF as a 'brake' on VEGF-mediated angiogenic sprouting. Taken together, these data reveal Dll4 as a novel negative regulator of vascular sprouting and vessel branching that is required for normal vascular network formation during development.

MicroRNA-3178 ameliorates inflammation and gastric carcinogenesis promoted by Helicobacter pylori new toxin, Tip-α, by targeting TRAF3.

PubMed

Zou, Meijuan; Wang, Fang; Jiang, Aiqin; Xia, Anliang; Kong, Siya; Gong, Chun; Zhu, Mingxia; Zhou, Xin; Zhu, Jun; Zhu, Wei; Cheng, Wenfang

2017-04-01

Helicobacter pylori infection is the main cause of chronic gastritis, peptic ulcer, and gastric cancer. Tip-α is a newly identified carcinogenic factor present in H. pylori. TRAF3 can activate NF-κB by both canonical and noncanonical signaling pathways. In this study, we found that the expression of TRAF3 and NF-κB was upregulated, while microRNA-3178 (miR-3178) was decreased in H. pylori-positive gastric tissues but not in H. pylori-negative tissues. GES-1 cells were incubated with 12.5 μg/mL recombinant Tip-α (rTip-α) in RPMI1640 for 2 hours. After another 24 hours, the supernatant medium was designed as inflammatory-conditioned medium (ICM) and that from the untreated control cells was designed as untreated control medium. The release of proinflammatory cytokines from GES-1 cells and proliferation of gastric cancer cells was determined by ELISA and CCK-8 kits. Cells were transfected with the mimic, inhibitor, negative control of miR-3178, or TRAF3 siRNA control siRNA. The medium was then replaced with RPMI1640, 12.5 μg/mL rTip-α, and collected, and the total cellular RNA and protein were extracted for the following detection. MiR-3178 mimic prevented the increasement of TRAF3 and hence decreased activation of NF-κB signals, whereas miR-3178 inhibitor could not, in GES-1 cells with Tip-α treatment. The condition medium from miR-3178 mimic transfected GES-1 cells could inhibit proliferation and induce apoptosis of inflammation-related gastric cancer cells SGC7901 and MGC803 by decreasing the production of inflammatory cytokines TNF-α and IL-6, which were secreted by GES-1 cells. Taken all together, Tip-α might activate NF-κB to promote inflammation and carcinogenesis by inhibiting miR-3178 expression, which directly targeting TRAF3, during H. pylori infection in gastric mucosal epithelial cells. © 2016 John Wiley & Sons Ltd.

Inducible nuclear translocation of a STAT protein in Dictyostelium prespore cells: implications for morphogenesis and cell-type regulation.

PubMed

Dormann, D; Abe, T; Weijer, C J; Williams, J

2001-04-01

Dd-STATa, the Dictyostelium STAT (signal transducer and activator of transcription) protein, is selectively localised in the nuclei of a small subset of prestalk cells located in the slug tip. Injection of cAMP into the extracellular spaces in the rear of the slug induces rapid nuclear translocation of a Dd-GFP:STATa fusion protein in prespore cells surrounding the site of injection. This suggests that cAMP signals that emanate from the tip direct the localised nuclear accumulation of Dd-STATa. It also shows that prespore cells are competent to respond to cAMP, by Dd-STATa activation, and it implies that cAMP signalling is in some way limiting in the rear of the slug. Co-injection of a specific inhibitor of the cAR1 serpentine cAMP receptor almost completely prevents the cAMP-induced nuclear translocation, showing that most or all of the cAMP signal is transduced by cAR1. Dd-GFP:STATa also rapidly translocates into the nuclei of cells adjoining the front and back cut edges when a slug is bisected. Less severe mechanical disturbances, such as pricking the rear of a slug with an unfilled micropipette, also cause a more limited nuclear translocation of Dd-GFP:STATa. We propose that these signalling events form part of a repair mechanism that is activated when the migrating slug suffers mechanical damage.

Ultrastructural studies of regenerating spines of the sea urchin Strongylocentrotus purpuratus. I. Cell types without spherules.

PubMed

Heatfield, B M; Travis, D F

1975-01-01

The fine structure of regenerating tips of spines of the sea urchin Strongylocentrotus purpuratus was investigated. Each conical tip consisted of an inner dermis, which deposits and contains the calcite skeleton, and an external layer of epidermis. Although cell types termed spherulecytes containing large, intracellular membrane bound spherules were also present in spine tissues, only epidermal and dermal cell types lacking such spherules are described in this paper. The epidermis was composed largely of free cells representing several functional types. Over the apical portion of the tip these cells occurred in groups, while proximally they were distributed within longitudinal grooves present along the periphery of the spine from the base to the tip. The terminal portions of apical processes extending from some of the epidermal cells formed a thin, contiguous outer layer consisting of small individual islands of cytoplasm bearing microvilli. Adjacent islands were connected around the periphery by a junctional complex extending roughly 200 A in depth in which the opposing plasma membranes were separated by a narrow gap about 145 A in width bridged by amorphous material. Other epidermal cells were closely associated with the basal lamina, which was 900 A in thickness and delineated the dermoepidermal junction; some of these cells appeared to synthesize the lamina, while others may be sensory nerve cells. The dermis at the spine tip also consisted of several functional types of free cells; the most interesting of these was the calcoblast, which deposits the skeleton. Calcoblasts extended a thin, cytoplasmic skeletal sheath which surrounded the tips and adjacent proximal portions of each of the longitudinally oriented microspines comprising the regenerating skeleton, and distally, formed a conical extracellular channel ahead of the mineralizing tip. The intimate relationship between calcoblasts and the growing mineral surface strongly suggests that these cells directly control both the kinetics of mineral deposition and morphogenesis of the skeleton. Other cell types in the dermis were precalcoblasts and phagocytes. Precalcoblasts may function as fibroblasts and are possible precursors of calcoblasts. Closely associated with the basal lamina at the dermoepidermal junction were extracellular unbanded anchoring fi0rils 150 A to 200 A51 in diameter. Scattered proximally among dermal cells were other extracellular fibrils, presumably collagenous, about 300 A in diameter wit

The Notch ligand Delta-like 4 negatively regulates endothelial tip cell formation and vessel branching

PubMed Central

Suchting, Steven; Freitas, Catarina; le Noble, Ferdinand; Benedito, Rui; Bréant, Christiane; Duarte, Antonio; Eichmann, Anne

2007-01-01

Delta-like 4 (Dll4) is a transmembrane ligand for Notch receptors that is expressed in arterial blood vessels and sprouting endothelial cells. Here we show that Dll4 regulates vessel branching during development by inhibiting endothelial tip cell formation. Heterozygous deletion of dll4 or pharmacological inhibition of Notch signaling using γ-secretase inhibitor revealed a striking vascular phenotype, with greatly increased numbers of filopodia-extending endothelial tip cells and increased expression of tip cell marker genes compared with controls. Filopodia extension in dll4+/− retinal vessels required the vascular growth factor VEGF and was inhibited when VEGF signaling was blocked. Although VEGF expression was not significantly altered in dll4+/− retinas, dll4+/− vessels showed increased expression of VEGF receptor 2 and decreased expression of VEGF receptor 1 compared with wild-type, suggesting they could be more responsive to VEGF stimulation. In addition, expression of dll4 in wild-type tip cells was itself decreased when VEGF signaling was blocked, indicating that dll4 may act downstream of VEGF as a “brake” on VEGF-mediated angiogenic sprouting. Taken together, these data reveal Dll4 as a negative regulator of vascular sprouting and vessel branching that is required for normal vascular network formation during development. PMID:17296941

The Notch ligand Delta-like 4 negatively regulates endothelial tip cell formation and vessel branching.

PubMed

Suchting, Steven; Freitas, Catarina; le Noble, Ferdinand; Benedito, Rui; Bréant, Christiane; Duarte, Antonio; Eichmann, Anne

2007-02-27

Delta-like 4 (Dll4) is a transmembrane ligand for Notch receptors that is expressed in arterial blood vessels and sprouting endothelial cells. Here we show that Dll4 regulates vessel branching during development by inhibiting endothelial tip cell formation. Heterozygous deletion of dll4 or pharmacological inhibition of Notch signaling using gamma-secretase inhibitor revealed a striking vascular phenotype, with greatly increased numbers of filopodia-extending endothelial tip cells and increased expression of tip cell marker genes compared with controls. Filopodia extension in dll4(+/-) retinal vessels required the vascular growth factor VEGF and was inhibited when VEGF signaling was blocked. Although VEGF expression was not significantly altered in dll4(+/-) retinas, dll4(+/-) vessels showed increased expression of VEGF receptor 2 and decreased expression of VEGF receptor 1 compared with wild-type, suggesting they could be more responsive to VEGF stimulation. In addition, expression of dll4 in wild-type tip cells was itself decreased when VEGF signaling was blocked, indicating that dll4 may act downstream of VEGF as a "brake" on VEGF-mediated angiogenic sprouting. Taken together, these data reveal Dll4 as a negative regulator of vascular sprouting and vessel branching that is required for normal vascular network formation during development.

Desiccation and osmotic stress increase the abundance of mRNA of the tonoplast aquaporin BobTIP26-1 in cauliflower cells.

PubMed

Barrieu, F; Marty-Mazars, D; Thomas, D; Chaumont, F; Charbonnier, M; Marty, F

1999-07-01

Changes in vacuolar structure and the expression at the RNA level of a tonoplast aquaporin (BobTIP26-1) were examined in cauliflower (Brassicaoleracea L. var. botrytis) under water-stress conditions. Gradual drying out of slices of cauliflower floret tissue caused its collapse, with a shrinkage in tissue and cell volumes and an apparent vesiculation of the central vacuole, whereas osmotic stress resulted in plasmolysis with a collapse of the cytoplasm and the central vacuole within. Osmotic stress caused a rapid and substantial increase in BobTIP26 mRNA in slices of floret tissue. Exposure of tissue slices to a regime of desiccation showed a slower but equally large rise in BobTIP26 mRNA followed by a rapid decline upon rehydration. In situ hybridization showed that BobTIP26-2 mRNA is expressed most highly in meristematic and expanding cells of the cauliflower florets and that desiccation strongly increased the expression in those cells and in differentiated cells near the xylem vessels. These data indicate that under water-deficit conditions, expression of the tonoplast aquaporin gene in cauliflower is subject to a precise regulation that can be correlated with important cytological changes in the cells.

Restaurant Dining: Seven Tips for Staying Gluten Free

MedlinePlus

GiG Education Bulletin Restaurant Dining: Seven Tips for Staying Gluten-Free Updated May 2014 Tips for Dining Away from Home 1. Selection of eating establishment. • Your success at gluten-free dining will be determined by a number of factors, including the ...

[Effectivity and Safety of a Modified Tip Design in Torsional Phacoemulsification].

PubMed

Schmidt, Sabine; Hubich, Sophie; Vetter, Jan Markus; Wirbelauer, Christopher

2018-02-16

Torsional mode phacoemulsification results in more effective fragmentation of the nucleus due to a different movement of the phacotip. In this clinical study, we investigated the influence of a modified tip design and active fluidics on the efficacy of phacoemulsification and safety for the corneal endothelium. We conducted a prospective randomized 2 : 1 study in which 40 patients were operated on with the mini-flared Kelman Tip using the Infiniti ® System (group 1), and 20 patients were operated on with the Intrepid ® Balanced Tip and the Centurion ® System. We analyzed the intraoperative cumulative dissipated energy and also the density of the corneal endothelium measured with an endothelial microscope (CEM 530, Nidek) pre- and postoperatively. Both groups did not differ preoperatively in age, sex, axial length of the globe or corneal endothelium cell density nor cataract density (LOCS3). All surgeries were uneventful. The cumulative dissipated energy in group 1 (mini-flared Kelman tip, Infiniti System) was 38% higher than in group 2 (balanced tip, Centurion System; p < 0.05). The endothelial cell loss was 8% in group 1 and 10.3% in group 2 (p > 0.05). The cell size (polymegathism) increased in both groups significantly with + 37 µm in group 1 (p < 0.05) und + 54 µm in group 2 (p < 0.05). There was no statistically significant difference between both groups (p > 0.05). The number of hexagonal cells (pleomorphism) and corneal thickness did not differ in both groups either pre- nor postoperatively. Compared to torsional phacoemulsification with a mini-flared Kelman Tip and gravity fluidics, torsional phacoemulsification with a modified tip design and active fluidics is 38% more effective regarding the cumulative dissipated energy. Endothelial cell loss occurs to a similar extend using both systems. The postoperative changes in cell size (polymegathism), number of hexagonal cells (pleomorphism) and corneal thickness (pachymetry) were similar among both systems. We conclude, that the intraoperative stress on the endothelium is equivalent with both systems used. Georg Thieme Verlag KG Stuttgart · New York.

Autoradiography and the Cell Cycle.

ERIC Educational Resources Information Center

Jones, C. Weldon

1992-01-01

Outlines the stages of a cell biology "pulse-chase" experiment in which the students apply autoradiography techniques to learn about the concept of the cell cycle. Includes (1) seed germination and plant growth; (2) radioactive labeling and fixation of root tips; (3) feulgen staining of root tips; (4) preparation of autoradiograms; and…

Control of the Water Transport Activity of Barley HvTIP3;1 Specifically Expressed in Seeds.

PubMed

Utsugi, Shigeko; Shibasaka, Mineo; Maekawa, Masahiko; Katsuhara, Maki

2015-09-01

Tonoplast intrinsic proteins (TIPs) are involved in the transport and storage of water, and control intracellular osmotic pressure by transporting material related to the water potential of cells. In the present study, we focused on HvTIP3;1 during the periods of seed development and desiccation in barley. HvTIP3;1 was specifically expressed in seeds. An immunochemical analysis showed that HvTIP3;1 strongly accumulated in the aleurone layers and outer layers of barley seeds. The water transport activities of HvTIP3;1 and HvTIP1;2, which also accumulated in seeds, were measured in the heterologous expression system of Xenopus oocytes. When they were expressed individually, HvTIP1;2 transported water, whereas HvTIP3;1 did not. However, HvTIP3;1 exhibited water transport activity when co-expressed with HvTIP1;2 in oocytes, and this activity was higher than when HvTIP1;2 was expressed alone. This is the first report to demonstrate that the water permeability of a TIP aquaporin was activated when co-expressed with another TIP. The split-yellow fluorescent protein (YFP) system in onion cells revealed that HvTIP3;1 interacted with HvTIP1;2 to form a heterotetramer in plants. These results suggest that HvTIP3;1 functions as an active water channel to regulate water movement through tissues during the periods of seed development and desiccation. © The Author 2015. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

Kinesins have a dual function in organizing microtubules during both tip growth and cytokinesis in Physcomitrella patens.

PubMed

Hiwatashi, Yuji; Sato, Yoshikatsu; Doonan, John H

2014-03-01

Microtubules (MTs) play a crucial role in the anisotropic deposition of cell wall material, thereby affecting the direction of growth. A wide range of tip-growing cells display highly polarized cell growth, and MTs have been implicated in regulating directionality and expansion. However, the molecular machinery underlying MT dynamics in tip-growing plant cells remains unclear. Here, we show that highly dynamic MT bundles form cyclically in the polarized expansion zone of the moss Physcomitrella patens caulonemal cells through the coalescence of growing MT plus ends. Furthermore, the plant-specific kinesins (KINID1) that are is essential for the proper MT organization at cytokinesis also regulate the turnover of the tip MT bundles as well as the directionality and rate of cell growth. The plus ends of MTs grow toward the expansion zone, and KINID1 is necessary for the stability of a single coherent focus of MTs in the center of the zone, whose formation coincides with the accumulation of KINID1. We propose that KINID-dependent MT bundling is essential for the correct directionality of growth as well as for promoting growth per se. Our findings indicate that two localized cell wall deposition processes, tip growth and cytokinesis, previously believed to be functionally and evolutionarily distinct, share common and plant-specific MT regulatory components.

Evolution of a double amino acid substitution in the 5-enolpyruvylshikimate-3-phosphate synthase in Eleusine indica conferring high-level glyphosate resistance.

PubMed

Yu, Qin; Jalaludin, Adam; Han, Heping; Chen, Ming; Sammons, R Douglas; Powles, Stephen B

2015-04-01

Glyphosate is the most important and widely used herbicide in world agriculture. Intensive glyphosate selection has resulted in the widespread evolution of glyphosate-resistant weed populations, threatening the sustainability of this valuable once-in-a-century agrochemical. Field-evolved glyphosate resistance due to known resistance mechanisms is generally low to modest. Here, working with a highly glyphosate-resistant Eleusine indica population, we identified a double amino acid substitution (T102I+P106S [TIPS]) in the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene in glyphosate-resistant individuals. This TIPS mutation recreates the biotechnology-engineered commercial first generation glyphosate-tolerant EPSPS in corn (Zea mays) and now in other crops. In E. indica, the naturally evolved TIPS mutants are highly (more than 180-fold) resistant to glyphosate compared with the wild type and more resistant (more than 32-fold) than the previously known P106S mutants. The E. indica TIPS EPSPS showed very high-level (2,647-fold) in vitro resistance to glyphosate relative to the wild type and is more resistant (600-fold) than the P106S variant. The evolution of the TIPS mutation in crop fields under glyphosate selection is likely a sequential event, with the P106S mutation being selected first and fixed, followed by the T102I mutation to create the highly resistant TIPS EPSPS. The sequential evolution of the TIPS mutation endowing high-level glyphosate resistance is an important mechanism by which plants adapt to intense herbicide selection and a dramatic example of evolution in action. © 2015 American Society of Plant Biologists. All Rights Reserved.

Evolution of a Double Amino Acid Substitution in the 5-Enolpyruvylshikimate-3-Phosphate Synthase in Eleusine indica Conferring High-Level Glyphosate Resistance1

PubMed Central

Yu, Qin; Jalaludin, Adam; Han, Heping; Chen, Ming; Sammons, R. Douglas; Powles, Stephen B.

2015-01-01

Glyphosate is the most important and widely used herbicide in world agriculture. Intensive glyphosate selection has resulted in the widespread evolution of glyphosate-resistant weed populations, threatening the sustainability of this valuable once-in-a-century agrochemical. Field-evolved glyphosate resistance due to known resistance mechanisms is generally low to modest. Here, working with a highly glyphosate-resistant Eleusine indica population, we identified a double amino acid substitution (T102I + P106S [TIPS]) in the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene in glyphosate-resistant individuals. This TIPS mutation recreates the biotechnology-engineered commercial first generation glyphosate-tolerant EPSPS in corn (Zea mays) and now in other crops. In E. indica, the naturally evolved TIPS mutants are highly (more than 180-fold) resistant to glyphosate compared with the wild type and more resistant (more than 32-fold) than the previously known P106S mutants. The E. indica TIPS EPSPS showed very high-level (2,647-fold) in vitro resistance to glyphosate relative to the wild type and is more resistant (600-fold) than the P106S variant. The evolution of the TIPS mutation in crop fields under glyphosate selection is likely a sequential event, with the P106S mutation being selected first and fixed, followed by the T102I mutation to create the highly resistant TIPS EPSPS. The sequential evolution of the TIPS mutation endowing high-level glyphosate resistance is an important mechanism by which plants adapt to intense herbicide selection and a dramatic example of evolution in action. PMID:25717039

Systemic control of cell division and endoreduplication by NAA and BAP by modulating CDKs in root tip cells of Allium cepa.

PubMed

Tank, Jigna G; Thaker, Vrinda S

2014-01-01

Molecular mechanism regulated by auxin and cytokinin during endoreduplication, cell division, and elongation process is studied by using Allium cepa roots as a model system. The activity of CDK genes modulated by auxin and cytokinin during cell division, elongation, and endoreduplication process is explained in this research work. To study the significance of auxin and cytokinin in the management of cell division and endoreduplication process in plant meristematic cells at molecular level endoreduplication was developed in root tips of Allium cepa by giving colchicine treatment. There were inhibition of vegetative growth, formation of c-tumor at root tip, and development of endoreduplicated cells after colchicine treatment. This c-tumor was further treated with NAA and BAP to reinitiate vegetative growth in roots. BAP gave positive response in reinitiation of vegetative growth of roots from center of c-tumor. However, NAA gave negative response in reinitiation of vegetative growth of roots from c-tumor. Further, CDKs gene expression analysis from normal, endoreduplicated, and phytohormone (NAA or BAP) treated root tip was done and remarkable changes in transcription level of CDK genes in normal, endoreduplicated, and phytohormones treated cells were observed.

Systemic Control of Cell Division and Endoreduplication by NAA and BAP by Modulating CDKs in Root Tip Cells of Allium cepa

PubMed Central

Tank, Jigna G.; Thaker, Vrinda S.

2014-01-01

Molecular mechanism regulated by auxin and cytokinin during endoreduplication, cell division, and elongation process is studied by using Allium cepa roots as a model system. The activity of CDK genes modulated by auxin and cytokinin during cell division, elongation, and endoreduplication process is explained in this research work. To study the significance of auxin and cytokinin in the management of cell division and endoreduplication process in plant meristematic cells at molecular level endoreduplication was developed in root tips of Allium cepa by giving colchicine treatment. There were inhibition of vegetative growth, formation of c-tumor at root tip, and development of endoreduplicated cells after colchicine treatment. This c-tumor was further treated with NAA and BAP to reinitiate vegetative growth in roots. BAP gave positive response in reinitiation of vegetative growth of roots from center of c-tumor. However, NAA gave negative response in reinitiation of vegetative growth of roots from c-tumor. Further, CDKs gene expression analysis from normal, endoreduplicated, and phytohormone (NAA or BAP) treated root tip was done and remarkable changes in transcription level of CDK genes in normal, endoreduplicated, and phytohormones treated cells were observed. PMID:24955358

Dislocation-free Ge Nano-crystals via Pattern Independent Selective Ge Heteroepitaxy on Si Nano-Tip Wafers.

PubMed

Niu, Gang; Capellini, Giovanni; Schubert, Markus Andreas; Niermann, Tore; Zaumseil, Peter; Katzer, Jens; Krause, Hans-Michael; Skibitzki, Oliver; Lehmann, Michael; Xie, Ya-Hong; von Känel, Hans; Schroeder, Thomas

2016-03-04

The integration of dislocation-free Ge nano-islands was realized via selective molecular beam epitaxy on Si nano-tip patterned substrates. The Si-tip wafers feature a rectangular array of nanometer sized Si tips with (001) facet exposed among a SiO2 matrix. These wafers were fabricated by complementary metal-oxide-semiconductor (CMOS) compatible nanotechnology. Calculations based on nucleation theory predict that the selective growth occurs close to thermodynamic equilibrium, where condensation of Ge adatoms on SiO2 is disfavored due to the extremely short re-evaporation time and diffusion length. The growth selectivity is ensured by the desorption-limited growth regime leading to the observed pattern independence, i.e. the absence of loading effect commonly encountered in chemical vapor deposition. The growth condition of high temperature and low deposition rate is responsible for the observed high crystalline quality of the Ge islands which is also associated with negligible Si-Ge intermixing owing to geometric hindrance by the Si nano-tip approach. Single island as well as area-averaged characterization methods demonstrate that Ge islands are dislocation-free and heteroepitaxial strain is fully relaxed. Such well-ordered high quality Ge islands present a step towards the achievement of materials suitable for optical applications.

Dislocation-free Ge Nano-crystals via Pattern Independent Selective Ge Heteroepitaxy on Si Nano-Tip Wafers

PubMed Central

Niu, Gang; Capellini, Giovanni; Schubert, Markus Andreas; Niermann, Tore; Zaumseil, Peter; Katzer, Jens; Krause, Hans-Michael; Skibitzki, Oliver; Lehmann, Michael; Xie, Ya-Hong; von Känel, Hans; Schroeder, Thomas

2016-01-01

The integration of dislocation-free Ge nano-islands was realized via selective molecular beam epitaxy on Si nano-tip patterned substrates. The Si-tip wafers feature a rectangular array of nanometer sized Si tips with (001) facet exposed among a SiO2 matrix. These wafers were fabricated by complementary metal-oxide-semiconductor (CMOS) compatible nanotechnology. Calculations based on nucleation theory predict that the selective growth occurs close to thermodynamic equilibrium, where condensation of Ge adatoms on SiO2 is disfavored due to the extremely short re-evaporation time and diffusion length. The growth selectivity is ensured by the desorption-limited growth regime leading to the observed pattern independence, i.e. the absence of loading effect commonly encountered in chemical vapor deposition. The growth condition of high temperature and low deposition rate is responsible for the observed high crystalline quality of the Ge islands which is also associated with negligible Si-Ge intermixing owing to geometric hindrance by the Si nano-tip approach. Single island as well as area-averaged characterization methods demonstrate that Ge islands are dislocation-free and heteroepitaxial strain is fully relaxed. Such well-ordered high quality Ge islands present a step towards the achievement of materials suitable for optical applications. PMID:26940260

Positively selected FimH residues enhance virulence during urinary tract infection by altering FimH conformation.

PubMed

Schwartz, Drew J; Kalas, Vasilios; Pinkner, Jerome S; Chen, Swaine L; Spaulding, Caitlin N; Dodson, Karen W; Hultgren, Scott J

2013-09-24

Chaperone-usher pathway pili are a widespread family of extracellular, Gram-negative bacterial fibers with important roles in bacterial pathogenesis. Type 1 pili are important virulence factors in uropathogenic Escherichia coli (UPEC), which cause the majority of urinary tract infections (UTI). FimH, the type 1 adhesin, binds mannosylated glycoproteins on the surface of human and murine bladder cells, facilitating bacterial colonization, invasion, and formation of biofilm-like intracellular bacterial communities. The mannose-binding pocket of FimH is invariant among UPEC. We discovered that pathoadaptive alleles of FimH with variant residues outside the binding pocket affect FimH-mediated acute and chronic pathogenesis of two commonly studied UPEC strains, UTI89 and CFT073. In vitro binding studies revealed that, whereas all pathoadaptive variants tested displayed the same high affinity for mannose when bound by the chaperone FimC, affinities varied when FimH was incorporated into pilus tip-like, FimCGH complexes. Structural studies have shown that FimH adopts an elongated conformation when complexed with FimC, but, when incorporated into the pilus tip, FimH can adopt a compact conformation. We hypothesize that the propensity of FimH to adopt the elongated conformation in the tip corresponds to its mannose binding affinity. Interestingly, FimH variants, which maintain a high-affinity conformation in the FimCGH tip-like structure, were attenuated during chronic bladder infection, implying that FimH's ability to switch between conformations is important in pathogenesis. Our studies argue that positively selected residues modulate fitness during UTI by affecting FimH conformation and function, providing an example of evolutionary tuning of structural dynamics impacting in vivo survival.

Cellular and dendritic growth in a binary melt - A marginal stability approach

NASA Technical Reports Server (NTRS)

Laxmanan, V.

1986-01-01

A simple model for the constrained growth of an array of cells or dendrites in a binary alloy in the presence of an imposed positive temperature gradient in the liquid is proposed, with the dendritic or cell tip radius calculated using the marginal stability criterion of Langer and Muller-Krumbhaar (1977). This approach, an approach adopting the ad hoc assumption of minimum undercooling at the cell or dendrite tip, and an approach based on the stability criterion of Trivedi (1980) all predict tip radii to within 30 percent of each other, and yield a simple relationship between the tip radius and the growth conditions. Good agreement is found between predictions and data obtained in a succinonitrile-acetone system, and under the present experimental conditions, the dendritic tip stability parameter value is found to be twice that obtained previously, possibly due to a transition in morphology from a cellular structure with just a few side branches, to a more fully developed dendritic structure.

Genome-Wide Characterization of Major Intrinsic Proteins in Four Grass Plants and Their Non-Aqua Transport Selectivity Profiles with Comparative Perspective

PubMed Central

Azad, Abul Kalam; Ahmed, Jahed; Alum, Md. Asraful; Hasan, Md. Mahbub; Ishikawa, Takahiro; Sawa, Yoshihiro; Katsuhara, Maki

2016-01-01

Major intrinsic proteins (MIPs), commonly known as aquaporins, transport not only water in plants but also other substrates of physiological significance and heavy metals. In most of the higher plants, MIPs are divided into five subfamilies (PIPs, TIPs, NIPs, SIPs and XIPs). Herein, we identified 68, 42, 38 and 28 full-length MIPs, respectively in the genomes of four monocot grass plants, specifically Panicum virgatum, Setaria italica, Sorghum bicolor and Brachypodium distachyon. Phylogenetic analysis showed that the grass plants had only four MIP subfamilies including PIPs, TIPs, NIPs and SIPs without XIPs. Based on structural analysis of the homology models and comparing the primary selectivity-related motifs [two NPA regions, aromatic/arginine (ar/R) selectivity filter and Froger's positions (FPs)] of all plant MIPs that have been experimentally proven to transport non-aqua substrates, we predicted the transport profiles of all MIPs in the four grass plants and also in eight other plants. Groups of MIP subfamilies based on ar/R selectivity filter and FPs were linked to the non-aqua transport profiles. We further deciphered the substrate selectivity profiles of the MIPs in the four grass plants and compared them with their counterparts in rice, maize, soybean, poplar, cotton, Arabidopsis thaliana, Physcomitrella patens and Selaginella moellendorffii. In addition to two NPA regions, ar/R filter and FPs, certain residues, especially in loops B and C, contribute to the functional distinctiveness of MIP groups. Expression analysis of transcripts in different organs indicated that non-aqua transport was related to expression of MIPs since most of the unexpressed MIPs were not predicted to facilitate the transport of non-aqua molecules. Among all MIPs in every plant, TIP (BdTIP1;1, SiTIP1;2, SbTIP2;1 and PvTIP1;2) had the overall highest mean expression. Our study generates significant information for understanding the diversity, evolution, non-aqua transport profiles and insight into comparative transport selectivity of plant MIPs, and provides tools for the development of transgenic plants. PMID:27327960

Genome-Wide Characterization of Major Intrinsic Proteins in Four Grass Plants and Their Non-Aqua Transport Selectivity Profiles with Comparative Perspective.

PubMed

Azad, Abul Kalam; Ahmed, Jahed; Alum, Md Asraful; Hasan, Md Mahbub; Ishikawa, Takahiro; Sawa, Yoshihiro; Katsuhara, Maki

2016-01-01

Major intrinsic proteins (MIPs), commonly known as aquaporins, transport not only water in plants but also other substrates of physiological significance and heavy metals. In most of the higher plants, MIPs are divided into five subfamilies (PIPs, TIPs, NIPs, SIPs and XIPs). Herein, we identified 68, 42, 38 and 28 full-length MIPs, respectively in the genomes of four monocot grass plants, specifically Panicum virgatum, Setaria italica, Sorghum bicolor and Brachypodium distachyon. Phylogenetic analysis showed that the grass plants had only four MIP subfamilies including PIPs, TIPs, NIPs and SIPs without XIPs. Based on structural analysis of the homology models and comparing the primary selectivity-related motifs [two NPA regions, aromatic/arginine (ar/R) selectivity filter and Froger's positions (FPs)] of all plant MIPs that have been experimentally proven to transport non-aqua substrates, we predicted the transport profiles of all MIPs in the four grass plants and also in eight other plants. Groups of MIP subfamilies based on ar/R selectivity filter and FPs were linked to the non-aqua transport profiles. We further deciphered the substrate selectivity profiles of the MIPs in the four grass plants and compared them with their counterparts in rice, maize, soybean, poplar, cotton, Arabidopsis thaliana, Physcomitrella patens and Selaginella moellendorffii. In addition to two NPA regions, ar/R filter and FPs, certain residues, especially in loops B and C, contribute to the functional distinctiveness of MIP groups. Expression analysis of transcripts in different organs indicated that non-aqua transport was related to expression of MIPs since most of the unexpressed MIPs were not predicted to facilitate the transport of non-aqua molecules. Among all MIPs in every plant, TIP (BdTIP1;1, SiTIP1;2, SbTIP2;1 and PvTIP1;2) had the overall highest mean expression. Our study generates significant information for understanding the diversity, evolution, non-aqua transport profiles and insight into comparative transport selectivity of plant MIPs, and provides tools for the development of transgenic plants.

Actin polymerization drives polar growth in Arabidopsis root hair cells.

PubMed

Vazquez, Luis Alfredo Bañuelos; Sanchez, Rosana; Hernandez-Barrera, Alejandra; Zepeda-Jazo, Isaac; Sánchez, Federico; Quinto, Carmen; Torres, Luis Cárdenas

2014-01-01

In plants, the actin cytoskeleton is a prime regulator of cell polarity, growth, and cytoplasmic streaming. Tip growth, as observed in root hairs, caulonema, and pollen tubes, is governed by many factors, including calcium gradients, exocytosis and endocytosis, reactive oxygen species, and the cytoskeleton. Several studies indicate that the polymerization of G-actin into F-actin also contributes to tip growth. The structure and function of F-actin within the apical dome is variable, ranging from a dense meshwork to sparse single filaments. The presence of multiple F-actin structures in the elongating apices of tip-growing cells suggests that this cytoskeletal array is tightly regulated. We recently reported that sublethal concentrations of fluorescently labeled cytochalasin could be used to visualize the distribution of microfilament plus ends using fluorescence microscopy, and found that the tip region of the growing root hair cells of a legume plant exhibits a clear response to the nodulation factors secreted by Rhizobium. (1) In this current work, we expanded our analysis using confocal microscopy and demonstrated the existence of highly dynamic fluorescent foci along Arabidopsis root hair cells. Furthermore, we show that the strongest fluorescence signal accumulates in the tip dome of the growing root hair and seems to be in close proximity to the apical plasma membrane. Based on these findings, we propose that actin polymerization within the dome of growing root hair cells regulates polar growth.

Cell-Biological Studies of Osmotic Shock Response in Streptomyces spp.

PubMed

Fuchino, Katsuya; Flärdh, Klas; Dyson, Paul; Ausmees, Nora

2017-01-01

Most bacteria are likely to face osmotic challenges, but there is yet much to learn about how such environmental changes affect the architecture of bacterial cells. Here, we report a cell-biological study in model organisms of the genus Streptomyces, which are actinobacteria that grow in a highly polarized fashion to form branching hyphae. The characteristic apical growth of Streptomyces hyphae is orchestrated by protein assemblies, called polarisomes, which contain coiled-coil proteins DivIVA and Scy, and recruit cell wall synthesis complexes and the stress-bearing cytoskeleton of FilP to the tip regions of the hyphae. We monitored cell growth and cell-architectural changes by time-lapse microscopy in osmotic upshift experiments. Hyperosmotic shock caused arrest of growth, loss of turgor, and hypercondensation of chromosomes. The recovery period was protracted, presumably due to the dehydrated state of the cytoplasm, before hyphae could restore their turgor and start to grow again. In most hyphae, this regrowth did not take place at the original hyphal tips. Instead, cell polarity was reprogrammed, and polarisomes were redistributed to new sites, leading to the emergence of multiple lateral branches from which growth occurred. Factors known to regulate the branching pattern of Streptomyces hyphae, such as the serine/threonine kinase AfsK and Scy, were not involved in reprogramming of cell polarity, indicating that different mechanisms may act under different environmental conditions to control hyphal branching. Our observations of hyphal morphology during the stress response indicate that turgor and sufficient hydration of cytoplasm are required for Streptomyces tip growth. Polar growth is an intricate manner of growth for accomplishing a complicated morphology, employed by a wide range of organisms across the kingdoms of life. The tip extension of Streptomyces hyphae is one of the most pronounced examples of polar growth among bacteria. The expansion of the cell wall by tip extension is thought to be facilitated by the turgor pressure, but it was unknown how external osmotic change influences Streptomyces tip growth. We report here that severe hyperosmotic stress causes cessation of growth, followed by reprogramming of cell polarity and rearrangement of growth zones to promote lateral hyphal branching. This phenomenon may represent a strategy of hyphal organisms to avoid osmotic stress encountered by the growing hyphal tip. Copyright © 2016 American Society for Microbiology.

The Stress-responsive Gene ATF3 Mediates Dichotomous UV Responses by Regulating the Tip60 and p53 Proteins*

PubMed Central

Cui, Hongmei; Li, Xingyao; Han, Chunhua; Wang, Qi-En; Wang, Hongbo; Ding, Han-Fei; Zhang, Junran; Yan, Chunhong

2016-01-01

The response to UV irradiation is important for a cell to maintain its genetic integrity when challenged by environmental genotoxins. An immediate early response to UV irradiation is the rapid induction of activating transcription factor 3 (ATF3) expression. Although emerging evidence has linked ATF3 to stress pathways regulated by the tumor suppressor p53 and the histone acetyltransferase Tip60, the role of ATF3 in the UV response remains largely unclear. Here, we report that ATF3 mediated dichotomous UV responses. Although UV irradiation enhanced the binding of ATF3 to Tip60, knockdown of ATF3 expression decreased Tip60 stability, thereby impairing Tip60 induction by UV irradiation. In line with the role of Tip60 in mediating UV-induced apoptosis, ATF3 promoted the death of p53-defective cells in response to UV irradiation. However, ATF3 could also activate p53 and promote p53-mediated DNA repair, mainly through altering histone modifications that could facilitate recruitment of DNA repair proteins (such as DDB2) to damaged DNA sites. As a result, ATF3 rather protected the p53 wild-type cells from UV-induced apoptosis. Our results thus indicate that ATF3 regulates cell fates upon UV irradiation in a p53-dependent manner. PMID:26994140

Aluminium-induced reduction of plant growth in alfalfa (Medicago sativa) is mediated by interrupting auxin transport and accumulation in roots.

PubMed

Wang, Shengyin; Ren, Xiaoyan; Huang, Bingru; Wang, Ge; Zhou, Peng; An, Yuan

2016-07-20

The objective of this study was to investigate Al(3+)-induced IAA transport, distribution, and the relation of these two processes to Al(3+)-inhibition of root growth in alfalfa. Alfalfa seedlings with or without apical buds were exposed to 0 or 100 μM AlCl3 and were foliar sprayed with water or 6 mg L(-1) IAA. Aluminium stress resulted in disordered arrangement of cells, deformed cell shapes, altered cell structure, and a shorter length of the meristematic zone in root tips. Aluminium stress significantly decreased the IAA concentration in apical buds and root tips. The distribution of IAA fluorescence signals in root tips was disturbed, and the IAA transportation from shoot base to root tip was inhibited. The highest intensity of fluorescence signals was detected in the apical meristematic zone. Exogenous application of IAA markedly alleviated the Al(3+)-induced inhibition of root growth by increasing IAA accumulation and recovering the damaged cell structure in root tips. In addition, Al(3+) stress up-regulated expression of AUX1 and PIN2 genes. These results indicate that Al(3+)-induced reduction of root growth could be associated with the inhibitions of IAA synthesis in apical buds and IAA transportation in roots, as well as the imbalance of IAA distribution in root tips.

Aluminium-induced reduction of plant growth in alfalfa (Medicago sativa) is mediated by interrupting auxin transport and accumulation in roots

PubMed Central

Wang, Shengyin; Ren, Xiaoyan; Huang, Bingru; Wang, Ge; Zhou, Peng; An, Yuan

2016-01-01

The objective of this study was to investigate Al3+-induced IAA transport, distribution, and the relation of these two processes to Al3+-inhibition of root growth in alfalfa. Alfalfa seedlings with or without apical buds were exposed to 0 or 100 μM AlCl3 and were foliar sprayed with water or 6 mg L−1 IAA. Aluminium stress resulted in disordered arrangement of cells, deformed cell shapes, altered cell structure, and a shorter length of the meristematic zone in root tips. Aluminium stress significantly decreased the IAA concentration in apical buds and root tips. The distribution of IAA fluorescence signals in root tips was disturbed, and the IAA transportation from shoot base to root tip was inhibited. The highest intensity of fluorescence signals was detected in the apical meristematic zone. Exogenous application of IAA markedly alleviated the Al3+-induced inhibition of root growth by increasing IAA accumulation and recovering the damaged cell structure in root tips. In addition, Al3+ stress up-regulated expression of AUX1 and PIN2 genes. These results indicate that Al3+-induced reduction of root growth could be associated with the inhibitions of IAA synthesis in apical buds and IAA transportation in roots, as well as the imbalance of IAA distribution in root tips. PMID:27435109

Invited Review Article: Tip modification methods for tip-enhanced Raman spectroscopy (TERS) and colloidal probe technique: A 10 year update (2006-2016) review

NASA Astrophysics Data System (ADS)

Yuan, C. C.; Zhang, D.; Gan, Y.

2017-03-01

Engineering atomic force microscopy tips for reliable tip enhanced Raman spectroscopy (TERS) and colloidal probe technique are becoming routine practices in many labs. In this 10 year update review, various new tip modification methods developed over the past decade are briefly reviewed to help researchers select the appropriate method. The perspective is put in a large context to discuss the opportunities and challenges in this area, including novel combinations of seemingly different methods, potential applications of some methods which were not originally intended for TERS tip fabrication, and the problems of high cost and poor reproducibility of tip fabrication.

Root tips moving through soil

PubMed Central

Curlango-Rivera, Gilberto

2011-01-01

Root elongation occurs by the generation of new cells from meristematic tissue within the apical 1–2 mm region of root tips. Therefore penetration of the soil environment is carried out by newly synthesized plant tissue, whose cells are inherently vulnerable to invasion by pathogens. This conundrum, on its face, would seem to reflect an intolerable risk to the successful establishment of root systems needed for plant life. Yet root tip regions housing the meristematic tissues repeatedly have been found to be free of microbial infection and colonization. Even when spore germination, chemotaxis, and/or growth of pathogens are stimulated by signals from the root tip, the underlying root tissue can escape invasion. Recent insights into the functions of root border cells, and the regulation of their production by transient exposure to external signals, may shed light on long-standing observations. PMID:21455030

Luminal mitosis drives epithelial cell dispersal within the branching ureteric bud

PubMed Central

Packard, Adam; Georgas, Kylie; Michos, Odyssé; Riccio, Paul; Cebrian, Cristina; Combes, Alexander N.; Ju, Adler; Ferrer-Vaquer, Anna; Hadjantonakis, Anna-Katerina; Zong, Hui; Little, Melissa H.; Costantini, Frank

2013-01-01

Summary The ureteric bud is an epithelial tube that undergoes branching morphogenesis to form the renal collecting system. Though development of a normal kidney depends on proper ureteric bud morphogenesis, the cellular events underlying this process remain obscure. Here, we used time-lapse microscopy together with several genetic labeling methods to observe ureteric bud cell behaviors in developing mouse kidneys. We observed an unexpected cell behavior in the branching tips of the ureteric bud, which we term “mitosis-associated cell dispersal”. Pre-mitotic ureteric tip cells delaminate from the epithelium and divide within the lumen; while one daughter cell retains a basal process, allowing it to reinsert into the epithelium at the site of origin, the other daughter cell reinserts at a position one to three cell diameters away. Given the high rate of cell division in ureteric tips, this cellular behavior causes extensive epithelial cell rearrangements that may contribute to renal branching morphogenesis. PMID:24183650

Ten Timeless Tips for Keeping on Top of Teaching Technology

ERIC Educational Resources Information Center

Poling, Devereaux A.; LoSchiavo, Frank M.

2014-01-01

We provide tips for helping psychology faculty effectively seek, select, and place new technology into pedagogical practice. We also provide tips to help psychology departments position themselves for a future that includes teaching technologies that have not yet been created. Instead of discussing today's top innovations in teaching (which…

Advanced ceramic material for high temperature turbine tip seals

NASA Technical Reports Server (NTRS)

Solomon, N. G.; Vogan, J. W.

1978-01-01

Ceramic material systems are being considered for potential use as turbine blade tip gas path seals at temperatures up to 1370 1/4 C. Silicon carbide and silicon nitride structures were selected for study since an initial analysis of the problem gave these materials the greatest potential for development into a successful materials system. Segments of silicon nitride and silicon carbide materials over a range of densities, processed by various methods, a honeycomb structure of silicon nitride and ceramic blade tip inserts fabricated from both materials by hot pressing were tested singly and in combination. The evaluations included wear under simulated engine blade tip rub conditions, thermal stability, impact resistance, machinability, hot gas erosion and feasibility of fabrication into engine components. The silicon nitride honeycomb and low-density silicon carbide using a selected grain size distribution gave the most promising results as rub-tolerant shroud liners. Ceramic blade tip inserts made from hot-pressed silicon nitride gave excellent test results. Their behavior closely simulated metal tips. Wear was similar to that of metals but reduced by a factor of six.

Rapid Polymer Sequencer

NASA Technical Reports Server (NTRS)

Stolc, Viktor (Inventor); Brock, Matthew W (Inventor)

2013-01-01

Method and system for rapid and accurate determination of each of a sequence of unknown polymer components, such as nucleic acid components. A self-assembling monolayer of a selected substance is optionally provided on an interior surface of a pipette tip, and the interior surface is immersed in a selected liquid. A selected electrical field is impressed in a longitudinal direction, or in a transverse direction, in the tip region, a polymer sequence is passed through the tip region, and a change in an electrical current signal is measured as each polymer component passes through the tip region. Each of the measured changes in electrical current signals is compared with a database of reference electrical change signals, with each reference signal corresponding to an identified polymer component, to identify the unknown polymer component with a reference polymer component. The nanopore preferably has a pore inner diameter of no more than about 40 nm and is prepared by heating and pulling a very small section of a glass tubing.

Electromicroinjection of particles into living cells

DOEpatents

Ray, F. Andrew; Cram, L. Scott; Galey, William R.

1988-01-01

Method and apparatus for introducing particles into living cells. Fluorescently-stained human chromosomes are introduced into cultured, mitotic Chinese hamster cells using electromicroinjection. The recipient cells frequently survived the physiological perturbation imposed by a successful chromosome injection. Successfully injected recipient cells maintained viability as evidenced by their ability to be expanded. The technique relies on the surface charge of fluorescently stained chromosomes and their ability to be attracted and repelled to and from the tip of a micropipette. The apparatus includes a micropipette having a tip suitable for piercing the membrane of a target cell and an electrode inserted into the lumen thereof. The target cells and suspended particles are located in an electrically conducted solution, and the lumen of the micropipette is filled with an electrically conducting solution which contacts the electrode located therein. A second electrode is also located in the conducting solution containing the target cells and particles. Voltages applied to the electrode within the micropipette attract the particles to the region of the tip thereof. The particles adhere to the surface of the micropipette with sufficient force that insertion of the micropipette tip and attached particle through the membrane of a target cell will not dislodge the particle. By applying a voltage having the opposite polarity of the attraction voltage, the particles are expelled from the micropipette to which is then withdrawn from the cell body.

Interface Symbiotic Membrane Formation in Root Nodules of Medicago truncatula: the Role of Synaptotagmins MtSyt1, MtSyt2 and MtSyt3

PubMed Central

Gavrin, Aleksandr; Kulikova, Olga; Bisseling, Ton; Fedorova, Elena E.

2017-01-01

Symbiotic bacteria (rhizobia) are maintained and conditioned to fix atmospheric nitrogen in infected cells of legume root nodules. Rhizobia are confined to the asymmetrical protrusions of plasma membrane (PM): infection threads (IT), cell wall-free unwalled droplets and symbiosomes. These compartments rapidly increase in surface and volume due to the microsymbiont expansion, and remarkably, the membrane resources of the host cells are targeted to interface membrane quite precisely. We hypothesized that the change in the membrane tension around the expanding microsymbionts creates a vector for membrane traffic toward the symbiotic interface. To test this hypothesis, we selected calcium sensors from the group of synaptotagmins: MtSyt1, Medicago truncatula homolog of AtSYT1 from Arabidopsis thaliana known to be involved in membrane repair, and two other homologs expressed in root nodules: MtSyt2 and MtSyt3. Here we show that MtSyt1, MtSyt2, and MtSyt3 are expressed in the expanding cells of the meristem, zone of infection and proximal cell layers of zone of nitrogen fixation (MtSyt1, MtSyt3). All three GFP-tagged proteins delineate the interface membrane of IT and unwalled droplets and create a subcompartments of PM surrounding these structures. The localization of MtSyt1 by EM immunogold labeling has shown the signal on symbiosome membrane and endoplasmic reticulum (ER). To specify the role of synaptotagmins in interface membrane formation, we compared the localization of MtSyt1, MtSyt3 and exocyst subunit EXO70i, involved in the tethering of post-Golgi secretory vesicles and operational in tip growth. The localization of EXO70i in root nodules and arbusculated roots was strictly associated with the tips of IT and the tips of arbuscular fine branches, but the distribution of synaptotagmins on membrane subcompartments was broader and includes lateral parts of IT, the membrane of unwalled droplets as well as the symbiosomes. The double silencing of synaptotagmins caused a delay in rhizobia release and blocks symbiosome maturation confirming the functional role of synaptotagmins. In conclusion: synaptotagmin-dependent membrane fusion along with tip-targeted exocytosis is operational in the formation of symbiotic interface. PMID:28265280

Elevated levels of N-lauroylethanolamine, an endogenous constituent of desiccated seeds, disrupt normal root development in Arabidopsis thaliana seedlings

NASA Technical Reports Server (NTRS)

Blancaflor, Elison B.; Hou, Guichuan; Chapman, Kent D.

2003-01-01

N-Acylethanolamines (NAEs) are prevalent in desiccated seeds of various plant species, and their levels decline substantially during seed imbibition and germination. Here, seeds of Arabidopsis thaliana (L.) Heynh. were germinated in, and seedlings maintained on, micromolar concentrations of N-lauroylethanolamine (NAE 12:0). NAE 12:0 inhibited root elongation, increased radial swelling of root tips, and reduced root hair numbers in a highly selective and concentration-dependent manner. These effects were reversible when seedlings were transferred to NAE-free medium. Older seedlings (14 days old) acclimated to exogenous NAE by increased formation of lateral roots, and generally, these lateral roots did not exhibit the severe symptoms observed in primary roots. Cells of NAE-treated primary roots were swollen and irregular in shape, and in many cases showed evidence, at the light- and electron-microscope levels, of improper cell wall formation. Microtubule arrangement was disrupted in severely distorted cells close to the root tip, and endoplasmic reticulum (ER)-localized green fluorescent protein (mGFP5-ER) was more abundant, aggregated and distributed differently in NAE-treated root cells, suggesting disruption of proper cell division, endomembrane organization and vesicle trafficking. These results suggest that NAE 12:0 likely influences normal cell expansion in roots by interfering with intracellular membrane trafficking to and/or from the cell surface. The rapid metabolism of NAEs during seed imbibition/germination may be a mechanism to remove this endogenous class of lipid mediators to allow for synchronized membrane reorganization associated with cell expansion.

Are fingernails a key to unlocking the puzzle of mammalian limb regeneration?

PubMed

Lehoczky, Jessica A

2017-06-01

Some mammalian digit tips, including those of mice and human children, can regenerate following amputation, whereas mammalian limb regeneration does not occur. One major difference between the digit tip and the rest of the limb is the presence of the nail, which is necessary for this type of regeneration. This couples well with the finding that canonical Wnt signalling and Lgr6, an agonist of Wnt signalling that marks nail stem cells, are necessary for digit tip regeneration. This viewpoint essay discusses the role of the nail in digit tip regeneration and explores whether nail stem cells and their presumptive niche can be solely accountable for why regeneration is possible in the digit tip, but not the rest of the limb. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

The aquatic hyphomycete Heliscus lugdunensis protects its hyphae tip cells from cadmium: A micro X-ray fluorescence and X-ray absorption near edge structure spectroscopy study

NASA Astrophysics Data System (ADS)

Isaure, Marie-Pierre; Leyh, Benjamin; Salomé, Murielle; Krauss, Gerd-Joachim; Schaumlöffel, Dirk; Dobritzsch, Dirk

2017-11-01

Aquatic fungi can be used to evaluate the functioning of natural ecosystems. Heliscus lugdunensis is an early colonizer of allochthone leafs. Since this aquatic hyphomycete is able to develop in metal contaminated habitats and tolerates cadmium, it appears to be a good candidate to investigate adaptation to metal pollution. This study aimed at examining the sequestration of Cd in the hyphae of H. lugdunensis, and particularly the role of the tip cells. For that, H. lugdunensis growth was evaluated under various Cd concentrations, and a combination of synchrotron micro X-ray fluorescence and X-ray absorption near edge structure spectroscopy was carried out to determine the compartments of Cd accumulation and the Cd chemical species, respectively. Results showed that the hyphal tip cells were depleted in Cd, and that the metal was stored in older cells. Cd was mainly associated with sulfur ligands and to a lesser extent bound to phosphates and carboxyl/hydroxyl groups from cell wall and/or organic acids. Finally, the aquatic fungus was able to maintain the tip cell as a functional system, thus allowing the colonization of contaminated environments.

Endothelial basement membrane limits tip cell formation by inducing Dll4/Notch signalling in vivo.

PubMed

Stenzel, Denise; Franco, Claudio A; Estrach, Soline; Mettouchi, Amel; Sauvaget, Dominique; Rosewell, Ian; Schertel, Andreas; Armer, Hannah; Domogatskaya, Anna; Rodin, Sergey; Tryggvason, Karl; Collinson, Lucy; Sorokin, Lydia; Gerhardt, Holger

2011-10-28

How individual components of the vascular basement membrane influence endothelial cell behaviour remains unclear. Here we show that laminin α4 (Lama4) regulates tip cell numbers and vascular density by inducing endothelial Dll4/Notch signalling in vivo. Lama4 deficiency leads to reduced Dll4 expression, excessive filopodia and tip cell formation in the mouse retina, phenocopying the effects of Dll4/Notch inhibition. Lama4-mediated Dll4 expression requires a combination of integrins in vitro and integrin β1 in vivo. We conclude that appropriate laminin/integrin-induced signalling is necessary to induce physiologically functional levels of Dll4 expression and regulate branching frequency during sprouting angiogenesis in vivo.

Endothelial basement membrane limits tip cell formation by inducing Dll4/Notch signalling in vivo

PubMed Central

Stenzel, Denise; Franco, Claudio A; Estrach, Soline; Mettouchi, Amel; Sauvaget, Dominique; Rosewell, Ian; Schertel, Andreas; Armer, Hannah; Domogatskaya, Anna; Rodin, Sergey; Tryggvason, Karl; Collinson, Lucy; Sorokin, Lydia; Gerhardt, Holger

2011-01-01

How individual components of the vascular basement membrane influence endothelial cell behaviour remains unclear. Here we show that laminin α4 (Lama4) regulates tip cell numbers and vascular density by inducing endothelial Dll4/Notch signalling in vivo. Lama4 deficiency leads to reduced Dll4 expression, excessive filopodia and tip cell formation in the mouse retina, phenocopying the effects of Dll4/Notch inhibition. Lama4-mediated Dll4 expression requires a combination of integrins in vitro and integrin β1 in vivo. We conclude that appropriate laminin/integrin-induced signalling is necessary to induce physiologically functional levels of Dll4 expression and regulate branching frequency during sprouting angiogenesis in vivo. PMID:21979816

Nanopipette exploring nanoworld.

PubMed

Takami, Tomohide; Park, Bae Ho; Kawai, Tomoji

2014-01-01

Nanopipettes, with tip orifices on the order of tens to hundreds of nanometers, have been utilized in the fields of analytical chemistry and nanophysiology. Nanopipettes make nanofabrication possible at liquid/solid interfaces. Moreover, they are utilized in time-resolved measurements and for imaging biological materials, e.g. , living cells, by using techniques such as scanning ion-conductance microscopy and scanning electrochemical microscopy. We have successfully fabricated ion-selective nanopipettes that can be used to identify targeted ions such as sodium and potassium in- and outside of living cells. In this review, we discuss the extent of utilization of nanopipettes in investigating the nanoworld. In addition, we discuss the potential applications of future nanopipettes.

Nanopipette exploring nanoworld

NASA Astrophysics Data System (ADS)

Takami, Tomohide; Park, Bae Ho; Kawai, Tomoji

2014-04-01

Nanopipettes, with tip orifices on the order of tens to hundreds of nanometers, have been utilized in the fields of analytical chemistry and nanophysiology. Nanopipettes make nanofabrication possible at liquid/solid interfaces. Moreover, they are utilized in time-resolved measurements and for imaging biological materials, e.g., living cells, by using techniques such as scanning ion-conductance microscopy and scanning electrochemical microscopy. We have successfully fabricated ion-selective nanopipettes that can be used to identify targeted ions such as sodium and potassium in- and outside of living cells. In this review, we discuss the extent of utilization of nanopipettes in investigating the nanoworld. In addition, we discuss the potential applications of future nanopipettes.

Thermo-solutal growth of an anisotropic dendrite with six-fold symmetry

NASA Astrophysics Data System (ADS)

Alexandrov, D. V.; Galenko, P. K.

2018-03-01

A stable growth of dendritic crystal with the six-fold crystalline anisotropy is analyzed in a binary nonisothermal mixture. A selection criterion representing a relationship between the dendrite tip velocity and its tip diameter is derived on the basis of morphological stability analysis and solvability theory. A complete set of nonlinear equations, consisting of the selection criterion and undercooling balance condition, which determines implicit dependencies of the dendrite tip velocity and tip diameter as functions of the total undercooling, is formulated. Exact analytical solutions of these nonlinear equations are found in a parametric form. Asymptotic solutions describing the crystal growth at small Péclet numbers are determined. Theoretical predictions are compared with experimental data obtained for ice dendrites growing in binary water-ethylenglycol solutions as well as in pure water.

A spindle pole antigen gene MoSPA2 is important for polar cell growth of vegetative hyphae and conidia, but is dispensable for pathogenicity in Magnaporthe oryzae.

PubMed

Li, Chao; Yang, Jun; Zhou, Wei; Chen, Xiao-Lin; Huang, Jin-Guang; Cheng, Zhi-Hua; Zhao, Wen-Sheng; Zhang, Yan; Peng, You-Liang

2014-11-01

Spa2 is an important component of the multiprotein complex polarisome, which is involved in the establishment, maintenance, termination of polarized cell growth and is important for defining tip growth of filamentous fungi. In this study, we isolated an insertional mutant of the rice blast fungus Magnaporthe oryzae that formed smaller colony and conidia compared with the wild type. In the mutant, a spindle pole antigen gene MoSPA2 was disrupted by the integration of an exogenous plasmid. Targeted gene deletion and complementation assays demonstrated the gene disruption was responsible for the defects of the insertional mutant. Interestingly, the MoSpa2-GFP fusion protein was found to accumulate as a spot at hyphal tips, septa of hyphae and conidial tip cells where germ tubes are usually produced, but not in appressoria, infection hyphae or at the septa of conidia. Furthermore, the deletion mutants of MoSPA2 exhibited slower hyphal tip growth, more hyphal branches, and smaller size of conidial tip cells. However, MoSPA2 is not required for plant infection. These results indicate that MoSPA2 is required for vegetative hyphal growth and maintaining conidium morphology and that spotted accumulation of MoSpa2 is important for its functions during cell polar growth.

ICK is essential for cell type-specific ciliogenesis and the regulation of ciliary transport.

PubMed

Chaya, Taro; Omori, Yoshihiro; Kuwahara, Ryusuke; Furukawa, Takahisa

2014-06-02

Cilia and flagella are formed and maintained by intraflagellar transport (IFT) and play important roles in sensing and moving across species. At the distal tip of the cilia/flagella, IFT complexes turn around to switch from anterograde to retrograde transport; however, the underlying regulatory mechanism is unclear. Here, we identified ICK localization at the tip of cilia as a regulator of ciliary transport. In ICK-deficient mice, we found ciliary defects in neuronal progenitor cells with Hedgehog signal defects. ICK-deficient cells formed cilia with mislocalized Hedgehog signaling components. Loss of ICK caused the accumulation of IFT-A, IFT-B, and BBSome components at the ciliary tips. In contrast, overexpression of ICK induced the strong accumulation of IFT-B, but not IFT-A or BBSome components at ciliary tips. In addition, ICK directly phosphorylated Kif3a, while inhibition of this Kif3a phosphorylation affected ciliary formation. Our results suggest that ICK is a Kif3a kinase and essential for proper ciliogenesis in development by regulating ciliary transport at the tip of cilia. © 2014 The Authors.

ICK is essential for cell type-specific ciliogenesis and the regulation of ciliary transport

PubMed Central

Chaya, Taro; Omori, Yoshihiro; Kuwahara, Ryusuke; Furukawa, Takahisa

2014-01-01

Cilia and flagella are formed and maintained by intraflagellar transport (IFT) and play important roles in sensing and moving across species. At the distal tip of the cilia/flagella, IFT complexes turn around to switch from anterograde to retrograde transport; however, the underlying regulatory mechanism is unclear. Here, we identified ICK localization at the tip of cilia as a regulator of ciliary transport. In ICK-deficient mice, we found ciliary defects in neuronal progenitor cells with Hedgehog signal defects. ICK-deficient cells formed cilia with mislocalized Hedgehog signaling components. Loss of ICK caused the accumulation of IFT-A, IFT-B, and BBSome components at the ciliary tips. In contrast, overexpression of ICK induced the strong accumulation of IFT-B, but not IFT-A or BBSome components at ciliary tips. In addition, ICK directly phosphorylated Kif3a, while inhibition of this Kif3a phosphorylation affected ciliary formation. Our results suggest that ICK is a Kif3a kinase and essential for proper ciliogenesis in development by regulating ciliary transport at the tip of cilia. PMID:24797473

S. pombe CLASP needs dynein, not EB1 or CLIP170, to induce microtubule instability and slows polymerization rates at cell tips in a dynein-dependent manner

PubMed Central

Grallert, Agnes; Beuter, Christoph; Craven, Rachel A.; Bagley, Steve; Wilks, Deepti; Fleig, Ursula; Hagan, Iain M.

2006-01-01

The Schizosaccharomyces pombe CLIP170-associated protein (CLASP) Peg1 was identified in a screen for mutants with spindle formation defects and a screen for molecules that antagonized EB1 function. The conditional peg1.1 mutant enabled us to identify key features of Peg1 function. First, Peg1 was required to form a spindle and astral microtubules, yet destabilized interphase microtubules. Second, Peg1 was required to slow the polymerization rate of interphase microtubules that establish end-on contact with the cortex at cell tips. Third, Peg1 antagonized the action of S. pombe CLIP170 (Tip1) and EB1 (Mal3). Fourth, although Peg1 resembled higher eukaryotic CLASPs by physically associating with both Mal3 and Tip1, neither Tip1 nor Mal3 was required for Peg1 to destabilize interphase microtubules or for it to associate with microtubules. Conversely, neither Mal3 nor Tip1 required Peg1 to associate with microtubules or cell tips. Consistently, while mal3.Δ and tip1.Δ disrupted linear growth, corrupting peg1 + did not. Fifth, peg1.1 phenotypes resembled those arising from deletion of the single heavy or both light chains of fission yeast dynein. Furthermore, all interphase phenotypes arising from peg1 + manipulation relied on dynein function. Thus, the impact of S. pombe CLASP on interphase microtubule behavior is more closely aligned to dynein than EB1 or CLIP170. PMID:16951255

CDC-42 Orients Cell Migration during Epithelial Intercalation in the Caenorhabditis elegans Epidermis.

PubMed

Walck-Shannon, Elise; Lucas, Bethany; Chin-Sang, Ian; Reiner, David; Kumfer, Kraig; Cochran, Hunter; Bothfeld, William; Hardin, Jeff

2016-11-01

Cell intercalation is a highly directed cell rearrangement that is essential for animal morphogenesis. As such, intercalation requires orchestration of cell polarity across the plane of the tissue. CDC-42 is a Rho family GTPase with key functions in cell polarity, yet its role during epithelial intercalation has not been established because its roles early in embryogenesis have historically made it difficult to study. To circumvent these early requirements, in this paper we use tissue-specific and conditional loss-of-function approaches to identify a role for CDC-42 during intercalation of the Caenorhabditis elegans dorsal embryonic epidermis. CDC-42 activity is enriched in the medial tips of intercalating cells, which extend as cells migrate past one another. Moreover, CDC-42 is involved in both the efficient formation and orientation of cell tips during cell rearrangement. Using conditional loss-of-function we also show that the PAR complex functions in tip formation and orientation. Additionally, we find that the sole C. elegans Eph receptor, VAB-1, functions during this process in an Ephrin-independent manner. Using epistasis analysis, we find that vab-1 lies in the same genetic pathway as cdc-42 and is responsible for polarizing CDC-42 activity to the medial tip. Together, these data establish a previously uncharacterized role for polarized CDC-42, in conjunction with PAR-6, PAR-3 and an Eph receptor, during epithelial intercalation.

CDC-42 Orients Cell Migration during Epithelial Intercalation in the Caenorhabditis elegans Epidermis

PubMed Central

Lucas, Bethany; Chin-Sang, Ian; Reiner, David; Kumfer, Kraig

2016-01-01

Cell intercalation is a highly directed cell rearrangement that is essential for animal morphogenesis. As such, intercalation requires orchestration of cell polarity across the plane of the tissue. CDC-42 is a Rho family GTPase with key functions in cell polarity, yet its role during epithelial intercalation has not been established because its roles early in embryogenesis have historically made it difficult to study. To circumvent these early requirements, in this paper we use tissue-specific and conditional loss-of-function approaches to identify a role for CDC-42 during intercalation of the Caenorhabditis elegans dorsal embryonic epidermis. CDC-42 activity is enriched in the medial tips of intercalating cells, which extend as cells migrate past one another. Moreover, CDC-42 is involved in both the efficient formation and orientation of cell tips during cell rearrangement. Using conditional loss-of-function we also show that the PAR complex functions in tip formation and orientation. Additionally, we find that the sole C. elegans Eph receptor, VAB-1, functions during this process in an Ephrin-independent manner. Using epistasis analysis, we find that vab-1 lies in the same genetic pathway as cdc-42 and is responsible for polarizing CDC-42 activity to the medial tip. Together, these data establish a previously uncharacterized role for polarized CDC-42, in conjunction with PAR-6, PAR-3 and an Eph receptor, during epithelial intercalation. PMID:27861585

Roughness-dependent friction force of the tarsal claw system in the beetle Pachnoda marginata (Coleoptera, Scarabaeidae).

PubMed

Dai, Zhendong; Gorb, Stanislav N; Schwarz, Uli

2002-08-01

This paper studies slide-resisting forces generated by claws in the free-walking beetle Pachnoda marginata (Coleoptera, Scarabaeoidea) with emphasis on the relationship between the dimension of the claw tip and the substrate texture. To evaluate the force range by which the claw can interact with a substrate, forces generated by the freely moving legs were measured using a load cell force transducer. To obtain information about material properties of the claw, its mechanical strength was tested in a fracture experiment, and the internal structure of the fractured claw material was studied by scanning electron microscopy. The bending stress of the claw was evaluated as 143.4-684.2 MPa, depending on the cross-section model selected. Data from these different approaches led us to propose a model explaining the saturation of friction force with increased texture roughness. The forces are determined by the relative size of the surface roughness R(a) (or an average particle diameter) and the diameter of the claw tip. When surface roughness is much bigger than the claw tip diameter, the beetle can grasp surface irregularities and generate a high degree of attachment due to mechanical interlocking with substrate texture. When R(a) is lower than or comparable to the claw tip diameter, the frictional properties of the contact between claw and substrate particles play a key role in the generation of the friction force.

All-in-One Nanowire-Decorated Multifunctional Membrane for Rapid Cell Lysis and Direct DNA Isolation

PubMed Central

2015-01-01

This paper describes a handheld device that uses an all-in-one membrane for continuous mechanical cell lysis and rapid DNA isolation without the assistance of power sources, lysis reagents, and routine centrifugation. This nanowire-decorated multifunctional membrane was fabricated to isolate DNA by selective adsorption to silica surface immediately after disruption of nucleus membranes by ultrasharp tips of nanowires for a rapid cell lysis, and it can be directly assembled with commercial syringe filter holders. The membrane was fabricated by photoelectrochemical etching to create microchannel arrays followed by hydrothermal synthesis of nanowires and deposition of silica. The proposed membrane successfully purifies high-quality DNA within 5 min, whereas a commercial purification kit needs more than an hour. PMID:25420232

Recovery patterns, histological observations and genetic integrity in Malus shoot tips cryopreserved using droplet-vitrification and encapsulation-dehydration procedures.

PubMed

Li, Bai-Quan; Feng, Chao-Hong; Wang, Min-Rui; Hu, Ling-Yun; Volk, Gayle; Wang, Qiao-Chun

2015-11-20

A droplet-vitrification procedure is described for cryopreservation of Malus shoot tips. Survival patterns, recovery types, histological observations, and genetic integrity were compared for Malus shoot tips cryopreserved using this droplet-vitrification procedure and an encapsulation-dehydration procedure that was previously reported by us. In both procedures, three types of shoot tip recovery were observed following cryopreservation: callus formation without shoot regrowth, leaf formation without shoot regrowth, and shoot regrowth. Three categories of histological observations were also identified in cross-sections of shoot tips recovered after cryopreservation using the two cryogenic procedures. In category 1, almost all of the cells (94-95%) in the apical dome (AD) were damaged or killed and only some cells (30-32%) in the leaf primordia (LPs) survived. In category 2, only a few cells (18-20%) in the AD and some cells (30-31%) in the LPs survived. In category 3, majority of the cells (60-62%) in the AD and some cells (30-33%) in the LPs survived. These data suggest that shoot regrowth is correlated to the presence of a majority of surviving cells in the AD after liquid nitrogen exposure. No polymorphic bands were detected by inter-simple sequence repeats or by random amplified polymorphic DNA assessments, and ploidy levels analyzed by flow cytometry were unchanged when plants recovered after cryoexposure were compared to controls. The droplet-vitrification procedure appears to be robust since seven genotypes representing four Malus species and one hybrid recovered shoots following cryopreservation. Mean shoot regrowth levels of these seven genotypes were 48% in the droplet-vitrification method, which were lower than those (61%) in the encapsulation-dehydration procedure reported in our previous study, suggesting the latter may be preferred for routine cryobanking applications for Malus shoot tips. Copyright © 2015 Elsevier B.V. All rights reserved.

Cut-cell method based large-eddy simulation of tip-leakage flow

NASA Astrophysics Data System (ADS)

Pogorelov, Alexej; Meinke, Matthias; Schröder, Wolfgang

2015-07-01

The turbulent low Mach number flow through an axial fan at a Reynolds number of 9.36 × 105 based on the outer casing diameter is investigated by large-eddy simulation. A finite-volume flow solver in an unstructured hierarchical Cartesian setup for the compressible Navier-Stokes equations is used. To account for sharp edges, a fully conservative cut-cell approach is applied. A newly developed rotational periodic boundary condition for Cartesian meshes is introduced such that the simulations are performed just for a 72° segment, i.e., the flow field over one out of five axial blades is resolved. The focus of this numerical analysis is on the development of the vortical flow structures in the tip-gap region. A detailed grid convergence study is performed on four computational grids with 50 × 106, 250 × 106, 1 × 109, and 1.6 × 109 cells. Results of the instantaneous and the mean fan flow field are thoroughly analyzed based on the solution with 1 × 109 cells. High levels of turbulent kinetic energy and pressure fluctuations are generated by a tip-gap vortex upstream of the blade, the separating vortices inside the tip gap, and a counter-rotating vortex on the outer casing wall. An intermittent interaction of the turbulent wake, generated by the tip-gap vortex, with the downstream blade, leads to a cyclic transition with high pressure fluctuations on the suction side of the blade and a decay of the tip-gap vortex. The disturbance of the tip-gap vortex results in an unsteady behavior of the turbulent wake causing the intermittent interaction. For this interaction and the cyclic transition, two dominant frequencies are identified which perfectly match with the characteristic frequencies in the experimental sound power level and therefore explain their physical origin.

Filopodial retraction force is generated by cortical actin dynamics and controlled by reversible tethering at the tip

PubMed Central

Bornschlögl, Thomas; Romero, Stéphane; Vestergaard, Christian L.; Joanny, Jean-François; Van Nhieu, Guy Tran; Bassereau, Patricia

2013-01-01

Filopodia are dynamic, finger-like plasma membrane protrusions that sense the mechanical and chemical surroundings of the cell. Here, we show in epithelial cells that the dynamics of filopodial extension and retraction are determined by the difference between the actin polymerization rate at the tip and the retrograde flow at the base of the filopodium. Adhesion of a bead to the filopodial tip locally reduces actin polymerization and leads to retraction via retrograde flow, reminiscent of a process used by pathogens to invade cells. Using optical tweezers, we show that filopodial retraction occurs at a constant speed against counteracting forces up to 50 pN. Our measurements point toward retrograde flow in the cortex together with frictional coupling between the filopodial and cortical actin networks as the main retraction-force generator for filopodia. The force exerted by filopodial retraction, however, is limited by the connection between filopodial actin filaments and the membrane at the tip. Upon mechanical rupture of the tip connection, filopodia exert a passive retraction force of 15 pN via their plasma membrane. Transient reconnection at the tip allows filopodia to continuously probe their surroundings in a load-and-fail manner within a well-defined force range. PMID:24198333

Carbon Nanoelectrodes for Single-Cell Probing

PubMed Central

Anderson, Sean E.; Bau, Haim H.

2015-01-01

Carbon nanoelectrodes with tip diameters ranging from tens to hundreds of nm are fabricated by pyrolitic deposition of carbon films along the entire inner surfaces of pulled-glass pipettes. The pulled end of each glass pipette is then etched to expose a desired length (typically, a few µm) of carbon pipe. The carbon film provides an electrically conductive path from the nanoscopic carbon tip to the distal, macroscopic end of the pipette, bridging between the nanoscale tip and the macroscale handle, without a need for assembly. We used our nanoelectrodes to penetrate into individual cells and cell nuclei and measured the variations in the electrode impedance upon cell and nucleus penetration as well as the electrode impedance as a function of cell penetration depth. Theoretical predictions based on a simple circuit model were in good agreement with experimental data. PMID:25876625

Plant Evo-Devo: How Tip Growth Evolved.

PubMed

Rensing, Stefan A

2016-12-05

Apical elongation of polarized plant cells (tip growth) occurs in root hairs of flowering plants and in rhizoids of bryophytes. A new report shows that the formation of these cells relies on genes already present in the first land plants. Copyright © 2016 Elsevier Ltd. All rights reserved.

Alk1 and Alk5 inhibition by Nrp1 controls vascular sprouting downstream of Notch.

PubMed

Aspalter, Irene Maria; Gordon, Emma; Dubrac, Alexandre; Ragab, Anan; Narloch, Jarek; Vizán, Pedro; Geudens, Ilse; Collins, Russell Thomas; Franco, Claudio Areias; Abrahams, Cristina Luna; Thurston, Gavin; Fruttiger, Marcus; Rosewell, Ian; Eichmann, Anne; Gerhardt, Holger

2015-06-17

Sprouting angiogenesis drives blood vessel growth in healthy and diseased tissues. Vegf and Dll4/Notch signalling cooperate in a negative feedback loop that specifies endothelial tip and stalk cells to ensure adequate vessel branching and function. Current concepts posit that endothelial cells default to the tip-cell phenotype when Notch is inactive. Here we identify instead that the stalk-cell phenotype needs to be actively repressed to allow tip-cell formation. We show this is a key endothelial function of neuropilin-1 (Nrp1), which suppresses the stalk-cell phenotype by limiting Smad2/3 activation through Alk1 and Alk5. Notch downregulates Nrp1, thus relieving the inhibition of Alk1 and Alk5, thereby driving stalk-cell behaviour. Conceptually, our work shows that the heterogeneity between neighbouring endothelial cells established by the lateral feedback loop of Dll4/Notch utilizes Nrp1 levels as the pivot, which in turn establishes differential responsiveness to TGF-β/BMP signalling.

Alk1 and Alk5 inhibition by Nrp1 controls vascular sprouting downstream of Notch

PubMed Central

Aspalter, Irene Maria; Gordon, Emma; Dubrac, Alexandre; Ragab, Anan; Narloch, Jarek; Vizán, Pedro; Geudens, Ilse; Collins, Russell Thomas; Franco, Claudio Areias; Abrahams, Cristina Luna; Thurston, Gavin; Fruttiger, Marcus; Rosewell, Ian; Eichmann, Anne; Gerhardt, Holger

2015-01-01

Sprouting angiogenesis drives blood vessel growth in healthy and diseased tissues. Vegf and Dll4/Notch signalling cooperate in a negative feedback loop that specifies endothelial tip and stalk cells to ensure adequate vessel branching and function. Current concepts posit that endothelial cells default to the tip-cell phenotype when Notch is inactive. Here we identify instead that the stalk-cell phenotype needs to be actively repressed to allow tip-cell formation. We show this is a key endothelial function of neuropilin-1 (Nrp1), which suppresses the stalk-cell phenotype by limiting Smad2/3 activation through Alk1 and Alk5. Notch downregulates Nrp1, thus relieving the inhibition of Alk1 and Alk5, thereby driving stalk-cell behaviour. Conceptually, our work shows that the heterogeneity between neighbouring endothelial cells established by the lateral feedback loop of Dll4/Notch utilizes Nrp1 levels as the pivot, which in turn establishes differential responsiveness to TGF-β/BMP signalling. PMID:26081042

The touch dome defines an epidermal niche specialized for mechanosensory signaling

PubMed Central

Doucet, Yanne S.; Woo, Seung-Hyun; Ruiz, Marlon E.; Owens, David M.

2013-01-01

Summary In mammalian skin, Merkel cells are mechanoreceptor cells that are required for the perception of gentle touch. Recent evidence indicates that mature Merkel cells descend from the proliferative layer of skin epidermis; however, the stem cell niche for Merkel cell homeostasis has not been reported. Here, we provide the first genetic evidence for maintenance of mature Merkel cells during homeostasis by Krt17+ stem cells located in epidermal touch domes of hairy skin and in the tips of the rete ridges of glabrous skin. Lineage tracing analysis indicated that the entire pool of mature Merkel cells is turned over every 7–8 weeks in adult epidermis and that Krt17+ stem cells also maintain squamous differentiation in the touch dome and in glabrous skin. Finally, selective genetic ablation of Krt17+ touch dome keratinocytes indicates that these cells, and not mature Merkel cells, are primarily responsible for maintaining innervation of the Merkel cell-neurite complex. PMID:23727240

Correlation between spectral-domain OCT findings and visual acuity in X-linked retinoschisis.

PubMed

Yang, Hyun Seung; Lee, Jung Bok; Yoon, Young Hee; Lee, Joo Yong

2014-05-08

To investigate the tomographic characteristics of the outer retina and choroid and their relationship with visual acuity in X-linked juvenile retinoschisis (XLRS) patients using spectral-domain optical coherence tomography (SD-OCT). In this retrospective, observational, case-control study, we analyzed 20 eyes of 10 patients with XLRS using SD-OCT. The clinical and tomographic features of the outer retina, including the external limiting membrane (ELM), inner segment/outer segment (IS/OS) junction, cone cell outer segment tips (COST) line, photoreceptor outer segment (PROS) length, and choroid, were evaluated. As controls, 40 age-, sex-, and refraction-matched healthy eyes (1:2 matched) were randomly selected and imaged in parallel. The most prevalent area of abnormality in the outer retina layer of our patients was the outer plexiform layer (OPL; 60% of all affected eyes) and COST line (75% of all affected eyes). On average, the subfoveal choroid and PROS lengths were 35 μm thicker and 19 μm thinner, respectively, in XLRS patients (P = 0.084 and P < 0.001, respectively). A dominant IS/OS junction, COST line defects, and PROS length were related to patient best-corrected visual acuity (BCVA; P = 0.029, P = 0.001, and P < 0.001, respectively) by univariate analysis. Cone cell outer segment tips line defect and PROS length were the only factors related to BCVA in multivariate analysis (P = 0.028 and 0.003, respectively). Outer plexiform layer and photoreceptor microstructure defects are frequent in XLRS patients. Cone cell outer segment tips line defects and shortened PROS lengths as well as other photoreceptor microstructure defects may be closely related to poor vision in XLRS.

Fabrication of silver tips for scanning tunneling microscope induced luminescence.

PubMed

Zhang, C; Gao, B; Chen, L G; Meng, Q S; Yang, H; Zhang, R; Tao, X; Gao, H Y; Liao, Y; Dong, Z C

2011-08-01

We describe a reliable fabrication procedure of silver tips for scanning tunneling microscope (STM) induced luminescence experiments. The tip was first etched electrochemically to yield a sharp cone shape using selected electrolyte solutions and then sputter cleaned in ultrahigh vacuum to remove surface oxidation. The tip status, in particular the tip induced plasmon mode and its emission intensity, can be further tuned through field emission and voltage pulse. The quality of silver tips thus fabricated not only offers atomically resolved STM imaging, but more importantly, also allows us to perform challenging "color" photon mapping with emission spectra taken at each pixel simultaneously during the STM scan under relatively small tunnel currents and relatively short exposure time.

Neuroligin-1 overexpression in newborn granule cells in vivo.

PubMed

Schnell, Eric; Bensen, Aesoon L; Washburn, Eric K; Westbrook, Gary L

2012-01-01

Adult-born dentate granule cells integrate into the hippocampal network, extend neurites and form synapses in otherwise mature tissue. Excitatory and inhibitory inputs innervate these new granule cells in a stereotyped, temporally segregated manner, which presents a unique opportunity to study synapse development in the adult brain. To examine the role of neuroligins as synapse-inducing molecules in vivo, we infected dividing neural precursors in adult mice with a retroviral construct that increased neuroligin-1 levels during granule cell differentiation. By 21 days post-mitosis, exogenous neuroligin-1 was expressed at the tips of dendritic spines and increased the number of dendritic spines. Neuroligin-1-overexpressing cells showed a selective increase in functional excitatory synapses and connection multiplicity by single afferent fibers, as well as an increase in the synaptic AMPA/NMDA receptor ratio. In contrast to its synapse-inducing ability in vitro, neuroligin-1 overexpression did not induce precocious synapse formation in adult-born neurons. However, the dendrites of neuroligin-1-overexpressing cells did have more thin protrusions during an early period of dendritic outgrowth, suggesting enhanced filopodium formation or stabilization. Our results indicate that neuroligin-1 expression selectively increases the degree, but not the onset, of excitatory synapse formation in adult-born neurons.

Intracellular Ca(2+) and K(+) concentration in Brassica oleracea leaf induces differential expression of transporter and stress-related genes.

PubMed

Lee, Jeongyeo; Kim, Jungeun; Choi, Jae-Pil; Lee, MiYe; Kim, Min Keun; Lee, Young Han; Hur, Yoonkang; Nou, Ill-Sup; Park, Sang Un; Min, Sung Ran; Kim, HyeRan

2016-03-09

One of the most important members of the genus Brassica, cabbage, requires a relatively high level of calcium for normal growth (Plant Cell Environ 7: 397-405, 1984; Plant Physiol 60: 854-856, 1977). Localized Ca(2+) deficiency in cabbage leaves causes tip-burn, bringing about serious economic losses (Euphytica 9:203-208, 1960; Ann Bot 43:363-372, 1979; Sci Hortic 14:131-138, 1981). Although it has been known that the occurrence of tip-burn is related to Ca(2+) deficiency, there is limited information on the underlying mechanisms of tip-burn or the relationship between Ca(2+) and tip-burn incidence. To obtain more information on the genetic control of tip-burn symptoms, we focused on the identification of genes differentially expressed in response to increasing intracellular Ca(2+) and K(+) concentrations in B. oleracea lines derived from tip-burn susceptible, tip-burn resistant cabbages (B. oleracea var. capitata), and kale (B. oleracea var. acephala). We compared the levels of major macronutrient cations, including Ca(2+) and K(+), in three leaf segments, the leaf apex (LA), middle of leaf (LM), and leaf base (LB), of tip-burn susceptible, tip-burn resistant cabbages, and kale. Ca(2+) and K(+) concentrations were highest in kale, followed by tip-burn resistant and then tip-burn susceptible cabbages. These cations generally accumulated to a greater extent in the LB than in the LA. Transcriptome analysis identified 58,096 loci as putative non-redundant genes in the three leaf segments of the three B. oleracea lines and showed significant changes in expression of 27,876 loci based on Ca(2+) and K(+) levels. Among these, 1844 loci were identified as tip-burn related phenotype-specific genes. Tip-burn resistant cabbage and kale-specific genes were largely related to stress and transport activity based on GO annotation. Tip-burn resistant cabbage and kale plants showed phenotypes clearly indicative of heat-shock, freezing, and drought stress tolerance compared to tip-burn susceptible cabbages, demonstrating a correlation between intracellular Ca(2+) and K(+) concentrations and tolerance of abiotic stress with differential gene expression. We selected 165 genes that were up- or down-regulated in response to increasing Ca(2+) and K(+) concentrations in the three leaf segments of the three plant lines. Gene ontology enrichment analysis indicated that these genes participated in regulatory metabolic processes or stress responses. Our results indicate that the genes involved in regulatory metabolic processes or stress responses were differentially expressed in response to increasing Ca(2+) and K(+) concentrations in the B. oleracea leaf. Our transcriptome data and the genes identified may serve as a starting point for understanding the mechanisms underlying essential macronutrient deficiencies in plants, as well as the features of tip-burn in cabbage and other Brassica species.

Small-tip fast recovery imaging using non-slice-selective tailored tip-up pulses and RF-spoiling

PubMed Central

Nielsen, Jon-Fredrik; Yoon, Daehyun; Noll, Douglas C.

2012-01-01

Small-tip fast recovery (STFR) imaging is a new steady-state imaging sequence that is a potential alternative to balanced steady-state free precession (bSSFP). Under ideal imaging conditions, STFR may provide comparable signal-to-noise ratio (SNR) and image contrast as bSSFP, but without signal variations due to resonance offset. STFR relies on a tailored “tip-up”, or “fast recovery”, RF pulse to align the spins with the longitudinal axis after each data readout segment. The design of the tip-up pulse is based on the acquisition of a separate off-resonance (B0) map. Unfortunately, the design of fast (a few ms) slice- or slab-selective RF pulses that accurately tailor the excitation pattern to the local B0 inhomogeneity over the entire imaging volume remains a challenging and unsolved problem. We introduce a novel implementation of STFR imaging based on non-slice-selective tip-up pulses, which simplifies the RF design problem significantly. Out-of-slice magnetization pathways are suppressed using RF-spoiling. Brain images obtained with this technique show excellent gray/white matter contrast, and point to the possibility of rapid steady-state T2/T1-weighted imaging with intrinsic suppression of cerebrospinal fluid, through-plane vessel signal, and off-resonance artifacts. In the future we expect STFR imaging to benefit significantly from parallel excitation hardware and high-order gradient shim systems. PMID:22511367

Microstructural examination of fatigue crack tip in high strength steel

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

Fukuoka, C.; Yoshizawa, H.; Nakagawa, Y.G.

1993-10-01

Fatigue tests were performed to examine how microstructural conditioning influences crack initiation and propagation in SA508 class 3 low-carbon steel. A 3-mm-long crack was introduced in compact tension (CT) fatigue test specimens under four different loads in order to obtain crack tip plastic zones at different stress intensity factor ranges, [Delta]K = 18, 36, 54, and 72 MPa[radical]m. The microstructure of the plastic zones around the crack tip were examined by transmission electron microscopy (TEM) and selected area electron diffraction (SAD). Micro-orientation of the dislocation cells in the plastic zones of all of the CT samples increased to 4 degmore » from the level of an as-received sample. Four-point bending fatigue tests were performed for plate shape samples with a large cyclic strain range. The SAD value of the bending samples was also 4 deg in the damaged area where cracks already initiated at an early stage of the fatigue process. These test results indicate that the microstructural conditioning is a prerequisite for the fatigue crack initiation and propagation in SA508. These observations may lead to better understanding of how fatigue initiation processes transit to cracks.« less

In Vitro Propagation and Branching Morphogenesis from Single Ureteric Bud Cells.

PubMed

Yuri, Shunsuke; Nishikawa, Masaki; Yanagawa, Naomi; Jo, Oak D; Yanagawa, Norimoto

2017-02-14

A method to maintain and rebuild ureteric bud (UB)-like structures from UB cells in vitro could provide a useful tool for kidney regeneration. We aimed in our present study to establish a serum-free culture system that enables the expansion of UB progenitor cells, i.e., UB tip cells, and reconstruction of UB-like structures. We found that fibroblast growth factors or retinoic acid (RA) was sufficient for the survival of UB cells in serum-free condition, while the proliferation and maintenance of UB tip cells required glial cell-derived neurotrophic factor together with signaling from either WNT-β-catenin pathway or RA. The activation of WNT-β-catenin signaling in UB cells by endogenous WNT proteins required R-spondins. Together with Rho kinase inhibitor, our culture system facilitated the expansion of UB tip cells to form UB-like structures from dispersed single cells. The UB-like structures thus formed retained the original UB characteristics and integrated into the native embryonic kidneys. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

Characterization of size-dependent mechanical properties of tip-growing cells using a lab-on-chip device.

PubMed

Hu, Chengzhi; Munglani, Gautam; Vogler, Hannes; Ndinyanka Fabrice, Tohnyui; Shamsudhin, Naveen; Wittel, Falk K; Ringli, Christoph; Grossniklaus, Ueli; Herrmann, Hans J; Nelson, Bradley J

2016-12-20

Quantification of mechanical properties of tissues, living cells, and cellular components is crucial for the modeling of plant developmental processes such as mechanotransduction. Pollen tubes are tip-growing cells that provide an ideal system to study the mechanical properties at the single cell level. In this article, a lab-on-a-chip (LOC) device is developed to quantitatively measure the biomechanical properties of lily (Lilium longiflorum) pollen tubes. A single pollen tube is fixed inside the microfluidic chip at a specific orientation and subjected to compression by a soft membrane. By comparing the deformation of the pollen tube at a given external load (compressibility) and the effect of turgor pressure on the tube diameter (stretch ratio) with finite element modeling, its mechanical properties are determined. The turgor pressure and wall stiffness of the pollen tubes are found to decrease considerably with increasing initial diameter of the pollen tubes. This observation supports the hypothesis that tip-growth is regulated by a delicate balance between turgor pressure and wall stiffness. The LOC device is modular and adaptable to a variety of cells that exhibit tip-growth, allowing for the straightforward measurement of mechanical properties.

Autonomy and Non-autonomy of Angiogenic Cell Movements Revealed by Experiment-Driven Mathematical Modeling.

PubMed

Sugihara, Kei; Nishiyama, Koichi; Fukuhara, Shigetomo; Uemura, Akiyoshi; Arima, Satoshi; Kobayashi, Ryo; Köhn-Luque, Alvaro; Mochizuki, Naoki; Suda, Toshio; Ogawa, Hisao; Kurihara, Hiroki

2015-12-01

Angiogenesis is a multicellular phenomenon driven by morphogenetic cell movements. We recently reported morphogenetic vascular endothelial cell (EC) behaviors to be dynamic and complex. However, the principal mechanisms orchestrating individual EC movements in angiogenic morphogenesis remain largely unknown. Here we present an experiment-driven mathematical model that enables us to systematically dissect cellular mechanisms in branch elongation. We found that cell-autonomous and coordinated actions governed these multicellular behaviors, and a cell-autonomous process sufficiently illustrated essential features of the morphogenetic EC dynamics at both the single-cell and cell-population levels. Through refining our model and experimental verification, we further identified a coordinated mode of tip EC behaviors regulated via a spatial relationship between tip and follower ECs, which facilitates the forward motility of tip ECs. These findings provide insights that enhance our mechanistic understanding of not only angiogenic morphogenesis, but also other types of multicellular phenomenon. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

In Vitro Comparison of a Novel Single Probe Dual-Energy Lithotripter to Current Devices.

PubMed

Carlos, Evan C; Wollin, Daniel A; Winship, Brenton B; Jiang, Ruiyang; Radvak, Daniela; Chew, Ben H; Gustafson, Michael R; Simmons, W Neal; Zhong, Pei; Preminger, Glenn M; Lipkin, Michael E

2018-06-01

The LithoClast Trilogy is a novel single probe, dual-energy lithotripter with ultrasonic (US) vibration and electromagnetic impact forces. ShockPulse and LithoClast Select are existing lithotripters that also use a combination of US and mechanical impact energies. We compared the efficacy and tip motion of these devices in an in vitro setting. Begostones, in the ratio 15:3, were used in all trials. Test groups were Trilogy, ShockPulse, Select ultrasound (US) only, and Select ultrasound with pneumatic (USP). For clearance testing, a single investigator facile with each lithotripter fragmented 10 stones per device. For drill testing, a hands-free apparatus with a submerged balance was used to apply 1 or 2 lbs of pressure on a stone in contact with the device tip. High-speed photography was used to assess Trilogy and ShockPulse's probe tip motion. Select-USP was slowest and Trilogy fastest on clearance testing (p < 0.01). On 1 lbs drill testing, Select-US was slowest (p = 0.001). At 2 lbs, ShockPulse was faster than Select US (p = 0.027), but did not significantly outpace Trilogy nor Select-USP. At either weight, there was no significant difference between Trilogy and ShockPulse. During its US function, Trilogy's maximum downward tip displacement was 0.041 mm relative to 0.0025 mm with ShockPulse. Trilogy had 0.25 mm of maximum downward displacement during its impactor function while ShockPulse had 0.01 mm. Single probe dual-energy devices, such as Trilogy and ShockPulse, represent the next generation of lithotripters. Trilogy more efficiently cleared stone than currently available devices, which could be explained by its larger probe diameter and greater downward tip displacement during both US and impactor functions.

Susceptibility of loblolly x slash pine interspecific F1 hybrids to tip moth infestation and fusiform rust infection in a south Mississippi planting

Treesearch

M.T. Highsmith; L.H. Lott; C.D. Nelson

2005-01-01

Tip moth damage and fusiform rust incidence among families of three loblolly pine (Pinus taeda) parent trees from Mississippi, Louisiana, and Texas that were selected for southern pine bark beetle resistance and three slash pines (Pinus elliotti var. elliotti) selected for different levels of fusiform rust resistance, and five of their interspecific...

Phosphorylation of Tip60 by GSK-3 determines the induction of PUMA and apoptosis by p53

PubMed Central

Charvet, Céline; Wissler, Manuela; Brauns-Schubert, Prisca; Wang, Shang-Jui; Tang, Yi; Sigloch, Florian C.; Mellert, Hestia; Brandenburg, Martin; Lindner, Silke E.; Breit, Bernhard; Green, Douglas R.; McMahon, Steven B.; Borner, Christoph; Gu, Wei; Maurer, Ulrich

2011-01-01

Summary Activation of p53 by DNA damage results in either cell cycle arrest, allowing DNA repair and cell survival, or induction of apoptosis. As these opposite outcomes are both mediated by p53 stabilization, additional mechanisms to determine this decision must exist. Here we show that glycogen synthase kinase-3 (GSK-3) is required for the p53-mediated induction of the pro-apoptotic BH3 only-protein PUMA, an essential mediator of p53-induced apoptosis. Inhibition of GSK-3 protected from cell death induced by DNA damage and promoted increased long-term cell survival. We demonstrate that GSK-3 phosphorylates serine 86 of the p53-acetyltransferase Tip60. A Tip60S86A mutant was less active to induce p53 K120 acetylation, Histone 4 acetylation and expression of PUMA. Our data suggest that GSK-3 mediated Tip60S86-phosphorylation provides a link between PI3K signaling and the choice for or against apoptosis induction by p53. PMID:21658600

Effects of Mg{sup 2+}, Co{sup 2+}, and Hg{sup 2+} on the nucleus and nucleolus in root tip cells of allium cepa

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

Liu, D.; Jiang, W.; Wang, W.

Metal toxicity in plants has been known for a long time. Much importance has increasingly been attached to the problems of metal pollution with the development of modern industry and agriculture. If metals in plants are accumulated to a large extent, it might seriously affect them. The cytological effects of cobalt and mercury have been studied in Allium cepa by documentation of c-mitosis. Also, the quantification of chromosome aberration in Vicia faba root-tip cells treated by magnesium sulphate and in Allium cepa by metyl mercury chloride and mercuric chloride has been reported. Cytological research on the poisoning effects of Mg,more » Co and Hg on the nuclei and nucleoli in root-tip cells of plants has hardly been reported. The aim of this study was to determine the effects of different concentrations of magnesium, cobalt and mercury ions on root growth, and on the nuclei and nucleoli of root tip cells of Allium-cepa. 20 refs., 3 figs.« less

The tip-link antigen, a protein associated with the transduction complex of sensory hair cells, is protocadherin-15.

PubMed

Ahmed, Zubair M; Goodyear, Richard; Riazuddin, Saima; Lagziel, Ayala; Legan, P Kevin; Behra, Martine; Burgess, Shawn M; Lilley, Kathryn S; Wilcox, Edward R; Riazuddin, Sheikh; Griffith, Andrew J; Frolenkov, Gregory I; Belyantseva, Inna A; Richardson, Guy P; Friedman, Thomas B

2006-06-28

Sound and acceleration are detected by hair bundles, mechanosensory structures located at the apical pole of hair cells in the inner ear. The different elements of the hair bundle, the stereocilia and a kinocilium, are interconnected by a variety of link types. One of these links, the tip link, connects the top of a shorter stereocilium with the lateral membrane of an adjacent taller stereocilium and may gate the mechanotransducer channel of the hair cell. Mass spectrometric and Western blot analyses identify the tip-link antigen, a hitherto unidentified antigen specifically associated with the tip and kinocilial links of sensory hair bundles in the inner ear and the ciliary calyx of photoreceptors in the eye, as an avian ortholog of human protocadherin-15, a product of the gene for the deaf/blindness Usher syndrome type 1F/DFNB23 locus. Multiple protocadherin-15 transcripts are shown to be expressed in the mouse inner ear, and these define four major isoform classes, two with entirely novel, previously unidentified cytoplasmic domains. Antibodies to the three cytoplasmic domain-containing isoform classes reveal that each has a different spatiotemporal expression pattern in the developing and mature inner ear. Two isoforms are distributed in a manner compatible for association with the tip-link complex. An isoform located at the tips of stereocilia is sensitive to calcium chelation and proteolysis with subtilisin and reappears at the tips of stereocilia as transduction recovers after the removal of calcium chelators. Protocadherin-15 is therefore associated with the tip-link complex and may be an integral component of this structure and/or required for its formation.

Genotoxicity effects of silver nanoparticles on wheat (Triticum aestivum L.) root tip cells.

PubMed

Abdelsalam, Nader R; Abdel-Megeed, Ahmed; Ali, Hayssam M; Salem, Mohamed Z M; Al-Hayali, Muwafaq F A; Elshikh, Mohamed S

2018-07-15

The distribution and use of nanoparticles have rapidly increased over recent years, but the available knowledge regarding their mode of action, ecological tolerance and biodegradability remains insufficient. Wheat (Triticum aestivum L.) is the most important crop worldwide. In the current study, the effects of silver nanoparticles (AgNPs) obtained from two different sources, namely, green and chemical syntheses, on chromosomal aberrations and cell division were investigated. Wheat root tips were treated with four different AgNP concentrations (10, 20, 40 and 50 ppm) for three different exposure durations (8, 16 and 24 h), and the different concentrations of the nanoparticles were added to the tested grains until the root lengths reached 1.5-2 cm. For each concentration, the mitotic indexes (%) were obtained from an analysis of ~ 2000 cells. The treated root-tip cells exhibited various types of chromosomal aberrations, such as incorrect orientation at metaphase, chromosomal breakage, metaphasic plate distortion, spindle dysfunction, stickiness, aberrant movement at metaphase, fragmentation, scattering, unequal separation, scattering, chromosomal gaps, multipolar anaphase, erosion, and distributed and lagging chromosomes. These results demonstrate that the root tip cells of wheat can readily internalize the AgNPs and that the internalized AgNPs can interfere with the cells' normal function. Copyright © 2018 Elsevier Inc. All rights reserved.

Electrochemical Visualization of Intracellular Hydrogen Peroxide at Single Cells.

PubMed

He, Ruiqin; Tang, Huifen; Jiang, Dechen; Chen, Hong-yuan

2016-02-16

In this Letter, the electrochemical visualization of hydrogen peroxide inside one cell was achieved first using a comprehensive Au-luminol-microelectrode and electrochemiluminescence. The capillary with a tip opening of 1-2 μm was filled with the mixture of chitosan and luminol, which was coated with the thin layers of polyvinyl chloride/nitrophenyloctyl ether (PVC/NPOE) and gold as the microelectrode. Upon contact with the aqueous hydrogen peroxide, hydrogen peroxide and luminol in contact with the gold layer were oxidized under the positive potential resulting in luminescence for the imaging. Due to the small diameter of the electrode, the microelectrode tip was inserted into one cell and the bright luminescence observed at the tip confirmed the visualization of intracellular hydrogen peroxide. The further coupling of oxidase on the electrode surface could open the field in the electrochemical imaging of intracellular biomolecules at single cells, which benefited the single cell electrochemical detection.

Cyclic Nucleotide-gated Channel α-3 (CNGA3) Interacts with Stereocilia Tip-Link Cadherin 23 + Exon 68 or Alternatively with Myosin VIIa, Two Proteins Required for Hair Cell Mechanotransduction*

PubMed Central

Selvakumar, Dakshnamurthy; Drescher, Marian J.; Drescher, Dennis G.

2013-01-01

Previously, we obtained evidence for a photoreceptor/olfactory type of CNGA3 transcript in a purified teleost vestibular hair cell preparation with immunolocalization of CNGA3 protein to stereocilia of teleost vestibular and mammalian cochlear hair cells. The carboxyl terminus of highly Ca2+-permeable CNGA3 expressed in the mammalian organ of Corti and saccular hair cells was found to interact with an intracellular domain of microfibril interface-located protein 1 (EMILIN 1), a member of the elastin superfamily, also immunolocalizd to hair cell stereocilia (Selvakumar, D., Drescher, M. J., Dowdall, J. R., Khan, K. M., Hatfield, J. S., Ramakrishnan, N. A., and Drescher, D. G. (2012) Biochem. J. 443, 463–476). Here, we provide evidence for organ of Corti proteins, of Ca2+-dependent binding of the amino terminus of CNGA3 specifically to the carboxyl terminus of stereocilia tip-link protein CDH23 +68 (cadherin 23 with expressed exon 68) by yeast two-hybrid mating and co-transformation protocols, pulldown assays, and surface plasmon resonance analysis. Myosin VIIa, required for adaptation of hair cell mechanotransduction (MET) channel(s), competed with CDH23 +68, with direct Ca2+-dependent binding to the amino terminus of CNGA3. Based upon the premise that hair cell stereocilia tip-link proteins are closely coupled with MET, these results are consistent with the possibility that CNGA3 participates in hair-cell MET. Together with the demonstration of protein-protein interaction between HCN1 and tip-link protein protocadherin 15 CD3 (Ramakrishnan, N. A., Drescher, M. J., Barretto, R. L., Beisel, K. W., Hatfield, J. S., and Drescher, D. G. (2009) J. Biol. Chem. 284, 3227–3238; Ramakrishnan, N. A., Drescher, M. J., Khan, K. M., Hatfield, J. S., and Drescher, D. G. (2012) J. Biol. Chem. 287, 37628–37646), a protein-protein interaction for CNGA3 and a second tip-link protein, CDH23 +68, further suggests possible association of two different channels with a single stereocilia tip link. PMID:23329832

Cyclic nucleotide-gated channel α-3 (CNGA3) interacts with stereocilia tip-link cadherin 23 + exon 68 or alternatively with myosin VIIa, two proteins required for hair cell mechanotransduction.

PubMed

Selvakumar, Dakshnamurthy; Drescher, Marian J; Drescher, Dennis G

2013-03-08

Previously, we obtained evidence for a photoreceptor/olfactory type of CNGA3 transcript in a purified teleost vestibular hair cell preparation with immunolocalization of CNGA3 protein to stereocilia of teleost vestibular and mammalian cochlear hair cells. The carboxyl terminus of highly Ca(2+)-permeable CNGA3 expressed in the mammalian organ of Corti and saccular hair cells was found to interact with an intracellular domain of microfibril interface-located protein 1 (EMILIN 1), a member of the elastin superfamily, also immunolocalizd to hair cell stereocilia (Selvakumar, D., Drescher, M. J., Dowdall, J. R., Khan, K. M., Hatfield, J. S., Ramakrishnan, N. A., and Drescher, D. G. (2012) Biochem. J. 443, 463-476). Here, we provide evidence for organ of Corti proteins, of Ca(2+)-dependent binding of the amino terminus of CNGA3 specifically to the carboxyl terminus of stereocilia tip-link protein CDH23 +68 (cadherin 23 with expressed exon 68) by yeast two-hybrid mating and co-transformation protocols, pulldown assays, and surface plasmon resonance analysis. Myosin VIIa, required for adaptation of hair cell mechanotransduction (MET) channel(s), competed with CDH23 +68, with direct Ca(2+)-dependent binding to the amino terminus of CNGA3. Based upon the premise that hair cell stereocilia tip-link proteins are closely coupled with MET, these results are consistent with the possibility that CNGA3 participates in hair-cell MET. Together with the demonstration of protein-protein interaction between HCN1 and tip-link protein protocadherin 15 CD3 (Ramakrishnan, N. A., Drescher, M. J., Barretto, R. L., Beisel, K. W., Hatfield, J. S., and Drescher, D. G. (2009) J. Biol. Chem. 284, 3227-3238; Ramakrishnan, N. A., Drescher, M. J., Khan, K. M., Hatfield, J. S., and Drescher, D. G. (2012) J. Biol. Chem. 287, 37628-37646), a protein-protein interaction for CNGA3 and a second tip-link protein, CDH23 +68, further suggests possible association of two different channels with a single stereocilia tip link.

Recombineering reveals a diverse collection of ribosomal proteins L4 and L22 that confer resistance to macrolide antibiotics

PubMed Central

Diner, Elie J.; Hayes, Christopher S.

2009-01-01

Summary Mutations in ribosomal proteins L4 and L22 confer resistance to erythromycin and other macrolide antibiotics in a variety of bacteria. L4 and L22 have elongated loops whose tips converge in the peptide exit tunnel near the macrolide binding site, and resistance mutations typically affect residues within these loops. Here, we use bacteriophage λ Red-mediated recombination, or “recombineering”, to uncover new L4 and L22 alleles that confer macrolide resistance in Escherichia coli. We randomized residues at the tips of the L4 and L22 loops using recombineered oligonucleotide libraries, and selected the mutagenized cells for erythromycin-resistant mutants. These experiments led to the identification of 341 different resistance mutations encoding 278 unique L4 and L22 proteins – the overwhelming majority of which are novel. Many resistance mutations were complex, involving multiple missense mutations, in-frame deletions, and insertions. Transfer of L4 and L22 mutations into wild-type cells by phage P1-mediated transduction demonstrated that each allele was sufficient to confer macrolide resistance. Although L4 and L22 mutants are typically resistant to most macrolides, selections carried out on different antibiotics revealed macrolide-specific resistance mutations. L22 Lys90Trp is one such allele, which confers resistance to erythromycin, but not tylosin or spiramycin. Purified L22 Lys90Trp ribosomes show reduced erythromycin binding, but have the same affinity for tylosin as wild-type ribosomes. Moreover, DMS methylation protection assays demonstrated that L22 Lys90Trp ribosomes bind tylosin more readily than erythromycin in vivo. This work underscores the exceptional functional plasticity of the L4 and L22 proteins, and highlights the utility of Red-mediated recombination in targeted genetic selections. PMID:19150357

Selected mode of dendritic growth with n-fold symmetry in the presence of a forced flow

NASA Astrophysics Data System (ADS)

Alexandrov, D. V.; Galenko, P. K.

2017-07-01

The effect of n-fold crystal symmetry is investigated for a two-dimensional stable dendritic growth in the presence of a forced convective flow. We consider dendritic growth in a one-component undercooled liquid. The theory is developed for the parabolic solid-liquid surface of dendrite growing at arbitrary growth Péclet numbers keeping in mind small anisotropies of surface energy and growth kinetics. The selection criterion determining the stable growth velocity of the dendritic tip and its stable tip diameter is found on the basis of solvability analysis. The obtained criterion includes previously developed theories of thermally and kinetically controlled dendritic growth with convection for the case of four-fold crystal symmetry. The obtained nonlinear system of equations (representing the selection criterion and undercooling balance) for the determination of dendrite tip velocity and dendrite tip diameter is analytically solved in a parametric form. These exact solutions clearly demonstrate a transition between thermally and kinetically controlled growth regimes. In addition, we show that the dendrites with larger crystal symmetry grow faster than those with smaller symmetry.

Cellular functions of TIP60.

PubMed

Sapountzi, Vasileia; Logan, Ian R; Robson, Craig N

2006-01-01

TIP60 was originally identified as a cellular acetyltransferase protein that interacts with HIV-1 Tat. As a consequence, the role of TIP60 in transcriptional regulation has been investigated intensively. Recent data suggest that TIP60 has more divergent functions than originally thought and roles for TIP60 in many processes, such as cellular signalling, DNA damage repair, cell cycle and checkpoint control and apoptosis are emerging. TIP60 is a tightly regulated transcriptional coregulator, acting in a large multiprotein complex for a range of transcription factors including androgen receptor, Myc, STAT3, NF-kappaB, E2F1 and p53. This usually involves recruitment of TIP60 acetyltransferase activities to chromatin. Additionally, in response to DNA double strand breaks, TIP60 is recruited to DNA lesions where it participates both in the initial as well as the final stages of repair. Here, we describe how TIP60 is a multifunctional enzyme involved in multiple nuclear transactions.

Spearhead Nanometric Field-Effect Transistor Sensors for Single-Cell Analysis.

PubMed

Zhang, Yanjun; Clausmeyer, Jan; Babakinejad, Babak; Córdoba, Ainara López; Ali, Tayyibah; Shevchuk, Andrew; Takahashi, Yasufumi; Novak, Pavel; Edwards, Christopher; Lab, Max; Gopal, Sahana; Chiappini, Ciro; Anand, Uma; Magnani, Luca; Coombes, R Charles; Gorelik, Julia; Matsue, Tomokazu; Schuhmann, Wolfgang; Klenerman, David; Sviderskaya, Elena V; Korchev, Yuri

2016-03-22

Nanometric field-effect-transistor (FET) sensors are made on the tip of spear-shaped dual carbon nanoelectrodes derived from carbon deposition inside double-barrel nanopipettes. The easy fabrication route allows deposition of semiconductors or conducting polymers to comprise the transistor channel. A channel from electrodeposited poly pyrrole (PPy) exhibits high sensitivity toward pH changes. This property is exploited by immobilizing hexokinase on PPy nano-FETs to give rise to a selective ATP biosensor. Extracellular pH and ATP gradients are key biochemical constituents in the microenvironment of living cells; we monitor their real-time changes in relation to cancer cells and cardiomyocytes. The highly localized detection is possible because of the high aspect ratio and the spear-like design of the nano-FET probes. The accurately positioned nano-FET sensors can detect concentration gradients in three-dimensional space, identify biochemical properties of a single living cell, and after cell membrane penetration perform intracellular measurements.

Spearhead Nanometric Field-Effect Transistor Sensors for Single-Cell Analysis

PubMed Central

Córdoba, Ainara López; Ali, Tayyibah; Shevchuk, Andrew; Takahashi, Yasufumi; Novak, Pavel; Edwards, Christopher; Lab, Max; Gopal, Sahana; Chiappini, Ciro; Anand, Uma; Magnani, Luca; Coombes, R. Charles; Gorelik, Julia; Matsue, Tomokazu; Schuhmann, Wolfgang; Klenerman, David; Sviderskaya, Elena V.; Korchev, Yuri

2016-01-01

Nanometric field-effect-transistor (FET) sensors are made on the tip of spear-shaped dual carbon nanoelectrodes derived from carbon deposition inside double-barrel nanopipettes. The easy fabrication route allows deposition of semiconductors or conducting polymers to comprise the transistor channel. A channel from electrodeposited poly pyrrole (PPy) exhibits high sensitivity toward pH changes. This property is exploited by immobilizing hexokinase on PPy nano-FETs to give rise to a selective ATP biosensor. Extracellular pH and ATP gradients are key biochemical constituents in the microenvironment of living cells; we monitor their real-time changes in relation to cancer cells and cardiomyocytes. The highly localized detection is possible because of the high aspect ratio and the spear-like design of the nano-FET probes. The accurately positioned nano-FET sensors can detect concentration gradients in three-dimensional space, identify biochemical properties of a single living cell, and after cell membrane penetration perform intracellular measurements. PMID:26816294

The influence of tip shape on bending force during needle insertion

PubMed Central

van de Berg, Nick J.; de Jong, Tonke L.; van Gerwen, Dennis J.; Dankelman, Jenny; van den Dobbelsteen, John J.

2017-01-01

Steering of needles involves the planning and timely modifying of instrument-tissue force interactions to allow for controlled deflections during the insertion in tissue. In this work, the effect of tip shape on these forces was studied using 10 mm diameter needle tips. Six different tips were selected, including beveled and conical versions, with or without pre-bend or pre-curve. A six-degree-of-freedom force/torque sensor measured the loads during indentations in tissue simulants. The increased insertion (axial) and bending (radial) forces with insertion depth — the force-displacement slopes — were analyzed. Results showed that the ratio between radial and axial forces was not always proportional. This means that the tip load does not have a constant orientation, as is often assumed in mechanics-based steering models. For all tip types, the tip-load assumed a more radial orientation with increased axial load. This effect was larger for straight tips than for pre-bent or pre-curved tips. In addition, the force-displacement slopes were consistently higher for (1) increased tip angles, and for (2) beveled tips compared to conical tips. Needles with a bent or curved tip allow for an increased bending force and a decreased variability of the tip load vector orientation. PMID:28074939

Rhodamine B induces long nucleoplasmic bridges and other nuclear anomalies in Allium cepa root tip cells.

PubMed

Tan, Dehong; Bai, Bing; Jiang, Donghua; Shi, Lin; Cheng, Shunchang; Tao, Dongbing; Ji, Shujuan

2014-03-01

The cytogenetic toxicity of rhodamine B on root tip cells of Allium cepa was investigated. A. cepa were cultured in water (negative control), 10 ppm methyl methanesulfonate (positive control), and three concentrations of rhodamine B (200, 100, and 50 ppm) for 7 days. Rhodamine B inhibited mitotic activity; increased nuclear anomalies, including micronuclei, nuclear buds, and bridged nuclei; and induced oxidative stress in A. cepa root tissues. Furthermore, a substantial amount of long nucleoplasmic bridges were entangled together, and some nuclei were simultaneously linked to several other nuclei and to nuclear buds with nucleoplasmic bridges in rhodamine B-treated cells. In conclusion, rhodamine B induced cytogenetic effects in A. cepa root tip cells, which suggests that the A. cepa root is an ideal model system for detecting cellular interactions.

LRIT3 is essential to localize TRPM1 to the dendritic tips of depolarizing bipolar cells and may play a role in cone synapse formation

PubMed Central

Neuillé, Marion; Morgans, Catherine W.; Cao, Yan; Orhan, Elise; Michiels, Christelle; Sahel, José-Alain; Audo, Isabelle; Duvoisin, Robert M.; Martemyanov, Kirill A.; Zeitz, Christina

2016-01-01

Mutations in LRIT3 lead to complete congenital stationary night blindness (cCSNB). The exact role of LRIT3 in ON-bipolar cell signaling cascade remains to be elucidated. Recently, we have characterized a novel mouse model lacking Lrit3 (no b-wave 6, (Lrit3nob6/nob6)), which displays similar abnormalities as patients with cCSNB with LRIT3 mutations. Here we compare the localization of components of the ON-bipolar cell signaling cascade in wild-type and Lrit3nob6/nob6 retinal sections by immunofluorescence confocal microscopy. An anti-LRIT3 antibody was generated. Immunofluorescent staining of LRIT3 in wild-type mice revealed a specific punctate labeling in the outer plexiform layer (OPL), which was absent in Lrit3nob6/nob6 mice. LRIT3 did not colocalize with ribeye or calbindin but colocalized with mGluR6. TRPM1 staining was severely decreased at the dendritic tips of all depolarizing bipolar cells in Lrit3nob6/nob6 mice. mGluR6, GPR179, RGS7, RGS11 and Gβ5 immunofluorescence was absent at the dendritic tips of cone ON-bipolar cells in Lrit3nob6/nob6 mice, while it was present at the dendritic tips of rod bipolar cells. Furthermore, PNA labeling was severely reduced in the OPL in Lrit3nob6/nob6 mice. This study confirmed the localization of LRIT3 at the dendritic tips of depolarizing bipolar cells in mouse retina and demonstrated the dependence of TRPM1 localization on the presence of LRIT3. Since tested components of the ON-bipolar cell signaling cascade and PNA revealed disrupted localization, an additional function of LRIT3 in cone synapse formation is suggested. These results point to a possibly different regulation of the mGluR6 signaling cascade between rod and cone ON-bipolar cells. PMID:25997951

Superresolution and pulse-chase imaging reveal the role of vesicle transport in polar growth of fungal cells

PubMed Central

Zhou, Lu; Evangelinos, Minoas; Wernet, Valentin; Eckert, Antonia F.; Ishitsuka, Yuji; Fischer, Reinhard; Nienhaus, G. Ulrich; Takeshita, Norio

2018-01-01

Polarized growth of filamentous fungi requires continuous transport of biomolecules to the hyphal tip. To this end, construction materials are packaged in vesicles and transported by motor proteins along microtubules and actin filaments. We have studied these processes with quantitative superresolution localization microscopy of live Aspergillus nidulans cells expressing the photoconvertible protein mEosFPthermo fused to the chitin synthase ChsB. ChsB is mainly located at the Spitzenkörper near the hyphal tip and produces chitin, a key component of the cell wall. We have visualized the pulsatory dynamics of the Spitzenkörper, reflecting vesicle accumulation before exocytosis and their subsequent fusion with the apical plasma membrane. Furthermore, high-speed pulse-chase imaging after photoconversion of mEosFPthermo in a tightly focused spot revealed that ChsB is transported with two different speeds from the cell body to the hyphal tip and vice versa. Comparative analysis using motor protein deletion mutants allowed us to assign the fast movements (7 to 10 μm s−1) to transport of secretory vesicles by kinesin-1, and the slower ones (2 to 7 μm s−1) to transport by kinesin-3 on early endosomes. Our results show how motor proteins ensure the supply of vesicles to the hyphal tip, where temporally regulated exocytosis results in stepwise tip extension. PMID:29387789

Structure of a force-conveying cadherin bond essential for inner-ear mechanotransduction.

PubMed

Sotomayor, Marcos; Weihofen, Wilhelm A; Gaudet, Rachelle; Corey, David P

2012-12-06

Hearing and balance use hair cells in the inner ear to transform mechanical stimuli into electrical signals. Mechanical force from sound waves or head movements is conveyed to hair-cell transduction channels by tip links, fine filaments formed by two atypical cadherins known as protocadherin 15 and cadherin 23 (refs 4, 5). These two proteins are involved in inherited deafness and feature long extracellular domains that interact tip-to-tip in a Ca(2+)-dependent manner. However, the molecular architecture of this complex is unknown. Here we combine crystallography, molecular dynamics simulations and binding experiments to characterize the protocadherin 15-cadherin 23 bond. We find a unique cadherin interaction mechanism, in which the two most amino-terminal cadherin repeats (extracellular cadherin repeats 1 and 2) of each protein interact to form an overlapped, antiparallel heterodimer. Simulations predict that this tip-link bond is mechanically strong enough to resist forces in hair cells. In addition, the complex is shown to become unstable in response to Ca(2+) removal owing to increased flexure of Ca(2+)-free cadherin repeats. Finally, we use structures and biochemical measurements to study the molecular mechanisms by which deafness mutations disrupt tip-link function. Overall, our results shed light on the molecular mechanics of hair-cell sensory transduction and on new interaction mechanisms for cadherins, a large protein family implicated in tissue and organ morphogenesis, neural connectivity and cancer.

Fabrication of near-field optical apertures in aluminium by a highly selective corrosion process in the evanescent field.

PubMed

Haefliger, D; Stemmer, A

2003-03-01

A simple, one-step process to fabricate high-quality apertures for scanning near-field optical microscope probes based on aluminium-coated silicon nitride cantilevers is presented. A thin evanescent optical field at a glass-water interface was used to heat the aluminium at the tip apex due to light absorption. The heat induced a breakdown of the passivating oxide layer and local corrosion of the metal, which selectively exposed the front-most part of the probe tip from the aluminium. Apertures with a protruding silicon nitride tip up to 72 nm in height were fabricated. The height of the protrusion was controlled by the extent of the evanescent field, whereas the diameter depended on the geometry of the probe substrate. The corrosion process proved to be self-terminating, yielding highly reproducible tip heights. Near-field optical resolution in a transmission mode of 85 nm was demonstrated.

Effects of inversion on plastid position and gravitropism in Ceratodon protonemata

NASA Technical Reports Server (NTRS)

Schwuchow, J.; Sack, F. D.

1993-01-01

When dark-grown tip cells of protonemata of the moss Ceratodon purpureus are turned to the horizontal, plastids first sediment towards gravity in a specific zone and then the tip curves upward. To determine whether gravitropism and plastid sedimentation occur in other orientations, protonemata were reoriented to angles other than 90 degrees. Qualitative and quantitative light microscopic observations show that plastid sedimentation along the cell axis occurs in both upright and inverted cells. However, only some plastids fall and sedimentation is incomplete; plastids remain distributed throughout the length of the cell, and those plastids that sediment do not fall all the way to the bottom of the cell. Tip cells are gravitropic regardless of stimulation angle, and as in higher plants, the maximal rate of initial curvature is in response to a 120 degrees reorientation. Infrared videomicroscopy, time-lapse studies of living, inverted protonemata indicate that amyloplast sedimentation precedes upward curvature. Together, these data further support (i) the hypothesis that amyloplast sedimentation functions in gravitropic sensing in these cells, and (ii) the idea that gravity affected the evolution of cell organization.

Design and synthesis of diverse functional kinked nanowire structures for nanoelectronic bioprobes.

PubMed

Xu, Lin; Jiang, Zhe; Qing, Quan; Mai, Liqiang; Zhang, Qingjie; Lieber, Charles M

2013-02-13

Functional kinked nanowires (KNWs) represent a new class of nanowire building blocks, in which functional devices, for example, nanoscale field-effect transistors (nanoFETs), are encoded in geometrically controlled nanowire superstructures during synthesis. The bottom-up control of both structure and function of KNWs enables construction of spatially isolated point-like nanoelectronic probes that are especially useful for monitoring biological systems where finely tuned feature size and structure are highly desired. Here we present three new types of functional KNWs including (1) the zero-degree KNW structures with two parallel heavily doped arms of U-shaped structures with a nanoFET at the tip of the "U", (2) series multiplexed functional KNW integrating multi-nanoFETs along the arm and at the tips of V-shaped structures, and (3) parallel multiplexed KNWs integrating nanoFETs at the two tips of W-shaped structures. First, U-shaped KNWs were synthesized with separations as small as 650 nm between the parallel arms and used to fabricate three-dimensional nanoFET probes at least 3 times smaller than previous V-shaped designs. In addition, multiple nanoFETs were encoded during synthesis in one of the arms/tip of V-shaped and distinct arms/tips of W-shaped KNWs. These new multiplexed KNW structures were structurally verified by optical and electron microscopy of dopant-selective etched samples and electrically characterized using scanning gate microscopy and transport measurements. The facile design and bottom-up synthesis of these diverse functional KNWs provides a growing toolbox of building blocks for fabricating highly compact and multiplexed three-dimensional nanoprobes for applications in life sciences, including intracellular and deep tissue/cell recordings.

Structured Post-IQ Domain Governs Selectivity of Myosin X for Fascin-Actin Bundles*

PubMed Central

Nagy, Stanislav; Rock, Ronald S.

2010-01-01

Without guidance cues, cytoskeletal motors would traffic components to the wrong destination with disastrous consequences for the cell. Recently, we identified a motor protein, myosin X, that identifies bundled actin filaments for transport. These bundles direct myosin X to a unique destination, the tips of cellular filopodia. Because the structural and kinetic features that drive bundle selection are unknown, we employed a domain-swapping approach with the nonselective myosin V to identify the selectivity module of myosin X. We found a surprising role of the myosin X tail region (post-IQ) in supporting long runs on bundles. Moreover, the myosin X head is adapted for initiating processive runs on bundles. We found that the tail is structured and biases the orientation of the two myosin X heads because a targeted insertion that introduces flexibility in the tail abolishes selectivity. Together, these results suggest how myosin motors may manage to read cellular addresses. PMID:20538587

Rational design and validation of a Tip60 histone acetyltransferase inhibitor

NASA Astrophysics Data System (ADS)

Gao, Chunxia; Bourke, Emer; Scobie, Martin; Famme, Melina Arcos; Koolmeister, Tobias; Helleday, Thomas; Eriksson, Leif A.; Lowndes, Noel F.; Brown, James A. L.

2014-06-01

Histone acetylation is required for many aspects of gene regulation, genome maintenance and metabolism and dysfunctional acetylation is implicated in numerous diseases, including cancer. Acetylation is regulated by histone acetyltransferases (HATs) and histone deacetylases and currently, few general HAT inhibitors have been described. We identified the HAT Tip60 as an excellent candidate for targeted drug development, as Tip60 is a key mediator of the DNA damage response and transcriptional co-activator. Our modeling of Tip60 indicated that the active binding pocket possesses opposite charges at each end, with the positive charges attributed to two specific side chains. We used structure based drug design to develop a novel Tip60 inhibitor, TH1834, to fit this specific pocket. We demonstrate that TH1834 significantly inhibits Tip60 activity in vitro and treating cells with TH1834 results in apoptosis and increased unrepaired DNA damage (following ionizing radiation treatment) in breast cancer but not control cell lines. Furthermore, TH1834 did not affect the activity of related HAT MOF, as indicated by H4K16Ac, demonstrating specificity. The modeling and validation of the small molecule inhibitor TH1834 represents a first step towards developing additional specific, targeted inhibitors of Tip60 that may lead to further improvements in the treatment of breast cancer.

Proliferative activity and branching morphogenesis in the human prostate: a closer look at pre- and postnatal prostate growth.

PubMed

Xue, Y; Sonke, G; Schoots, C; Schalken, J; Verhofstad, A; de la Rosette, J; Smedts, F

2001-10-01

To gain further insight into the molecular cell biologic features of prostate development, we investigated the proliferative activity of prostate epithelial and stromal cells and their topographic relationship with neuroendocrine (NE) cell distribution and regional heterogeneity. Consecutive sections from 43 prostates taken during autopsy representing fetuses (12-38 weeks of gestation), infants, prepubertal males and adults were double stained for chromogranin A and MIB-1. MIB-1 labeling index (LI) was calculated in the budding tips, forming acini, major collecting ducts, adjacent and non-adjacent stromal compartments. Furthermore, the topographic relationship between proliferating cells and NE cells was evaluated. In the first half of gestation, cell proliferation as revealed by MIB-1 LI was significantly higher in epithelial structures and stroma than in older fetuses and other age groups. MIB-1 LI was higher in budding tips than in other epithelial regions. MIB-1 LI in stroma adjacent to budding tips was not higher than that adjacent to other epithelial branching segments. Co-expression of chromogranin A and MIB-1 staining was not observed. MIB-1 LI was lower in cells in the direct vicinity of chromogranin A positive NE cells than at a distance from NE cells. Prostate development in the first half of gestation is explosive. Thereafter, the prostate basically is a slow-growing organ. Budding tips are the major growth foci during early prostate development, while stromal growth is evenly distributed throughout the prostate, probably indicating that stromal-epithelial interactions do not manifest in enhanced proliferation at their interface. NE cells may have an inhibitory effect on proliferation of exocrine epithelial cells and are probably only associated with differentiation of prostate exocrine cells in the prostate. Copyright 2001 Wiley-Liss, Inc.

Aeroelastic considerations for torsionally soft rotors

NASA Technical Reports Server (NTRS)

Mantay, W. R.; Yeager, W. T., Jr.

1985-01-01

A research study was initiated to systematically determine the impact of selected blade tip geometric parameters on conformable rotor performance and loads characteristics. The model articulated rotors included baseline and torsionally soft blades with interchangeable tips. Seven blade tip designs were evaluated on the baseline rotor and six tip designs were tested on the torsionally soft blades. The designs incorporated a systemmatic variation in geometric parameters including sweep, taper, and anhedral. The rotors were evaluated in the NASA Langley Transonic Dynamics Tunnel at several advance ratios, lift and propulsive force values, and tip Mach numbers. A track sensitivity study was also conducted at several advance ratios for both rotors. Based on the test results, tip parameter variations generated significant rotor performance and loads differences for both baseline and torsionally soft blades.

Reactive oxygen species triggering systemic programmed cell death process via elevation of nuclear calcium ion level in tomatoes resisting tobacco mosaic virus.

PubMed

Li, Yang; Li, Qi; Hong, Qiang; Lin, Yichun; Mao, Wang; Zhou, Shumin

2018-05-01

Programmed cell death (PCD) plays a positive role in the systemic response of plants to pathogen resistance. It has been confirmed that local tobacco mosaic virus (TMV) infecting tomato leaves can induce systemic PCD process in root-tip tissues. But up to now the underlying physiological mechanisms are poorly understood. This study focused on the detailed investigation of the physiological responses of root-tip cells during the initiation of systemic PCD. Physiological, biochemical examination and cytological observation showed that 1 day post-inoculation (dpi) of TMV inoculation there was an increase in calcium fluorescence intensity in root tip tissue cells. Then at 2 dpi, 4 dpi, 8 dpi and 15 dpi, the fluorescence intensity of calcium ion continued to increase. However, at 5 dpi, the reactive oxygen species (ROS) began to accumulate in the root-tip cells. And finally at 20 dpi, the obvious PCD reaction was detected. In addition, the experimental results also showed that the above process involved the elevation of two types of intracellular Ca 2+ , including cytoplasmic calcium ([Ca 2+ ] cyt ) and nuclear calcium ([Ca 2+ ] nuc ). The [Ca 2+ ] cyt , as a pilot signal could lead to the subsequent elevation of intracellular ROS concentration. Then, the high levels of ROS stimulated an increase of [Ca 2+ ] nuc and eventually caused PCD reactions in the root-tip tissues. In particular, the high level of nuclear calcium is an essential mediator in systemic PCD of plants. Copyright © 2018 Elsevier B.V. All rights reserved.

KCH kinesin drives nuclear transport and cytoskeletal coalescence for tip cell growth.

PubMed

Yamada, Moé; Goshima, Gohta

2018-06-07

Long-distance transport along microtubules (MTs) is critical for intracellular organisation. In animals, antagonistic motor proteins kinesin (plus end-directed) and dynein (minus end-directed) drive cargo transport. In land plants, however, the identity of motors responsible for transport is poorly understood, as genes encoding cytoplasmic dynein are absent in plant genomes. How other functions of dynein are brought about in plants also remains unknown. Here, we show that a subclass of the kinesin-14 family, KCH (kinesin with calponin homology domain)-which can also bind actin-drives MT minus end-directed nuclear transport in the moss Physcomitrella patens. When all four KCH genes were deleted, the nucleus was not maintained in the cell centre, but was translocated to the apical end of protonemal cells. In the knockout (KO) line, apical cell tip growth was also severely suppressed. KCH was localized to MTs, including at the MT focal point near the tip of protonemal cells, where MT plus ends coalesced with actin filaments. MT focus was not stably maintained in KCH KO lines, whereas actin destabilisation also disrupted the MT focus in wild-type lines despite KCH remaining on unfocused MTs. KCH had distinct functions in nuclear transport and tip growth, as a truncated KCH construct restored nuclear transport activity, but not tip growth retardation of the KO line. Thus, our study identified KCH as a long-distance retrograde transporter as well as a MT crosslinker, reminiscent of the versatile animal dynein. © 2018 American Society of Plant Biologists. All rights reserved.

Small diameter carbon nanopipettes

NASA Astrophysics Data System (ADS)

Singhal, Riju; Bhattacharyya, Sayan; Orynbayeva, Zulfiya; Vitol, Elina; Friedman, Gary; Gogotsi, Yury

2010-01-01

Nanoscale multifunctional carbon probes facilitate cellular studies due to their small size, which makes it possible to interrogate organelles within living cells in a minimally invasive fashion. However, connecting nanotubes to macroscopic devices and constructing an integrated system for the purpose of fluid and electrical signal transfer is challenging, as is often the case with nanoscale components. We describe a non-catalytic chemical vapor deposition based method for batch fabrication of integrated multifunctional carbon nanopipettes (CNPs) with tip diameters much smaller (10-30 nm) than previously reported (200 nm and above) and approaching those observed for multiwalled carbon nanotubes. This eliminates the need for complicated attachment/assembly of nanotubes into nanofluidic devices. Variable tip geometries and structures were obtained by controlled deposition of carbon inside and outside quartz pipettes. We have shown that the capillary length and gas flow rate have a marked effect on the carbon deposition. This gives us a flexible protocol, useful for growing carbon layers of different thicknesses at selective locations on a glass pipette to yield a large variety of cellular probes in bulk quantities. The CNPs possess an open channel for fluid transfer with the carbon deposited inside at 875 °C behaving like an amorphous semiconductor. Vacuum annealing of the CNP tips at temperatures up to 2000 °C yields graphitic carbon structures with an increase in conductivity of two orders of magnitude. Penetration of the integrated carbon nanoprobes into cells was shown to produce minimal Ca2+ signals, fast recovery of basal Ca2+ levels and no adverse activation of the cellular metabolism during interrogation times as long as 0.5-1 h.

Small diameter carbon nanopipettes.

PubMed

Singhal, Riju; Bhattacharyya, Sayan; Orynbayeva, Zulfiya; Vitol, Elina; Friedman, Gary; Gogotsi, Yury

2010-01-08

Nanoscale multifunctional carbon probes facilitate cellular studies due to their small size, which makes it possible to interrogate organelles within living cells in a minimally invasive fashion. However, connecting nanotubes to macroscopic devices and constructing an integrated system for the purpose of fluid and electrical signal transfer is challenging, as is often the case with nanoscale components. We describe a non-catalytic chemical vapor deposition based method for batch fabrication of integrated multifunctional carbon nanopipettes (CNPs) with tip diameters much smaller (10-30 nm) than previously reported (200 nm and above) and approaching those observed for multiwalled carbon nanotubes. This eliminates the need for complicated attachment/assembly of nanotubes into nanofluidic devices. Variable tip geometries and structures were obtained by controlled deposition of carbon inside and outside quartz pipettes. We have shown that the capillary length and gas flow rate have a marked effect on the carbon deposition. This gives us a flexible protocol, useful for growing carbon layers of different thicknesses at selective locations on a glass pipette to yield a large variety of cellular probes in bulk quantities. The CNPs possess an open channel for fluid transfer with the carbon deposited inside at 875 degrees C behaving like an amorphous semiconductor. Vacuum annealing of the CNP tips at temperatures up to 2000 degrees C yields graphitic carbon structures with an increase in conductivity of two orders of magnitude. Penetration of the integrated carbon nanoprobes into cells was shown to produce minimal Ca(2+) signals, fast recovery of basal Ca(2+) levels and no adverse activation of the cellular metabolism during interrogation times as long as 0.5-1 h.

Using ToxCast™ Data to Reconstruct Dynamic Cell State Trajectories and Estimate Toxicological Points of Departure

PubMed Central

Shah, Imran; Setzer, R. Woodrow; Jack, John; Houck, Keith A.; Judson, Richard S.; Knudsen, Thomas B.; Liu, Jie; Martin, Matthew T.; Reif, David M.; Richard, Ann M.; Thomas, Russell S.; Crofton, Kevin M.; Dix, David J.; Kavlock, Robert J.

2015-01-01

Background: High-content imaging (HCI) allows simultaneous measurement of multiple cellular phenotypic changes and is an important tool for evaluating the biological activity of chemicals. Objectives: Our goal was to analyze dynamic cellular changes using HCI to identify the “tipping point” at which the cells did not show recovery towards a normal phenotypic state. Methods: HCI was used to evaluate the effects of 967 chemicals (in concentrations ranging from 0.4 to 200 μM) on HepG2 cells over a 72-hr exposure period. The HCI end points included p53, c-Jun, histone H2A.x, α-tubulin, histone H3, alpha tubulin, mitochondrial membrane potential, mitochondrial mass, cell cycle arrest, nuclear size, and cell number. A computational model was developed to interpret HCI responses as cell-state trajectories. Results: Analysis of cell-state trajectories showed that 336 chemicals produced tipping points and that HepG2 cells were resilient to the effects of 334 chemicals up to the highest concentration (200 μM) and duration (72 hr) tested. Tipping points were identified as concentration-dependent transitions in system recovery, and the corresponding critical concentrations were generally between 5 and 15 times (25th and 75th percentiles, respectively) lower than the concentration that produced any significant effect on HepG2 cells. The remaining 297 chemicals require more data before they can be placed in either of these categories. Conclusions: These findings show the utility of HCI data for reconstructing cell state trajectories and provide insight into the adaptation and resilience of in vitro cellular systems based on tipping points. Cellular tipping points could be used to define a point of departure for risk-based prioritization of environmental chemicals. Citation: Shah I, Setzer RW, Jack J, Houck KA, Judson RS, Knudsen TB, Liu J, Martin MT, Reif DM, Richard AM, Thomas RS, Crofton KM, Dix DJ, Kavlock RJ. 2016. Using ToxCast™ data to reconstruct dynamic cell state trajectories and estimate toxicological points of departure. Environ Health Perspect 124:910–919; http://dx.doi.org/10.1289/ehp.1409029 PMID:26473631

Transduction channels' gating can control friction on vibrating hair-cell bundles in the ear.

PubMed

Bormuth, Volker; Barral, Jérémie; Joanny, Jean-François; Jülicher, Frank; Martin, Pascal

2014-05-20

Hearing starts when sound-evoked mechanical vibrations of the hair-cell bundle activate mechanosensitive ion channels, giving birth to an electrical signal. As for any mechanical system, friction impedes movements of the hair bundle and thus constrains the sensitivity and frequency selectivity of auditory transduction. Friction is generally thought to result mainly from viscous drag by the surrounding fluid. We demonstrate here that the opening and closing of the transduction channels produce internal frictional forces that can dominate viscous drag on the micrometer-sized hair bundle. We characterized friction by analyzing hysteresis in the force-displacement relation of single hair-cell bundles in response to periodic triangular stimuli. For bundle velocities high enough to outrun adaptation, we found that frictional forces were maximal within the narrow region of deflections that elicited significant channel gating, plummeted upon application of a channel blocker, and displayed a sublinear growth for increasing bundle velocity. At low velocity, the slope of the relation between the frictional force and velocity was nearly fivefold larger than the hydrodynamic friction coefficient that was measured when the transduction machinery was decoupled from bundle motion by severing tip links. A theoretical analysis reveals that channel friction arises from coupling the dynamics of the conformational change associated with channel gating to tip-link tension. Varying channel properties affects friction, with faster channels producing smaller friction. We propose that this intrinsic source of friction may contribute to the process that sets the hair cell's characteristic frequency of responsiveness.

[Introduction of hexaploid of Chinese narcissus and analysis of its chromosome change].

PubMed

Wang, Rui; Zhang, Ya Nan; Wang, Ya Ying; Tian, Hui Qiao

2007-06-01

Anthers of Chinese narcissus (Narcissus tazetta L. var chinesis Roem) were used as explants for callus induction and plant regeneration. About 80% anthers produced callus and 28% of the callus differentiated out bulbs, making a good experiment system of tissue culture of Chinese narcissus for further cellular and gene engineering. The 700 callus were treated by 0.5% colchicin for 5-6 days and then transformed into a MS medium containing 3 mg/L 6-BA to induce differentiation. 90 bulbs were obtained and 55 bulbs among them were checked the chromosome number from their root tips for three times. 29 bulbs (53%, 29/55) still kept triploidy and the most cells of root tips contained 30 chromosomes. 22 bulbs (40%, 22/55) displayed aneuploidy and the most cells of its root tips contained 10-50 chromosomes. 4 bulbs displayed hexaploidy and contained 60 chromosomes. After three months growing, the cells of root tips containing aneuploidy chromosomes disappeared, and the bulbs became triploidy. The chromosomes of 4 hexaploidy bulbs did not changed during three checks. The origin and disappearance of aneuploidy cells of Chinese narcissus after treated by colchicin were discussed.

Aluminum-induced cell death of barley-root border cells is correlated with peroxidase- and oxalate oxidase-mediated hydrogen peroxide production.

PubMed

Tamás, L; Budíková, S; Huttová, J; Mistrík, I; Simonovicová, M; Siroká, B

2005-06-01

The function of root border cells (RBC) during aluminum (Al) stress and the involvement of oxalate oxidase, peroxidase and H(2)O(2) generation in Al toxicity were studied in barley roots. Our results suggest that RBC effectively protect the barley root tip from Al relative to the situation in roots cultivated in hydroponics where RBC are not sustained in the area surrounding the root tip. The removal of RBC from Al-treated roots increased root growth inhibition, Al and Evans blue uptake, inhibition of RBC production, the level of dead RBC, peroxidase and oxalate oxidase activity and the production of H(2)O(2). Our results suggest that even though RBC actively produce active oxygen species during Al stress, their role in the protection of root tips against Al toxicity is to chelate Al in their dead cell body.

ZIFL1.1 transporter modulates polar auxin transport by stabilizing membrane abundance of multiple PINs in Arabidopsis root tip

PubMed Central

Remy, Estelle; Baster, Pawel; Friml, Jiří; Duque, Paula

2013-01-01

Cell-to-cell directional flow of the phytohormone auxin is primarily established by polar localization of the PIN auxin transporters, a process tightly regulated at multiple levels by auxin itself. We recently reported that, in the context of strong auxin flows, activity of the vacuolar ZIFL1.1 transporter is required for fine-tuning of polar auxin transport rates in the Arabidopsis root. In particular, ZIFL1.1 function protects plasma-membrane stability of the PIN2 carrier in epidermal root tip cells under conditions normally triggering PIN2 degradation. Here, we show that ZIFL1.1 activity at the root tip also promotes PIN1 plasma-membrane abundance in central cylinder cells, thus supporting the notion that ZIFL1.1 acts as a general positive modulator of polar auxin transport in roots. PMID:23857365

Fifteen Tips for Improving Your Grounds This Summer.

ERIC Educational Resources Information Center

Sturgeon, Julie

1998-01-01

Provides tips on how to improve a university's or college's campus grounds maintenance program that can enhance a school's appearance and improve the odds of increasing enrollment. Grass selection, tree and shrub management, and the use of flowers are examined. (GR)

Induction of numerical chromosomal aberrations during DNA synthesis using the fungicides nimrod and rubigan-4 in root tips of Vicia faba L.

PubMed

Shahin, S A; el-Amoodi, K H

1991-11-01

The 2 fungicides nimrod and rubigan-4 were tested for genotoxicity using Vicia faba root tips as the biological test system. Treating lateral roots with different concentrations of each fungicide for different periods showed that both fungicides were able to produce numerical but not structural chromosomal aberrations. The percentage of total aberrations in root tips exposed to nimrod reached 54.39% at 250 ppm for 4 h, and 64.69% in root tips exposed to rubigan-4 at 250 ppm for 6 h. The types of numerical chromosomal aberrations produced by both fungicides included: binucleate cells, c-metaphases, sticky chromosomes, polyploid cells, and laggards. Recovery experiments for 24, 48, and 96 h showed no significant differences between the percentage of total aberrations in treated and control groups.

Association of Spectrin-Like Proteins with the Actin-Organized Aggregate of Endoplasmic Reticulum in the Spitzenkörper of Gravitropically Tip-Growing Plant Cells1

PubMed Central

Braun, Markus

2001-01-01

Spectrin-like epitopes were immunochemically detected and immunofluorescently localized in gravitropically tip-growing rhizoids and protonemata of characean algae. Antiserum against spectrin from chicken erythrocytes showed cross-reactivity with rhizoid proteins at molecular masses of about 170 and 195 kD. Confocal microscopy revealed a distinct spherical labeling of spectrin-like proteins in the apices of both cell types tightly associated with an apical actin array and a specific subdomain of endoplasmic reticulum (ER), the ER aggregate. The presence of spectrin-like epitopes, the ER aggregate, and the actin cytoskeleton are strictly correlated with active tip growth. Application of cytochalasin D and A23187 has shown that interfering with actin or with the calcium gradient, which cause the disintegration of the ER aggregate and abolish tip growth, inhibits labeling of spectrin-like proteins. At the beginning of the graviresponse in rhizoids the labeling of spectrin-like proteins remained in its symmetrical position at the cell tip, but was clearly displaced to the upper flank in gravistimulated protonemata. These findings support the hypothesis that a displacement of the Spitzenkörper is required for the negative gravitropic response in protonemata, but not for the positive gravitropic response in rhizoids. It is evident that the actin/spectrin system plays a role in maintaining the organization of the ER aggregate and represents an essential part in the mechanism of gravitropic tip growth. PMID:11299343

A Comparison of Three Different Thick Epinucleus Removal Techniques in Cataract Surgery.

PubMed

Hwang, Ho Sik; Lim, Byung-Su; Kim, Man Soo; Kim, Eun Chul

2017-01-01

To compare the outcomes of cataract surgery performed with three different types of the epinucleus removal techniques (safe boat, infusion/aspiration (I/A) cannulas, and phacoemulsification tip). Ninety eyes with thick adhesive epinuclei were randomly subdivided into three groups according to epinucleus removal technique: epinucleus floating (safe boat) technique, 30 patients; I/A tip, 30 patients; and phaco tip, 30 patients. Intraoperative measurements included ultrasound time (UST), mean cumulative dissipated ultrasound energy (CDE), and balanced salt solution (BSS) use. Clinical measurements were made preoperatively, and at one day, one month and two months postoperatively, including the best corrected visual acuity (BCVA), the central corneal thickness (CCT), and the endothelial cell count (ECC). Intraoperative measurements showed significantly less UST, CDE, and BSS use in the safe boat group than in the phaco tip groups (p < 0.05). The percentage of endothelial cell loss in the safe boat group was significantly lower than that in the phaco tip groups at two months post-cataract surgery (p < 0.05). The safe boat technique is a safer and more effective epinucleus removal technique than phaco tip techniques in cases with thick epinucleus.

A role for Mfb1p in region-specific anchorage of high-functioning mitochondria and lifespan in Saccharomyces cerevisiae

PubMed Central

Pernice, Wolfgang M.; Vevea, Jason D.; Pon, Liza A.

2016-01-01

Previous studies indicate that replicative lifespan in daughter cells of Sacchraromyces cerevisiae depends on the preferential inheritance of young, high-functioning mitochondria. We report here that mitochondria are functionally segregated even within single mother cells in S. cerevisiae. A high-functioning population of mitochondria accumulates at the tip of the mother cell distal to the bud. We find that the mitochondrial F-box protein (Mfb1p) localizes to mitochondria in the mother tip and is required for mitochondrial anchorage at that site, independent of the previously identified anchorage protein Num1p. Deletion of MFB1 results in loss of the mother-tip-localized mitochondrial population, defects in mitochondrial function and premature replicative ageing. Inhibiting mitochondrial inheritance to buds, by deletion of MMR1, in mfb1Δ cells restores mitochondrial distribution, promotes mitochondrial function and extends replicative lifespan. Our results identify a mechanism that retains a reservoir of high-functioning mitochondria in mother cells and thereby preserves maternal reproductive capacity. PMID:26839174

Quantification of cellular penetrative forces using lab-on-a-chip technology and finite element modeling

PubMed Central

Sanati Nezhad, Amir; Naghavi, Mahsa; Packirisamy, Muthukumaran; Bhat, Rama; Geitmann, Anja

2013-01-01

Tip-growing cells have the unique property of invading living tissues and abiotic growth matrices. To do so, they exert significant penetrative forces. In plant and fungal cells, these forces are generated by the hydrostatic turgor pressure. Using the TipChip, a microfluidic lab-on-a-chip device developed for tip-growing cells, we tested the ability to exert penetrative forces generated in pollen tubes, the fastest-growing plant cells. The tubes were guided to grow through microscopic gaps made of elastic polydimethylsiloxane material. Based on the deformation of the gaps, the force exerted by the elongating tubes to permit passage was determined using finite element methods. The data revealed that increasing mechanical impedance was met by the pollen tubes through modulation of the cell wall compliance and, thus, a change in the force acting on the obstacle. Tubes that successfully passed a narrow gap frequently burst, raising questions about the sperm discharge mechanism in the flowering plants. PMID:23630253

Lifeact-mEGFP Reveals a Dynamic Apical F-Actin Network in Tip Growing Plant Cells

PubMed Central

Hepler, Peter K.; Bezanilla, Magdalena

2009-01-01

Background Actin is essential for tip growth in plants. However, imaging actin in live plant cells has heretofore presented challenges. In previous studies, fluorescent probes derived from actin-binding proteins often alter growth, cause actin bundling and fail to resolve actin microfilaments. Methodology/Principal Findings In this report we use Lifeact-mEGFP, an actin probe that does not affect the dynamics of actin, to visualize actin in the moss Physcomitrella patens and pollen tubes from Lilium formosanum and Nicotiana tobaccum. Lifeact-mEGFP robustly labels actin microfilaments, particularly in the apex, in both moss protonemata and pollen tubes. Lifeact-mEGFP also labels filamentous actin structures in other moss cell types, including cells of the gametophore. Conclusions/Significance Lifeact-mEGFP, when expressed at optimal levels does not alter moss protonemal or pollen tube growth. We suggest that Lifeact-mEGFP represents an exciting new versatile probe for further studies of actin's role in tip growing plant cells. PMID:19478943

Effects of ginsenosides Rg1 and Rb1 of Panax ginseng on mitosis in root tip cells of Allium cepa.

PubMed

Ng, W Y; Chao, C Y

1981-01-01

The effects of ginsenosides Rg1 and Rb1 of Panax ginseng on mitosis in the onion root tip cells as well as on the rate of DNA synthesis in onion seedlings were studied. Results obtained from the concentration and time course study in bulb and seeding root tip cells indicate that Rg1 promotes and Rb1 inhibits mitosis, both being dose-dependent. The promoting effect of Rg1 on the rate of DNA synthesis was observed at the peak hour which occurs at the same time as that of the control. Rb1 was found to shift the peak hour of DNA synthesis to a later period of the experiment. These results are in agreement with the results obtained from the study of the cell cycle by pulse labeling and autoradiography, which show that Rg1 shortens the mitotic cell cycle and S period while Rb1 lengthens them. They in turn increase and decrease the mitotic indices respectively.

An Osmotic Model of the Growing Pollen Tube

PubMed Central

Hill, Adrian E.; Shachar-Hill, Bruria; Skepper, Jeremy N.; Powell, Janet; Shachar-Hill, Yair

2012-01-01

Pollen tube growth is central to the sexual reproduction of plants and is a longstanding model for cellular tip growth. For rapid tip growth, cell wall deposition and hardening must balance the rate of osmotic water uptake, and this involves the control of turgor pressure. Pressure contributes directly to both the driving force for water entry and tip expansion causing thinning of wall material. Understanding tip growth requires an analysis of the coordination of these processes and their regulation. Here we develop a quantitative physiological model which includes water entry by osmosis, the incorporation of cell wall material and the spreading of that material as a film at the tip. Parameters of the model have been determined from the literature and from measurements, by light, confocal and electron microscopy, together with results from experiments made on dye entry and plasmolysis in Lilium longiflorum. The model yields values of variables such as osmotic and turgor pressure, growth rates and wall thickness. The model and its predictive capacity were tested by comparing programmed simulations with experimental observations following perturbations of the growth medium. The model explains the role of turgor pressure and its observed constancy during oscillations; the stability of wall thickness under different conditions, without which the cell would burst; and some surprising properties such as the need for restricting osmotic permeability to a constant area near the tip, which was experimentally confirmed. To achieve both constancy of pressure and wall thickness under the range of conditions observed in steady-state growth the model reveals the need for a sensor that detects the driving potential for water entry and controls the deposition rate of wall material at the tip. PMID:22615784

Epigenetic Regulation of Axonal Growth of Drosophila Pacemaker Cells by Histone Acetyltransferase Tip60 Controls Sleep

PubMed Central

Pirooznia, Sheila K.; Chiu, Kellie; Chan, May T.; Zimmerman, John E.; Elefant, Felice

2012-01-01

Tip60 is a histone acetyltransferase (HAT) enzyme that epigenetically regulates genes enriched for neuronal functions through interaction with the amyloid precursor protein (APP) intracellular domain. However, whether Tip60-mediated epigenetic dysregulation affects specific neuronal processes in vivo and contributes to neurodegeneration remains unclear. Here, we show that Tip60 HAT activity mediates axonal growth of the Drosophila pacemaker cells, termed “small ventrolateral neurons” (sLNvs), and their production of the neuropeptide pigment-dispersing factor (PDF) that functions to stabilize Drosophila sleep–wake cycles. Using genetic approaches, we show that loss of Tip60 HAT activity in the presence of the Alzheimer’s disease-associated APP affects PDF expression and causes retraction of the sLNv synaptic arbor required for presynaptic release of PDF. Functional consequence of these effects is evidenced by disruption of the sleep–wake cycle in these flies. Notably, overexpression of Tip60 in conjunction with APP rescues these sleep–wake disturbances by inducing overelaboration of the sLNv synaptic terminals and increasing PDF levels, supporting a neuroprotective role for dTip60 in sLNv growth and function under APP-induced neurodegenerative conditions. Our findings reveal a novel mechanism for Tip60 mediated sleep–wake regulation via control of axonal growth and PDF levels within the sLNv-encompassing neural network and provide insight into epigenetic-based regulation of sleep disturbances observed in neurodegenerative diseases like Alzheimer’s disease. PMID:22982579

MIF Promotes Classical Activation and Conversion of Inflammatory Ly6Chigh Monocytes into TipDCs during Murine Toxoplasmosis

PubMed Central

Ruiz-Rosado, Juan de Dios; Olguín, Jonadab E.; Juárez-Avelar, Imelda; Saavedra, Rafael; Terrazas, Luis I.; Robledo-Avila, Frank H.; Vazquez-Mendoza, Alicia; Fernández, Jacquelina; Satoskar, Abhay R.; Partida-Sánchez, Santiago; Rodriguez-Sosa, Miriam

2016-01-01

Macrophage migration inhibitory factor (MIF) mediates immunity against Toxoplasma gondii infection by inducing inflammatory cytokines required to control the parasite replication. However, the role of this inflammatory mediator in the cell-mediated immune response against this infection is still poorly understood. Here, we used T. gondii-infected WT and Mif −/− mice to analyze the role of MIF in the maturation of CD11b+ and CD8α + dendritic cells (DCs). We found that MIF promotes maturation of CD11b+ but not CD8α + DCs, by inducing IL-12p70 production and CD86 expression. Infected Mif −/− mice showed significantly lower numbers of TNF and inducible nitric oxide synthase- (iNOS-) producing DCs (TipDCs) compared to infected WT mice. The adoptive transfer of Ly6Chigh monocytes into infected WT or Mif −/− mice demonstrated that MIF participates in the differentiation of Ly6Chigh monocytes into TipDCs. In addition, infected Mif −/− mice display a lower percentage of IFN-γ-producing natural killer (NK) cells compared to WT mice, which is associated with reducing numbers of TipDCs in Mif −/− mice. Furthermore, administration of recombinant MIF (rMIF) into T. gondii-infected Mif −/− mice restored the numbers of TipDCs and reversed the susceptible phenotype of Mif −/− mice. Collectively, these results demonstrate an important role for MIF inducing cell-mediated immunity to T. gondii infection. PMID:27057101

Identification and validation of reference genes for quantification of target gene expression with quantitative real-time PCR for tall fescue under four abiotic stresses.

PubMed

Yang, Zhimin; Chen, Yu; Hu, Baoyun; Tan, Zhiqun; Huang, Bingru

2015-01-01

Tall fescue (Festuca arundinacea Schreb.) is widely utilized as a major forage and turfgrass species in the temperate regions of the world and is a valuable plant material for studying molecular mechanisms of grass stress tolerance due to its superior drought and heat tolerance among cool-season species. Selection of suitable reference genes for quantification of target gene expression is important for the discovery of molecular mechanisms underlying improved growth traits and stress tolerance. The stability of nine potential reference genes (ACT, TUB, EF1a, GAPDH, SAND, CACS, F-box, PEPKR1 and TIP41) was evaluated using four programs, GeNorm, NormFinder, BestKeeper, and RefFinder. The combinations of SAND and TUB or TIP41 and TUB were most stably expressed in salt-treated roots or leaves. The combinations of GAPDH with TIP41 or TUB were stable in roots and leaves under drought stress. TIP41 and PEPKR1 exhibited stable expression in cold-treated roots, and the combination of F-box, TIP41 and TUB was also stable in cold-treated leaves. CACS and TUB were the two most stable reference genes in heat-stressed roots. TIP41 combined with TUB and ACT was stably expressed in heat-stressed leaves. Finally, quantitative real-time polymerase chain reaction (qRT-PCR) assays of the target gene FaWRKY1 using the identified most stable reference genes confirmed the reliability of selected reference genes. The selection of suitable reference genes in tall fescue will allow for more accurate identification of stress-tolerance genes and molecular mechanisms conferring stress tolerance in this stress-tolerant species.

Generation and multiplication of plantlets from callus derived from Haplopappus gracilus (Nutt.) Gray and their karyotype analysis

NASA Technical Reports Server (NTRS)

Kann, R. P.; O'Connor, S. A.; Levine, H. G.; Krikorian, A. D.

1991-01-01

Unopened flower heads of Haplopappus gracilis (2n = 4) provided primary explants for callus production and subsequent induction of organized growth. Callus was initiated from small (3-5 mm in length) floral buds with benzylaminopurine (BAP) (44.4 micromoles; 10 mg/l) and naphthalene acetic acid (NAA) (0.54 micromole; 0.1 mg/l). Lowering the BAP level to 4.44 micromoles (1 mg/l) but maintaining the NAA level, gave rise to organized but highly compressed shoot growing points from an otherwise undifferentiated callus mass. Shoots selected from such cultures were maintainable and could be proliferated by growing 1-1.5-cm stem tip cuttings on Murashige and Skoog basal medium (solidified with agar) containing 0.444 micromole (0.1 mg/l) BAP and 0.054 micromole (0.01 mg/l) NAA. The stem tip multiplication rates obtainable by these means permit reliable strategies for shoot multiplication or production of rooted plantlets. Prolonged subculture and maintenance of shoots on growth regulator-free medium leads to in vitro flowering and greatly reduces rooting capacity. Karyotype analysis of chromosomes from root tip cells at metaphase and chromosome measurements show that karyologically uniform plantlets (based on chromosome number and morphology) can be obtained.

[Influence of Four Kinds of PPCPs on Micronucleus Rate of the Root-Tip Cells of Vicia-faba and Garlic].

PubMed

Wang, Lan-jun; Wang, Jin-hua; Zhu, Lu-sheng; Wang, Jun; Zhao, Xiang

2016-04-15

In order to determine the degree of biological genetic injury induced by PPCPs, the genotoxic effects of the doxycycline (DOX), ciprofloxacin (CIP), triclocarban (TCC) and carbamazepine (CBZ) in the concentration range of 12.5-100 mg · L⁻¹ were studied using micronucleus rate and micronucleus index of Vicia-fabe and garlic. The results showed that: (1) When the Vicia-faba root- tip cells were exposed to DOX, CIP, TCC and CBZ, micronucleus rates were higher than 1.67 ‰ (CK₁), it was significantly different from that of the control group (P < 0.05), and the micronucleus index was even greater than 3.5; With the increasing concentrations of the PPCPs, the micronucleus rates first increased and then decreased. (2) When the garlic root tip cells were exposed to DOX, CIP, TCC and CBZ respectively, the micronucleus rates were less than those of the Vicia-faba, while in most treatments significantly higher than that of the control group (0.67‰). The micronucleus index was higher than 3.5 in the groups exposed to CIP with concentrations of 25, 50, 100 mg · L⁻¹ and TCC and CBZ with concentrations of 25 mg · L⁻¹; With the increase of exposure concentrations, the micronucleus rate showed a trend of first increasing and then decreasing as well. (3) Under the same experimental conditions, the cells micronucleus rates of the garlic cells caused by the four tested compounds were significantly lower than those of Vicia-faba. (4) The micronucleus index of the root tip cells of Vicia-faba and garlic treated with the four kinds of compounds followed the order of CIP > CBZ > TCC > DOX. These results demonstrated that the four compounds caused biological genetic injury to root-tip cells of Vicia-faba and garlic, and the genetic damage caused to garlic was significantly lower than that to Vicia-faba. The damages caused by the four kinds of different compounds were also different.

Dynamics of Micropipette Vibration During Piezo-assisted Microinjection

NASA Astrophysics Data System (ADS)

Karzar-Jeddi, Mehdi; Olgac, Nejat; Fan, Tai-Hsi

Microinjection is a well-accepted method to introduce materials such as sperm, DNA materials, or nucleus into a living cell for biomedical applications. The conventional microinjection procedure consists of immobilizing the cell by applying suction through a holding pipette, and then an injecting micropipette penetrates through the cell membrane and introduces the materials into the cell. To assist the penetration process a piezo-generated pulse train is applied to the injecting pipette, which causes an undesirable lateral vibration at the injecting pipette tip. In this research we provide an analytical model to study the response of micropipette to the piezo-pulse train using the Duhamel integral method. Our results show that filling the micropipette tip with mercury causes a larger amplitude stroke vibration in micropipette tip than that of empty micropipette when it is submerged in the viscous medium surrounding the cell. The mercury introduced larger stroke vibration can cause a larger shear force and assist the penetration of micropipette through the cell membrane. This work is supported by NSF CBET-0828733 and NIH R24RR018934-01.

Model of Fission Yeast Cell Shape Driven by Membrane-Bound Growth Factors and the Cytoskeleton

PubMed Central

Drake, Tyler; Vavylonis, Dimitrios

2013-01-01

Fission yeast serves as a model for how cellular polarization machinery consisting of signaling molecules and the actin and microtubule cytoskeleton regulates cell shape. In this work, we develop mathematical models to investigate how these cells maintain a tubular shape of approximately constant diameter. Many studies identify active Cdc42, found in a cap at the inner membrane of growing cell tips, as an important regulator of local cell wall remodeling, likely through control of exocyst tethering and the targeting of other polarity-enhancing structures. First, we show that a computational model with Cdc42-dependent local cell wall remodeling under turgor pressure predicts a relationship between spatial extent of growth signal and cell diameter that is in agreement with prior experiments. Second, we model the consequences of feedback between cell shape and distribution of Cdc42 growth signal at cell tips. We show that stability of cell diameter over successive cell divisions places restrictions on their mutual dependence. We argue that simple models where the spatial extent of the tip growth signal relies solely on geometrical alignment of confined microtubules might lead to unstable width regulation. Third, we study a computational model that combines a growth signal distributed over a characteristic length scale (as, for example, by a reaction-diffusion mechanism) with an axis-sensing microtubules system that places landmarks at positions where microtubule tips touch the cortex. A two-dimensional implementation of this model leads to stable cell diameter for a wide range of parameters. Changes to the parameters of this model reproduce straight, bent, and bulged cell shapes, and we discuss how this model is consistent with other observed cell shapes in mutants. Our work provides an initial quantitative framework for understanding the regulation of cell shape in fission yeast, and a scaffold for understanding this process on a more molecular level in the future. PMID:24146607

Reciprocal trade of Carbon and Nitrogen at the root-fungus interface in ectomycorrhizal beech plants

NASA Astrophysics Data System (ADS)

Kaiser, Christina; Mayerhofer, Werner; Dietrich, Marlies; Gorka, Stefan; Schintlmeister, Arno; Reipert, Siegfried; Schweiger, Peter; Weidinger, Marieluise; Wiesenbauer, Julia; Martin, Victoria; Richter, Andreas; Woebken, Dagmar

2017-04-01

Plants deliver recently assimilated carbon (C) to mycorrhizal fungi, and receive nutrients, such as N and P, in exchange. A reciprocal exchange of C and nutrients between plants and mycorrhizal fungi (i.e., fungi which deliver more nutrients receive more plant C in return and vice versa) has been suggested for arbuscular mycorrhizal symbioses by some studies, but challenged by others. For ectomycorrhizal associations even less is known on how the exchange of C for nutrients is regulated, and whether it is based on reciprocity, or other controls. The aim of this study was to test the concept of reciprocal rewards between beech (Fagus sylvatica) and their associated ectomycorrhizal fungi on different scales, namely (a) across associations between individual root tips of beech and different fungal partners, and (b) at the subcellular scale at the plant-fungus interface. We exposed young beech trees associated with natural mycorrhizal fungal communities to a 13CO2 atmosphere and added 15N-labelled amino acids to a 'litter compartment', that mycorrhizal hyphae, but not plant roots could access. Plants were harvested within 2 days after application of 15N and less than one day after applying 13CO2. If the trading of C for N was reciprocal, we expect that 13C would be correlated to 15N across individual plant-fungal connections and at the subcellular scale within one mycorrhizal root tip, respectively. We collected individual mycorrhizal root-tips from 8 plants right after harvest, analyzed their 13C and 15N content by isotope-ratio mass spectrometry (EA-IRMS) and performed ITS sequencing to identify fungal communities associated with individual root tips. Selected mycorrhizal root tips were also prepared for nano-scale secondary ion mass spectrometry (NanoSIMS) to visualize the spatial distribution of 13C and 15N in cross-sections of mycorrhizal root-tips at the subcellular scale. Our results showed a significant, albeit weak correlation between 13C and 15N across collected mycorrhizal root-tips, the variability of which was seemingly influenced by fungal colonization pattern. Within a cross-section of an individual root-tip, however, NanoSIMS imaging revealed not only a high spatial heterogeneity of 13C and 15N across plant and fungal cells, but also a strong spatial correlation between 13C and 15N in both, plant cells and fungal cells of the Hartig Net, the fungal mantle and external hyphae. Intriguingly, individual 'hotspots' of external fungal hyphae that were highly enriched in 15N (delivering high amounts of the added 15N to the plant), were also always extraordinarily enriched in 13C (receiving more 13C in return). Our results provide first evidence for a reciprocal exchange of C for N between plants and ectomycorrhizal fungi at the subcellular scale. This indicates that a mechanism at the cellular level exists, that (i) either allows plants to direct their C flow into N-delivering parts of the mycorrhizal hyphal network or (ii) allow the fungus to 'draw' more C from the plant (develop a higher sink strength) when it has access to N. While such a mechanism still remains to be elucidated, our study shows, for the first time, direct evidence for its existence.

The PDZ protein tax-interacting protein-1 inhibits beta-catenin transcriptional activity and growth of colorectal cancer cells.

PubMed

Kanamori, Mutsumi; Sandy, Peter; Marzinotto, Stefania; Benetti, Roberta; Kai, Chikatoshi; Hayashizaki, Yoshihide; Schneider, Claudio; Suzuki, Harukazu

2003-10-03

Wnt signaling is essential during development while deregulation of this pathway frequently leads to the formation of various tumors including colorectal carcinomas. A key component of the pathway is beta-catenin that, in association with TCF-4, directly regulates the expression of Wnt-responsive genes. To identify novel binding partners of beta-catenin that may control its transcriptional activity, we performed a mammalian two-hybrid screen and isolated the Tax-interacting protein (TIP-1). The in vivo complex formation between beta-catenin and TIP-1 was verified by coimmunoprecipitation, and a direct physical association was revealed by glutathione S-transferase pull-down experiments in vitro. By using a panel of deletion mutants of both proteins, we demonstrate that the interaction is mediated by the PDZ (PSD-95/DLG/ZO-1 homology) domain of TIP-1 and requires primarily the last four amino acids of beta-catenin. TIP-1 overexpression resulted in a dose-dependent decrease in the transcriptional activity of beta-catenin when tested on the TOP/FOPFLASH reporter system. Conversely, siRNA-mediated knock-down of endogenous TIP-1 slightly increased endogenous beta-catenin transactivation function. Moreover, we show that overexpression of TIP-1 reduced the proliferation and anchorage-independent growth of colorectal cancer cells. These data suggest that TIP-1 may represent a novel regulatory element in the Wnt/beta-catenin signaling pathway.

Cu2O-tipped ZnO nanorods with enhanced photoelectrochemical performance for CO2 photoreduction

NASA Astrophysics Data System (ADS)

Iqbal, Muzaffar; Wang, Yanjie; Hu, Haifeng; He, Meng; Hassan Shah, Aamir; Lin, Lin; Li, Pan; Shao, Kunjuan; Reda Woldu, Abebe; He, Tao

2018-06-01

The design of Cu2O-tipped ZnO nanorods is proposed here aiming at enhanced photoelectrochemical properties. The tip-selective deposition of Cu2O is confirmed by scanning transmission electron microscopy (STEM). The photoinduced charge behavior like charge generation, separation and transport has been thoroughly studied by UV-vis absorption analysis and different photoelectrochemical characterizations, including transient photocurrent, incident photon-to-current efficiency (IPCE), electrochemical impedance spectroscopy (EIS), intensity-modulated photocurrent spectroscopy (IMPS), and Mott-Schottky measurements. The photoelectrochemical characterizations clearly indicate that ZnO/Cu2O structures exhibit much higher performance than pristine ZnO, due to the formation of p-n junction, as well as the tip selective growth of Cu2O on ZnO. Photocatalytic CO2 reduction in aqueous solution under UV-visible light illumination shows that CO is the main product, and with the increase of the Cu2O content in the heterostructure, the CO yield increases. This work shows that Cu2O-tipped ZnO nanorods possess improved behavior of charge generation, separation and transport, which may work as a potential candidate for photocatalytic CO2 reduction.

In vivo Monitoring of Serotonin by Nanomaterial Functionalized Acupuncture Needle

NASA Astrophysics Data System (ADS)

Li, Yu-Tao; Tang, Li-Na; Ning, Yong; Shu, Qing; Liang, Feng-Xia; Wang, Hua; Zhang, Guo-Jun

2016-06-01

Acupuncture treatment is amazing but controversial. Up to now, the mechanism of treating diseases by acupuncture and moxibustion is still unclear, especially the occurrence of the molecular events in local acupoints. Herein, we report an extremely stable microsensor by modifying carbon nanotube (CNT) to the tip surface of acupuncture needle and applying this CNT-modified acupuncture needle for real time monitoring of serotonin (5-HT) in vivo. To stabilize CNT modification on the needle tip surface, poly(3,4-ethylenedioxythiophene)(PEDOT) was employed as glue water to stick CNT on the needle. The detection limit of the CNT-modified needle was found to be approximately 50 nM and 78 nM in the PBS and the cell medium, respectively. In addition, the needle showed good selectivity to some inflammatory mediators and some electroactive molecules. For the first time, the CNT-modified needle could be directly probed into rat body for real time monitoring of 5-HT in vivo, showing a great potential for better understanding the mechanism of acupuncture treatment.

Cell wall evolution and diversity

PubMed Central

Fangel, Jonatan U.; Ulvskov, Peter; Knox, J. P.; Mikkelsen, Maria D.; Harholt, Jesper; Popper, Zoë A.; Willats, William G.T.

2012-01-01

Plant cell walls display a considerable degree of diversity in their compositions and molecular architectures. In some cases the functional significance of a particular cell wall type appears to be easy to discern: secondary cells walls are often reinforced with lignin that provides durability; the thin cell walls of pollen tubes have particular compositions that enable their tip growth; lupin seed cell walls are characteristically thickened with galactan used as a storage polysaccharide. However, more frequently the evolutionary mechanisms and selection pressures that underpin cell wall diversity and evolution are unclear. For diverse green plants (chlorophytes and streptophytes) the rapidly increasing availability of transcriptome and genome data sets, the development of methods for cell wall analyses which require less material for analysis, and expansion of molecular probe sets, are providing new insights into the diversity and occurrence of cell wall polysaccharides and associated biosynthetic genes. Such research is important for refining our understanding of some of the fundamental processes that enabled plants to colonize land and to subsequently radiate so comprehensively. The study of cell wall structural diversity is also an important aspect of the industrial utilization of global polysaccharide bio-resources. PMID:22783271

Gravisensing in single-celled systems - update on characean rhizoids and protonemata

NASA Astrophysics Data System (ADS)

Braun, M.; Limbach, C.

Single-celled and tip-growing rhizoids and protonemata of the characean algae have been intensively studied and there is considerable progress in the understanding of the molecular and cellular mechanisms underlying gravisensing and gravity-dependent growth. In higher plant statocytes, the role of actin in both processes is still a matter of intense debate, but there is clear evidence that actin coordinates both processes in characean rhizoids and protonemata. The multiple functions and dynamic nature of the actin cytoskeleton in these cells are based on the concerted action of a variety of actin-binding proteins. Profilin, actin-depolymerizing factor, a spectrin-like protein, villin and fimbrin have been detected which control apical actin polymerization and regulate the dynamic remodeling of the actin arrangement. An actomyosin-based system was shown to (i) mediate the transport of secretory vesicles to the growing tip, (ii) establish the incorporation of cell wall material and (iii) coordinate the tip-focussed distribution of calcium channels which establish the tip-high calcium gradient for local exocytosis. Experiments performed in microgravity have shown that the actomyosin system precisely coordinates the position of statoliths in rhizoids and protonemata and, upon a change in orientation, directs sedimenting statoliths to specific areas at the plasma membrane where physical contact with gravisensor molecules initiates growth reorientation. The upward growth response of protonemata was shown to be preceded by a statolith-induced and actin-dependent relocalization of the Ca2+-gradient to the upper flank that does not occur in positively gravitropic rhizoids, in which sedimented statoliths cause differential growth of the opposite subapical cell flank. Thus, constant actin polymerization in the growing tip and the spatiotemporal control of actin remodeling by numerous actin-binding proteins are essential for gravity sensing and polarized growth of characean rhizoids and protonemata.

Mercury-sensitive water channels as possible sensors of water potentials in pollen

PubMed Central

Hill, Adrian E.

2013-01-01

The growing pollen tube is central to plant reproduction and is a long-standing model for cellular tip growth in biology. Rapid osmotically driven growth is maintained under variable conditions, which requires osmosensing and regulation. This study explores the mechanism of water entry and the potential role of osmosensory regulation in maintaining pollen growth. The osmotic permeability of the plasmalemma of Lilium pollen tubes was measured from plasmolysis rates to be 1.32±0.31×10–3 cm s–1. Mercuric ions reduce this permeability by 65%. Simulations using an osmotic model of pollen tube growth predict that an osmosensor at the cell membrane controls pectin deposition at the cell tip; inhibiting the sensor is predicted to cause tip bursting due to cell wall thinning. It was found that adding mercury to growing pollen tubes caused such a bursting of the tips. The model indicates that lowering the osmotic permeability per se does not lead to bursting but rather to thickening of the tip. The time course of induced bursting showed no time lag and was independent of mercury concentration, compatible with a surface site of action. The submaximal bursting response to intermediate mercuric ion concentration was independent of the concentration of calcium ions, showing that bursting is not due to a competitive inhibition of calcium binding or entry. Bursting with the same time course was also shown by cells growing on potassium-free media, indicating that potassium channels (implicated in mechanosensing) are not involved in the bursting response. The possible involvement of mercury-sensitive water channels as osmosensors and current knowledge of these in pollen cells are discussed. PMID:24098048

Subpolar addition of new cell wall is directed by DivIVA in mycobacteria

PubMed Central

Meniche, Xavier; Otten, Renee; Siegrist, M. Sloan; Baer, Christina E.; Murphy, Kenan C.; Bertozzi, Carolyn R.; Sassetti, Christopher M.

2014-01-01

Mycobacteria are surrounded by a complex multilayered envelope and elongate at the poles. The principles that organize the coordinated addition of chemically diverse cell wall layers during polar extension remain unclear. We show that enzymes mediating the terminal cytosolic steps of peptidoglycan, arabinogalactan, and mycolic acid synthesis colocalize at sites of cell growth or division. The tropomyosin-like protein, DivIVA, is targeted to the negative curvature of the pole, is enriched at the growing end, and determines cell shape from this site. In contrast, cell wall synthetic complexes are concentrated at a distinct subpolar location. When viewed at subdiffraction resolution, new peptidoglycan is deposited at this subpolar site, and inert cell wall covers the DivIVA-marked tip. The differentiation between polar tip and cell wall synthetic complexes is also apparent at the biochemical level. Enzymes that generate mycolate precursors interact with DivIVA, but the final condensation of mycolic acids occurs in a distinct protein complex at the site of nascent cell wall addition. We propose an ultrastructural model of mycobacterial polar growth where new cell wall is added in an annular zone below the cell tip. This model may be broadly applicable to other bacterial and fungal organisms that grow via polar extension. PMID:25049412

Chemotaxis without Conventional Two-Component System, Based on Cell Polarity and Aerobic Conditions in Helicity-Switching Swimming of Spiroplasma eriocheiris

PubMed Central

Liu, Peng; Zheng, Huajun; Meng, Qingguo; Terahara, Natsuho; Gu, Wei; Wang, Shengyue; Zhao, Guoping; Nakane, Daisuke; Wang, Wen; Miyata, Makoto

2017-01-01

Spiroplasma eriocheiris is a pathogen that causes mass mortality in Chinese mitten crab, Eriocheir sinensis. S. eriocheiris causes tremor disease and infects almost all of the artificial breeding crustaceans, resulting in disastrous effects on the aquaculture economy in China. S. eriocheiris is a wall-less helical bacterium, measuring 2.0 to 10.0 μm long, and can swim up to 5 μm per second in a viscous medium without flagella by switching the cell helicity at a kink traveling from the front to the tail. In this study, we showed that S. eriocheiris performs chemotaxis without the conventional two-component system, a system commonly found in bacterial chemotaxis. The chemotaxis of S. eriocheiris was observed more clearly when the cells were cultivated under anaerobic conditions. The cells were polarized as evidenced by a tip structure, swimming in the direction of the tip, and were shown to reverse their swimming direction in response to attractants. Triton X-100 treatment revealed the internal structure, a dumbbell-shaped core in the tip that is connected by a flat ribbon, which traces the shortest line in the helical cell shape from the tip to the other pole. Sixteen proteins were identified as the components of the internal structure by mass spectrometry, including Fibril protein and four types of MreB proteins. PMID:28217108

Novel histone deacetylase inhibitor N25 exerts anti-tumor effects and induces autophagy in human glioma cells by inhibiting HDAC3

PubMed Central

Sun, Xin-Yuan; Qu, Yue; Ni, An-Ran; Wang, Gui-Xiang; Huang, Wei-Bin; Chen, Zhong-Ping; Lv, Zhu-Fen; Zhang, Song; Lindsay, Holly; Zhao, Sibo; Li, Xiao-Nan; Feng, Bing-Hong

2017-01-01

N25, a novel histone deacetylase inhibitor, was created through structural modification of suberoylanilide hydroxamic acid. To evaluate the anti-tumor activity of N25 and clarify its molecular mechanism of inducing autophagy in glioma cells, we investigated its in vitro anti-proliferative effect and in vivo anticancer effect. Moreover, we detected whether N25 induces autophagy in glioma cells by transmission electron microscope and analyzed the protein expression level of HDAC3, Tip60, LC3 in glioma samples by western blot. We additionally analyzed the protein expression level of HDAC3, Tip60, ULK1 (Atg1), and Beclin-1 (Atg6) after treatment with N25 in glioma cells. Our results showed that the anti-tumor activity of N25 in glioma cells is slightly stronger than SAHA both in vitro and in vivo. We found that N25 induced autophagy, and HDAC3 was significantly elevated and Tip60 and LC3 significantly decreased in glioma samples compared with normal brain tissues. Nevertheless, N25 inhibited HDAC3 and up-regulated the protein expression of Tip60, ULK1 (Atg1), and Beclin-1 (Atg6) after treatment of glioma cells with N25. In conclusion, these data suggest that N25 has striking anti-tumor activity in part due to inhibition of HDAC3. Additionally, N25 may induce autophagy through inhibiting HDAC3. PMID:29088860

Nitric Oxide Inhibits Al-Induced Programmed Cell Death in Root Tips of Peanut (Arachis hypogaea L.) by Affecting Physiological Properties of Antioxidants Systems and Cell Wall

PubMed Central

Pan, Chun-Liu; Yao, Shao-Chang; Xiong, Wei-Jiao; Luo, Shu-Zhen; Wang, Ya-Lun; Wang, Ai-Qin; Xiao, Dong; Zhan, Jie; He, Long-Fei

2017-01-01

It has been reported that nitric oxide (NO) is a negative regulator of aluminum (Al)-induced programmed cell death (PCD) in peanut root tips. However, the inhibiting mechanism of NO on Al-induced PCD is unclear. In order to investigate the mechanism by which NO inhibits Al-induced PCD, the effects of co-treatment Al with the exogenous NO donor or the NO-specific scavenger on peanut root tips, the physiological properties of antioxidants systems and cell wall (CW) in root tip cells of NO inhibiting Al-induced PCD were studied with two peanut cultivars. The results showed that Al exposure induced endogenous NO accumulation, and endogenous NO burst increased antioxidant enzyme activity in response to Al stress. The addition of NO donor sodium nitroprusside (SNP) relieved Al-induced root elongation inhibition, cell death and Al adsorption in CW, as well as oxidative damage and ROS accumulation. Furthermore, co-treatment with the exogenous NO donor decreased MDA content, LOX activity and pectin methylesterase (PME) activity, increased xyloglucan endotransglucosylase (XET) activity and relative expression of the xyloglucan endotransglucosylase/hydrolase (XTH-32) gene. Taken together, exogenous NO alleviated Al-induced PCD by inhibiting Al adsorption in CW, enhancing antioxidant defense and reducing peroxidation of membrane lipids, alleviating the inhibition of Al on root elongation by maintaining the extensibility of CW, decreasing PME activity, and increasing XET activity and relative XTH-32 expression of CW. PMID:29311970

Nitric Oxide Inhibits Al-Induced Programmed Cell Death in Root Tips of Peanut (Arachis hypogaea L.) by Affecting Physiological Properties of Antioxidants Systems and Cell Wall.

PubMed

Pan, Chun-Liu; Yao, Shao-Chang; Xiong, Wei-Jiao; Luo, Shu-Zhen; Wang, Ya-Lun; Wang, Ai-Qin; Xiao, Dong; Zhan, Jie; He, Long-Fei

2017-01-01

It has been reported that nitric oxide (NO) is a negative regulator of aluminum (Al)-induced programmed cell death (PCD) in peanut root tips. However, the inhibiting mechanism of NO on Al-induced PCD is unclear. In order to investigate the mechanism by which NO inhibits Al-induced PCD, the effects of co-treatment Al with the exogenous NO donor or the NO-specific scavenger on peanut root tips, the physiological properties of antioxidants systems and cell wall (CW) in root tip cells of NO inhibiting Al-induced PCD were studied with two peanut cultivars. The results showed that Al exposure induced endogenous NO accumulation, and endogenous NO burst increased antioxidant enzyme activity in response to Al stress. The addition of NO donor sodium nitroprusside (SNP) relieved Al-induced root elongation inhibition, cell death and Al adsorption in CW, as well as oxidative damage and ROS accumulation. Furthermore, co-treatment with the exogenous NO donor decreased MDA content, LOX activity and pectin methylesterase (PME) activity, increased xyloglucan endotransglucosylase (XET) activity and relative expression of the xyloglucan endotransglucosylase/hydrolase ( XTH-32 ) gene. Taken together, exogenous NO alleviated Al-induced PCD by inhibiting Al adsorption in CW, enhancing antioxidant defense and reducing peroxidation of membrane lipids, alleviating the inhibition of Al on root elongation by maintaining the extensibility of CW, decreasing PME activity, and increasing XET activity and relative XTH-32 expression of CW.

β-(1,3)-Glucan Unmasking in Some Candida albicans Mutants Correlates with Increases in Cell Wall Surface Roughness and Decreases in Cell Wall Elasticity

DOE PAGES

Hasim, Sahar; Allison, David P.; Retterer, Scott T.; ...

2016-11-14

Candida albicans is among the most common human fungal pathogens, causing a broad range of infections, including life-threatening systemic infections. The cell wall of C. albicans is the interface between the fungus and the innate immune system. The cell wall is composed of an outer layer enriched in mannosylated glycoproteins (mannan) and an inner layer enriched in β-(1,3)-glucan and chitin. Detection of C. albicans by Dectin-1, a C-type signaling lectin specific for β-(1,3)-glucan, is important for the innate immune system to recognize systemic fungal infections. Increased exposure of β-(1,3)-glucan to the immune system occurs when the mannan layer is alteredmore » or removed in a process called unmasking. Nanoscale changes to the cell wall during unmasking were explored in this paper in live cells with atomic force microscopy (AFM). Two mutants, the cho1Δ/Δ and kre5Δ/Δ mutants, were selected as representatives that exhibit modest and strong unmasking, respectively. Comparisons of the cho1Δ/Δ and kre5Δ/Δ mutants to the wild type reveal morphological changes in their cell walls that correlate with decreases in cell wall elasticity. In addition, AFM tips functionalized with Dectin-1 revealed that the forces of binding of Dectin-1 to all of the strains were similar, but the frequency of binding was highest for the kre5Δ/Δ mutant, decreased for the cho1Δ/Δ mutant, and rare for the wild type. These data show that nanoscale changes in surface topology are correlated with increased Dectin-1 adhesion and decreased cell wall elasticity. Finally, AFM, using tips functionalized with immunologically relevant molecules, can map epitopes of the cell wall and increase our understanding of pathogen recognition by the immune system.« less

β-(1,3)-Glucan Unmasking in Some Candida albicans Mutants Correlates with Increases in Cell Wall Surface Roughness and Decreases in Cell Wall Elasticity

PubMed Central

Hasim, Sahar; Allison, David P.; Retterer, Scott T.; Hopke, Alex; Wheeler, Robert T.; Doktycz, Mitchel J.

2016-01-01

ABSTRACT Candida albicans is among the most common human fungal pathogens, causing a broad range of infections, including life-threatening systemic infections. The cell wall of C. albicans is the interface between the fungus and the innate immune system. The cell wall is composed of an outer layer enriched in mannosylated glycoproteins (mannan) and an inner layer enriched in β-(1,3)-glucan and chitin. Detection of C. albicans by Dectin-1, a C-type signaling lectin specific for β-(1,3)-glucan, is important for the innate immune system to recognize systemic fungal infections. Increased exposure of β-(1,3)-glucan to the immune system occurs when the mannan layer is altered or removed in a process called unmasking. Nanoscale changes to the cell wall during unmasking were explored in live cells with atomic force microscopy (AFM). Two mutants, the cho1Δ/Δ and kre5Δ/Δ mutants, were selected as representatives that exhibit modest and strong unmasking, respectively. Comparisons of the cho1Δ/Δ and kre5Δ/Δ mutants to the wild type reveal morphological changes in their cell walls that correlate with decreases in cell wall elasticity. In addition, AFM tips functionalized with Dectin-1 revealed that the forces of binding of Dectin-1 to all of the strains were similar, but the frequency of binding was highest for the kre5Δ/Δ mutant, decreased for the cho1Δ/Δ mutant, and rare for the wild type. These data show that nanoscale changes in surface topology are correlated with increased Dectin-1 adhesion and decreased cell wall elasticity. AFM, using tips functionalized with immunologically relevant molecules, can map epitopes of the cell wall and increase our understanding of pathogen recognition by the immune system. PMID:27849179

β-(1,3)-Glucan Unmasking in Some Candida albicans Mutants Correlates with Increases in Cell Wall Surface Roughness and Decreases in Cell Wall Elasticity

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

Hasim, Sahar; Allison, David P.; Retterer, Scott T.

Candida albicans is among the most common human fungal pathogens, causing a broad range of infections, including life-threatening systemic infections. The cell wall of C. albicans is the interface between the fungus and the innate immune system. The cell wall is composed of an outer layer enriched in mannosylated glycoproteins (mannan) and an inner layer enriched in β-(1,3)-glucan and chitin. Detection of C. albicans by Dectin-1, a C-type signaling lectin specific for β-(1,3)-glucan, is important for the innate immune system to recognize systemic fungal infections. Increased exposure of β-(1,3)-glucan to the immune system occurs when the mannan layer is alteredmore » or removed in a process called unmasking. Nanoscale changes to the cell wall during unmasking were explored in this paper in live cells with atomic force microscopy (AFM). Two mutants, the cho1Δ/Δ and kre5Δ/Δ mutants, were selected as representatives that exhibit modest and strong unmasking, respectively. Comparisons of the cho1Δ/Δ and kre5Δ/Δ mutants to the wild type reveal morphological changes in their cell walls that correlate with decreases in cell wall elasticity. In addition, AFM tips functionalized with Dectin-1 revealed that the forces of binding of Dectin-1 to all of the strains were similar, but the frequency of binding was highest for the kre5Δ/Δ mutant, decreased for the cho1Δ/Δ mutant, and rare for the wild type. These data show that nanoscale changes in surface topology are correlated with increased Dectin-1 adhesion and decreased cell wall elasticity. Finally, AFM, using tips functionalized with immunologically relevant molecules, can map epitopes of the cell wall and increase our understanding of pathogen recognition by the immune system.« less

A viscoplastic study of crack-tip deformation and crack growth in a nickel-based superalloy at elevated temperature

NASA Astrophysics Data System (ADS)

Zhao, L. G.; Tong, J.

Viscoplastic crack-tip deformation behaviour in a nickel-based superalloy at elevated temperature has been studied for both stationary and growing cracks in a compact tension (CT) specimen using the finite element method. The material behaviour was described by a unified viscoplastic constitutive model with non-linear kinematic and isotropic hardening rules, and implemented in the finite element software ABAQUS via a user-defined material subroutine (UMAT). Finite element analyses for stationary cracks showed distinctive strain ratchetting behaviour near the crack tip at selected load ratios, leading to progressive accumulation of tensile strain normal to the crack-growth plane. Results also showed that low frequencies and superimposed hold periods at peak loads significantly enhanced strain accumulation at crack tip. Finite element simulation of crack growth was carried out under a constant Δ K-controlled loading condition, again ratchetting was observed ahead of the crack tip, similar to that for stationary cracks. A crack-growth criterion based on strain accumulation is proposed where a crack is assumed to grow when the accumulated strain ahead of the crack tip reaches a critical value over a characteristic distance. The criterion has been utilized in the prediction of crack-growth rates in a CT specimen at selected loading ranges, frequencies and dwell periods, and the predictions were compared with the experimental results.

Solvent selection for explaining the morphology of nitroguanidine crystal by molecular dynamics simulation

NASA Astrophysics Data System (ADS)

Song, Liang; Chen, Lizhen; Cao, Duanlin; Wang, Jianlong

2018-02-01

In this article, a method was performed to predict the morphology of needle-shaped crystals by analyzing the growth mechanisms for the various crystal faces. As an example, the crystal morphology of a nitroguanidine (NQ) was investigated via molecular dynamics simulations. The modified attachment energy (MEA) model was constructed by introducing surface chemistry terms and the relevant morphology of the habit crystal faces. The results indicate that the growth morphology of NQ in vacuum is dominated by {2 2 0}, {0 4 0}, {1 1 1}, {1 3 1} and {3 1 1} faces. The {2 2 0} and {0 4 0} faces are parallel to the elongation direction of the crystal, while the other faces are at the needle tips direction. The atoms or atomic groups exposed in crystal surface were used to analyze the relationship between structure and morphology. Compared to the surrounding faces, the needle tip faces have a large number of polar atoms or atomic groups. The needle tip faces have a high electronegativity on N, O atoms via molecular electrostatic potential (ESP) analysis. Furthermore, the protic solvent was used to reduce the attachment energy of the tip surfaces for achieving the purpose of inhibiting the growth of needle tips. Gamma-butyrolactone as the selected solvent inhibited effectively the growth of the needle tip faces. The predicted result is serviceable for the formation design.

Sequoia establishes tip-cell number in Drosophila trachea by regulating FGF levels.

PubMed

Araújo, Sofia J; Casanova, Jordi

2011-07-15

Competition and determination of leading and trailing cells during collective cell migration is a widespread phenomenon in development, wound healing and tumour invasion. Here, we analyse this issue during in vivo ganglionic branch cell migration in the Drosophila tracheal system. We identify Sequoia (Seq) as a negative transcriptional regulator of Branchless (Bnl), a Drosophila FGF homologue, and observe that modulation of Bnl levels determines how many cells will lead this migrating cluster, regardless of Notch lateral inhibition. Our results show that becoming a tip cell does not prevent others in the branch taking the same position, suggesting that leader choice does not depend only on sensing relative amounts of FGF receptor activity.

Tip-alpha (hp0596 gene product) is a highly immunogenic Helicobacter pylori protein involved in colonization of mouse gastric mucosa.

PubMed

Godlewska, Renata; Pawlowski, Marcin; Dzwonek, Artur; Mikula, Michal; Ostrowski, Jerzy; Drela, Nadzieja; Jagusztyn-Krynicka, Elzbieta K

2008-03-01

A product of the Helicobacter pylori hp0596 gene (Tip-alpha) is a highly immunogenic homodimeric protein, unique for this bacterium. Cell fractionation experiments indicate that Tip-alpha is anchored to the inner membrane. In contrast, the three-dimensional model of the protein suggests that Tip-alpha is soluble or, at least, largely exposed to the solvent. hp0596 gene knockout resulted in a significant decrease in the level of H. pylori colonization as measured by real-time PCR assay. In addition, the Tip-alpha recombinant protein was determined to stimulate macrophage to produce IL-1alpha and TNF-alpha. Both results imply that Tip-alpha is rather loosely connected to the inner membrane and potentially released during infection.

Regulation of Cell Diameter, For3p Localization, and Cell Symmetry by Fission Yeast Rho-GAP Rga4p

PubMed Central

Das, Maitreyi; Wiley, David J.; Medina, Saskia; Vincent, Helen A.; Larrea, Michelle; Oriolo, Andrea

2007-01-01

Control of cellular dimensions and cell symmetry are critical for development and differentiation. Here we provide evidence that the putative Rho-GAP Rga4p of Schizosaccharomyces pombe controls cellular dimensions. rga4Δ cells are wider in diameter and shorter in length, whereas Rga4p overexpression leads to reduced diameter of the growing cell tip. Consistent with a negative role in cell growth control, Rga4p protein localizes to the cell sides in a “corset” pattern, and to the nongrowing cell tips. Additionally, rga4Δ cells show an altered growth pattern similar to that observed in mutants of the formin homology protein For3p. Consistent with these observations, Rga4p is required for normal localization of For3p and for normal distribution of the actin cytoskeleton. We show that different domains of the Rga4p protein mediate diverse morphological functions. The C-terminal GAP domain mediates For3p localization to the cell tips and maintains cell diameter. Conversely, overexpression of the N-terminal LIM homology domain of Rga4p promotes actin cable formation in a For3p-dependent manner. Our studies indicate that Rga4p functionally interacts with For3p and has a novel function in the control of cell diameter and cell growth. PMID:17377067

Energy reduction for the spot welding process in the automotive industry

NASA Astrophysics Data System (ADS)

Cullen, J. D.; Athi, N.; Al-Jader, M. A.; Shaw, A.; Al-Shamma'a, A. I.

2007-07-01

When performing spot welding on galvanised metals, higher welding force and current are required than on uncoated steels. This has implications for the energy usage when creating each spot weld, of which there are approximately 4300 in each passenger car. The paper presented is an overview of electrode current selection and its variance over the lifetime of the electrode tip. This also describes the proposed analysis system for the selection of welding parameters for the spot welding process, as the electrode tip wears.

Salicylic Acid Regulates Pollen Tip Growth through an NPR3/NPR4-Independent Pathway.

PubMed

Rong, Duoyan; Luo, Nan; Mollet, Jean Claude; Liu, Xuanming; Yang, Zhenbiao

2016-11-07

Tip growth is a common strategy for the rapid elongation of cells to forage the environment and/or to target to long-distance destinations. In the model tip growth system of Arabidopsis pollen tubes, several small-molecule hormones regulate their elongation, but how these rapidly diffusing molecules control extremely localized growth remains mysterious. Here we show that the interconvertible salicylic acid (SA) and methylated SA (MeSA), well characterized for their roles in plant defense, oppositely regulate Arabidopsis pollen tip growth with SA being inhibitory and MeSA stimulatory. The effect of SA and MeSA was independent of known NPR3/NPR4 SA receptor-mediated signaling pathways. SA inhibited clathrin-mediated endocytosis in pollen tubes associated with an increased accumulation of less stretchable demethylated pectin in the apical wall, whereas MeSA did the opposite. Furthermore, SA and MeSA alter the apical activation of ROP1 GTPase, a key regulator of tip growth in pollen tubes, in an opposite manner. Interestingly, both MeSA methylesterase and SA methyltransferase, which catalyze the interconversion between SA and MeSA, are localized at the apical region of pollen tubes, indicating of the tip-localized production of SA and MeSA and consistent with their effects on the apical cellular activities. These findings suggest that local generation of a highly diffusible signal can regulate polarized cell growth, providing a novel mechanism of cell polarity control apart from the one involving protein and mRNA polarization. Copyright © 2016 The Author. Published by Elsevier Inc. All rights reserved.

Gravisensing in single-celled systems

NASA Astrophysics Data System (ADS)

Braun, M.; Limbach, C.

Single-celled systems are favourable cell types for studying several aspects of gravisensing and gravitropic responses. Whether and how actin is involved in both processes in higher plant statocytes is still a matter of intensive debate. In single-celled and tip-growing characean rhizoids and protonemata, however, there is clear evidence that actin is a central keyplayer controlling polarized growth and the mechanisms of gravity sensing and growth reorientation. Both cell types exhibit a unique actin polymerization in the extending tip, strictly colocalized with the prominent ER-aggregate in the center of the Spitzenkoerper. The local accumulation of ADF and profilin in this central array suggest that actin polymerization is controlled by these actin-binding proteins, which can be regulated by calcium, pH and a variety of other parameters. Distinct actin filaments extend even into the outermost tip and form a dense meshwork in the apical and subapical region, before they become bundled by villin to form two populations of thick actin cables that generate rotational cytoplasmic streaming in the basal region. Actomyosin not only mediates the delivery of secretory vesicles to the growing tip and controls the incorporation pattern of cell wall material, but also coordinates the tip-focused distribution pattern of calcium channels in the apical membrane. They establish the tip-high calcium gradient, a prerequisite for exocytosis. Microgravity experiments have added much to our understanding that both cell types use an efficient actomyosin-based system to control and correct the position of their statoliths and to direct sedimenting statoliths to confined graviperception sites at the plasma membrane. Actin's involvement in the graviresponses is more indirect. The upward growth of negatively gravitropic protonemata was shown to be preceded by a statolith-induced relocalization the Ca2+-calcium gradient to the upper flank that does not occur in positively gravitropic (downward growing) rhizoids, in which statoliths sedimentation is followed by differential flank growth. Based on these results, it is evident that polymerization, dynamic reorganization and the diverse functions of actin spatiotemporally controlled by numerous actin-binding proteins are fundamental for the processes of gravity sensing and gravity-oriented polarized growth. Financial support by Deutsches Zentrum für Luft- und Raumfahrt (DLR) on behalf of the Bundesministerium für Bildung und Forschung (50WB9998).

A high-yield procedure for isolation of metaphase chromosomes from root tips of Vicia faba L.

PubMed

Doležel, J; Cíhalíková, J; Lucretti, S

1992-08-01

A new method is described for the isolation of large quantities of Vicia faba metaphase chromosomes. Roots were treated with 2.5 mM hydroxyurea for 18 h to accumulate meristem tip cells at the G1/S interface. After release from the block, the cells re-entered the cell cycle with a high degree of synchrony. A treatment with 2.5 μM amiprophos-methyl (APM) was used to accumulate mitotic cells in metaphase. The highest metaphase index (53.9%) was achieved when, 6 h after the release from the hydroxyurea block, the roots were exposed to APM for 4 h. The chromosomes were released from formaldehyde-fixed root tips by chopping with a scalpel in LB01 lysis buffer. Both the quality and the quantity of isolated chromosomes, examined microscopically and by flow cytometry, depended on the extent of the fixation. The best results were achieved after fixation with 6% formaldehyde for 30 min. Under these conditions, 1 · 10(6) chromosomes were routinely obtained from 30 root tips. The chromosomes were morphologically intact and suitable both for high-resolution chromosome studies and for flow-cytometric analysis and sorting. After the addition of hexylene glycol, the chromosome suspensions could be stored at 4° C for six months without any signs of deterioration.

Catch and Patch: A Pipette-Based Approach for Automating Patch Clamp That Enables Cell Selection and Fast Compound Application.

PubMed

Danker, Timm; Braun, Franziska; Silbernagl, Nikole; Guenther, Elke

2016-03-01

Manual patch clamp, the gold standard of electrophysiology, represents a powerful and versatile toolbox to stimulate, modulate, and record ion channel activity from membrane fragments and whole cells. The electrophysiological readout can be combined with fluorescent or optogenetic methods and allows for ultrafast solution exchanges using specialized microfluidic tools. A hallmark of manual patch clamp is the intentional selection of individual cells for recording, often an essential prerequisite to generate meaningful data. So far, available automation solutions rely on random cell usage in the closed environment of a chip and thus sacrifice much of this versatility by design. To parallelize and automate the traditional patch clamp technique while perpetuating the full versatility of the method, we developed an approach to automation, which is based on active cell handling and targeted electrode placement rather than on random processes. This is achieved through an automated pipette positioning system, which guides the tips of recording pipettes with micrometer precision to a microfluidic cell handling device. Using a patch pipette array mounted on a conventional micromanipulator, our automated patch clamp process mimics the original manual patch clamp as closely as possible, yet achieving a configuration where recordings are obtained from many patch electrodes in parallel. In addition, our implementation is extensible by design to allow the easy integration of specialized equipment such as ultrafast compound application tools. The resulting system offers fully automated patch clamp on purposely selected cells and combines high-quality gigaseal recordings with solution switching in the millisecond timescale.

Simulation of CNT-AFM tip based on finite element analysis for targeted probe of the biological cell

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

Yousefi, Amin Termeh, E-mail: at.tyousefi@gmail.com; Miyake, Mikio, E-mail: miyakejaist@gmail.com; Ikeda, Shoichiro, E-mail: sho16.ikeda@gmail.com

Carbon nanotubes (CNTs) are potentially ideal tips for atomic force microscopy (AFM) due to the robust mechanical properties, nano scale diameter and also their ability to be functionalized by chemical and biological components at the tip ends. This contribution develops the idea of using CNTs as an AFM tip in computational analysis of the biological cell’s. Finite element analysis employed for each section and displacement of the nodes located in the contact area was monitored by using an output database (ODB). This reliable integration of CNT-AFM tip process provides a new class of high performance nanoprobes for single biological cellmore » analysis.« less

Analytical correlation of centrifugal compressor design geometry for maximum efficiency with specific speed

NASA Technical Reports Server (NTRS)

Galvas, M. R.

1972-01-01

Centrifugal compressor performance was examined analytically to determine optimum geometry for various applications as characterized by specific speed. Seven specific losses were calculated for various combinations of inlet tip-exit diameter ratio, inlet hub-tip diameter ratio, blade exit backsweep, and inlet-tip absolute tangential velocity for solid body prewhirl. The losses considered were inlet guide vane loss, blade loading loss, skin friction loss, recirculation loss, disk friction loss, vaneless diffuser loss, and vaned diffuser loss. Maximum total efficiencies ranged from 0.497 to 0.868 for a specific speed range of 0.257 to 1.346. Curves of rotor exit absolute flow angle, inlet tip-exit diameter ratio, inlet hub-tip diameter ratio, head coefficient and blade exit backsweep are presented over a range of specific speeds for various inducer tip speeds to permit rapid selection of optimum compressor size and shape for a variety of applications.

A small protein inhibits proliferating cell nuclear antigen by breaking the DNA clamp

DOE PAGES

Altieri, Amanda S.; Ladner, Jane E.; Li, Zhuo; ...

2016-05-03

Here, proliferating cell nuclear antigen (PCNA) forms a trimeric ring that encircles duplex DNA and acts as an anchor for a number of proteins involved in DNA metabolic processes. PCNA has two structurally similar domains (I and II) linked by a long loop (inter-domain connector loop, IDCL) on the outside of each monomer of the trimeric structure that makes up the DNA clamp. All proteins that bind to PCNA do so via a PCNA-interacting peptide (PIP) motif that binds near the IDCL. A small protein, called TIP, binds to PCNA and inhibits PCNA-dependent activities although it does not contain amore » canonical PIP motif. The X-ray crystal structure of TIP bound to PCNA reveals that TIP binds to the canonical PIP interaction site, but also extends beyond it through a helix that relocates the IDCL. TIP alters the relationship between domains I and II within the PCNA monomer such that the trimeric ring structure is broken, while the individual domains largely retain their native structure. Small angle X-ray scattering (SAXS) confirms the disruption of the PCNA trimer upon addition of the TIP protein in solution and together with the X-ray crystal data, provides a structural basis for the mechanism of PCNA inhibition by TIP.« less

TACC3 is a microtubule plus end–tracking protein that promotes axon elongation and also regulates microtubule plus end dynamics in multiple embryonic cell types

PubMed Central

Nwagbara, Belinda U.; Faris, Anna E.; Bearce, Elizabeth A.; Erdogan, Burcu; Ebbert, Patrick T.; Evans, Matthew F.; Rutherford, Erin L.; Enzenbacher, Tiffany B.; Lowery, Laura Anne

2014-01-01

Microtubule plus end dynamics are regulated by a conserved family of proteins called plus end–tracking proteins (+TIPs). It is unclear how various +TIPs interact with each other and with plus ends to control microtubule behavior. The centrosome-associated protein TACC3, a member of the transforming acidic coiled-coil (TACC) domain family, has been implicated in regulating several aspects of microtubule dynamics. However, TACC3 has not been shown to function as a +TIP in vertebrates. Here we show that TACC3 promotes axon outgrowth and regulates microtubule dynamics by increasing microtubule plus end velocities in vivo. We also demonstrate that TACC3 acts as a +TIP in multiple embryonic cell types and that this requires the conserved C-terminal TACC domain. Using high-resolution live-imaging data on tagged +TIPs, we show that TACC3 localizes to the extreme microtubule plus end, where it lies distal to the microtubule polymerization marker EB1 and directly overlaps with the microtubule polymerase XMAP215. TACC3 also plays a role in regulating XMAP215 stability and localizing XMAP215 to microtubule plus ends. Taken together, our results implicate TACC3 as a +TIP that functions with XMAP215 to regulate microtubule plus end dynamics. PMID:25187649

Inducible growth mode switches influence Valonia rhizoid differentiation.

PubMed

Elvira, Paul Rommel; Sekida, Satoko; Okuda, Kazuo

2013-02-01

Cell differentiation and cell type commitment are an integral part of plant growth and development. Investigations on how environmental conditions affect the formation of shoots, roots, and rhizoids can help illustrate how plants determine cell fate and overall morphology. In this study, we evaluated the role of substratum and light on rhizoid differentiation in the coenocytic green alga, Valonia aegagropila. Elongating rhizoids displayed varying growth modes and cell shape upon exposure to different substrata and light conditions. It was found that soft substrata and dark incubation promoted rhizoid elongation via tip growth while subsequent exposure to light prevented tip growth and instead induced swelling in the apical region of rhizoids. Swelling was accompanied by the accumulation of protoplasm in the rhizoid tip through expansion of the cell wall and uninhibited cytoplasmic streaming. Subsequent diffuse growth led to the transformation from slender, rod-shaped rhizoids into spherical thallus-like structures that required photosynthesis. Further manipulation of light regimes caused vacillating cell growth redirections. An elongating V. aegagropila rhizoid cell thus appears capable of growth mode switching that is regulated by immediate environmental conditions thereby influencing ultimate cell shape and function. This is the first description of inducible, multiple growth mode shifts in a single intact plant cell that directly impact its differentiation.

Cytokinesis-Based Constraints on Polarized Cell Growth in Fission Yeast

PubMed Central

Bohnert, K. Adam; Gould, Kathleen L.

2012-01-01

The rod-shaped fission yeast Schizosaccharomyces pombe, which undergoes cycles of monopolar-to-bipolar tip growth, is an attractive organism for studying cell-cycle regulation of polarity establishment. While previous research has described factors mediating this process from interphase cell tips, we found that division site signaling also impacts the re-establishment of bipolar cell growth in the ensuing cell cycle. Complete loss or targeted disruption of the non-essential cytokinesis protein Fic1 at the division site, but not at interphase cell tips, resulted in many cells failing to grow at new ends created by cell division. This appeared due to faulty disassembly and abnormal persistence of the cell division machinery at new ends of fic1Δ cells. Moreover, additional mutants defective in the final stages of cytokinesis exhibited analogous growth polarity defects, supporting that robust completion of cell division contributes to new end-growth competency. To test this model, we genetically manipulated S. pombe cells to undergo new end take-off immediately after cell division. Intriguingly, such cells elongated constitutively at new ends unless cytokinesis was perturbed. Thus, cell division imposes constraints that partially override positive controls on growth. We posit that such constraints facilitate invasive fungal growth, as cytokinesis mutants displaying bipolar growth defects formed numerous pseudohyphae. Collectively, these data highlight a role for previous cell cycles in defining a cell's capacity to polarize at specific sites, and they additionally provide insight into how a unicellular yeast can transition into a quasi-multicellular state. PMID:23093943

Endothelial Notch signalling limits angiogenesis via control of artery formation

PubMed Central

Hasan, Sana S.; Tsaryk, Roman; Lange, Martin; Wisniewski, Laura; Moore, John C.; Lawson, Nathan D.; Wojciechowska, Karolina; Schnittler, Hans; Siekmann, Arndt F.

2017-01-01

Angiogenic sprouting needs to be tightly controlled. It has been suggested that the Notch ligand dll4 expressed in leading tip cells restricts angiogenesis by activating Notch signalling in trailing stalk cells. Here, we show using live imaging in zebrafish that activation of Notch signalling is rather required in tip cells. Notch activation initially triggers expression of the chemokine receptor cxcr4a. This allows for proper tip cell migration and connection to the pre-existing arterial circulation, ultimately establishing functional arterial-venous blood flow patterns. Subsequently, Notch signalling reduces cxcr4a expression, thereby preventing excessive blood vessel growth. Finally, we find that Notch signalling is dispensable for limiting blood vessel growth during venous plexus formation that does not generate arteries. Together, these findings link the role of Notch signalling in limiting angiogenesis to its role during artery formation and provide a framework for our understanding of the mechanisms underlying blood vessel network expansion and maturation. PMID:28714969

The effect of cyanide compounds, fluorides, aluminum, and inorganic oxides present in spent pot liner on germination and root tip cells of Lactuca sativa.

PubMed

Andrade, L F; Davide, L C; Gedraite, L S

2010-05-01

SPL (spent pot liner) is a solid waste produced by the aluminum industry. This waste has a highly variable composition, consisting of cyanides, fluorides, organics, and metals. The aim of this work was to study the effect of SPL on root tips of Lactuca sativa using current plant bioassays. We observed a decrease in the germination rate with increasing concentrations of SPL. In addition, SPL was found to reduce root growth, which is correlated with a decrease in the mitotic index. Nevertheless, we noticed a significant enhancement in the percentage of stickiness, c-metaphase, anaphase bridges, and laggard chromosomes in dividing cells and also an increase in the number of cells with condensed nuclei. Moreover, SPL was found to alter the root tip surface, resulting in a reduction in the amount of root hair. These results demonstrate that SPL is a toxic agent that leads to cell damage and disturbance. Copyright 2009 Elsevier Inc. All rights reserved.

Microinjector for blasocysts

DOEpatents

Remenyik, Carl J.; Woychik, Richard P.; Patek, David R.; Hawk, James A.; Turner, John C.

1999-01-01

An electromechanical device for driving the tip of a microinjection cannula, or needle, through the outer barrier of a blastocyst, cell, or cell nucleus for the injection of cells or other bioactive materials. Either a flexible frame or a ram moving within a base member is employed. Cannula motion is achieved by means of a piezoelectric stack and spring return system. The thrust motion over a predetermined microscopic distance is achieved without cannula setback prior to the thrust movement. Instead of specially prepared beveled and tipped needles, standard unimproved cannulas or needles can be used.

Microinjector for blasocysts

DOEpatents

Remenyik, C.J.; Woychik, R.P.; Patek, D.R.; Hawk, J.A.; Turner, J.C.

1999-03-02

An electromechanical device is disclosed for driving the tip of a microinjection cannula, or needle, through the outer barrier of a blastocyst, cell, or cell nucleus for the injection of cells or other bioactive materials. Either a flexible frame or a ram moving within a base member is employed. Cannula motion is achieved by means of a piezoelectric stack and spring return system. The thrust motion over a predetermined microscopic distance is achieved without cannula setback prior to the thrust movement. Instead of specially prepared beveled and tipped needles, standard unimproved cannulas or needles can be used. 6 figs.

Conflicting Selection Pressures Will Constrain Viral Escape from Interfering Particles: Principles for Designing Resistance-Proof Antivirals.

PubMed

Rast, Luke I; Rouzine, Igor M; Rozhnova, Ganna; Bishop, Lisa; Weinberger, Ariel D; Weinberger, Leor S

2016-05-01

The rapid evolution of RNA-encoded viruses such as HIV presents a major barrier to infectious disease control using conventional pharmaceuticals and vaccines. Previously, it was proposed that defective interfering particles could be developed to indefinitely control the HIV/AIDS pandemic; in individual patients, these engineered molecular parasites were further predicted to be refractory to HIV's mutational escape (i.e., be 'resistance-proof'). However, an outstanding question has been whether these engineered interfering particles-termed Therapeutic Interfering Particles (TIPs)-would remain resistance-proof at the population-scale, where TIP-resistant HIV mutants may transmit more efficiently by reaching higher viral loads in the TIP-treated subpopulation. Here, we develop a multi-scale model to test whether TIPs will maintain indefinite control of HIV at the population-scale, as HIV ('unilaterally') evolves toward TIP resistance by limiting the production of viral proteins available for TIPs to parasitize. Model results capture the existence of two intrinsic evolutionary tradeoffs that collectively prevent the spread of TIP-resistant HIV mutants in a population. First, despite their increased transmission rates in TIP-treated sub-populations, unilateral TIP-resistant mutants are shown to have reduced transmission rates in TIP-untreated sub-populations. Second, these TIP-resistant mutants are shown to have reduced growth rates (i.e., replicative fitness) in both TIP-treated and TIP-untreated individuals. As a result of these tradeoffs, the model finds that TIP-susceptible HIV strains continually outcompete TIP-resistant HIV mutants at both patient and population scales when TIPs are engineered to express >3-fold more genomic RNA than HIV expresses. Thus, the results provide design constraints for engineering population-scale therapies that may be refractory to the acquisition of antiviral resistance.

Analysis of Statoliths Displacement in Chara Rhizoids for Validating the Microgravity-Simulation Quality of Clinorotation Modes

NASA Astrophysics Data System (ADS)

Krause, Lars; Braun, Markus; Hauslage, Jens; Hemmersbach, Ruth

2018-05-01

In single-celled rhizoids of the green algae Chara, positively gravitropic growth is governed by statoliths kept in a dynamically stable position 10-25 μ m above the cell tip by a complex interaction of gravity and actomyosin forces. Any deviation of the tube-like cells from the tip-downward orientation causes statoliths to sediment onto the gravisensitive subapical cell flank which initiates a gravitropic curvature response. Microgravity experiments have shown that abolishing the net tip-directed gravity force results in an actomyosin-mediated axial displacement of statoliths away from the cell tip. The present study was performed to critically assess the quality of microgravity simulation provided by different operational modes of a Random Positioning Machine (RPM) running with one axis (2D mode) or two axes (3D mode) and different rotational speeds (2D), speed ranges and directions (3D). The effects of 2D and 3D rotation were compared with data from experiments in real microgravity conditions (MAXUS sounding rocket missions). Rotational speeds in the range of 60-85 rpm in 2D and 3D modes resulted in a similar kinetics of statolith displacement as compared to real microgravity data, while slower clinorotation (2-11 rpm) caused a reduced axial displacement and a more dispersed arrangement of statoliths closer to the cell tip. Increasing the complexity of rotation by adding a second rotation axis in case of 3D clinorotation did not increase the quality of microgravity simulation, however, increased side effects such as the level of vibrations resulting in a more dispersed arrangement of statoliths. In conclusion, fast 2D clinorotation provides the most appropriate microgravity simulation for investigating the graviperception mechanism in Chara rhizoids, whereas slower clinorotation speeds and rotating samples around two axes do not improve the quality of microgravity simulation.

Analysis of Statoliths Displacement in Chara Rhizoids for Validating the Microgravity-Simulation Quality of Clinorotation Modes

NASA Astrophysics Data System (ADS)

Krause, Lars; Braun, Markus; Hauslage, Jens; Hemmersbach, Ruth

2018-01-01

In single-celled rhizoids of the green algae Chara, positively gravitropic growth is governed by statoliths kept in a dynamically stable position 10-25 μ m above the cell tip by a complex interaction of gravity and actomyosin forces. Any deviation of the tube-like cells from the tip-downward orientation causes statoliths to sediment onto the gravisensitive subapical cell flank which initiates a gravitropic curvature response. Microgravity experiments have shown that abolishing the net tip-directed gravity force results in an actomyosin-mediated axial displacement of statoliths away from the cell tip. The present study was performed to critically assess the quality of microgravity simulation provided by different operational modes of a Random Positioning Machine (RPM) running with one axis (2D mode) or two axes (3D mode) and different rotational speeds (2D), speed ranges and directions (3D). The effects of 2D and 3D rotation were compared with data from experiments in real microgravity conditions (MAXUS sounding rocket missions). Rotational speeds in the range of 60-85 rpm in 2D and 3D modes resulted in a similar kinetics of statolith displacement as compared to real microgravity data, while slower clinorotation (2-11 rpm) caused a reduced axial displacement and a more dispersed arrangement of statoliths closer to the cell tip. Increasing the complexity of rotation by adding a second rotation axis in case of 3D clinorotation did not increase the quality of microgravity simulation, however, increased side effects such as the level of vibrations resulting in a more dispersed arrangement of statoliths. In conclusion, fast 2D clinorotation provides the most appropriate microgravity simulation for investigating the graviperception mechanism in Chara rhizoids, whereas slower clinorotation speeds and rotating samples around two axes do not improve the quality of microgravity simulation.

General Tips Concerning What Has Been Learned About Cotton Processing in Traditional Textile Manufacturing

USDA-ARS?s Scientific Manuscript database

This article, under the following sub-headings, briefly describes a few important practical tips involving processing of cotton in traditional textile manufacturing: (1)Bale Selection and Fiber Mixing, (2) Fiber Opening and Cleaning, (3) Carding, (4) Drawing, (5) Combing (if necessary), (6) Roving, ...

Development of end-selective functionalized carbon nanotubes for biomedical applications

NASA Astrophysics Data System (ADS)

Lee, Seung Ho; Kim, Wan Sun; Lee, Ha Rim; Park, Kyu Chang; Lee, Chang Hoon; Park, Hun Kuk; Kim, Kyung Sook

2015-12-01

Carbon nanotube (CNT) is a type of carbon allotrope with excellent physical and electrical properties, including high thermal conductivity, mechanical strength, and thermal stability. Therefore, applications of CNT have been considered for a variety of fields, including biosensors, molecular electronics, X-ray, and fuel cells. However, the application of CNT to biomedicine is limited because this material is cytotoxic and inhomogeneous. In particular, the irregularity in the structural properties of paste or bundle-type CNTs causes an uncontrolled modification in biomolecules. Therefore, using CNT as biosensors to obtain quantitative analyses is difficult. In this study, we developed a new method to perform end-selective functionalization of CNT in order to enable quantitative analysis for biomedical applications. The process was as follows: (1) etching the tip of vertically-aligned CNTs under optimum conditions, (2) oxidation of exposed CNTs, and (3) end-selective linkage of functionalized CNTs with biomolecules (dsDNA).

Crystal Structure of an Ammonia-Permeable Aquaporin

PubMed Central

Kirscht, Andreas; Kaptan, Shreyas S.; Bienert, Gerd Patrick; Chaumont, François; Nissen, Poul; de Groot, Bert L.; Kjellbom, Per; Gourdon, Pontus; Johanson, Urban

2016-01-01

Aquaporins of the TIP subfamily (Tonoplast Intrinsic Proteins) have been suggested to facilitate permeation of water and ammonia across the vacuolar membrane of plants, allowing the vacuole to efficiently sequester ammonium ions and counteract cytosolic fluctuations of ammonia. Here, we report the structure determined at 1.18 Å resolution from twinned crystals of Arabidopsis thaliana aquaporin AtTIP2;1 and confirm water and ammonia permeability of the purified protein reconstituted in proteoliposomes as further substantiated by molecular dynamics simulations. The structure of AtTIP2;1 reveals an extended selectivity filter with the conserved arginine of the filter adopting a unique unpredicted position. The relatively wide pore and the polar nature of the selectivity filter clarify the ammonia permeability. By mutational studies, we show that the identified determinants in the extended selectivity filter region are sufficient to convert a strictly water-specific human aquaporin into an AtTIP2;1-like ammonia channel. A flexible histidine and a novel water-filled side pore are speculated to deprotonate ammonium ions, thereby possibly increasing permeation of ammonia. The molecular understanding of how aquaporins facilitate ammonia flux across membranes could potentially be used to modulate ammonia losses over the plasma membrane to the atmosphere, e.g., during photorespiration, and thereby to modify the nitrogen use efficiency of plants. PMID:27028365

The Brassicaceae species Heliophila coronopifolia produces root border-like cells that protect the root tip and secrete defensin peptides.

PubMed

Weiller, Florent; Moore, John P; Young, Philip; Driouich, Azeddine; Vivier, Melané A

2017-03-01

Root border cells and border-like cells (BLCs), the latter originally described in Arabidopsis thaliana , have been described as cells released at the root tips of the species in which they occur. BLCs are thought to provide protection to root meristems similar to classical root border cells. In addition, four defensin peptides (Hc-AFP1-4) have previously been characterized from Heliophila coronopifolia , a South African semi-desert flower, and found to be strongly antifungal. This provided an opportunity to evaluate if the BLCs of H. coronopifolia indeed produce these defensins, which would provide evidence towards a defence role for BLCs. Fluorescence microscopy, using live-cell-imaging technology, was used to characterize the BLCs of H. coronopifolia . Quantitative real-time PCR (qRT-PCR) analysis and immunofluorescence microscopy was used to characterize these defensin peptides. BLCs originated at the root apical meristem and formed a protective sheath at the tip and along the sides as the root elongated in solid medium. BLCs have a cellulose-enriched cell wall, intact nuclei and are embedded in a layer of pectin-rich mucilage. Pectinase treatments led to the dissolution of the sheath and dissociation of the root BLCs. Hc-AFP1-4 genes were all expressed in root tissues, but Hc-AFP3 transcripts were the most abundant in these tissues as measured by qRT-PCR. A polyclonal antibody that was cross-reactive with all four defensins, and probably recognizing a general plant defensin epitope, was used in fluorescence microscopy analysis to examine the presence of the peptides in the root tip and BLCs. Data confirmed the peptides present in the root tip tissues, the mucilage sheath and the BLCs. This study provides a link between defensin peptides and BLCs, both embedded in a protective pectin mucilage sheath, during normal plant growth and development. The presence of the Hc-AFP3 defensin peptides in the BLCs suggests a role for these cells in root protection. © 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

The Brassicaceae species Heliophila coronopifolia produces root border-like cells that protect the root tip and secrete defensin peptides

PubMed Central

Weiller, Florent; Young, Philip; Driouich, Azeddine; Vivier, Melané A.

2017-01-01

Background and Aims Root border cells and border-like cells (BLCs), the latter originally described in Arabidopsis thaliana, have been described as cells released at the root tips of the species in which they occur. BLCs are thought to provide protection to root meristems similar to classical root border cells. In addition, four defensin peptides (Hc-AFP1–4) have previously been characterized from Heliophila coronopifolia, a South African semi-desert flower, and found to be strongly antifungal. This provided an opportunity to evaluate if the BLCs of H. coronopifolia indeed produce these defensins, which would provide evidence towards a defence role for BLCs. Methods Fluorescence microscopy, using live-cell-imaging technology, was used to characterize the BLCs of H. coronopifolia. Quantitative real-time PCR (qRT-PCR) analysis and immunofluorescence microscopy was used to characterize these defensin peptides. Key Results BLCs originated at the root apical meristem and formed a protective sheath at the tip and along the sides as the root elongated in solid medium. BLCs have a cellulose-enriched cell wall, intact nuclei and are embedded in a layer of pectin-rich mucilage. Pectinase treatments led to the dissolution of the sheath and dissociation of the root BLCs. Hc-AFP1–4 genes were all expressed in root tissues, but Hc-AFP3 transcripts were the most abundant in these tissues as measured by qRT-PCR. A polyclonal antibody that was cross-reactive with all four defensins, and probably recognizing a general plant defensin epitope, was used in fluorescence microscopy analysis to examine the presence of the peptides in the root tip and BLCs. Data confirmed the peptides present in the root tip tissues, the mucilage sheath and the BLCs. Conclusions This study provides a link between defensin peptides and BLCs, both embedded in a protective pectin mucilage sheath, during normal plant growth and development. The presence of the Hc-AFP3 defensin peptides in the BLCs suggests a role for these cells in root protection. PMID:27481828

Particle trapping in 3-D using a single fiber probe with an annular light distribution.

PubMed

Taylor, R; Hnatovsky, C

2003-10-20

A single optical fiber probe has been used to trap a solid 2 ìm diameter glass bead in 3-D in water. Optical confinement in 2-D was produced by the annular light distribution emerging from a selectively chemically etched, tapered, hollow tipped metalized fiber probe. Confinement of the bead in 3-D was achieved by balancing an electrostatic force of attraction towards the tip and the optical scattering force pushing the particle away from the tip.

NASA Tech Briefs, February 2008

NASA Technical Reports Server (NTRS)

2008-01-01

Topics discussed include: Optical Measurement of Mass Flow of a Two-Phase Fluid; Selectable-Tip Corrosion-Testing Electrochemical Cell; Piezoelectric Bolt Breakers and Bolt Fatigue Testers; Improved Measurement of B(sub 22) of Macromolecules in a Flow Cell; Measurements by a Vector Network Analyzer at 325 to 508 GHz; Using Light to Treat Mucositis and Help Wounds Heal; Increasing Discharge Capacities of Li-(CF)(sub n) Cells; Dot-in-Well Quantum-Dot Infrared Photodetectors; Integrated Microbatteries for Implantable Medical Devices; Oxidation Behavior of Carbon Fiber-Reinforced Composites; GIDEP Batching Tool; Generic Spacecraft Model for Real-Time Simulation; Parallel-Processing Software for Creating Mosaic Images; Software for Verifying Image-Correlation Tie Points; Flexcam Image Capture Viewing and Spot Tracking; Low-Pt-Content Anode Catalyst for Direct Methanol Fuel Cells; Graphite/Cyanate Ester Face Sheets for Adaptive Optics; Atomized BaF2-CaF7 for Better-Flowing Plasma-Spray Feedstock; Nanophase Nickel-Zirconium Alloys for Fuel Cells; Vacuum Packaging of MEMS With Multiple Internal Seal Rings; Compact Two-Dimensional Spectrometer Optics; and Fault-Tolerant Coding for State Machines.

Live-cell imaging of actin dynamics reveals mechanisms of stereocilia length regulation in the inner ear

PubMed Central

Drummond, Meghan C.; Barzik, Melanie; Bird, Jonathan E.; Zhang, Duan-Sun; Lechene, Claude P.; Corey, David P.; Cunningham, Lisa L.; Friedman, Thomas B.

2015-01-01

The maintenance of sensory hair cell stereocilia is critical for lifelong hearing; however, mechanisms of structural homeostasis remain poorly understood. Conflicting models propose that stereocilia F-actin cores are either continually renewed every 24–48 h via a treadmill or are stable, exceptionally long-lived structures. Here to distinguish between these models, we perform an unbiased survey of stereocilia actin dynamics in more than 500 utricle hair cells. Live-imaging EGFP-β-actin or dendra2-β-actin reveal stable F-actin cores with turnover and elongation restricted to stereocilia tips. Fixed-cell microscopy of wild-type and mutant β-actin demonstrates that incorporation of actin monomers into filaments is required for localization to stereocilia tips. Multi-isotope imaging mass spectrometry and live imaging of single differentiating hair cells capture stereociliogenesis and explain uniform incorporation of 15N-labelled protein and EGFP-β-actin into nascent stereocilia. Collectively, our analyses support a model in which stereocilia actin cores are stable structures that incorporate new F-actin only at the distal tips. PMID:25898120

Ergonomic factors related to drop-off detection with the long cane: effects of cane tips and techniques.

PubMed

Kim, Dae Shik; Emerson, Robert S Wall; Curtis, Amy B

2010-06-01

This study examined the effect of cane tips and cane techniques on drop-off detection with the long cane. Blind pedestrians depend on a long cane to detect drop-offs. Missing a drop-off may result in falls or collision with moving vehicles in the street. Although cane tips appear to affect a cane user's ability to detect drop-offs, few experimental studies have examined such effect. A repeated-measures design with block randomization was used for the study. Participants were 17 adults who were legally blind and had no other disabilities. Participants attempted to detect the drop-offs of varied depths using different cane tips and cane techniques. Drop-off detection rates were similar between the marshmallow tip (77.0%) and the marshmallow roller tip (79.4%) when both tips were used with the constant contact technique, p = .294. However, participants detected drop-offs at a significantly higher percentage when they used the constant contact technique with the marshmallow roller tip (79.4%) than when they used the two-point touch technique with the marshmallow tip (63.2%), p < .001. The constant contact technique used with a marshmallow roller tip (perceived as a less advantageous tip) was more effective than the two-point touch technique used with a marshmallow tip (perceived as a more advantageous tip) in detecting drop-offs. The findings of the study may help cane users and orientation and mobility specialists select appropriate cane techniques and cane tips in accordance with the cane user's characteristics and the nature of the travel environment.

Stem Cell Basics

MedlinePlus

... Tips Info Center Research Topics Federal Policy Glossary Stem Cell Information General Information Clinical Trials Funding Information Current ... Basics » Stem Cell Basics I. Back to top Stem Cell Basics I. Introduction: What are stem cells, and ...

Fe65 is required for Tip60-directed histone H4 acetylation at DNA strand breaks

PubMed Central

Stante, Maria; Minopoli, Giuseppina; Passaro, Fabiana; Raia, Maddalena; Vecchio, Luigi Del; Russo, Tommaso

2009-01-01

Fe65 is a binding partner of the Alzheimer's β-amyloid precursor protein APP. The possible involvement of this protein in the cellular response to DNA damage was suggested by the observation that Fe65 null mice are more sensitive to genotoxic stress than WT counterpart. Fe65 associated with chromatin under basal conditions and its involvement in DNA damage repair requires this association. A known partner of Fe65 is the histone acetyltransferase Tip60. Considering the crucial role of Tip60 in DNA repair, we explored the hypothesis that the phenotype of Fe65 null cells depended on its interaction with Tip60. We demonstrated that Fe65 knockdown impaired recruitment of Tip60-TRRAP complex to DNA double strand breaks and decreased histone H4 acetylation. Accordingly, the efficiency of DNA repair was decreased upon Fe65 suppression. To explore whether APP has a role in this mechanism, we analyzed a Fe65 mutant unable to bind to APP. This mutant failed to rescue the phenotypes of Fe65 null cells; furthermore, APP/APLP2 suppression results in the impairment of recruitment of Tip60-TRRAP complex to DNA double strand breaks, decreased histone H4 acetylation and repair efficiency. On these bases, we propose that Fe65 and its interaction with APP play an important role in the response to DNA damage by assisting the recruitment of Tip60-TRRAP to DNA damage sites. PMID:19282473

NOTCH1 Inhibits Activation of ATM by Impairing the Formation of an ATM-FOXO3a-KAT5/Tip60 Complex.

PubMed

Adamowicz, Marek; Vermezovic, Jelena; d'Adda di Fagagna, Fabrizio

2016-08-23

The DNA damage response (DDR) signal transduction pathway is responsible for sensing DNA damage and further relaying this signal into the cell. ATM is an apical DDR kinase that orchestrates the activation and the recruitment of downstream DDR factors to induce cell-cycle arrest and repair. We have previously shown that NOTCH1 inhibits ATM activation upon DNA damage, but the underlying mechanism remains unclear. Here, we show that NOTCH1 does not impair ATM recruitment to DNA double-strand breaks (DSBs). Rather, NOTCH1 prevents binding of FOXO3a and KAT5/Tip60 to ATM through a mechanism in which NOTCH1 competes with FOXO3a for ATM binding. Lack of FOXO3a binding to ATM leads to the loss of KAT5/Tip60 association with ATM. Moreover, expression of NOTCH1 or depletion of ATM impairs the formation of the FOXO3a-KAT5/Tip60 protein complex. Finally, we show that pharmacological induction of FOXO3a nuclear localization sensitizes NOTCH1-driven cancers to DNA-damage-induced cell death. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

Tip-localized receptors control pollen tube growth and LURE sensing in Arabidopsis.

PubMed

Takeuchi, Hidenori; Higashiyama, Tetsuya

2016-03-10

Directional control of tip-growing cells is essential for proper tissue organization and cell-to-cell communication in animals and plants. In the sexual reproduction of flowering plants, the tip growth of the male gametophyte, the pollen tube, is precisely guided by female cues to achieve fertilization. Several female-secreted peptides have recently been identified as species-specific attractants that directly control the direction of pollen tube growth. However, the method by which pollen tubes precisely and promptly respond to the guidance signal from their own species is unknown. Here we show that tip-localized pollen-specific receptor-like kinase 6 (PRK6) with an extracellular leucine-rich repeat domain is an essential receptor for sensing of the LURE1 attractant peptide in Arabidopsis thaliana under semi-in-vivo conditions, and is important for ovule targeting in the pistil. PRK6 interacted with pollen-expressed ROPGEFs (Rho of plant guanine nucleotide-exchange factors), which are important for pollen tube growth through activation of the signalling switch Rho GTPase ROP1 (refs 7, 8). PRK6 conferred responsiveness to AtLURE1 in pollen tubes of the related species Capsella rubella. Furthermore, our genetic and physiological data suggest that PRK6 signalling through ROPGEFs and sensing of AtLURE1 are achieved in cooperation with the other PRK family receptors, PRK1, PRK3 and PRK8. Notably, the tip-focused PRK6 accumulated asymmetrically towards an external AtLURE1 source before reorientation of pollen tube tip growth. These results demonstrate that PRK6 acts as a key membrane receptor for external AtLURE1 attractants, and recruits the core tip-growth machinery, including ROP signalling proteins. This work provides insights into the orchestration of efficient pollen tube growth and species-specific pollen tube attraction by multiple receptors during male-female communication.

Transduction channels’ gating can control friction on vibrating hair-cell bundles in the ear

PubMed Central

Bormuth, Volker; Barral, Jérémie; Joanny, Jean-François; Jülicher, Frank; Martin, Pascal

2014-01-01

Hearing starts when sound-evoked mechanical vibrations of the hair-cell bundle activate mechanosensitive ion channels, giving birth to an electrical signal. As for any mechanical system, friction impedes movements of the hair bundle and thus constrains the sensitivity and frequency selectivity of auditory transduction. Friction is generally thought to result mainly from viscous drag by the surrounding fluid. We demonstrate here that the opening and closing of the transduction channels produce internal frictional forces that can dominate viscous drag on the micrometer-sized hair bundle. We characterized friction by analyzing hysteresis in the force–displacement relation of single hair-cell bundles in response to periodic triangular stimuli. For bundle velocities high enough to outrun adaptation, we found that frictional forces were maximal within the narrow region of deflections that elicited significant channel gating, plummeted upon application of a channel blocker, and displayed a sublinear growth for increasing bundle velocity. At low velocity, the slope of the relation between the frictional force and velocity was nearly fivefold larger than the hydrodynamic friction coefficient that was measured when the transduction machinery was decoupled from bundle motion by severing tip links. A theoretical analysis reveals that channel friction arises from coupling the dynamics of the conformational change associated with channel gating to tip-link tension. Varying channel properties affects friction, with faster channels producing smaller friction. We propose that this intrinsic source of friction may contribute to the process that sets the hair cell’s characteristic frequency of responsiveness. PMID:24799674

Ion transport: Tipping a cell's ionic balance

NASA Astrophysics Data System (ADS)

Davis, Jeffery T.

2014-10-01

A synthetic compound that transports chloride across membranes can kill both normal cells and cancer cells in vitro. The transporter works together with sodium channels to move NaCl into the cells, which triggers cell death.

Mathematical modeling and experimental validation of the spatial distribution of boron in the root of Arabidopsis thaliana identify high boron accumulation in the tip and predict a distinct root tip uptake function.

PubMed

Shimotohno, Akie; Sotta, Naoyuki; Sato, Takafumi; De Ruvo, Micol; Marée, Athanasius F M; Grieneisen, Verônica A; Fujiwara, Toru

2015-04-01

Boron, an essential micronutrient, is transported in roots of Arabidopsis thaliana mainly by two different types of transporters, BORs and NIPs (nodulin26-like intrinsic proteins). Both are plasma membrane localized, but have distinct transport properties and patterns of cell type-specific accumulation with different polar localizations, which are likely to affect boron distribution. Here, we used mathematical modeling and an experimental determination to address boron distributions in the root. A computational model of the root is created at the cellular level, describing the boron transporters as observed experimentally. Boron is allowed to diffuse into roots, in cells and cell walls, and to be transported over plasma membranes, reflecting the properties of the different transporters. The model predicts that a region around the quiescent center has a higher concentration of soluble boron than other portions. To evaluate this prediction experimentally, we determined the boron distribution in roots using laser ablation-inductivity coupled plasma-mass spectrometry. The analysis indicated that the boron concentration is highest near the tip and is lower in the more proximal region of the meristem zone, similar to the pattern of soluble boron distribution predicted by the model. Our model also predicts that upward boron flux does not continuously increase from the root tip toward the mature region, indicating that boron taken up in the root tip is not efficiently transported to shoots. This suggests that root tip-absorbed boron is probably used for local root growth, and that instead it is the more mature root regions which have a greater role in transporting boron toward the shoots. © The Author 2015. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists.

Mathematical Modeling and Experimental Validation of the Spatial Distribution of Boron in the Root of Arabidopsis thaliana Identify High Boron Accumulation in the Tip and Predict a Distinct Root Tip Uptake Function

PubMed Central

Shimotohno, Akie; Sotta, Naoyuki; Sato, Takafumi; De Ruvo, Micol; Marée, Athanasius F.M.; Grieneisen, Verônica A.; Fujiwara, Toru

2015-01-01

Boron, an essential micronutrient, is transported in roots of Arabidopsis thaliana mainly by two different types of transporters, BORs and NIPs (nodulin26-like intrinsic proteins). Both are plasma membrane localized, but have distinct transport properties and patterns of cell type-specific accumulation with different polar localizations, which are likely to affect boron distribution. Here, we used mathematical modeling and an experimental determination to address boron distributions in the root. A computational model of the root is created at the cellular level, describing the boron transporters as observed experimentally. Boron is allowed to diffuse into roots, in cells and cell walls, and to be transported over plasma membranes, reflecting the properties of the different transporters. The model predicts that a region around the quiescent center has a higher concentration of soluble boron than other portions. To evaluate this prediction experimentally, we determined the boron distribution in roots using laser ablation-inductivity coupled plasma-mass spectrometry. The analysis indicated that the boron concentration is highest near the tip and is lower in the more proximal region of the meristem zone, similar to the pattern of soluble boron distribution predicted by the model. Our model also predicts that upward boron flux does not continuously increase from the root tip toward the mature region, indicating that boron taken up in the root tip is not efficiently transported to shoots. This suggests that root tip-absorbed boron is probably used for local root growth, and that instead it is the more mature root regions which have a greater role in transporting boron toward the shoots. PMID:25670713

Mitochondrial anchorage and fusion contribute to mitochondrial inheritance and quality control in the budding yeast Saccharomyces cerevisiae.

PubMed

Higuchi-Sanabria, Ryo; Charalel, Joseph K; Viana, Matheus P; Garcia, Enrique J; Sing, Cierra N; Koenigsberg, Andrea; Swayne, Theresa C; Vevea, Jason D; Boldogh, Istvan R; Rafelski, Susanne M; Pon, Liza A

2016-03-01

Higher-functioning mitochondria that are more reduced and have less ROS are anchored in the yeast bud tip by the Dsl1-family protein Mmr1p. Here we report a role for mitochondrial fusion in bud-tip anchorage of mitochondria. Fluorescence loss in photobleaching (FLIP) and network analysis experiments revealed that mitochondria in large buds are a continuous reticulum that is physically distinct from mitochondria in mother cells. FLIP studies also showed that mitochondria that enter the bud can fuse with mitochondria that are anchored in the bud tip. In addition, loss of fusion and mitochondrial DNA (mtDNA) by deletion of mitochondrial outer or inner membrane fusion proteins (Fzo1p or Mgm1p) leads to decreased accumulation of mitochondria at the bud tip and inheritance of fitter mitochondria by buds compared with cells with no mtDNA. Conversely, increasing the accumulation and anchorage of mitochondria in the bud tip by overexpression of MMR1 results in inheritance of less-fit mitochondria by buds and decreased replicative lifespan and healthspan. Thus quantity and quality of mitochondrial inheritance are ensured by two opposing processes: bud-tip anchorage by mitochondrial fusion and Mmr1p, which favors bulk inheritance; and quality control mechanisms that promote segregation of fitter mitochondria to the bud. © 2016 Higuchi-Sanabria et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

Antibodies derived from an enterotoxigenic Escherichia coli (ETEC) adhesin tip MEFA (multiepitope fusion antigen) against adherence of nine ETEC adhesins: CFA/I, CS1, CS2, CS3, CS4, CS5, CS6, CS21 and EtpA.

PubMed

Nandre, Rahul M; Ruan, Xiaosai; Duan, Qiangde; Sack, David A; Zhang, Weiping

2016-06-30

Diarrhea continues to be a leading cause of death in children younger than 5 years in developing countries. Enterotoxigenic Escherichia coli (ETEC) is a leading bacterial cause of children's diarrhea and travelers' diarrhea. ETEC bacteria initiate diarrheal disease by attaching to host receptors at epithelial cells and colonizing in small intestine. Therefore, preventing ETEC attachment has been considered the first line of defense against ETEC diarrhea. However, developing vaccines effectively against ETEC bacterial attachment encounters challenge because ETEC strains produce over 23 immunologically heterogeneous adhesins. In this study, we applied MEFA (multiepitope fusion antigen) approach to integrate epitopes from adhesin tips or adhesive subunits of CFA/I, CS1, CS2, CS3, CS4, CS5, CS6, CS21 and EtpA adhesins and to construct an adhesin tip MEFA peptide. We then examined immunogenicity of this tip MEFA in mouse immunization, and assessed potential application of this tip MEFA for ETEC vaccine development. Data showed that mice intraperitoneally immunized with this adhesin tip MEFA developed IgG antibody responses to all nine ETEC adhesins. Moreover, ETEC and E. coli bacteria expressing these nine adhesins, after incubation with serum of the immunized mice, exhibited significant reduction in attachment to Caco-2 cells. These results indicated that anti-adhesin antibodies induced by this adhesin tip MEFA blocked adherence of the most important ETEC adhesins, suggesting this multivalent tip MEFA may be useful for developing a broadly protective anti-adhesin vaccine against ETEC diarrhea. Copyright © 2016 Elsevier Ltd. All rights reserved.

A design approach and selected wind tunnel results at high subsonic speeds for wing-tip mounted winglets

NASA Technical Reports Server (NTRS)

Whitcomb, R. T.

1976-01-01

Winglets, which are small, nearly vertical, winglike surfaces, substantially reduce drag coefficients at lifting conditions. The primary winglet surfaces are rearward above the wing tips; secondary surfaces are forward below the wing tips. This report presents a discussion of the considerations involved in the design of the winglets; measured effects of these surfaces on the aerodynamic forces, moments, and loads for a representative first generation, narrow body jet transport wing; and a comparison of these effects with those for a wing tip extension which results in approximately the same increase in bending moment at the wing-fuselage juncture as did the addition of the winglets.

Thidiazuron Triggers Morphogenesis in Rosa canina L. Protocorm-Like Bodies by Changing Incipient Cell Fate.

PubMed

Kou, Yaping; Yuan, Cunquan; Zhao, Qingcui; Liu, Guoqin; Nie, Jing; Ma, Zhimin; Cheng, Chenxia; Teixeira da Silva, Jaime A; Zhao, Liangjun

2016-01-01

Thidiazuron (N-phenyl-N'-1,2,3-thiadiazol-5-ylurea; TDZ) is an artificial plant growth regulator that is widely used in plant tissue culture. Protocorm-like bodies (PLBs) induced by TDZ serve as an efficient and rapid in vitro regeneration system in Rosa species. Despite this, the mechanism of PLB induction remains relatively unclear. TDZ, which can affect the level of endogenous auxins and cytokinins, converts the cell fate of rhizoid tips and triggers PLB formation and plantlet regeneration in Rosa canina L. In callus-rhizoids, which are rhizoids that co-develop from callus, auxin and a Z-type cytokinin accumulated after applying TDZ, and transcription of the auxin transporter gene RcPIN1 was repressed. The expression of RcARF4, RcRR1, RcCKX2, RcCKX3, and RcLOG1 increased in callus-rhizoids and rhizoid tips while the transcription of an auxin response factor (RcARF1) and auxin transport proteins (RcPIN2, RcPIN3) decreased in callus-rhizoids but increased in rhizoid tips. In situ hybridization of rhizoids showed that RcWUS and RcSERK1 were highly expressed in columella cells and root stem cells resulting in the conversion of cell fate into shoot apical meristems or embryogenic callus. In addition, transgenic XVE::RcWUS lines showed repressed RcWUS overexpression while RcWUS had no effect on PLB morphogenesis. Furthermore, higher expression of the root stem cell marker RcWOX5 and root stem cell maintenance regulator genes RcPLT1 and RcPLT2 indicated the presence of a dedifferentiation developmental pathway in the stem cell niche of rhizoids. Viewed together, our results indicate that different cells in rhizoid tips acquired regeneration competence after induction by TDZ. A novel developmental pathway containing different cell types during PLB formation was identified by analyzing the endogenous auxin and cytokinin content. This study also provides a deeper understanding of the mechanisms underlying in vitro regeneration in Rosa.

Portfolio Preparation Tips for Teachers, Part 2

ERIC Educational Resources Information Center

Hunter-Lombardi, Brooke

2009-01-01

In the August/September 2008 issue of "SchoolArts," the author talked about strategies for helping students develop content and good working habits to support making portfolio-quality pieces. In this article, she offers some tips which focus on the importance of critique, presentation, and selection of images to end up with a polished,…

Card Access Know-How.

ERIC Educational Resources Information Center

College Planning & Management, 2000

2000-01-01

Provides tips for expanding a school's card access system and making it more productive for all concerned. Tips include the use of electronic locks for better security; the need for state-of-the-art card reader system to save money in the long run; selecting vendors to one that manufactures, installs, and supports the card system; and having…

Polarization Control with Plasmonic Antenna Tips: A Universal Approach to Optical Nanocrystallography and Vector-Field Imaging

NASA Astrophysics Data System (ADS)

Park, Kyoung-Duck; Raschke, Markus B.

2018-05-01

Controlling the propagation and polarization vectors in linear and nonlinear optical spectroscopy enables to probe the anisotropy of optical responses providing structural symmetry selective contrast in optical imaging. Here we present a novel tilted antenna-tip approach to control the optical vector-field by breaking the axial symmetry of the nano-probe in tip-enhanced near-field microscopy. This gives rise to a localized plasmonic antenna effect with significantly enhanced optical field vectors with control of both \\textit{in-plane} and \\textit{out-of-plane} components. We use the resulting vector-field specificity in the symmetry selective nonlinear optical response of second-harmonic generation (SHG) for a generalized approach to optical nano-crystallography and -imaging. In tip-enhanced SHG imaging of monolayer MoS$_2$ films and single-crystalline ferroelectric YMnO$_3$, we reveal nano-crystallographic details of domain boundaries and domain topology with enhanced sensitivity and nanoscale spatial resolution. The approach is applicable to any anisotropic linear and nonlinear optical response, and provides for optical nano-crystallographic imaging of molecular or quantum materials.

Tip aerodynamics and acoustics test: A report and data survey

NASA Technical Reports Server (NTRS)

Cross, Jeffrey L.; Watts, Michael E.

1988-01-01

In a continuing effort to understand helicopter rotor tip aerodynamics and acoustics, a flight test was conducted by NASA Ames Research Center. The test was performed using the NASA White Cobra and a set of highly instrumented blades. All aspects of the flight test instrumentation and test procedures are explained. Additionally, complete data sets for selected test points are presented and analyzed. Because of the high volume of data acquired, only selected data points are presented. However, access to the entire data set is available to the researcher on request.

Apertureless near-field optical microscopy

NASA Astrophysics Data System (ADS)

Kazantsev, D. V.; Kuznetsov, E. V.; Timofeev, S. V.; Shelaev, A. V.; Kazantseva, E. A.

2017-05-01

We discuss the operating principles of the apertureless scanning near-field optical microscope (ASNOM), in which the probe acts as a rod antenna and its electromagnetic radiation plays the role of the registered signal. The phase and amplitude of the emitted wave vary depending on the ‘grounding conditions’ of the antenna tip at the sample point under study. Weak radiation from a tiny (2-15 μm long) tip is detected using optical homo- and heterodyning and the nonlinear dependence of the tip polarizability on the tip-surface distance. The lateral resolution of ASNOMs is determined by the tip curvature radius (1- 20 nm), regardless of the wavelength (500 nm-100 μm). ASNOMs are shown to be capable of providing a surface optical map with nanometer resolution and carrying out spectral- and time-resolved measurements at a selected point on the surface.

Catalyst-induced growth of carbon nanotubes on tips of cantilevers and nanowires

DOEpatents

Lee, James Weifu; Lowndes, Douglas H.; Merkulov, Vladimir I.; Eres, Gyula; Wei, Yayi; Greenbaum, Elias; Lee, Ida

2004-06-29

A method is described for catalyst-induced growth of carbon nanotubes, nanofibers, and other nanostructures on the tips of nanowires, cantilevers, conductive micro/nanometer structures, wafers and the like. The method can be used for production of carbon nanotube-anchored cantilevers that can significantly improve the performance of scaning probe microscopy (AFM, EFM etc). The invention can also be used in many other processes of micro and/or nanofabrication with carbon nanotubes/fibers. Key elements of this invention include: (1) Proper selection of a metal catalyst and programmable pulsed electrolytic deposition of the desired specific catalyst precisely at the tip of a substrate, (2) Catalyst-induced growth of carbon nanotubes/fibers at the catalyst-deposited tips, (3) Control of carbon nanotube/fiber growth pattern by manipulation of tip shape and growth conditions, and (4) Automation for mass production.

Enhanced biomimic bactericidal surfaces by coating with positively-charged ZIF nano-dagger arrays.

PubMed

Yuan, Yuan; Zhang, Yugen

2017-10-01

Cicada wing surfaces are covered with dense patterns of nano-pillar structure that prevent bacterial growth by rupturing adhered microbial cells. To mimic the natural nano-pillar structure, we developed a general and simple method to grow metal organic framework (MOF) nano-dagger arrays on a wide range of surfaces. These nano-daggers possess high bactericidal activity, with log reduction >7 for Escherichia coli and Staphylococcus aureus. It was hypothesized that the positively-charged ZIF-L nano-dagger surfaces enhance bacterial cell adhesion, facilitating selective and efficient bacteria killing by the rigid and sharp nano-dagger tips. This research provides a safe and clean antimicrobial surface technology which does not require external chemicals and will not cause drug resistance. Copyright © 2017 Elsevier Inc. All rights reserved.

Trans-jugular intrahepatic porto-systemic shunt placement for refractory ascites: a ‘real-world’ UK health economic evaluation

PubMed Central

Parker, Matthew J; Guha, Neil; Stedman, Brian; Hacking, Nigel; Wright, Mark

2013-01-01

Objective To assess the benefit of trans-jugular intrahepatic porto-systemic shunt (TIPS) placement for refractory ascites. Design A retrospective observational study of all patients undergoing TIPS for refractory ascites in our hospital between 2003 and 2012. Setting Secondary care. Patients Cirrhotic patients with refractory ascites. Main outcome measures We examined direct real-world (National Health Service) health related costs in the year before and after the TIPS procedure took place. Data were collected relating to the need for reintervention and hepatic encephalopathy. Results Data were available for 24 patients who underwent TIPS for refractory ascites (86% of eligible patients). TIPS was technically successful in all cases. Mean number of bed days in the year prior to TIPS was 30.3 and 14.3 in the year following (p=0.005). No patient had ascites at the end of the year after the TIPS with less requirement for paracentesis over the course of the year (p<0.001). Mean reduction in cost was £2759 per patient. TIPS was especially cost-effective in patients requiring between 6 and 12 drains per year with a mean saving of £9204 per patient. Conclusions TIPS is both a clinically effective and economically advantageous therapeutic option for selected patients with refractory ascites. PMID:28839725

Trans-jugular intrahepatic porto-systemic shunt placement for refractory ascites: a 'real-world' UK health economic evaluation.

PubMed

Parker, Matthew J; Guha, Neil; Stedman, Brian; Hacking, Nigel; Wright, Mark

2013-07-01

To assess the benefit of trans-jugular intrahepatic porto-systemic shunt (TIPS) placement for refractory ascites. A retrospective observational study of all patients undergoing TIPS for refractory ascites in our hospital between 2003 and 2012. Secondary care. Cirrhotic patients with refractory ascites. We examined direct real-world (National Health Service) health related costs in the year before and after the TIPS procedure took place. Data were collected relating to the need for reintervention and hepatic encephalopathy. Data were available for 24 patients who underwent TIPS for refractory ascites (86% of eligible patients). TIPS was technically successful in all cases. Mean number of bed days in the year prior to TIPS was 30.3 and 14.3 in the year following (p=0.005). No patient had ascites at the end of the year after the TIPS with less requirement for paracentesis over the course of the year (p<0.001). Mean reduction in cost was £2759 per patient. TIPS was especially cost-effective in patients requiring between 6 and 12 drains per year with a mean saving of £9204 per patient. TIPS is both a clinically effective and economically advantageous therapeutic option for selected patients with refractory ascites.

Stable transformation via particle bombardment in two different soybean regeneration systems.

PubMed

Sato, S; Newell, C; Kolacz, K; Tredo, L; Finer, J; Hinchee, M

1993-05-01

The Biolistics(®) particle delivery system for the transformation of soybean (Glycine max L. Merr.) was evaluated in two different regeneration systems. The first system was multiple shoot proliferation from shoot tips obtained from immature zygotic embryos of the cultivar Williams 82, and the second was somatic embryogenesis from a long term proliferative suspension culture of the cultivar Fayette. Bombardment of shoot tips with tungsten particles, coated with precipitated DNA containing the gene for β-glucuronidase (GUS), produced GUS-positive sectors in 30% of the regenerated shoots. However, none of the regenerants which developed into plants continued to produce GUS positive tissue. Bombardment of embryogenic suspension cultures produced GUS positive globular somatic embryos which proliferated into GUS positive somatic embryos and plants. An average of 4 independent transgenic lines were generated per bombarded flask of an embryogenic suspension. Particle bombardment delivered particles into the first two cell layers of either shoot tips or somatic embryos. Histological analysis indicated that shoot organogenesis appeared to involve more than the first two superficial cell layers of a shoot tip, while somatic embryo proliferation occurred from the first cell layer of existing somatic embryos. The different transformation results obtained with these two systems appeared to be directly related to differences in the cell types which were responsible for regeneration and their accessibility to particle penetration.

Genotoxicity of fenpropathrin and fenitrothion on root tip cells of Vicia faba.

PubMed

Bu, N; Wang, S H; Yu, C M; Zhang, Y; Ma, C Y; Li, X M; Ma, L J

2011-11-01

The genotoxicity of fenpropathrin and fenitrothion on root tip cells of Vicia faba was studied. The symptoms were investigated about the mitotic index, the micronucleus frequency and chromosomal aberration frequency of root tip cells of Vicia faba which were induced by different concentrations of fenpropathrin and fenitrothion (1 × 10(-10)-1 × 10(-2) g L(-1)). Results showed that fenpropathrin and fenitrothion could induce the micronucleus of root tip cells of Vicia faba. It occurred in a dose-dependent manner. Peaks were observed at 1 × 10( -6) g L(-1) fenpropathrin and 1 × 10(-4) g L(-1) fenitrothion, and micronucleus frequency reached 14.587 ± 1.511‰ and 14.164 ± 1.623‰, respectively. From 1 × 10(-10) g L(-1) to 1 × 10( -6) g L(-1) fenpropathrin and 1 × 10(-4) g L(-1) fenitrothion, the micronucleus frequency increased with the increase of the concentrations, but beyond this range, the micronucleus frequency decreased with the further increase of the concentrations. A similar trend was observed for mitotic index. Moreover, fenpropathrin and fenitrothion could induce various types of chromosome aberration, such as lagging chromosomes, chromosome fragment, chromosome bridge, multipolar, nuclear buds, karyorrhexis, etc.

Experimental study of main rotor tip geometry and tail rotor interactions in hover. Volume 1. Text and figures

NASA Technical Reports Server (NTRS)

Balch, D. T.; Lombardi, J.

1985-01-01

A model scale hover test was conducted in the Sikorsky Aircraft Model rotor hover Facility to identify and quantify the impact of the tail rotor on the demonstrated advantages of advanced geometry tip configurations. The test was conducted using the Basic Model Test Rig and two scaled main rotor systems, one representing a 1/5.727 scale UH-60A BLACK HAWK and the others a 1/4.71 scale S-76. Eight alternate rotor tip configurations were tested, 3 on the BLACK HAWK rotor and 6 on the S-76 rotor. Four of these tips were then selected for testing in close proximity to an operating tail rotor (operating in both tractor and pusher modes) to determine if the performance advantages that could be obtained from the use of advanced geometry tips in a main rotor only environment would still exist in the more complex flow field involving a tail rotor. The test showed that overall the tail rotor effects on the advanced tip configurations tested are not substantially different from the effects on conventional tips.

Aeroelastic considerations for torsionally soft rotors

NASA Technical Reports Server (NTRS)

Mantay, W. R.; Yeager, W. T., Jr.

1986-01-01

A research study was initiated to systematically determine the impact of selected blade tip geometric parameters on conformable rotor performance and loads characteristics. The model articulated rotors included baseline and torsionally soft blades with interchangeable tips. Seven blade tip designs were evaluated on the baseline rotor and six tip designs were tested on the torsionally soft blades. The designs incorporated a systemmatic variation in geometric parameters including sweep, taper, and anhedral. The rotors were evaluated in the NASA Langley Transonic Dynamics Tunnel at several advance ratios, lift and propulsive force values, and tip Mach numbers. A track sensitivity study was also conducted at several advance ratios for both rotors. Based on the test results, tip parameter variations generated significant rotor performance and loads differences for both baseline and torsionally soft blades. Azimuthal variation of elastic twist generated by variations in the tip parameters strongly correlated with rotor performance and loads, but the magnitude of advancing blade elastic twist did not. In addition, fixed system vibratory loads and rotor track for potential conformable rotor candidates appears very sensitive to parametric rotor changes.

Polymer scaffolds with no skin-effect for tissue engineering applications fabricated by thermally induced phase separation.

PubMed

Kasoju, Naresh; Kubies, Dana; Sedlačík, Tomáš; Janoušková, Olga; Koubková, Jana; Kumorek, Marta M; Rypáček, František

2016-01-11

Thermally induced phase separation (TIPS) based methods are widely used for the fabrication of porous scaffolds for tissue engineering and related applications. However, formation of a less-/non-porous layer at the scaffold's outer surface at the air-liquid interface, often known as the skin-effect, restricts the cell infiltration inside the scaffold and therefore limits its efficacy. To this end, we demonstrate a TIPS-based process involving the exposure of the just quenched poly(lactide-co-caprolactone):dioxane phases to the pure dioxane for a short time while still being under the quenching strength, herein after termed as the second quenching (2Q). Scanning electron microscopy, mercury intrusion porosimetry and contact angle analysis revealed a direct correlation between the time of 2Q and the gradual disappearance of the skin, followed by the widening of the outer pores and the formation of the fibrous filaments over the surface, with no effect on the internal pore architecture and the overall porosity of scaffolds. The experiments at various quenching temperatures and polymer concentrations revealed the versatility of 2Q in removing the skin. In addition, the in vitro cell culture studies with the human primary fibroblasts showed that the scaffolds prepared by the TIPS based 2Q process, with the optimal exposure time, resulted in a higher cell seeding and viability in contrast to the scaffolds prepared by the regular TIPS. Thus, TIPS including the 2Q step is a facile, versatile and innovative approach to fabricate the polymer scaffolds with a skin-free and fully open porous surface morphology for achieving a better cell response in tissue engineering and related applications.

Cell population structure prior to bifurcation predicts efficiency of directed differentiation in human induced pluripotent cells

PubMed Central

Bargaje, Rhishikesh; Trachana, Kalliopi; Shelton, Martin N.; McGinnis, Christopher S.; Zhou, Joseph X.; Chadick, Cora; Cook, Savannah; Cavanaugh, Christopher; Huang, Sui; Hood, Leroy

2017-01-01

Steering the differentiation of induced pluripotent stem cells (iPSCs) toward specific cell types is crucial for patient-specific disease modeling and drug testing. This effort requires the capacity to predict and control when and how multipotent progenitor cells commit to the desired cell fate. Cell fate commitment represents a critical state transition or “tipping point” at which complex systems undergo a sudden qualitative shift. To characterize such transitions during iPSC to cardiomyocyte differentiation, we analyzed the gene expression patterns of 96 developmental genes at single-cell resolution. We identified a bifurcation event early in the trajectory when a primitive streak-like cell population segregated into the mesodermal and endodermal lineages. Before this branching point, we could detect the signature of an imminent critical transition: increase in cell heterogeneity and coordination of gene expression. Correlation analysis of gene expression profiles at the tipping point indicates transcription factors that drive the state transition toward each alternative cell fate and their relationships with specific phenotypic readouts. The latter helps us to facilitate small molecule screening for differentiation efficiency. To this end, we set up an analysis of cell population structure at the tipping point after systematic variation of the protocol to bias the differentiation toward mesodermal or endodermal cell lineage. We were able to predict the proportion of cardiomyocytes many days before cells manifest the differentiated phenotype. The analysis of cell populations undergoing a critical state transition thus affords a tool to forecast cell fate outcomes and can be used to optimize differentiation protocols to obtain desired cell populations. PMID:28167799

Cell population structure prior to bifurcation predicts efficiency of directed differentiation in human induced pluripotent cells.

PubMed

Bargaje, Rhishikesh; Trachana, Kalliopi; Shelton, Martin N; McGinnis, Christopher S; Zhou, Joseph X; Chadick, Cora; Cook, Savannah; Cavanaugh, Christopher; Huang, Sui; Hood, Leroy

2017-02-28

Steering the differentiation of induced pluripotent stem cells (iPSCs) toward specific cell types is crucial for patient-specific disease modeling and drug testing. This effort requires the capacity to predict and control when and how multipotent progenitor cells commit to the desired cell fate. Cell fate commitment represents a critical state transition or "tipping point" at which complex systems undergo a sudden qualitative shift. To characterize such transitions during iPSC to cardiomyocyte differentiation, we analyzed the gene expression patterns of 96 developmental genes at single-cell resolution. We identified a bifurcation event early in the trajectory when a primitive streak-like cell population segregated into the mesodermal and endodermal lineages. Before this branching point, we could detect the signature of an imminent critical transition: increase in cell heterogeneity and coordination of gene expression. Correlation analysis of gene expression profiles at the tipping point indicates transcription factors that drive the state transition toward each alternative cell fate and their relationships with specific phenotypic readouts. The latter helps us to facilitate small molecule screening for differentiation efficiency. To this end, we set up an analysis of cell population structure at the tipping point after systematic variation of the protocol to bias the differentiation toward mesodermal or endodermal cell lineage. We were able to predict the proportion of cardiomyocytes many days before cells manifest the differentiated phenotype. The analysis of cell populations undergoing a critical state transition thus affords a tool to forecast cell fate outcomes and can be used to optimize differentiation protocols to obtain desired cell populations.

Ergonomic Factors Related to Drop-Off Detection With the Long Cane: Effects of Cane Tips and Techniques

PubMed Central

Kim, Dae Shik; Wall Emerson, Robert S.; Curtis, Amy B.

2010-01-01

Objective This study examined the effect of cane tips and cane techniques on drop-off detection with the long cane. Background Blind pedestrians depend on a long cane to detect drop-offs. Missing a drop-off may result in falls or collision with moving vehicles in the street. Although cane tips appear to affect a cane user’s ability to detect drop-offs, few experimental studies have examined such effect. Method A repeated-measures design with block randomization was used for the study. Participants were 17 adults who were legally blind and had no other disabilities. Participants attempted to detect the drop-offs of varied depths using different cane tips and cane techniques. Results Drop-off detection rates were similar between the marshmallow tip (77.0%) and the marshmallow roller tip (79.4%) when both tips were used with the constant contact technique, p = .294. However, participants detected drop-offs at a significantly higher percentage when they used the constant contact technique with the marshmallow roller tip (79.4%) than when they used the two-point touch technique with the marshmallow tip (63.2%), p < .001. Conclusion The constant contact technique used with a marshmallow roller tip (perceived as a less advantageous tip) was more effective than the two-point touch technique used with a marshmallow tip (perceived as a more advantageous tip) in detecting drop-offs. Application The findings of the study may help cane users and orientation and mobility specialists select appropriate cane techniques and cane tips in accordance with the cane user’s characteristics and the nature of the travel environment. PMID:21077566

Crack-tips enriched platinum-copper superlattice nanoflakes as highly efficient anode electrocatalysts for direct methanol fuel cells.

PubMed

Zheng, Lijun; Yang, Dachi; Chang, Rong; Wang, Chengwen; Zhang, Gaixia; Sun, Shuhui

2017-07-06

We have developed "crack-tips" and "superlattice" enriched Pt-Cu nanoflakes (NFs), benefiting from the synergetic effects of "crack-tips" and "superlattice crystals"; the Pt-Cu NFs exhibit 4 times higher mass activity, 6 times higher specific activity and 6 times higher stability than those of the commercial Pt/C catalyst, respectively. Meanwhile, the Pt-Cu NFs show more enhanced CO tolerance than the commercial Pt/C catalyst.

Proximal hypospadias: A persistent challenge. Single institution outcome analysis of three surgical techniques over a 10-year period.

PubMed

Pippi Salle, J L; Sayed, S; Salle, A; Bagli, D; Farhat, W; Koyle, M; Lorenzo, A J

2016-02-01

The optimal treatment of proximal hypospadias remains controversial. Several techniques have been described, but the best approach remains unsettled. To evaluate and compare the complication rates of proximal hypospadias with and without ventral curvature (VC), according to three different surgical techniques: tubularized incised plate (TIP) uretroplasty, dorsal inlay graft TIP (DIG), and staged preputial repair (SR). It was hypothesized that SR performs better than TIP and DIG for proximal hypospadias. Single-center, retrospective chart review of all patients with primary proximal hypospadias reconstructed between 2003 and 2013. The DIG was selectively employed in cases with narrow urethral plate (UP) and deficient spongiosum. Extensive urethral plate (UP) mobilization (UPM), dorsal plication (DP) and/or deep transverse incisions of tunica albuginea (DTITA) were selectively performed when attempting to spare transecting the UP. Division of UP and SR was favored in cases with severe VC (>50°), which was often concurrently managed with DTITA if intrinsic curvature was present. For SR, tubularization of the graft was performed 6 months later. A total of 140 patients were included. Tubularized incised plate (TIP), DIG, and SR techniques were performed in 57, 23, and 60 patients, respectively. The TIP and DIG techniques achieved similar success rates, although DIG was performed in cases of narrow and spongiosum-deficient plates. Reoperation rates with TIP and DIG techniques was 52.6% and 52.1% (NS). Urethro-cutaneous fistulas were seen in 31.5% and 13% of TIP and DIG techniques, respectively. Staged repair accomplished better results than both TIP and DIG techniques, despite being performed in the most unfavorable cases (reoperation rate 28%). After technical modifications, the DIG technique achieved similar outcomes of SR. Proximal hypospadias remains challenging, regardless of the technique utilized for its repair. Urethro-cutaneous fistulas were more commonly seen after long TIP repairs. Approximately half of the patients undergoing long TIP and DIG procedures needed re-intervention, although the percentage decreased significantly with late modifications in the DIG group. Recurrence of VC after TIP and DIG techniques seemed to be a significant and under-reported complication. Staged repairs, despite being performed for the most severe cases, resulted in overall better outcomes. Copyright © 2015 Journal of Pediatric Urology Company. Published by Elsevier Ltd. All rights reserved.

Contact-induced apical asymmetry drives the thigmotropic responses of Candida albicans hyphae.

PubMed

Thomson, Darren D; Wehmeier, Silvia; Byfield, FitzRoy J; Janmey, Paul A; Caballero-Lima, David; Crossley, Alison; Brand, Alexandra C

2015-03-01

Filamentous hyphae of the human pathogen, Candida albicans, invade mucosal layers and medical silicones. In vitro, hyphal tips reorient thigmotropically on contact with small obstacles. It is not known how surface topography is sensed but hyphae lacking the cortical marker, Rsr1/Bud1, are unresponsive. We show that, on surfaces, the morphology of hyphal tips and the position of internal polarity protein complexes are asymmetrically skewed towards the substratum and biased towards the softer of two surfaces. In nano-fabricated chambers, the Spitzenkörper (Spk) responded to touch by translocating across the apex towards the point of contact, where its stable maintenance correlated with contour-following growth. In the rsr1Δ mutant, the position of the Spk meandered and these responses were attenuated. Perpendicular collision caused lateral Spk oscillation within the tip until after establishment of a new growth axis, suggesting Spk position does not predict the direction of growth in C. albicans. Acute tip reorientation occurred only in cells where forward growth was countered by hyphal friction sufficient to generate a tip force of ∼ 8.7 μN (1.2 MPa), more than that required to penetrate host cell membranes. These findings suggest mechanisms through which the organization of hyphal tip growth in C. albicans facilitates the probing, penetration and invasion of host tissue. © 2014 The Authors. Cellular Microbiology published by John Wiley & Sons Ltd.

Contact-induced apical asymmetry drives the thigmotropic responses of Candida albicans hyphae

PubMed Central

Thomson, Darren D; Wehmeier, Silvia; Byfield, FitzRoy J; Janmey, Paul A; Caballero-Lima, David; Crossley, Alison; Brand, Alexandra C

2015-01-01

Filamentous hyphae of the human pathogen, Candida albicans, invade mucosal layers and medical silicones. In vitro, hyphal tips reorient thigmotropically on contact with small obstacles. It is not known how surface topography is sensed but hyphae lacking the cortical marker, Rsr1/Bud1, are unresponsive. We show that, on surfaces, the morphology of hyphal tips and the position of internal polarity protein complexes are asymmetrically skewed towards the substratum and biased towards the softer of two surfaces. In nano-fabricated chambers, the Spitzenkörper (Spk) responded to touch by translocating across the apex towards the point of contact, where its stable maintenance correlated with contour-following growth. In the rsr1Δ mutant, the position of the Spk meandered and these responses were attenuated. Perpendicular collision caused lateral Spk oscillation within the tip until after establishment of a new growth axis, suggesting Spk position does not predict the direction of growth in C. albicans. Acute tip reorientation occurred only in cells where forward growth was countered by hyphal friction sufficient to generate a tip force of ∼ 8.7 μN (1.2 MPa), more than that required to penetrate host cell membranes. These findings suggest mechanisms through which the organization of hyphal tip growth in C. albicans facilitates the probing, penetration and invasion of host tissue. PMID:25262778

Mitochondrial anchorage and fusion contribute to mitochondrial inheritance and quality control in the budding yeast Saccharomyces cerevisiae

PubMed Central

Higuchi-Sanabria, Ryo; Charalel, Joseph K.; Viana, Matheus P.; Garcia, Enrique J.; Sing, Cierra N.; Koenigsberg, Andrea; Swayne, Theresa C.; Vevea, Jason D.; Boldogh, Istvan R.; Rafelski, Susanne M.; Pon, Liza A.

2016-01-01

Higher-functioning mitochondria that are more reduced and have less ROS are anchored in the yeast bud tip by the Dsl1-family protein Mmr1p. Here we report a role for mitochondrial fusion in bud-tip anchorage of mitochondria. Fluorescence loss in photobleaching (FLIP) and network analysis experiments revealed that mitochondria in large buds are a continuous reticulum that is physically distinct from mitochondria in mother cells. FLIP studies also showed that mitochondria that enter the bud can fuse with mitochondria that are anchored in the bud tip. In addition, loss of fusion and mitochondrial DNA (mtDNA) by deletion of mitochondrial outer or inner membrane fusion proteins (Fzo1p or Mgm1p) leads to decreased accumulation of mitochondria at the bud tip and inheritance of fitter mitochondria by buds compared with cells with no mtDNA. Conversely, increasing the accumulation and anchorage of mitochondria in the bud tip by overexpression of MMR1 results in inheritance of less-fit mitochondria by buds and decreased replicative lifespan and healthspan. Thus quantity and quality of mitochondrial inheritance are ensured by two opposing processes: bud-tip anchorage by mitochondrial fusion and Mmr1p, which favors bulk inheritance; and quality control mechanisms that promote segregation of fitter mitochondria to the bud. PMID:26764088

Biometric ratio in estimating widths of maxillary anterior teeth derived after correlating anthropometric measurements with dental measurements.

PubMed

Kini, Ashwini Y; Angadi, Gangadhar S

2013-06-01

To correlate dental measurements i.e. combined mesiodistal width of six maxillary anterior teeth with facial measurements i.e. inner canthal distance, interpupillary distance and intercommissural width and acquire a biometric ratio to serve as a preliminary guide in selection of the maxillary anterior teeth. In the absence of pre-extraction records, the resultant denture can lead to patient dissatisfaction towards the aesthetic appeal of their dentures. The maxillary anterior teeth play a pivotal role in denture aesthetics. Various techniques and biometric ratios have been described in literature for selection of the maxillary anteriors. This study derives a biometric ratio for the same, obtained after correlating anthropometric measurements with dental measurements. Two standardized digital photographs of the face were generated; one, when the facial muscles were relaxed and the other, when the subject was smiling; thereby, revealing the maxillary anterior teeth upto the canine tip. Inner canthal distance, interpupillary distance, intercommissural distance, distance between the tips of the maxillary canines and distance between the distal surfaces of the canines were measured. On the cast, the distance between tips of maxillary canines and distance between distal surfaces of maxillary canines were noted. The data was analysed using Spearman's rank correlation coefficient. A high correlation was found between the intercommissural measurement with distance between the tips of the canines on the photograph and between the tips of the canines on the cast with the interpupillary distance, giving a biometric ratio of 1:1.35 and 1:1.41 respectively. The least correlation was between the inner canthal distance and the tips of the canines measured on the photograph. Extra oral anthropometric measurements of the interpupillary distances and the intercommissural distances with the help of standardised photographs can help us determine the combined widths of the anterior teeth accurately, thus aiding their selection in the absence of pre-extraction records. © 2012 John Wiley & Sons A/S and The Gerodontology Society. Published by John Wiley & Sons Ltd.

Role of orbital overlap in atomic manipulation

NASA Astrophysics Data System (ADS)

Jarvis, Sam; Sweetman, Adam; Bamidele, Joseph; Kantorovich, Lev; Moriarty, Philip

2012-06-01

We conduct ab initio simulations illustrating that the ability to achieve atomic manipulation using a dynamic force microscope depends on the precise orientation of the dangling bond(s) at the tip apex and their charge density with respect to those of surface atoms. Using the Si(100)-c(4×2) surface as a prototype, we demonstrate that it is possible to select tip apices capable of performing atomic manipulation tasks which are unachievable using another choice of apex. Specific tip apices can be identified via examination of F(z) curves taken at different lateral positions.

Methods for destriping Landsat Thematic Mapper images - A feasibility study for an online destriping process in the Thematic Mapper Image Processing System (TIPS)

NASA Technical Reports Server (NTRS)

Poros, D. J.; Peterson, C. J.

1985-01-01

Methods for destriping TM images and results of the application of these methods to selected TM scenes with sensor and scan striping, which was not removed by the radiometric correction during the TM Archive Generation Phase in TIPS, are presented. These methods correct only for gain and offset differences between detectors over many image lines and do not consider within-line effects. The feasibility of implementing a destriping process online in TIPS is also described.

Selected advanced aerodynamics and active controls technology concepts development on a derivative B-747 aircraft

NASA Technical Reports Server (NTRS)

1983-01-01

Analytical design and wind tunnel test evaluations covering the feasibility of applying wing tip extensions, winglets, and active control wing had alleviation to the model B747 are described. Aerodynamic improvement offered by wing tip extension and winglet individually, and the combined aerodynamic and weight improvements when wing load alleviation is combined with the tip extension or the winglet are evaluated. Results are presented in the form of incremental effects on weight mission range, fuel usage, cost, and airline operating economics.

Visible light-sensitive APTES-bound ZnO nanowire toward a potent nanoinjector sensing biomolecules in a living cell

NASA Astrophysics Data System (ADS)

Lee, Jooran; Choi, Sunyoung; Bae, Seon Joo; Yoon, Seok Min; Choi, Joon Sig; Yoon, Minjoong

2013-10-01

Nanoscale cell injection techniques combined with nanoscopic photoluminescence (PL) spectroscopy have been important issues in high-resolution optical biosensing, gene and drug delivery and single-cell endoscopy for medical diagnostics and therapeutics. However, the current nanoinjectors remain limited for optical biosensing and communication at the subwavelength level, requiring an optical probe such as semiconductor quantum dots, separately. Here, we show that waveguided red emission is observed at the tip of a single visible light-sensitive APTES-modified ZnO nanowire (APTES-ZnO NW) and it exhibits great enhancement upon interaction with a complementary sequence-based double stranded (ds) DNA, whereas it is not significantly affected by non-complementary ds DNA. Further, the tip of a single APTES-ZnO NW can be inserted into the subcellular region of living HEK 293 cells without significant toxicity, and it can also detect the enhancement of the tip emission from subcellular regions with high spatial resolution. These results indicate that the single APTES-ZnO NW would be useful as a potent nanoinjector which can guide visible light into intracellular compartments of mammalian cells, and can also detect nanoscopic optical signal changes induced by interaction with the subcellular specific target biomolecules without separate optical probes.Nanoscale cell injection techniques combined with nanoscopic photoluminescence (PL) spectroscopy have been important issues in high-resolution optical biosensing, gene and drug delivery and single-cell endoscopy for medical diagnostics and therapeutics. However, the current nanoinjectors remain limited for optical biosensing and communication at the subwavelength level, requiring an optical probe such as semiconductor quantum dots, separately. Here, we show that waveguided red emission is observed at the tip of a single visible light-sensitive APTES-modified ZnO nanowire (APTES-ZnO NW) and it exhibits great enhancement upon interaction with a complementary sequence-based double stranded (ds) DNA, whereas it is not significantly affected by non-complementary ds DNA. Further, the tip of a single APTES-ZnO NW can be inserted into the subcellular region of living HEK 293 cells without significant toxicity, and it can also detect the enhancement of the tip emission from subcellular regions with high spatial resolution. These results indicate that the single APTES-ZnO NW would be useful as a potent nanoinjector which can guide visible light into intracellular compartments of mammalian cells, and can also detect nanoscopic optical signal changes induced by interaction with the subcellular specific target biomolecules without separate optical probes. Electronic supplementary information (ESI) available: Synthesis of APTES-modified ZnO nanowires, DNA functionalization and spectroscopic measurements with additional fluorescence image ad fluorescence decay times, cell culture, injection of a single nanowire into living cells, subcellular imaging and determination of cytotoxicity. See DOI: 10.1039/c3nr03042c

The passive cable properties of hair cell stereocilia and their contribution to somatic capacitance measurements.

PubMed

Breneman, Kathryn D; Highstein, Stephen M; Boyle, Richard D; Rabbitt, Richard D

2009-01-01

Somatic measurements of whole-cell capacitance are routinely used to understand physiologic events occurring in remote portions of cells. These studies often assume the intracellular space is voltage-clamped. We questioned this assumption in auditory and vestibular hair cells with respect to their stereocilia based on earlier studies showing that neurons, with radial dimensions similar to stereocilia, are not always isopotential under voltage-clamp. To explore this, we modeled the stereocilia as passive cables with transduction channels located at their tips. We found that the input capacitance measured at the soma changes when the transduction channels at the tips of the stereocilia are open compared to when the channels are closed. The maximum capacitance is felt with the transducer closed but will decrease as the transducer opens due to a length-dependent voltage drop along the stereocilium length. This potential drop is proportional to the intracellular resistance and stereocilium tip conductance and can produce a maximum capacitance error on the order of fF for single stereocilia and pF for the bundle.

Composition and Antioxidant Activity of Water-Soluble Polysaccharides from Tuber indicum

PubMed Central

Luo, Qiang; Zhang, Jie; Yan, Liang; Tang, Yuanlin; Ding, Xiang; Yang, Zhirong

2011-01-01

Abstract Crude water-soluble Chinese truffle Tuber indicum polysaccharide (TIP) was extracted from the fruiting bodies with water and then successively purified by DEAE–cellulose 52 and Sephadex G-100 column chromatography, yielding two major polysaccharide fractions: TIP1-1 and TIP2-1. High-performance gel permeation chromatography analysis showed that the average molecular sizes of TIP1-1 and TIP2-1 were approximately 1.75×104 Da and 5.73×103 Da, respectively. Monosaccharide component analysis by gas chromatography indicated that TIP1-1 was composed of mannose, glucose, galactose, and rhamannose in the respective molar ratio of 3.93:1.24:0.75:1.26 and that TIP2-1 contained mannose, glucose, and arabinose in the respective molar ratio of 5.27:1.44:0.43. The antioxidant activity analyses revealed that TIP1-1 and TIP2-1 possessed considerable antioxidant activity. Compared with TIP1-1, which has a higher molecular weight and contains no uronic acid, TIP2-1 exhibited a protective effect on PC12 cells injured by H2O2 and a higher scavenging activity against free radicals. The relative effects of the lower molecular size, the presence of uronic acid, and the antioxidant activity of TIP2-1 appear to be significant. Accordingly, the Chinese truffle T. indicum might serve as an effective antioxidative healthcare food and source of natural antioxidants. PMID:21877953

SERCA directs cell migration and branching across species and germ layers

PubMed Central

Lansdale, Nick; Navarro, Sonia; Truong, Thai V.; Bower, Dan J.; Featherstone, Neil C.; Connell, Marilyn G.; Al Alam, Denise; Frey, Mark R.; Trinh, Le A.; Fernandez, G. Esteban; Warburton, David; Fraser, Scott E.; Bennett, Daimark; Jesudason, Edwin C.

2017-01-01

ABSTRACT Branching morphogenesis underlies organogenesis in vertebrates and invertebrates, yet is incompletely understood. Here, we show that the sarco-endoplasmic reticulum Ca2+ reuptake pump (SERCA) directs budding across germ layers and species. Clonal knockdown demonstrated a cell-autonomous role for SERCA in Drosophila air sac budding. Live imaging of Drosophila tracheogenesis revealed elevated Ca2+ levels in migratory tip cells as they form branches. SERCA blockade abolished this Ca2+ differential, aborting both cell migration and new branching. Activating protein kinase C (PKC) rescued Ca2+ in tip cells and restored cell migration and branching. Likewise, inhibiting SERCA abolished mammalian epithelial budding, PKC activation rescued budding, while morphogens did not. Mesoderm (zebrafish angiogenesis) and ectoderm (Drosophila nervous system) behaved similarly, suggesting a conserved requirement for cell-autonomous Ca2+ signaling, established by SERCA, in iterative budding. PMID:28821490

Efficacy of TACE in TIPS Patients: Comparison of Treatment Response to Chemoembolization for Hepatocellular Carcinoma in Patients With and Without a Transjugular Intrahepatic Portosystemic Shunt

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

Kuo, Yuo-Chen, E-mail: yuo-chen.kuo@ucsf.edu; Kohi, Maureen P., E-mail: maureen.kohi@ucsf.edu; Naeger, David M., E-mail: david.naeger@ucsf.edu

Purpose: To compare treatment response after transarterial chemoembolization (TACE) for hepatocellular carcinoma (HCC) in patients with and without a transjugular intrahepatic portosystemic shunt (TIPS). Materials and Methods: A retrospective review of patients who underwent conventional TACE for HCC between January 2005 and December 2009 identified 10 patients with patent TIPS. From the same time period, 23 patients without TIPS were selected to control for comparable Model for End-Stage Liver Disease and Child-Pugh-Turcotte scores. The two groups showed similar distribution of Barcelona Clinic Liver Cancer and United Network of Organ Sharing stages. Target HCC lesions were evaluated according to the modifiedmore » response evaluation criteria in solid tumors (mRECIST) guidelines. Transplantation rate, time to tumor progression, and overall survival (OS) were documented. Results: After TACE, the rate of complete response was significantly greater in non-TIPS patients compared with TIPS patients (74 vs. 30 %, p = 0.03). Objective response rate (complete and partial response) trended greater in the non-TIPS group (83 vs. 50 %, p = 0.09). The liver transplantation rate was 80 and 74 % in the TIPS and non-TIPS groups, respectively (p = 1.0). Time to tumor progression was similar (p = 0.47) between the two groups. OS favored the non-TIPS group (p = 0.01) when censored for liver transplantation. Conclusion: TACE is less effective in achieving complete or partial response using mRECIST criteria in TIPS patients compared with those without a TIPS. Nevertheless, similar clinical outcomes may be achieved, particularly in TIPS patients who are liver-transplantation candidates.« less

Probabilistic Analysis and Design of a Raked Wing Tip for a Commercial Transport

NASA Technical Reports Server (NTRS)

Mason Brian H.; Chen, Tzi-Kang; Padula, Sharon L.; Ransom, Jonathan B.; Stroud, W. Jefferson

2008-01-01

An approach for conducting reliability-based design and optimization (RBDO) of a Boeing 767 raked wing tip (RWT) is presented. The goal is to evaluate the benefits of RBDO for design of an aircraft substructure. A finite-element (FE) model that includes eight critical static load cases is used to evaluate the response of the wing tip. Thirteen design variables that describe the thickness of the composite skins and stiffeners are selected to minimize the weight of the wing tip. A strain-based margin of safety is used to evaluate the performance of the structure. The randomness in the load scale factor and in the strain limits is considered. Of the 13 variables, the wing-tip design was controlled primarily by the thickness of the thickest plies in the upper skins. The report includes an analysis of the optimization results and recommendations for future reliability-based studies.

Apoplastic ROS production upon pollination by RbohH and RbohJ in Arabidopsis

PubMed Central

Kaya, Hidetaka; Iwano, Megumi; Takeda, Seiji; Kanaoka, Masahiro M; Kimura, Sachie; Abe, Mitsutomo; Kuchitsu, Kazuyuki

2015-01-01

Reactive oxygen species (ROS) accumulate at the tip of growing pollen tubes. In Arabidopsis, NADPH oxidases RbohH and RbohJ are localized at the plasma membrane of pollen tube tip and produce ROS in a Ca2+-dependent manner. The ROS produced by Rbohs and Ca2+ presumably play a critical role in the positive feedback regulation that maintains the tip growth. Ultrastructural cytochemical analysis revealed ROS accumulation in the apoplast/cell wall of the pollen grains on the stigmatic papillae in the wild type, but not in the rbohH rbohJ double mutant, suggesting that apoplastic ROS derived from RbohH and RbohJ are involved in pollen tube elongation into the stigmatic papillae by affecting the cell wall metabolism. PMID:25751652

[Sheng's acupuncture manipulation at bone-nearby acupoints and the academic thoughts].

PubMed

Sheng, Ji-li; Jin, Xiao-qing

2014-11-01

Sheng's acupuncture manipulation at bone-nearby acupoints is a set of needling manipulation of the chief physician of TCM, SHENG Xie-sun, summarized through his over 50 years clinical experiences and on the basis of Internal Classic. Regarding this manipulation, on the premise of acupoint selection based on syndrome differentiation, the acupoints close to bone are possibly selected and punctured, with the needle tip toward bone edge, and followed by the technique to achieve reducing purpose. Clinically, the significant immediate analgesia can be achieved in pain disorders such as headache and toothache. Professor Sheng thought, corresponding to the location of needle insertion and needling depth, the tissue layers of needle tip passing through should be considered specially. The site of needle insertion should be changeable so as to ensure the needle tip reaching the bone. This manipulation for analgesia provides a certain guide for acupuncture study, especially for the mechanism study on acupuncture analgesia.

Vacuolar biogenesis and aquaporin expression at early germination of broad bean seeds.

PubMed

Novikova, Galina V; Tournaire-Roux, Colette; Sinkevich, Irina A; Lityagina, Snejana V; Maurel, Christophe; Obroucheva, Natalie

2014-09-01

A key event in seed germination is water uptake-mediated growth initiation in embryonic axes. Vicia faba var. minor (broad bean) seeds were used for studying cell growth, vacuolar biogenesis, expression and function of tonoplast water channel proteins (aquaporins) in embryonic axes during seed imbibition, radicle emergence and growth. Hypocotyl and radicle basal cells showed vacuole restoration from protein storage vacuoles, whereas de novo vacuole formation from provacuoles was observed in cells newly produced by root meristem. cDNA fragments of seven novel aquaporin isoforms including five Tonoplast Intrinsic Proteins (TIP) from three sub-types were amplified by PCR. The expression was probed using q-RT-PCR and when possible with isoform-specific antibodies. Decreased expression of TIP3s was associated to the transformation of protein storage vacuoles to vacuoles, whereas enhanced expression of a TIP2 homologue was closely linked to the fast cell elongation. Water channel functioning checked by inhibitory test with mercuric chloride showed closed water channels prior to growth initiation and active water transport into elongating cells. The data point to a crucial role of tonoplast aquaporins during germination, especially during growth of embryonic axes, due to accelerated water uptake and vacuole enlargement resulting in rapid cell elongation. Copyright © 2014 Elsevier Masson SAS. All rights reserved.

Actin cable distribution and dynamics arising from cross-linking, motor pulling, and filament turnover

PubMed Central

Tang, Haosu; Laporte, Damien; Vavylonis, Dimitrios

2014-01-01

The growth of fission yeast relies on the polymerization of actin filaments nucleated by formin For3p, which localizes at tip cortical sites. These actin filaments bundle to form actin cables that span the cell and guide the movement of vesicles toward the cell tips. A big challenge is to develop a quantitative understanding of these cellular actin structures. We used computer simulations to study the spatial and dynamical properties of actin cables. We simulated individual actin filaments as semiflexible polymers in three dimensions composed of beads connected with springs. Polymerization out of For3p cortical sites, bundling by cross-linkers, pulling by type V myosin, and severing by cofilin are simulated as growth, cross-linking, pulling, and turnover of the semiflexible polymers. With the foregoing mechanisms, the model generates actin cable structures and dynamics similar to those observed in live-cell experiments. Our simulations reproduce the particular actin cable structures in myoVΔ cells and predict the effect of increased myosin V pulling. Increasing cross-linking parameters generates thicker actin cables. It also leads to antiparallel and parallel phases with straight or curved cables, consistent with observations of cells overexpressing α-actinin. Finally, the model predicts that clustering of formins at cell tips promotes actin cable formation. PMID:25103242

Gravisensing in single-celled systems: characean rhizoids and protonemata

NASA Astrophysics Data System (ADS)

Braun, M.

Gravitropically tip-growing cell types are attractive unicellular model systems for investigating the mechanisms and the regulation of gravitropism. Especially useful for studying the mechanisms of positive and negative gravitropic tip-growth are characean rhizoids and protonemata. They originate from the same cell type, show the same overall cell shape, cytoplasmic zonation, arrangement of actin and microtubule cytoskeleton, use statoliths for gravisensing, but show opposite gravitropism. In both cell types, actin microfilaments are complexly organized in the apical dome, where a dense spherical actin array is colocalized with spectrin-like epitopes and a unique endoplasmic reticulum aggregate, the structural center of the Spitzenkörper. The opposite gravitropic responses seem to be based on differences in the actin-organized anchorage of the Spitzenkörper and the actin-mediated transport of statoliths. In negatively gravitropic (upward bending) protonemata, the statoliths-induced drastic upward shift of the cell tip is preceded by a relocalization of dihydropyridine-binding calcium channels and of the apical calcium gradient to the upper flank (bending by bulging). Such relocalizations have not been observed in positively gravitropically responding (downward growing) rhizoids in which statoliths sedimentation is followed by differential flank growth (bending by bowing). This paper reviews the current knowledge and hypotheses on the mechanisms of the opposite gravitropic responses in characean rhizoids and protonemata.

A cell-body groove housing the new flagellum tip suggests an adaptation of cellular morphogenesis for parasitism in the bloodstream form of Trypanosoma brucei.

PubMed

Hughes, Louise; Towers, Katie; Starborg, Tobias; Gull, Keith; Vaughan, Sue

2013-12-15

Flagella are highly conserved organelles present in a wide variety of species. In Trypanosoma brucei the single flagellum is necessary for morphogenesis, cell motility and pathogenesis, and is attached along the cell body. A new flagellum is formed alongside the old during the cell division cycle. In the (insect) procyclic form, the flagella connector (FC) attaches the tip of the new flagellum to the side of the old flagellum, ensuring faithful replication of cell architecture. The FC is not present in the bloodstream form of the parasite. We show here, using new imaging techniques including serial block-face scanning electron microscopy (SBF-SEM), that the distal tip of the new flagellum in the bloodstream form is embedded within an invagination in the cell body plasma membrane, named the groove. We suggest that the groove has a similar function to the flagella connector. The groove is a mobile junction located alongside the microtubule quartet (MtQ) and occurred within a gap in the subpellicular microtubule corset, causing significant modification of microtubules during elongation of the new flagellum. It appears likely that this novel form of morphogenetic structure has evolved to withstand the hostile immune response in the mammalian blood.

Evolution of multinucleated Ashbya gossypii hyphae from a budding yeast-like ancestor.

PubMed

Schmitz, Hans-Peter; Philippsen, Peter

2011-06-01

In the filamentous ascomycete Ashbya gossypii polarity establishment at sites of germ tube and lateral branch emergence depends on homologues of Saccharomyces cerevisiae factors controlling bud site selection and bud emergence. Maintenance of polar growth involves homologues of well-known polarity factors of budding yeast. To achieve the much higher rates of sustained polar surface expansion of hyphae compared to mainly non-polarly growing yeast buds five important alterations had to evolve. Permanent presence of the polarity machinery at a confined area in the rapidly expanding hyphal tip, increased cytoplasmic space with a much enlarged ER surface for generating secretory vesicles, efficient directed transport of secretory vesicles to and accumulation at the tip, increased capacity of the exocytosis system to process these vesicles, and an efficient endocytosis system for membrane and polarity factor recycling adjacent to the zone of exocytosis. Morphological, cell biological, and molecular aspects of this evolution are discussed based on experiments performed within the past 10 y. Copyright © 2011 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

Microstructural examination of

NASA Astrophysics Data System (ADS)

Fukuoka, C.; Yoshizawa, H.; Nakagawa, Y. G.; Lapides, M. E.

1993-10-01

Fatigue tests were performed to examine how microstructural conditioning influences crack initiation and propagation in SA508 class 3 low-carbon steel. A 3-mm-long crack was introduced in compact tension (CT) fatigue test specimens under four different loads in order to obtain crack tip plastic zones at different stress intensity factor ranges, ΔK = 18, 36, 54, and 72 MPa√m. The microstructure of the plastic zones around the crack tip were examined by trans- mission electron microscopy (TEM) and selected area electron diffraction (SAD). Micro- orientation of the dislocation cells in the plastic zones of all of the CT samples increased to 4 deg from the level of an as-received sample. Four-point bending fatigue tests were performed for plate shape samples with a large cyclic strain range. The SAD value of the bending samples was also 4 deg in the damaged area where cracks already initiated at an early stage of the fatigue process. These test results indicate that the microstructural conditioning is a prerequisite for the fatigue crack initiation and propagation in SA508. These observations may lead to better under- standing of how fatigue initiation processes transit to cracks.

Noncontact Viscoelastic Imaging of Living Cells Using a Long-Needle Atomic Force Microscope with Dual-Frequency Modulation

NASA Astrophysics Data System (ADS)

Guan, Dongshi; Charlaix, Elisabeth; Qi, Robert Z.; Tong, Penger

2017-10-01

Imaging of surface topography and elasticity of living cells can provide insight into the roles played by the cells' volumetric and mechanical properties and their response to external forces in regulating the essential cellular events and functions. Here, we report a unique technique of noncontact viscoelastic imaging of live cells using atomic force microscopy (AFM) with a long-needle glass probe. Because only the probe tip is placed in a liquid medium near the cell surface, the AFM cantilever in air functions well under dual-frequency modulation, retaining its high-quality resonant modes. The probe tip interacts with the cell surface through a minute hydrodynamic flow in the nanometer-thin gap region between them without physical contact. Quantitative measurements of the cell height, volume, and Young's modulus are conducted simultaneously. The experiment demonstrates that the long-needle AFM has a wide range of applications in the study of cell mechanics.

Dynamics of cell wall elasticity pattern shapes the cell during yeast mating morphogenesis

PubMed Central

Goldenbogen, Björn; Giese, Wolfgang; Hemmen, Marie; Uhlendorf, Jannis; Herrmann, Andreas

2016-01-01

The cell wall defines cell shape and maintains integrity of fungi and plants. When exposed to mating pheromone, Saccharomyces cerevisiae grows a mating projection and alters in morphology from spherical to shmoo form. Although structural and compositional alterations of the cell wall accompany shape transitions, their impact on cell wall elasticity is unknown. In a combined theoretical and experimental approach using finite-element modelling and atomic force microscopy (AFM), we investigated the influence of spatially and temporally varying material properties on mating morphogenesis. Time-resolved elasticity maps of shmooing yeast acquired with AFM in vivo revealed distinct patterns, with soft material at the emerging mating projection and stiff material at the tip. The observed cell wall softening in the protrusion region is necessary for the formation of the characteristic shmoo shape, and results in wider and longer mating projections. The approach is generally applicable to tip-growing fungi and plants cells. PMID:27605377

The GDNF Target Vsnl1 Marks the Ureteric Tip

PubMed Central

Ola, Roxana; Jakobson, Madis; Kvist, Jouni; Perälä, Nina; Kuure, Satu; Braunewell, Karl-Heinz; Bridgewater, Darren; Rosenblum, Norman D.; Chilov, Dmitri; Immonen, Tiina; Sainio, Kirsi

2011-01-01

Glial cell line-derived neurotrophic factor (GDNF) is indispensable for ureteric budding and branching. If applied exogenously, GDNF promotes ectopic ureteric buds from the Wolffian duct. Although several downstream effectors of GDNF are known, the identification of early response genes is incomplete. Here, microarray screening detected several GDNF-regulated genes in the Wolffian duct, including Visinin like 1 (Vsnl1), which encodes a neuronal calcium-sensor protein. We observed renal Vsnl1 expression exclusively in the ureteric epithelium, but not in Gdnf-null kidneys. In the tissue culture of Gdnf-deficient kidney primordium, exogenous GDNF and alternative bud inducers (FGF7 and follistatin) restored Vsnl1 expression. Hence, Vsnl1 characterizes the tip of the ureteric bud epithelium regardless of the inducer. In the tips, Vsnl1 showed a mosaic expression pattern that was mutually exclusive with β-catenin transcriptional activation. Vsnl1 was downregulated in both β-catenin-stabilized and β-catenin-deficient kidneys. Moreover, in a mouse collecting duct cell line, Vsnl1 compromised β-catenin stability, suggesting a counteracting relationship between Vsnl1 and β-catenin. In summary, Vsnl1 marks ureteric bud tips in embryonic kidneys, and its mosaic pattern demonstrates a heterogeneity of cell types that may be critical for normal ureteric branching. PMID:21289216

CLASPs are required for proper microtubule localization of End-binding proteins

PubMed Central

Grimaldi, Ashley D.; Maki, Takahisa; Fitton, Benjamin P.; Roth, Daniel; Yampolsky, Dmitry; Davidson, Michael W.; Svitkina, Tatyana; Straube, Anne; Hayashi, Ikuko; Kaverina, Irina

2014-01-01

Summary Microtubule (MT) plus-end tracking proteins (+TIPs) preferentially localize to MT plus-ends. End-binding proteins (EBs) are master regulators of the +TIP complex; however, it is unknown whether EBs are regulated by other +TIPs. Here, we show that Cytoplasmic linker associated proteins (CLASPs) modulate EB localization at MTs. In CLASP-depleted cells, EBs localized along the MT lattice in addition to plus-ends. The MT-binding region of CLASP was sufficient for restoring normal EB localization, while neither EB-CLASP interactions nor EB tail-binding proteins are involved. In vitro assays revealed that CLASP directly functions to remove EB from MTs. Importantly, this effect occurs specifically during MT polymerization, but not at pre-formed MTs. Increased GTP-tubulin content within MTs in CLASP-depleted cells suggests that CLASPs facilitate GTP-hydrolysis to reduce EB lattice binding. Together, these findings suggest that CLASPs influence the MT lattice itself to regulate EB and determine exclusive plus-end localization of EBs in cells. PMID:25117684

Sprouting angiogenesis in human midterm uterus and fallopian tube is guided by endothelial tip cells.

PubMed

Rusu, M C; Motoc, A G M; Pop, F; Folescu, R

2013-01-01

Five samples of human midterm fetal uterus and fallopian tube (four donor bodies) were used to assess whether or not processes of angiogenesis are guided by endothelial tip cells (ETCs), and if cytokine-receptors, such as CD117/c-kit and PDGFR-α, are expressed in the microenvironment of the endothelial tubes. CD34 labeled microvessels in the uterine wall (myometrium and endometrium) and in the wall of the uterine (fallopian) tube, and accurately identified ETCs in both organs. We conclude that sprouting angiogenesis in the developing human female tract is guided by ETCs. Moreover, CD117/c-kit antibodies labeled mural networks of pericytes, α-SMA-positive and desmin-negative, related to the endometrial (but not myometrial) microvessels, and similar labeling was identified in the wall of the uterine tube. PDGFR-α positive labeling, stromal and pericytary, was also found. Thus, sprouting angiogenesis in human fetal genital organs appears to be guided by tip cells and is influenced by tyrosine kinase receptor signaling.

A Fluorescence Reporter Model Defines “Tip-DCs” as the Cellular Source of Interferon β in Murine Listeriosis

PubMed Central

Dresing, Philipp; Borkens, Stephanie; Kocur, Magdalena; Kropp, Sonja; Scheu, Stefanie

2010-01-01

Production of type I interferons, consisting mainly of multiple IFNα subtypes and IFNβ, represents an essential part of the innate immune defense against invading pathogens. While in most situations, namely viral infections, this class of cytokines is indispensable for host survival they mediate a detrimental effect during infection with L. monocytogenes by rendering macrophages insensitive towards IFNγ signalling which leads to a lethal bacterial pathology in mice. Due to a lack of suitable analytic tools the precise identity of the cell population responsible for type I IFN production remains ill-defined and so far these cells have been described to be macrophages. As in general IFNβ is the first type I interferon to be produced, we took advantage of an IFNβ fluorescence reporter-knockin mouse model in which YFP is expressed from a bicistronic mRNA linked by an IRES to the endogenous ifnb mRNA to assess the IFNβ production on a single cell level in situ. Our results showed highest frequencies and absolute numbers of IFNβ+ cells in the spleen 24 h after infection with L. monocytogenes where they were located predominately in the white pulp within the foci of infection. Detailed FACS surface marker analyses, intracellular cytokine stainings and T cell proliferation assays revealed that the IFNβ+ cells were a phenotypically and functionally further specialized subpopulation of TNF and iNOS producing DCs (Tip-DCs) which are known to be essential for the early containment of L. monocytogenes infection. We proved that the IFNβ+ cells exhibited the hallmark characteristics of Tip-DCs as they produced iNOS and TNF and possessed T cell priming abilities. These results point to a yet unappreciated ambiguous role for a multi-effector, IFNβ producing subpopulation of Tip-DCs in controlling the balance between containment of L. monocytogenes infection and effects detrimental to the host driven by IFNβ. PMID:21179567

A fluorescence reporter model defines "Tip-DCs" as the cellular source of interferon β in murine listeriosis.

PubMed

Dresing, Philipp; Borkens, Stephanie; Kocur, Magdalena; Kropp, Sonja; Scheu, Stefanie

2010-12-16

Production of type I interferons, consisting mainly of multiple IFNα subtypes and IFNβ, represents an essential part of the innate immune defense against invading pathogens. While in most situations, namely viral infections, this class of cytokines is indispensable for host survival they mediate a detrimental effect during infection with L. monocytogenes by rendering macrophages insensitive towards IFNγ signalling which leads to a lethal bacterial pathology in mice. Due to a lack of suitable analytic tools the precise identity of the cell population responsible for type I IFN production remains ill-defined and so far these cells have been described to be macrophages. As in general IFNβ is the first type I interferon to be produced, we took advantage of an IFNβ fluorescence reporter-knockin mouse model in which YFP is expressed from a bicistronic mRNA linked by an IRES to the endogenous ifnb mRNA to assess the IFNβ production on a single cell level in situ. Our results showed highest frequencies and absolute numbers of IFNβ+ cells in the spleen 24 h after infection with L. monocytogenes where they were located predominately in the white pulp within the foci of infection. Detailed FACS surface marker analyses, intracellular cytokine stainings and T cell proliferation assays revealed that the IFNβ+ cells were a phenotypically and functionally further specialized subpopulation of TNF and iNOS producing DCs (Tip-DCs) which are known to be essential for the early containment of L. monocytogenes infection. We proved that the IFNβ+ cells exhibited the hallmark characteristics of Tip-DCs as they produced iNOS and TNF and possessed T cell priming abilities. These results point to a yet unappreciated ambiguous role for a multi-effector, IFNβ producing subpopulation of Tip-DCs in controlling the balance between containment of L. monocytogenes infection and effects detrimental to the host driven by IFNβ.

Excitation of surface plasmons in Al-coated SNOM tips

NASA Astrophysics Data System (ADS)

Palm, Viktor; Rähn, Mihkel; Jäme, Joonas; Hizhnyakov, Vladimir

2012-10-01

The mesoscopic effect of spectral modulation occurring due to the interference of two photonic fiber modes filtered out by a metal-coated SNOM tip is used to observe the surface plasmon polariton (SPP) excitation in SNOM tips. In a spectrum of the broadband light transmitted by a SNOM tip a region of highly regular spectral modulation can be found, indicating the spectral interval in which only two photonic modes (apparently HE11 and TM01) are transmitted with significant and comparable amplitudes. The modulation period yields the value of optical path difference (OPD) for this pair of modes. Due to the multimode fiber's inherent modal dispersion, this OPD value depends linearly on the fiber tail length l. An additional contribution to OPD can be generated in a metal-coated SNOM tip due to a mode-dependent photon-plasmon coupling strength resulting in generation of SPPs with different propagation velocities. For an Al-coated 200 nm SNOM tip spectra of transmitted light have been registered for ten different l values. An extrapolation of the linear OPD (l) dependence to l=0 yields a significant residual OPD value, indicating according to our theoretical considerations a mode-selective SPP excitation in the metal-coated tip. The modal dispersion is shown to switch its sign in the SNOM tip. First results of analogous experiments with an Al-coated 150 nm SNOM tip confirm our conclusions.

The role of Arabidopsis Actin-Related Protein 3 in amyloplast sedimentation and polar auxin transport in root gravitropism

PubMed Central

Zou, Jun-Jie; Zheng, Zhong-Yu; Xue, Shan; Li, Han-Hai; Wang, Yu-Ren; Le, Jie

2016-01-01

Gravitropism is vital for shaping directional plant growth in response to the forces of gravity. Signals perceived in the gravity-sensing cells can be converted into biochemical signals and transmitted. Sedimentation of amyloplasts in the columella cells triggers asymmetric auxin redistribution in root tips, leading to downward root growth. The actin cytoskeleton is thought to play an important role in root gravitropism, although the molecular mechanism has not been resolved. DISTORTED1 (DIS1) encodes the ARP3 subunit of the Arabidopsis Actin-Related Protein 2/3 (ARP2/3) complex, and the ARP3/DIS1 mutant dis1-1 showed delayed root curvature after gravity stimulation. Microrheological analysis revealed that the high apparent viscosity within dis1-1 central columella cells is closely associated with abnormal movement trajectories of amyloplasts. Analysis using a sensitive auxin input reporter DII-VENUS showed that asymmetric auxin redistribution was reduced in the root tips of dis1-1, and the actin-disrupting drug Latrunculin B increased the asymmetric auxin redistribution. An uptake assay using the membrane-selective dye FM4-64 indicated that endocytosis was decelerated in dis1-1 root epidermal cells. Treatment and wash-out with Brefeldin A, which inhibits protein transport from the endoplasmic reticulum to the Golgi apparatus, showed that cycling of the auxin-transporter PIN-FORMED (PIN) proteins to the plasma membrane was also suppressed in dis1-1 roots. The results reveal that ARP3/DIS1 acts in root gravitropism by affecting amyloplast sedimentation and PIN-mediated polar auxin transport through regulation of PIN protein trafficking. PMID:27473572

The role of Arabidopsis Actin-Related Protein 3 in amyloplast sedimentation and polar auxin transport in root gravitropism.

PubMed

Zou, Jun-Jie; Zheng, Zhong-Yu; Xue, Shan; Li, Han-Hai; Wang, Yu-Ren; Le, Jie

2016-10-01

Gravitropism is vital for shaping directional plant growth in response to the forces of gravity. Signals perceived in the gravity-sensing cells can be converted into biochemical signals and transmitted. Sedimentation of amyloplasts in the columella cells triggers asymmetric auxin redistribution in root tips, leading to downward root growth. The actin cytoskeleton is thought to play an important role in root gravitropism, although the molecular mechanism has not been resolved. DISTORTED1 (DIS1) encodes the ARP3 subunit of the Arabidopsis Actin-Related Protein 2/3 (ARP2/3) complex, and the ARP3/DIS1 mutant dis1-1 showed delayed root curvature after gravity stimulation. Microrheological analysis revealed that the high apparent viscosity within dis1-1 central columella cells is closely associated with abnormal movement trajectories of amyloplasts. Analysis using a sensitive auxin input reporter DII-VENUS showed that asymmetric auxin redistribution was reduced in the root tips of dis1-1, and the actin-disrupting drug Latrunculin B increased the asymmetric auxin redistribution. An uptake assay using the membrane-selective dye FM4-64 indicated that endocytosis was decelerated in dis1-1 root epidermal cells. Treatment and wash-out with Brefeldin A, which inhibits protein transport from the endoplasmic reticulum to the Golgi apparatus, showed that cycling of the auxin-transporter PIN-FORMED (PIN) proteins to the plasma membrane was also suppressed in dis1-1 roots. The results reveal that ARP3/DIS1 acts in root gravitropism by affecting amyloplast sedimentation and PIN-mediated polar auxin transport through regulation of PIN protein trafficking. © The Author 2016. Published by Oxford University Press on behalf of the Society for Experimental Biology.

High-throughput, automated extraction of DNA and RNA from clinical samples using TruTip technology on common liquid handling robots.

PubMed

Holmberg, Rebecca C; Gindlesperger, Alissa; Stokes, Tinsley; Brady, Dane; Thakore, Nitu; Belgrader, Philip; Cooney, Christopher G; Chandler, Darrell P

2013-06-11

TruTip is a simple nucleic acid extraction technology whereby a porous, monolithic binding matrix is inserted into a pipette tip. The geometry of the monolith can be adapted for specific pipette tips ranging in volume from 1.0 to 5.0 ml. The large porosity of the monolith enables viscous or complex samples to readily pass through it with minimal fluidic backpressure. Bi-directional flow maximizes residence time between the monolith and sample, and enables large sample volumes to be processed within a single TruTip. The fundamental steps, irrespective of sample volume or TruTip geometry, include cell lysis, nucleic acid binding to the inner pores of the TruTip monolith, washing away unbound sample components and lysis buffers, and eluting purified and concentrated nucleic acids into an appropriate buffer. The attributes and adaptability of TruTip are demonstrated in three automated clinical sample processing protocols using an Eppendorf epMotion 5070, Hamilton STAR and STARplus liquid handling robots, including RNA isolation from nasopharyngeal aspirate, genomic DNA isolation from whole blood, and fetal DNA extraction and enrichment from large volumes of maternal plasma (respectively).

The possible involvement of root-cap mucilage in gravitropism and calcium movement across root tips of Allium cepa L

NASA Technical Reports Server (NTRS)

Moore, R.; Fondren, W. M.

1986-01-01

Roots of Allium cepa L. grown in aerated water elongate rapidly, but are not graviresponsive. These roots (1) possess extensive columella tissues comprised of cells containing numerous sedimented amyloplasts, (2) lack mucilage on their tips, and (3) are characterized by a weakly polar movement of calcium (Ca) across their tips. Placing roots in humid air correlates positively with the (1) onset of gravicurvature, (2) appearance of mucilage on tips of the roots, and (3) onset of the ability to transport Ca polarly to the lower side of the root tip. Gravicurvature of roots previously submerged in aerated water is more rapid when roots are oriented vertically for 1-2 h in humid air prior to being oriented horizontally. The more rapid gravicurvature of these roots correlates positively with the accumulation of mucilage at the tips of roots during the time the roots are oriented vertically. Therefore, the onset of gravicurvature and the ability of roots to transport Ca to the lower sides of their tips correlate positively with the presence of mucilage at their tips. These results suggest that mucilage may be important for the transport of Ca across root caps.

Tip off the HAT– Epigenetic control of learning and memory by Drosophila Tip60

PubMed Central

Xu, Songjun; Elefant, Felice

2015-01-01

Disruption of epigenetic gene control mechanisms involving histone acetylation in the brain causes cognitive impairment, a debilitating hallmark of most neurodegenerative disorders. Histone acetylation regulates cognitive gene expression via chromatin packaging control in neurons. Unfortunately, the histone acetyltransferases (HATs) that generate such neural epigenetic signatures and their mechanisms of action remain unclear. Our recent findings provide insight into this question by demonstrating that Tip60 HAT action is critical for morphology and function of the mushroom body (MB), the learning and memory center in the Drosophila brain. We show that Tip60 is robustly produced in MB Kenyon cells and extending axonal lobes and that targeted MB Tip60 HAT loss results in axonal outgrowth disruption. Functional consequences of loss and gain of Tip60 HAT levels in the MB are evidenced by defects in memory. Tip60 ChIP-Seq analysis reveals enrichment for genes that function in cognitive processes and accordingly, key genes representing these pathways are misregulated in the Tip60 HAT mutant fly brain. Remarkably, increasing levels of Tip60 in the MB rescues learning and memory deficits resulting from Alzheimer's disease associated amyloid precursor protein (APP) induced neurodegeneration. Our studies highlight the potential of HAT activators as a therapeutic option for cognitive disorders. PMID:26327426

The evaluation of a number of prototypes for the free-tip rotor constant-moment controller

NASA Technical Reports Server (NTRS)

Young, L. A.

1986-01-01

The development of several prototypes of a constant moment controller, a critical component of the free-tip rotor (FTR) concept, is described. Also presented are the experimental results of a whirl test designed to select a final controller configuration to be included in a future wind-tunnel test of this innovative rotor system. A brief explanation of the FTR concept and its history are included. The paper documents the controller design constraints, each prototype's operating principle, the evaluation test, and the individual prototype test results. A recommended design is identified, along with the selection rationale.

Single Cell "Glucose Nanosensor" Verifies Elevated Glucose Levels in Individual Cancer Cells.

PubMed

Nascimento, Raphael A S; Özel, Rıfat Emrah; Mak, Wai Han; Mulato, Marcelo; Singaram, Bakthan; Pourmand, Nader

2016-02-10

Because the transition from oxidative phosphorylation to anaerobic glycolytic metabolism is a hallmark of cancer progression, approaches to identify single living cancer cells by their unique glucose metabolic signature would be useful. Here, we present nanopipettes specifically developed to measure glucose levels in single cells with temporal and spatial resolution, and we use this technology to verify the hypothesis that individual cancer cells can indeed display higher intracellular glucose levels. The nanopipettes were functionalized as glucose nanosensors by immobilizing glucose oxidase (GOx) covalently to the tip so that the interaction of glucose with GOx resulted in a catalytic oxidation of β-d-glucose to d-gluconic acid, which was measured as a change in impedance due to drop in pH of the medium at the nanopipette tip. Calibration studies showed a direct relationship between impedance changes at the tip and glucose concentration in solution. The glucose nanosensor quantified single cell intracellular glucose levels in human fibroblasts and the metastatic breast cancer lines MDA-MB-231 and MCF7 and revealed that the cancer cells expressed reproducible and reliable increases in glucose levels compared to the nonmalignant cells. Nanopipettes allow repeated sampling of the same cell, as cells remain viable during and after measurements. Therefore, nanopipette-based glucose sensors provide an approach to compare changes in glucose levels with changes in proliferative or metastatic state. The platform has great promise for mechanistic investigations, as a diagnostic tool to distinguish cancer cells from nonmalignant cells in heterogeneous tissue biopsies, as well as a tool for monitoring cancer progression in situ.

A Novel Method for Differentiation of Human Mesenchymal Stem Cells into Smooth Muscle-Like Cells on Clinically Deliverable Thermally Induced Phase Separation Microspheres

PubMed Central

Parmar, Nina; Ahmadi, Raheleh

2015-01-01

Muscle degeneration is a prevalent disease, particularly in aging societies where it has a huge impact on quality of life and incurs colossal health costs. Suitable donor sources of smooth muscle cells are limited and minimally invasive therapeutic approaches are sought that will augment muscle volume by delivering cells to damaged or degenerated areas of muscle. For the first time, we report the use of highly porous microcarriers produced using thermally induced phase separation (TIPS) to expand and differentiate adipose-derived mesenchymal stem cells (AdMSCs) into smooth muscle-like cells in a format that requires minimal manipulation before clinical delivery. AdMSCs readily attached to the surface of TIPS microcarriers and proliferated while maintained in suspension culture for 12 days. Switching the incubation medium to a differentiation medium containing 2 ng/mL transforming growth factor beta-1 resulted in a significant increase in both the mRNA and protein expression of cell contractile apparatus components caldesmon, calponin, and myosin heavy chains, indicative of a smooth muscle cell-like phenotype. Growth of smooth muscle cells on the surface of the microcarriers caused no change to the integrity of the polymer microspheres making them suitable for a cell-delivery vehicle. Our results indicate that TIPS microspheres provide an ideal substrate for the expansion and differentiation of AdMSCs into smooth muscle-like cells as well as a microcarrier delivery vehicle for the attached cells ready for therapeutic applications. PMID:25205072

Generation and Long-term Maintenance of Nerve-free Hydra.

PubMed

Tran, Cassidy M; Fu, Sharon; Rowe, Trevor; Collins, Eva-Maria S

2017-07-07

The interstitial cell lineage of Hydra includes multipotent stem cells, and their derivatives: gland cells, nematocytes, germ cells, and nerve cells. The interstitial cells can be eliminated through two consecutive treatments with colchicine, a plant-derived toxin that kills dividing cells, thus erasing the potential for renewal of the differentiated cells that are derived from the interstitial stem cells. This allows for the generation of Hydra that lack nerve cells. A nerve-free polyp cannot open its mouth to feed, egest, or regulate osmotic pressure. Such animals, however, can survive and be cultured indefinitely in the laboratory if regularly force-fed and burped. The lack of nerve cells allows for studies of the role of the nervous system in regulating animal behavior and regeneration. Previously published protocols for nerve-free Hydra maintenance involve outdated techniques such as mouth-pipetting with hand-pulled micropipette tips to feed and clean the Hydra. Here, an improved protocol for maintenance of nerve-free Hydra is introduced. Fine-tipped forceps are used to force open the mouth and insert freshly killed Artemia. Following force-feeding, the body cavity of the animal is flushed with fresh medium using a syringe and hypodermic needle to remove undigested material, referred to here as "burping". This new method of force-feeding and burping nerve-free Hydra through the use of forceps and syringes eliminates the need for mouth-pipetting using hand-pulled micropipette tips. It thus makes the process safer and significantly more time efficient. To ensure that the nerve cells in the hypostome have been eliminated, immunohistochemistry using anti-tyrosine-tubulin is conducted.

Involvement of actin filaments in rhizoid morphogenesis of Spirogyra.

PubMed

Yoshida, Katsuhisa; Shimmen, Teruo

2009-01-01

The role of actin filaments in rhizoid morphogenesis was studied in Spirogyra. When the algal filaments were severed, new terminal cells started tip growth and finally formed rhizoids. Actin inhibitors, latrunculin B and cytochalasin D, reversibly inhibited the process. A mesh-like structure of actin filaments (AFs) was formed at the tip region. Gd(3+) inhibited tip growth and decreased AFs in the tip region. Either a decrease in turgor pressure or lowering of the external Ca(2+) concentration also induced similar results. It was suggested that the mesh-like AF structure is indispensable for the elongation of rhizoids. A possible organization mechanism of the mesh-like AF structure was discussed.

Using ToxCast data to reconstruct dynamic cell state trajectories and estimate toxicological points of departure.

EPA Pesticide Factsheets

Background: High-content imaging (HCI) allows simultaneous measurement of multiple cellular phenotypic changes and is an important tool for evaluating the biological activity of chemicals.Objectives: Our goal was to analyze dynamic cellular changes using HCI to identify the ??tipping point?? at which the cells did not show recovery towards a normal phenotypic state.Methods: HCI was used to evaluate the effects of 967 chemicals (in concentrations ranging from 0.4 to 200 03bcM) on HepG2 cells over a 72-hr exposure period. The HCI end points included p53, c-Jun, histone H2A.x, 03b1-tubulin, histone H3, alpha tubulin, mitochondrial membrane potential, mitochondrial mass, cell cycle arrest, nuclear size, and cell number. A computational model was developed to interpret HCI responses as cell-state trajectories.Results: Analysis of cell-state trajectories showed that 336 chemicals produced tipping points and that HepG2 cells were resilient to the effects of 334 chemicals up to the highest concentration (200 03bcM) and duration (72 hr) tested. Tipping points were identified as concentration-dependent transitions in system recovery, and the corresponding critical concentrations were generally between 5 and 15 times (25th and 75th percentiles, respectively) lower than the concentration that produced any significant effect on HepG2 cells. The remaining 297 chemicals require more data before they can be placed in either of these categories.Conclusions: These findings show t

Single-stage experimental evaluation of tandem-airfoil rotor and stator blading for compressors. Part 5: Analysis and design of stages D and E

NASA Technical Reports Server (NTRS)

Brent, J. A.; Cheatham, J. G.; Clemmons, D. R.

1972-01-01

A conventional and a tandem bladed stage were designed for a comparative experimental evaluation in a 0.8 hub/tip ratio single-stage compressor. Based on a preliminary design study, a radially constant work input distribution was selected for the rotor designs. Velocity diagrams and blade leading and trailing edge angles selected for the conventional rotor and stator were used in the design of the tandem blading. The effects of axial velocity ratio and secondary flow on turning were included in the selection of blade leading and trailing edge angles. Design values of rotor tip velocity and stage pressure ratio were 757 ft/sec and 1.26, respectively.

Indentation of Graphene-Covered Atomic Force Microscopy Probe Across a Lipid Bilayer Membrane: Effect of Tip Shape, Size, and Surface Hydrophobicity.

PubMed

Lv, Kang; Li, Yinfeng

2018-06-21

Understanding the interaction of graphene with cell membranes is crucial to the development of graphene-based biological applications and the management of graphene safety issues. To help reveal the key factors controlling the interaction between graphene and cell membranes, here we adopt the dissipative particle dynamics method to analyze the evolution of interaction force and free energy as the graphene-covered atomic force microscopy (AFM) probe indents across a lipid bilayer. The simulation results show that the graphene-covered AFM probe can cause severe deformation of the cell membrane which drives the lipid molecule to adsorb and diffuse at the surface of graphene. The breakthrough force and free energy are calculated to study the effects of the tip shape, size, and surface hydrophobicity on the piercing behaviors of graphene-covered AFM. In addition, the deformation of cell membrane can decrease the dependency of the breakthrough force on the tip shape. The analysis of surface functionalization suggests that the horizontal patterns on graphene can change the preferred orientation in the penetration process, but the vertical patterns on graphene may disrupt the cell membrane. What's more, the bending stiffness of graphene has little influence on the penetration process as graphene pierces into the cell membrane. These results provide useful guidelines for the molecular design of graphene materials with controllable cell penetrability.

Effect of IOP based infusion system with and without balanced phaco tip on cumulative dissipated energy and estimated fluid usage in comparison to gravity fed infusion in torsional phacoemulsification.

PubMed

Malik, Praveen K; Dewan, Taru; Patidar, Arun Kr; Sain, Ekta

2017-01-01

To evaluate the effect of three different combinations of tip designs and infusion systems in torsional phacoemulsification (INFINITI and CENTURION) in patients with cataract. According to the manufacturer, two unique improvements in the Centurion are: active fluid dynamic management system and use of an intrepid balanced tip. The study specifically aimed to evaluate the beneficial effects, if any, of change in tip design and infusion system individually and in combination on both per-operative parameters as well as endothelial health over 6 months. One hundred and twenty six consenting patients of grade 4.0-6.9 senile cataract were randomized into three groups for phacoemulsification: Group A ( n  = 42): Gravity fed infusion system and 45 0 Kelman miniflared ABS phaco tip; Group B ( n  = 42): intraocular pressure (IOP) based infusion system and 45 0 Kelman miniflared ABS phaco tip; Group C ( n  = 42): IOP based infusion system and 45 0 Intrepid balanced phaco tip. The cumulative dissipated energy (CDE), estimated fluid usage (EFU) and total aspiration time (TAT) were compared peroperatively. The endothelial parameters were followed up postoperatively for six months. The three arms were matched for age ( p  = 0.525), gender ( p  = 0.96) and grade of cataract ( p  = 0.177). Group C was associated with significant reductions in CDE ( p  = 0.001), EFU ( p  < 0.0005) as well as TAT ( p  = 0.001) in comparison to the other groups. All three groups had comparable baseline endothelial cell density ( p  = 0.876) and central corneal thickness ( p  = 0.561). On post-operative evaluation, although all groups were comparable till 3 months, by 6 months, the percentage losses in endothelial cell density were significantly lower in group C as compared to the other groups. Use of an IOP based phacoemulsification system in association with use of the Intrepid balanced tip reduces the CDE, EFU and TAT in comparison to a gravity fed system with a mini flared tip or IOP based system with a mini flared tip while also providing better endothelial preservation thus favouring the use of an IOP fed system with a balanced tip. Trial registration No.: CTRI/2016/06/007022.

Stalk cell differentiation without polyketides in the cellular slime mold.

PubMed

Sato, Yukie G; Suarez, Teresa; Saito, Tamao

2016-07-01

Polyketides induce prestalk cell differentiation in Dictyostelium. In the double-knockout mutant of the SteelyA and B polyketide synthases, most of the pstA cells-the major part of the prestalk cells-are lost, and we show by whole mount in situ hybridization that expression of prestalk genes is also reduced. Treatment of the double-knockout mutant with the PKS inhibitor cerulenin gave a further reduction, but some pstA cells still remained in the tip region, suggesting the existence of a polyketide-independent subtype of pstA cells. The double-knockout mutant and cerulenin-treated parental Ax2 cells form fruiting bodies with fragile, single-cell layered stalks after cerulenin treatment. Our results indicate that most pstA cells are induced by polyketides, but the pstA cells at the very tip of the slug are induced in some other way. In addition, a fruiting body with a single-cell layered, vacuolated stalk can form without polyketides.

The SNARE Protein Syntaxin 3 Confers Specificity for Polarized Axonal Trafficking in Neurons

PubMed Central

Soo Hoo, Linda; Banna, Chris D.; Radeke, Carolyn M.; Sharma, Nikunj; Albertolle, Mary E.; Low, Seng Hui; Weimbs, Thomas; Vandenberg, Carol A.

2016-01-01

Cell polarity and precise subcellular protein localization are pivotal to neuronal function. The SNARE machinery underlies intracellular membrane fusion events, but its role in neuronal polarity and selective protein targeting remain unclear. Here we report that syntaxin 3 is involved in orchestrating polarized trafficking in cultured rat hippocampal neurons. We show that syntaxin 3 localizes to the axonal plasma membrane, particularly to axonal tips, whereas syntaxin 4 localizes to the somatodendritic plasma membrane. Disruption of a conserved N-terminal targeting motif, which causes mislocalization of syntaxin 3, results in coincident mistargeting of the axonal cargos neuron-glia cell adhesion molecule (NgCAM) and neurexin, but not transferrin receptor, a somatodendritic cargo. Similarly, RNAi-mediated knockdown of endogenous syntaxin 3 leads to partial mistargeting of NgCAM, demonstrating that syntaxin 3 plays an important role in its targeting. Additionally, overexpression of syntaxin 3 results in increased axonal growth. Our findings suggest an important role for syntaxin 3 in maintaining neuronal polarity and in the critical task of selective trafficking of membrane protein to axons. PMID:27662481

The SNARE Protein Syntaxin 3 Confers Specificity for Polarized Axonal Trafficking in Neurons.

PubMed

Soo Hoo, Linda; Banna, Chris D; Radeke, Carolyn M; Sharma, Nikunj; Albertolle, Mary E; Low, Seng Hui; Weimbs, Thomas; Vandenberg, Carol A

Cell polarity and precise subcellular protein localization are pivotal to neuronal function. The SNARE machinery underlies intracellular membrane fusion events, but its role in neuronal polarity and selective protein targeting remain unclear. Here we report that syntaxin 3 is involved in orchestrating polarized trafficking in cultured rat hippocampal neurons. We show that syntaxin 3 localizes to the axonal plasma membrane, particularly to axonal tips, whereas syntaxin 4 localizes to the somatodendritic plasma membrane. Disruption of a conserved N-terminal targeting motif, which causes mislocalization of syntaxin 3, results in coincident mistargeting of the axonal cargos neuron-glia cell adhesion molecule (NgCAM) and neurexin, but not transferrin receptor, a somatodendritic cargo. Similarly, RNAi-mediated knockdown of endogenous syntaxin 3 leads to partial mistargeting of NgCAM, demonstrating that syntaxin 3 plays an important role in its targeting. Additionally, overexpression of syntaxin 3 results in increased axonal growth. Our findings suggest an important role for syntaxin 3 in maintaining neuronal polarity and in the critical task of selective trafficking of membrane protein to axons.

Flip the tip: an automated, high quality, cost-effective patch clamp screen.

PubMed

Lepple-Wienhues, Albrecht; Ferlinz, Klaus; Seeger, Achim; Schäfer, Arvid

2003-01-01

The race for creating an automated patch clamp has begun. Here, we present a novel technology to produce true gigaseals and whole cell preparations at a high rate. Suspended cells are flushed toward the tip of glass micropipettes. Seal, whole-cell break-in, and pipette/liquid handling are fully automated. Extremely stable seals and access resistance guarantee high recording quality. Data obtained from different cell types sealed inside pipettes show long-term stability, voltage clamp and seal quality, as well as block by compounds in the pM range. A flexible array of independent electrode positions minimizes consumables consumption at maximal throughput. Pulled micropipettes guarantee a proven gigaseal substrate with ultra clean and smooth surface at low cost.

Hair cell transduction, tuning and synaptic transmission in the mammalian cochlea

PubMed Central

Fettiplace, Robert

2017-01-01

Sound pressure fluctuations striking the ear are conveyed to the cochlea, where they vibrate the basilar membrane on which sit hair cells, the mechanoreceptors of the inner ear. Recordings of hair cell electrical responses have shown that they transduce sound via sub-micrometer deflections of their hair bundles, which are arrays of interconnected stereocilia containing the mechanoelectrical transducer (MET) channels. MET channels are activated by tension in extracellular tip links bridging adjacent stereocilia, and they can respond within microseconds to nanometer displacements of the bundle, facilitated by multiple processes of Ca2+-dependent adaptation. Studies of mouse mutants have produced much detail about the molecular organization of the stereocilia, the tip links and their attachment sites, and the MET channels localized to the lower ends of each tip link. The mammalian cochlea contains two categories of hair cells. Inner hair cells relay acoustic information via multiple ribbon synapses that transmit rapidly without rundown. Outer hair cells are important for amplifying sound-evoked vibrations. The amplification mechanism primarily involves contractions of the outer hair cells, which are driven by changes in membrane potential and mediated by prestin, a motor protein in the outer hair cell lateral membrane. Different sound frequencies are separated along the cochlea, with each hair cell being tuned to a narrow frequency range; amplification sharpens the frequency resolution and augments sensitivity 100-fold around the cell’s characteristic frequency. Genetic mutations and environmental factors such as acoustic overstimulation cause hearing loss through irreversible damage to the hair cells or degeneration of inner hair cell synapses. PMID:28915323

Application of pressure probe and UV-MALDI-TOF MS for direct analysis of plant underivatized carbohydrates in subpicoliter single-cell cytoplasm extract.

PubMed

Gholipour, Yousef; Nonami, Hiroshi; Erra-Balsells, Rosa

2008-12-01

Single-cell cytoplasm sap (1-10 pL) was extracted by using a pressure probe glass microcapillary tip from tulip leaf and bulb and analyzed by UV-MALDI-TOF MS for free underivatized carbohydrate content. Three matrices including 2,5-dihydroxybenzoic acid (DHB), 2,4,6-trihydroxyacetophenone (THAP), and carbon nanotubes (CNTs) in positive ion mode were selected for analysis because of acceptable carbohydrate-related signal reproducibility. Disaccharide and oligosaccharide (up to 15 Hex when THAP was used, 11 Hex with DHB, and 7 Hex with CNTs) were detected in tulip bulb cell cytoplasm sample. When DHB was used as matrix, neutral carbohydrates were more abundantly detected as sodiated cations; the sugar-related signals, however, appeared as dominant potassiated cations when THAP and CNTs were used. Small amount of monosaccharide was also detected in bulb cell cytoplasm with CNTs as matrix. UV-MALDI-TOF MS of leaf cell extract resulted in high-resolution detection of hexose and disaccharide with DHB, THAP, and CNTs.

Interspecies somatic cell nucleus transfer with porcine oocytes as recipients: A novel bioassay system for assessing the competence of canine somatic cells to develop into embryos.

PubMed

Sugimura, S; Narita, K; Yamashiro, H; Sugawara, A; Shoji, T; Terashita, Y; Nishimori, K; Konno, T; Yoshida, M; Sato, E

2009-09-01

Interspecies somatic cell nucleus transfer (iSCNT) could be a useful bioassay system for assessing the ability of mammalian somatic cells to develop into embryos. To examine this possibility, we performed canine iSCNT using porcine oocytes, allowed to mature in vitro, as recipients. Canine fibroblasts from the tail tips and dewclaws of a female poodle (Fp) and a male poodle (Mp) were used as donors. We demonstrated that the use of porcine oocytes induced blastocyst formation in the iSCNT embryos cultured in porcine zygote medium-3. In Fp and Mp, the rate of blastocyst formation from cleaved embryos (Fp: 6.3% vs. 22.4%; and Mp: 26.1% vs. 52.4%) and the number of cells at the blastocyst stage (Fp: 30.7 vs. 60.0; and Mp: 27.2 vs. 40.1) were higher in the embryos derived from dewclaw cells than in those derived from tail-tip cells (P<0.05). The use of donor cells of any type in later passages decreased the rate of blastocyst formation. Treatment with trichostatin-A did not improve the rate of blastocyst formation from cleaved dewclaw cell-derived embryos but did so in the embryos derived from the tail-tip cells of Fp. Only blastocysts derived from dewclaw cells of Mp developed outgrowths. However, outgrowth formation was retrieved in the embryos derived from dewclaw cells of Fp by aggregation at the 4-cell stage. We inferred that iSCNT performed using porcine oocytes as recipients could represent a novel bioassay system for evaluating the developmental competence of canine somatic cells.

Survival rate of eukaryotic cells following electrophoretic nanoinjection.

PubMed

Simonis, Matthias; Hübner, Wolfgang; Wilking, Alice; Huser, Thomas; Hennig, Simon

2017-01-25

Insertion of foreign molecules such as functionalized fluorescent probes, antibodies, or plasmid DNA to living cells requires overcoming the plasma membrane barrier without harming the cell during the staining process. Many techniques such as electroporation, lipofection or microinjection have been developed to overcome the cellular plasma membrane, but they all result in reduced cell viability. A novel approach is the injection of cells with a nanopipette and using electrophoretic forces for the delivery of molecules. The tip size of these pipettes is approximately ten times smaller than typical microinjection pipettes and rather than pressure pulses as delivery method, moderate DC electric fields are used to drive charged molecules out of the tip. Here, we show that this approach leads to a significantly higher survival rate of nanoinjected cells and that injection with nanopipettes has a significantly lower impact on the proliferation behavior of injected cells. Thus, we propose that injection with nanopipettes using electrophoretic delivery is an excellent alternative when working with valuable and rare living cells, such as primary cells or stem cells.

A novel way to go whole-cell in patch-clamp experiments.

PubMed

Inayat, Samsoon; Zhao, Yan; Cantrell, Donal R; Dikin, Dmitryi; Pinto, Lawrence H; Troy, John B

2010-11-01

With a conventional patch-clamp electrode, an Ag/AgCl wire sits stationary inside the pipette. To move from the gigaseal cell-attached configuration to whole-cell recording, suction is applied inside the pipette. We have designed and developed a novel Pushpen patch-clamp electrode, in which a W wire insulated and wound with Ag/AgCl wire can move linearly inside the pipette. The W wire has a conical tip, which can protrude from the pipette tip like a push pen, a procedure we call the Pushpen Operation. We use the Pushpen operation to impale the cell membrane in cell-attached configuration to go whole-cell without disruption of the gigaseal. We successfully recorded whole-cell currents from chinese hamster ovarian cells expressing influenza A virus protein A/M2, after obtaining whole-cell configuration with the Pushpen operation. This novel method of achieving whole-cell configuration may have a higher success rate than is the case with the conventional patch clamp technique.

Near-tip-screenout hydraulic fracturing of oil wells in the Bach Ho field, offshore Vietnam

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

Hung, L.V.; San, N.T.; Shelomentsev, A.G.

1995-10-01

The first hydraulic fracturing of wells in Vietnam were successfully performed Offshore in the Bach Ho (White Tiger) Oil Field. Near-tip-screenouts rather than tip-screenout treatments were performed. The goal of the project was to improve production from existing wells rather than drill new wells and reduce the cost per barrel produced. This case study involves wells with multiple perforated zones completed in the Oligocene sandstone. Zones were selectively fractured in order to optimize production. A detailed description candidate selection, design, execution and evaluation processes are presented. The Bach Ho field has been producing for 8 years but not at itsmore » potential due to various reasons including drilling and completion fluid damage. Although acidizing was an option for damage removal, hydraulic fracturing was selected as a way to bypass near-wellbore damage and generate a negative skin. Production simulators were used to quantify post-frac production. Due to suspected high closure stress, high strength proppant was selected and ramped in a high temperature fracturing fluid. Calibration treatments were conducted on several wells to quantify fluid leak-off, fracture height and Young`s modulus. Based on the results of the calibration treatment, fracture designs were modified. As predicted by computer simulation, near-tip-screenouts occurred as planned. The treatments were performed using a work boat with skid pumping/blending equipment, a computer monitoring/operation center and a laboratory. Strict QC procedures were followed to ensure the quality of all products. Post-frac well tests results and production data are presented. Overall, the fracturing campaign was very successful with wells showing negative skins and up to a five fold increase of production in agreement with systems analysis predictions.« less

Thidiazuron Triggers Morphogenesis in Rosa canina L. Protocorm-Like Bodies by Changing Incipient Cell Fate

PubMed Central

Kou, Yaping; Yuan, Cunquan; Zhao, Qingcui; Liu, Guoqin; Nie, Jing; Ma, Zhimin; Cheng, Chenxia; Teixeira da Silva, Jaime A.; Zhao, Liangjun

2016-01-01

Thidiazuron (N-phenyl-N′-1,2,3-thiadiazol-5-ylurea; TDZ) is an artificial plant growth regulator that is widely used in plant tissue culture. Protocorm-like bodies (PLBs) induced by TDZ serve as an efficient and rapid in vitro regeneration system in Rosa species. Despite this, the mechanism of PLB induction remains relatively unclear. TDZ, which can affect the level of endogenous auxins and cytokinins, converts the cell fate of rhizoid tips and triggers PLB formation and plantlet regeneration in Rosa canina L. In callus-rhizoids, which are rhizoids that co-develop from callus, auxin and a Z-type cytokinin accumulated after applying TDZ, and transcription of the auxin transporter gene RcPIN1 was repressed. The expression of RcARF4, RcRR1, RcCKX2, RcCKX3, and RcLOG1 increased in callus-rhizoids and rhizoid tips while the transcription of an auxin response factor (RcARF1) and auxin transport proteins (RcPIN2, RcPIN3) decreased in callus-rhizoids but increased in rhizoid tips. In situ hybridization of rhizoids showed that RcWUS and RcSERK1 were highly expressed in columella cells and root stem cells resulting in the conversion of cell fate into shoot apical meristems or embryogenic callus. In addition, transgenic XVE::RcWUS lines showed repressed RcWUS overexpression while RcWUS had no effect on PLB morphogenesis. Furthermore, higher expression of the root stem cell marker RcWOX5 and root stem cell maintenance regulator genes RcPLT1 and RcPLT2 indicated the presence of a dedifferentiation developmental pathway in the stem cell niche of rhizoids. Viewed together, our results indicate that different cells in rhizoid tips acquired regeneration competence after induction by TDZ. A novel developmental pathway containing different cell types during PLB formation was identified by analyzing the endogenous auxin and cytokinin content. This study also provides a deeper understanding of the mechanisms underlying in vitro regeneration in Rosa. PMID:27200031

Symbiotic adaptations in the fungal cultivar of leaf-cutting ants.

PubMed

De Fine Licht, Henrik H; Boomsma, Jacobus J; Tunlid, Anders

2014-12-01

Centuries of artificial selection have dramatically improved the yield of human agriculture; however, strong directional selection also occurs in natural symbiotic interactions. Fungus-growing attine ants cultivate basidiomycete fungi for food. One cultivar lineage has evolved inflated hyphal tips (gongylidia) that grow in bundles called staphylae, to specifically feed the ants. Here we show extensive regulation and molecular signals of adaptive evolution in gene trancripts associated with gongylidia biosynthesis, morphogenesis and enzymatic plant cell wall degradation in the leaf-cutting ant cultivar Leucoagaricus gongylophorus. Comparative analysis of staphylae growth morphology and transcriptome-wide expressional and nucleotide divergence indicate that gongylidia provide leaf-cutting ants with essential amino acids and plant-degrading enzymes, and that they may have done so for 20-25 million years without much evolutionary change. These molecular traits and signatures of selection imply that staphylae are highly advanced coevolutionary organs that play pivotal roles in the mutualism between leaf-cutting ants and their fungal cultivars.

Statolith positioning by microfilaments in Chara rhizoids and protonemata

NASA Astrophysics Data System (ADS)

Hodick, Dieter; Buchen, Brigitte; Sievers, Andreas

The rhizoids of the green alga Chara are tip-growing cells with a precise positive gravitropism. In rhizoids growing downwards the statoliths never sediment upon the cell wall at the very tip but keep a minimal distance of approximately 10 μm from the cell vertex. It has been argued that this position is attained by a force acting upon the statoliths in the basal direction and that this force is generated by an interaction between actin microfilaments and myosin on the statolith membrane. This hypothesis received experimental support from (1) effects of the actin-attacking drug cytochalasin, (2) experiments under microgravity conditions, and (3) clinostat experiments. Using video-microscopy it is now shown that this basipetal force also acts on statoliths during sedimentation. As a result, many statoliths in Chara rhizoids do not simply fall along the plumb line while sedimenting during gravistimulation, but move basipetally. This statolith movement is compared to the ones occurring in the unicellular Chara protonemata during gravistimulation. Dark-grown protonemata morphologically closely resemble the rhizoids but respond negatively gravitropic. In contrast to the rhizoids a gravistimulation of the protonemata induces a transport of statoliths towards the tip. This transport is mainly along the cell axis and not parallel to the gravity vector. It is stressed that the sedimentation of statoliths in Chara rhizoids and protonemata as well as in gravity sensing cells in mosses and higher plants is accompanied by statolith movements based on interactions with the cytoskeleton. In tip-growing cells these movements direct the statoliths to a definite region of the cell where they can sediment and elicit a gravitropic curvature. In the statocytes of higher plants the interactions of the statoliths with the cytoskeleton probably do not serve primarily to move the statoliths but to transduce mechanical stresses from the sedimenting statoliths to the plasma membrane.

Statolith positioning by microfilaments in Chara rhizoids and protonemata.

PubMed

Hodick, D; Buchen, B; Sievers, A

1998-01-01

The rhizoids of the green alga Chara are tip-growing cells with a precise positive gravitropism. In rhizoids growing downwards the statoliths never sediment upon the cell wall at the very tip but keep a minimal distance of approximately 10 micrometers from the cell vertex. It has been argued that this position is attained by a force acting upon the statoliths in the basal direction and that this force is generated by an interaction between actin microfilaments and myosin on the statolith membrane. This hypothesis received experimental support from (1) effects of the actin-attacking drug cytochalasin, (2) experiments under microgravity conditions, and (3) clinostat experiments. Using video-microscopy it is now shown that this basipetal force also acts on statoliths during sedimentation. As a result, many statoliths in Chara rhizoids do not simply fall along the plumb line while sedimenting during gravistimulation, but move basipetally. This statolith movement is compared to the ones occurring in the unicellular Chara protonemata during gravistimulation. Dark-grown protonemata morphologically closely resemble the rhizoids but respond negatively gravitropic. In contrast to the rhizoids a gravistimulation of the protonemata induces a transport of statoliths towards the tip. This transport is mainly along the cell axis and not parallel to the gravity vector. It is stressed that the sedimentation of statoliths in Chara rhizoids and protonemata as well as in gravity sensing cells in mosses and higher plants is accompanied by statolith movements based on interactions with the cytoskeleton. In tip-growing cells these movements direct the statoliths to a definite region of the cell where they can sediment and elicit a gravitropic curvature. In the statocytes of higher plants the interactions of the statoliths with the cytoskeleton probably do not serve primarily to move the statoliths but to transduce mechanical stresses from the sedimenting statoliths to the plasma membrane.

Fungal Morphogenesis, from the Polarized Growth of Hyphae to Complex Reproduction and Infection Structures.

PubMed

Riquelme, Meritxell; Aguirre, Jesús; Bartnicki-García, Salomon; Braus, Gerhard H; Feldbrügge, Michael; Fleig, Ursula; Hansberg, Wilhelm; Herrera-Estrella, Alfredo; Kämper, Jörg; Kück, Ulrich; Mouriño-Pérez, Rosa R; Takeshita, Norio; Fischer, Reinhard

2018-06-01

Filamentous fungi constitute a large group of eukaryotic microorganisms that grow by forming simple tube-like hyphae that are capable of differentiating into more-complex morphological structures and distinct cell types. Hyphae form filamentous networks by extending at their tips while branching in subapical regions. Rapid tip elongation requires massive membrane insertion and extension of the rigid chitin-containing cell wall. This process is sustained by a continuous flow of secretory vesicles that depends on the coordinated action of the microtubule and actin cytoskeletons and the corresponding motors and associated proteins. Vesicles transport cell wall-synthesizing enzymes and accumulate in a special structure, the Spitzenkörper, before traveling further and fusing with the tip membrane. The place of vesicle fusion and growth direction are enabled and defined by the position of the Spitzenkörper, the so-called cell end markers, and other proteins involved in the exocytic process. Also important for tip extension is membrane recycling by endocytosis via early endosomes, which function as multipurpose transport vehicles for mRNA, septins, ribosomes, and peroxisomes. Cell integrity, hyphal branching, and morphogenesis are all processes that are largely dependent on vesicle and cytoskeleton dynamics. When hyphae differentiate structures for asexual or sexual reproduction or to mediate interspecies interactions, the hyphal basic cellular machinery may be reprogrammed through the synthesis of new proteins and/or the modification of protein activity. Although some transcriptional networks involved in such reprogramming of hyphae are well studied in several model filamentous fungi, clear connections between these networks and known determinants of hyphal morphogenesis are yet to be established. Copyright © 2018 American Society for Microbiology.

Integrated Metabolomics and Transcriptomics Reveal Enhanced Specialized Metabolism in Medicago truncatula Root Border Cells1[OPEN

PubMed Central

Watson, Bonnie S.; Bedair, Mohamed F.; Urbanczyk-Wochniak, Ewa; Huhman, David V.; Yang, Dong Sik; Allen, Stacy N.; Li, Wensheng; Tang, Yuhong; Sumner, Lloyd W.

2015-01-01

Integrated metabolomics and transcriptomics of Medicago truncatula seedling border cells and root tips revealed substantial metabolic differences between these distinct and spatially segregated root regions. Large differential increases in oxylipin-pathway lipoxygenases and auxin-responsive transcript levels in border cells corresponded to differences in phytohormone and volatile levels compared with adjacent root tips. Morphological examinations of border cells revealed the presence of significant starch deposits that serve as critical energy and carbon reserves, as documented through increased β-amylase transcript levels and associated starch hydrolysis metabolites. A substantial proportion of primary metabolism transcripts were decreased in border cells, while many flavonoid- and triterpenoid-related metabolite and transcript levels were increased dramatically. The cumulative data provide compounding evidence that primary and secondary metabolism are differentially programmed in border cells relative to root tips. Metabolic resources normally destined for growth and development are redirected toward elevated accumulation of specialized metabolites in border cells, resulting in constitutively elevated defense and signaling compounds needed to protect the delicate root cap and signal motile rhizobia required for symbiotic nitrogen fixation. Elevated levels of 7,4′-dihydroxyflavone were further increased in border cells of roots exposed to cotton root rot (Phymatotrichopsis omnivora), and the value of 7,4′-dihydroxyflavone as an antimicrobial compound was demonstrated using in vitro growth inhibition assays. The cumulative and pathway-specific data provide key insights into the metabolic programming of border cells that strongly implicate a more prominent mechanistic role for border cells in plant-microbe signaling, defense, and interactions than envisioned previously. PMID:25667316

Three major nucleolar proteins migrate from nucleolus to nucleoplasm and cytoplasm in root tip cells of Vicia faba L. exposed to aluminum.

PubMed

Qin, Rong; Zhang, Huaning; Li, Shaoshan; Jiang, Wusheng; Liu, Donghua

2014-09-01

Results from our previous investigation indicated that Al could affect the nucleolus and induce extrusion of silver-staining nucleolar particles containing argyrophilic proteins from the nucleolus into the cytoplasm in root tip cells of Vicia faba L. So far, the nucleolar proteins involved have not been identified. It is well known that nucleophosmin (B23), nucleolin (C23), and fibrillarin are three major and multifunctional nucleolar proteins. Therefore, effects of Al on B23, C23, and fibrillarin in root tip cells of V. faba exposed to 100 μM Al for 48 h were observed and analyzed using indirect immunofluorescence microscopy and Western blotting. The results from this work demonstrated that after 100 μM of Al treatment for 48 h, B23 and C23 migrated from the nucleolus to the cytoplasm and fibrillarin from the nucleolus to the nucleoplasm. In some cells, fibrillarin was present only in the cytoplasm. Western blotting data revealed higher expression of the three major nucleolar proteins in Al-treated roots compared with the control and that the B23 content increased markedly. These findings confirmed our previous observations.

Comparative Evaluation of Cell Viability Immediately After Osteotomy for Implants With Drills and Piezosurgery: Immunohistochemistry Analysis.

PubMed

Pereira, Cassiano Costa Silva; Batista, Fábio Roberto de Souza; Jacob, Ricardo Garcia Mureb; Nogueira, Lamis Meorin; Carvalho, Abrahão Cavalcante Gomes de Souza; Gealh, Walter Cristiano; Garcia-Júnior, Idelmo Rangel; Okamoto, Roberta

2018-05-08

To evaluate the effect of reusing drills and piezosurgery tips during implant osteotomy on immediate bone cell viability through immunohistochemical analysis. Six male rabbits were divided into 2 groups and then divided into 5 subgroups-correspond to drills and tips used 10, 20, 30, 40, and 50 times, respectively. All animals received 10 osteotomies in each tibia, by use of the classic drilling procedure in one group (G1) and the piezosurgery device in the other group (G2). For immunohistochemical technique were utilized the osteoprotegerin, RANKL, osteocalcin, and caspase 3. Control procedures were performed by omitting the primary antibodies (negative control). Bone formation and resorption responses presented in more intense way during the piezosurgery. The expression of osteocalcin had become quite intense in piezosurgery groups, but with reduced immunostaining from the 30th osteotomy. The caspase 3 showed the viability of the osteoblast from the 20th osteotomy with piezosurgery and remained constant until the 50th. Piezosurgery provides greater osteoblastic cell viability than the system of conventional drilling. This study will provide data so that the authors can recycle the drills and tips for implant placement, thus enabling a better cell viability for osseointegration.

Auxin increases the hydrogen peroxide (H2O2) concentration in tomato (Solanum lycopersicum) root tips while inhibiting root growth

PubMed Central

Ivanchenko, Maria G.; den Os, Désirée; Monshausen, Gabriele B.; Dubrovsky, Joseph G.; Bednářová, Andrea; Krishnan, Natraj

2013-01-01

Background and Aims The hormone auxin and reactive oxygen species (ROS) regulate root elongation, but the interactions between the two pathways are not well understood. The aim of this study was to investigate how auxin interacts with ROS in regulating root elongation in tomato, Solanum lycopersicum. Methods Wild-type and auxin-resistant mutant, diageotropica (dgt), of tomato (S. lycopersicum ‘Ailsa Craig’) were characterized in terms of root apical meristem and elongation zone histology, expression of the cell-cycle marker gene Sl-CycB1;1, accumulation of ROS, response to auxin and hydrogen peroxide (H2O2), and expression of ROS-related mRNAs. Key Results The dgt mutant exhibited histological defects in the root apical meristem and elongation zone and displayed a constitutively increased level of hydrogen peroxide (H2O2) in the root tip, part of which was detected in the apoplast. Treatments of wild-type with auxin increased the H2O2 concentration in the root tip in a dose-dependent manner. Auxin and H2O2 elicited similar inhibition of cell elongation while bringing forth differential responses in terms of meristem length and number of cells in the elongation zone. Auxin treatments affected the expression of mRNAs of ROS-scavenging enzymes and less significantly mRNAs related to antioxidant level. The dgt mutation resulted in resistance to both auxin and H2O2 and affected profoundly the expression of mRNAs related to antioxidant level. Conclusions The results indicate that auxin regulates the level of H2O2 in the root tip, so increasing the auxin level triggers accumulation of H2O2 leading to inhibition of root cell elongation and root growth. The dgt mutation affects this pathway by reducing the auxin responsiveness of tissues and by disrupting the H2O2 homeostasis in the root tip. PMID:23965615

10 Tips for Getting Grants to Keep Your Library Afloat

ERIC Educational Resources Information Center

McCune, Bonnie

2007-01-01

In this article, the author, taking the view of the foundation or the grant giver, shares the basic steps on how funders assess requests for grants. Each of these steps may be waived if a key member of the selection process, such as a board member, has a personal interest in the application. The following are 10 tips from the funders' point of…

Activation of Parathyroid Hormone 2 Receptor Induces Decorin Expression and Promotes Wound Repair

PubMed Central

Sato, Emi; Zhang, Ling-juan; Dorschner, Robert A.; Adase, Christopher A.; Choudhury, Biswa P.; Gallo, Richard L.

2018-01-01

In this study, we report that TIP39, a parathyroid hormone ligand family member that was recently identified to be expressed in the skin, can induce decorin expression and enhance wound repair. Topical treatment of mice with TIP39 accelerated wound repair, whereas TIP39-deficient mice had delayed repair that was associated with formation of abnormal collagen bundles. To study the potential mechanism responsible for the action of TIP39 in the dermis, fibroblasts were cultured in three-dimensional collagen gels, a process that results in enhanced decorin expression unless activated to differentiate to adipocytes, whereupon these cells reduce expression of several proteoglycans, including decorin. Small interfering RNA-mediated silencing of parathyroid hormone 2 receptor (PTH2R), the receptor for TIP39, suppressed the expression of extracellular matrix-related genes, including decorin, collagens, fibronectin, and matrix metalloproteases. Skin wounds in TIP39−/− mice had decreased decorin expression, and addition of TIP39 to cultured fibroblasts induced decorin and increased phosphorylation and nuclear translocation of CREB. Fibroblasts differentiated to adipocytes and treated with TIP39 also showed increased decorin and production of chondroitin sulfate. Furthermore, the skin of PTH2R−/− mice showed abnormal extracellular matrix structure, decreased decorin expression, and skin hardness. Thus, the TIP39-PTH2R system appears to be a previously unrecognized mechanism for regulation of extracellular matrix formation and wound repair. PMID:28454729

Rga6 is a fission yeast Rho GAP involved in Cdc42 regulation of polarized growth

PubMed Central

Revilla-Guarinos, M. T.; Martín-García, Rebeca; Villar-Tajadura, M. Antonia; Estravís, Miguel; Coll, Pedro M.; Pérez, Pilar

2016-01-01

Active Cdc42 is essential for the establishment of polarized growth. This GTPase is negatively regulated by the GTPase-activating proteins (GAPs), which are important for the spatial specificity of Cdc42 function. Rga4 is the only GAP described as negative regulator of fission yeast Cdc42. We report here that Rga6, another fission yeast Cdc42 GAP, shares some functions with Rga4. Cells lacking Rga6 are viable but slightly shorter and broader than wild type, and cells lacking Rga6 and Rga4 simultaneously are rounded. In these cells, active Cdc42 is observed all around the membrane. These additive effects indicate that both GAPs collaborate in the spatial regulation of active Cdc42. Rga6 localizes to the plasma membrane, forming clusters different from those formed by Rga4. A polybasic region at the Rga6 C-terminus is responsible for its membrane localization. Rga6-GFP fluorescence decreases considerably at the growing tips, and this decrease is dependent on the actin cables. Of note, in the absence of Rga6, the amplitude of active Cdc42 oscillations at the tips decreases, and less GTP-Cdc42 accumulates at the new end of the cells. We propose that Rga6 collaborates with Rga4 to spatially restrict active Cdc42 at the cell tips and maintain cell dimensions. PMID:26960792

Gravisensing in single-celled systems: characean rhizoids and protonemata.

PubMed

Braun, M

2001-01-01

Gravitropically tip-growing cell types are attractive unicellular model systems for investigating the mechanisms and the regulation of gravitropism. Especially useful for studying the mechanisms of positive and negative gravitropic tip-growth are characean rhizoids and protonemata. They originate from the same cell type, show the same overall cell shape, cytoplasmic zonation, arrangement of actin and microtubule cytoskeleton, use statoliths for gravisensing, but show opposite gravitropism. In both cell types, actin microfilaments are complexly organized in the apical dome,where a dense spherical actin array is colocalized with spectrin-like epitopes and a unique endoplasmic reticulum aggregate, the structural center of the Spitzenkörper. The opposite gravitropic responses seem to be based on differences in the actin-organized anchorage of the Spitzenkörper and the actin-mediated transport of statoliths. In negatively gravitropic (upward bending) protonemata, the statoliths-induced drastic upward shift of the cell tip is preceded by a relocalization of dihydropyridine-binding calcium channels and of the apical calcium gradient to the upper flank (bending by bulging). Such relocalizations have not been observed in positively gravitropically responding (downward growing) rhizoids in which statoliths sedimentation is followed by differential flank growth (bending by bowing). This paper reviews the current knowledge and hypotheses on the mechanisms of the opposite gravitropic responses in characean rhizoids and protonemata. c 2001 COSPAR. Published by Elsevier Science Ltd. All rights reserved.

Transjugular Intrahepatic Portosystemic Shunt Placement During Pregnancy: A Case Series of Five Patients

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

Ingraham, Christopher R., E-mail: cringra@uw.edu; Padia, Siddharth A., E-mail: spadia@uw.edu; Johnson, Guy E., E-mail: gej@uw.edu

Background and AimsComplications of portal hypertension, such as variceal hemorrhage and ascites, are associated with significant increases in both mortality and complications during pregnancy. Transjugular intrahepatic portosystemic shunt (TIPS) is a well-established procedure for treating portal hypertension, but the safety of TIPS during pregnancy is largely unknown. In this series, we review five patients who underwent TIPS placement while pregnant and describe their clinical outcomes.MethodsFive pregnant patients with cirrhosis and portal hypertension underwent elective TIPS for complications of portal hypertension (four for secondary prevention of variceal bleeding and one for refractory ascites). Outcomes measured were recurrent bleeding episodes or needmore » for further paracenteses during pregnancy, estimated radiation dose to the fetus and gestational age at delivery. All patients were followed after delivery to evaluate technical and clinical success of the procedure.ResultsAll five patients survived pregnancy and went on to deliver successfully. When TIPS was performed for secondary prevention of variceal bleeding (n = 4), no patients demonstrated variceal bleeding after TIPS placement. When TIPS was performed for refractory ascites (n = 1), no further paracenteses were required. All patients delivered successfully, albeit prematurely. Average radiation dose estimated to the fetus was 16.3 mGy.ConclusionsThis series suggests that TIPS can be performed in selective pregnant patients with portal hypertension, with little added risk to the mother or fetus.« less

The interaction of a gold atom with carbon nanohorn and carbon nanotube tips and their complexes with a CO molecule: A first principle calculation

NASA Astrophysics Data System (ADS)

Khongpracha, P.; Probst, M.; Limtrakul, J.

2008-07-01

The interactions of a gold atom with: (a) a single-wall carbon nanohorn (SWNH) conic tip; (b) with a single-wall carbon nanotube (SWNT) tip; and (c) their complexes with a CO molecule were studied using first-principle calculations based on density functional theory. The analysis of the pyramidalization angle (θp) as well as the π-orbital misalignment angles indicate that there should be many reactive carbon sites on the tips of SWNH and SWNT. It was found that SWNH provides reactive sites that can more selectively interact with the target atom. We identified five sites on both the SWNT tip and the nanohorn where attachment of a gold atom leads to a stable complex. This metal is found to be bi-coordinated with the tip of SWNH, while it is mono-coordinated with the SWNT tip. The largest interaction energies are -10.75 kcal/mol and -16.17 kcal/mol, respectively. The CO probe molecule binds to Au on the Au/SWNH or Au/SWNT tips with interaction energies of -22.34 and -18.29 kcal/mol, respectively. The main contributions of the interaction with both carbon nanostructures stems from σ-donation and π-backbonding. The results suggest that SWNHs could be one of the promising candidates for the development of high-specifity nanosensors.

Tip induced mechanical deformation of epitaxial graphene grown on reconstructed 6H-SiC(0001) surface during scanning tunneling and atomic force microscopy studies.

PubMed

Meza, José Antonio Morán; Lubin, Christophe; Thoyer, François; Cousty, Jacques

2015-01-26

The structural and mechanical properties of an epitaxial graphene (EG) monolayer thermally grown on top of a 6H-SiC(0001) surface were studied by combined dynamic scanning tunneling microscopy (STM) and frequency modulation atomic force microscopy (FM-AFM). Experimental STM, dynamic STM and AFM images of EG on 6H-SiC(0001) show a lattice with a 1.9 nm period corresponding to the (6 × 6) quasi-cell of the SiC surface. The corrugation amplitude of this (6 × 6) quasi-cell, measured from AFM topographies, increases with the setpoint value of the frequency shift Δf (15-20 Hz, repulsive interaction). Excitation variations map obtained simultaneously with the AFM topography shows that larger dissipation values are measured in between the topographical bumps of the (6 × 6) quasi-cell. These results demonstrate that the AFM tip deforms the graphene monolayer. During recording in dynamic STM mode, a frequency shift (Δf) map is obtained in which Δf values range from 41 to 47 Hz (repulsive interaction). As a result, we deduced that the STM tip, also, provokes local mechanical distortions of the graphene monolayer. The origin of these tip-induced distortions is discussed in terms of electronic and mechanical properties of EG on 6H-SiC(0001).

Preparation of Regular Specimens for Atom Probes

NASA Technical Reports Server (NTRS)

Kuhlman, Kim; Wishard, James

2003-01-01

A method of preparation of specimens of non-electropolishable materials for analysis by atom probes is being developed as a superior alternative to a prior method. In comparison with the prior method, the present method involves less processing time. Also, whereas the prior method yields irregularly shaped and sized specimens, the present developmental method offers the potential to prepare specimens of regular shape and size. The prior method is called the method of sharp shards because it involves crushing the material of interest and selecting microscopic sharp shards of the material for use as specimens. Each selected shard is oriented with its sharp tip facing away from the tip of a stainless-steel pin and is glued to the tip of the pin by use of silver epoxy. Then the shard is milled by use of a focused ion beam (FIB) to make the shard very thin (relative to its length) and to make its tip sharp enough for atom-probe analysis. The method of sharp shards is extremely time-consuming because the selection of shards must be performed with the help of a microscope, the shards must be positioned on the pins by use of micromanipulators, and the irregularity of size and shape necessitates many hours of FIB milling to sharpen each shard. In the present method, a flat slab of the material of interest (e.g., a polished sample of rock or a coated semiconductor wafer) is mounted in the sample holder of a dicing saw of the type conventionally used to cut individual integrated circuits out of the wafers on which they are fabricated in batches. A saw blade appropriate to the material of interest is selected. The depth of cut and the distance between successive parallel cuts is made such that what is left after the cuts is a series of thin, parallel ridges on a solid base. Then the workpiece is rotated 90 and the pattern of cuts is repeated, leaving behind a square array of square posts on the solid base. The posts can be made regular, long, and thin, as required for samples for atom-probe analysis. Because of their small volume and regularity, the amount of FIB-milling time can be much less than that of the method of sharp shards. Individual posts can be broken off for mounting in a manner similar to that of the method of sharp shards. Alternatively, the posts can be left intact on the base and the base can be cut to a small square (e.g., 3 by 3 mm) suitable for mounting in an atom probe of a type capable of accepting multiple-tip specimens. The advantage of multiple-tip specimens is the possibility of analyzing many tips without the time-consuming interchange of specimens.

Growth regulation in tip-growing cells that develop on the epidermis.

PubMed

Honkanen, Suvi; Dolan, Liam

2016-12-01

Plants develop tip-growing extensions-root hairs and rhizoids-that initiate as swellings on the outer surface of individual epidermal cells. A conserved genetic mechanism controls the earliest stages in the initiation of these swellings. The same mechanism controls the formation of multicellular structures that develop from swellings on epidermal cells in early diverging land plants. Details of the molecular events that regulate the positioning of the swellings involve sterols and phosphatidylinositol phosphates. The final length of root hairs is determined by the intensity of a pulse of transcription factor synthesis. Genes encoding similar transcription factors control root hair development in cereals and are potential targets for crop improvement. Copyright © 2016. Published by Elsevier Ltd.

Aluminium localization in root tips of the aluminium-accumulating plant species buckwheat (Fagopyrum esculentum Moench)

PubMed Central

Klug, Benjamin; Specht, André; Horst, Walter J.

2011-01-01

Aluminium (Al) uptake and transport in the root tip of buckwheat is not yet completely understood. For localization of Al in root tips, fluorescent dyes and laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) were compared. The staining of Al with morin is an appropriate means to study qualitatively the radial distribution along the root tip axis of Al which is complexed by oxalate and citrate in buckwheat roots. The results compare well with the distribution of total Al determined by LA-ICP-MS which could be reliably calibrated to compare with Al contents by conventional total Al determination using graphite furnace atomic absorption spectrometry. The Al localization in root cross-sections along the root tip showed that in buckwheat Al is highly mobile in the radial direction. The root apex predominantly accumulated Al in the cortex. The subapical root section showed a homogenous Al distribution across the whole section. In the following root section Al was located particularly in the pericycle and the xylem parenchyma cells. With further increasing distance from the root apex Al could be detected only in individual xylem vessels. The results support the view that the 10 mm apical root tip is the main site of Al uptake into the symplast of the cortex, while the subapical 10–20 mm zone is the main site of xylem loading through the pericycle and xylem parenchyma cells. Progress in the better molecular understanding of Al transport in buckwheat will depend on the consideration of the tissue specificity of Al transport and complexation. PMID:21831842

Expression and inhibition of aquaporins in germinating Arabidopsis seeds.

PubMed

Vander Willigen, Clare; Postaire, Olivier; Tournaire-Roux, Colette; Boursiac, Yann; Maurel, Christophe

2006-09-01

Extensive and kinetically well-defined water exchanges occur during germination of seeds. A putative role for aquaporins in this process was investigated in Arabidopsis. Macro-arrays carrying aquaporin gene-specific tags and antibodies raised against aquaporin subclasses revealed two distinct aquaporin expression programs between dry seeds and young seedlings. High expression levels of a restricted number of tonoplast intrinsic protein (TIP) isoforms (TIP3;1 and/or TIP3;2, and TIP5;1) together with a low expression of all 13 plasma membrane aquaporin (PIP) isoforms was observed in dry and germinating materials. In contrast, prevalent expression of aquaporins of the TIP1, TIP2 and PIP subgroups was induced during seedling establishment. Mercury (5 microM HgCl(2)), a general blocker of aquaporins in various organisms, reduced the speed of seed germination and induced a true delay in maternal seed coat (testa) rupture and radicle emergence, by 8-9 and 25-30 h, respectively. Most importantly, mercury did not alter seed lot homogeneity nor the seed germination developmental sequence, and its effects were largely reversed by addition of 2 mM dithiothreitol, suggesting that these effects were primarily due to oxidation of cell components, possibly aquaporins, without irreversible alteration of cell integrity. Measurements of water uptake in control and mercury-treated seeds suggested that aquaporin functions are not involved in early seed imbibition (phase I) but would rather be associated with a delayed initiation of phase III, i.e. water uptake accompanying expansion and growth of the embryo. A possible role for aquaporins in germinating seeds and more generally in plant tissue growth is discussed.

Hairlike Percutaneous Photochemical Sensors

NASA Technical Reports Server (NTRS)

George, Thomas; Loeb, Gerald

2004-01-01

Instrumentation systems based on hairlike fiber-optic photochemical sensors have been proposed as minimally invasive means of detecting biochemicals associated with cancer and other diseases. The fiber-optic sensors could be mass-produced as inexpensive, disposable components. The sensory tip of a fiber-optic sensor would be injected through the patient's skin into subcutaneous tissue. A biosensing material on the sensory tip would bind or otherwise react with the biochemical(s) of interest [the analyte(s)] to produce a change in optical properties that would be measured by use of an external photonic analyzer. After use, a fiber-optic sensor could be simply removed by plucking it out with tweezers. A fiber-optic sensor according to the proposal would be of the approximate size and shape of a human hair, and its sensory tip would resemble a follicle. Once inserted into a patient's subcutaneous tissue, the sensor would even more closely resemble a hair growing from a follicle (see Figure 1). The biosensing material on the sensory tip could consist of a chemical and/or cells cultured and modified for the purpose. The biosensing material would be contained within a membrane that would cover the tip. If the membrane were not permeable by an analyte, then it would be necessary to create pores in the membrane that would be large enough to allow analyte molecules to diffuse to the biosensing material, but not so large as to allow cells (if present as part of the biosensing material) to diffuse out. The end of the fiber-optic sensor opposite the sensory tip would be inserted in a fiberoptic socket in the photonic analyzer.

Diffusion-convection effects on drug distribution at the cell membrane level in a patch-clamp setup.

PubMed

Baran, Irina; Iftime, Adrian; Popescu, Anca

2010-01-01

We present a model-based method for estimating the effective concentration of the active drug applied by a pressure pulse to an individual cell in a patch-clamp setup, which could be of practical use in the analysis of ligand-induced whole-cell currents recorded in patch-clamp experiments. Our modelling results outline several important factors which may be involved in the high variability of the electric response of the cells, and indicate that with a pressure pulse duration of 1s and diameter of the perfusion tip of 600 μm, elevated amounts of drug can accumulate locally between the pipette tip and the cell. Hence, the effective agonist concentration at the cell membrane level can be consistently higher than the initial concentration inside the perfusion tubes. We performed finite-difference and finite-element simulations to investigate the diffusion/convection effects on the agonist distribution on the cell membrane. Our model can explain the delay between the commencement of acetylcholine application and the onset of the whole-cell current that we recorded on human rhabdomyosarcoma TE671 cells, and reproduce quantitatively the decrease of signal latency with the concentration of agonist in the pipette. Results also show that not only the geometry of the bath chamber and pipette tip, but also the transport parameters of the diffusive and convective phenomena in the bath solution are determinant for the amplitude and kinetics of the recorded currents and have to be accounted for when analyzing patch-clamp data. Copyright © 2010 Elsevier Ireland Ltd. All rights reserved.

EGF domain of coagulation factor IX is conducive to exposure of phosphatidylserine.

PubMed

Hidai, Chiaki; Fujiwara, Yusuke; Kokubun, Shinichiro; Kitano, Hisataka

2017-04-01

Lipid rafts are an initiation site for many different signals. Recently, we reported that an EGF domain in activated coagulation factor IX (EGF-F9) increases lipid raft formation and accelerates cell migration. However, the detailed mechanism is not well understood. This study aimed to evaluate the effects of EGF-F9 on the cell membrane. A431 cells (derived from human squamous cell carcinoma) were treated with recombinant EGF-F9. Cells were immunocytochemically stained with probes for lipid rafts or phosphatidylserine (PS). After 3 min of treatment with EGF-F9, cholera toxin subunit B (CTxB) binding domains emerged at the adhesive tips of filopodia. Subsequently, CTxB staining was observed on the filopodial shaft. Finally, large clusters of CTxB domains were observed at the edge of cell bodies. Markers for lipid rafts, such as caveolin-1 and a GPI anchored protein, co-localized with CTxB. Staining with annexin V and XII revealed that PS was exposed at the tips of filopodia, translocated on filopodial shafts, and co-localized with CTxB at the rafts. Immunocytochemistry showed that scramblase-1 protein was present at the filopodial tips. Our data indicates that EGF-F9 accelerates PS exposure around the filopodial adhesion complex and induces clustering of lipid rafts in the cell body. PS exposure is thought to occur on cells undergoing apoptosis. Further study of the function of the EGF-F9 motif in mediating signal transduction is necessary because it is shared by a number of proteins. © 2017 International Federation for Cell Biology.

Selective microrobot control using a thermally responsive microclamper for microparticle manipulation

NASA Astrophysics Data System (ADS)

Go, Gwangjun; Choi, Hyunchul; Jeong, Semi; Ko, Seong Young; Park, Jong-Oh; Park, Sukho

2016-03-01

Microparticle manipulation using a microrobot in an enclosed environment, such as a lab-on-a-chip, has been actively studied because an electromagnetic actuated microrobot can have accurate motility and wireless controllability. In most studies on electromagnetic actuated microrobots, only a single microrobot has been used to manipulate cells or microparticles. However, the use of a single microrobot can pose several limitations when performing multiple roles in microparticle manipulation. To overcome the limitations associated with using a single microrobot, we propose a new method for the control of multiple microrobots. Multiple microrobots can be controlled independently by an electromagnetic actuation system and multiple microclampers combined with microheaters. To select a specific microrobot among multiple microrobots, we propose a microclamper composed of a clamper structure using thermally responsive hydrogel and a microheater for controlling the microclamper. A fundamental test of the proposed microparticle manipulation system is performed by selecting a specific microrobot among multiple microrobots. Through the independent locomotion of multiple microrobots with U- and V-shaped tips, heterogeneous microparticle manipulation is demonstrated in the creation of a two-dimensional structure. In the future, our proposed multiple-microrobot system can be applied to tasks that are difficult to perform using a single microrobot, such as cell manipulation, cargo delivery, tissue assembly, and cloning.

Renal papillary tip extract stimulates BNP production and excretion from cardiomyocytes

PubMed Central

Hashizume, Ryotaro; Suzuki, Noboru; Ito, Rie; Yamanaka, Keiichi; Saito, Hiromitsu; Kiyonari, Hiroshi; Tawara, Isao; Kageyama, Yuki; Ogihara, Yoshito; Ali, Yusuf; Yamada, Norikazu; Katayama, Naoyuki; Ito, Masaaki

2018-01-01

Background Brain natriuretic peptide (BNP) is an important biomarker for patients with cardiovascular diseases, including heart failure, hypertension and cardiac hypertrophy. It is also known that BNP levels are relatively higher in patients with chronic kidney disease and no heart disease; however, the mechanism remains unclear. Methods and results We developed a BNP reporter mouse and occasionally found that this promoter was activated specifically in the papillary tip of the kidneys, and its activation was not accompanied by BNP mRNA expression. No evidence was found to support the existence of BNP isoforms or other nucleotide expression apart from BNP and tdTomato. The pBNP-tdTomato-positive cells were interstitial cells and were not proliferative. Unexpectedly, both the expression and secretion of BNP increased in primary cultured neonatal cardiomyocytes after their treatment with an extract of the renal papillary tip. Intraperitoneal injection of the extract of the papillary tips reduced blood pressure from 210 mmHg to 165 mmHg, the decrease being accompanied by an increase in serum BNP and urinary cGMP production in stroke-prone spontaneously hypertensive (SHR-SP) rats. Furthermore the induction of BNP by the papillary extract from rats with heart failure due to myocardial infarction was increased in cardiomyocytes. Conclusions These results suggested that the papillary tip express a substance that can stimulate BNP production and secretion from cardiomyocytes. PMID:29734386

Actin is an essential component of plant gravitropic signaling pathways

NASA Astrophysics Data System (ADS)

Braun, Markus; Hauslage, Jens; Limbach, Christoph

2003-08-01

A role of the actin cytoskeleton in the different phases of gravitropism in higher plant organs seems obvious, but experimental evidence is still inconclusive and contradictory. In gravitropically tip-growing rhizoids and protonemata, however, it is well documented that actin is an essential component of the tip-growth machinery and is involved either in the cellular mechanisms that lead to gravity sensing and in the processes of the graviresponses that result in the reorientation of the growth direction. All these processes depend on a complexly organized and highly dynamic organization of actin filaments whose diverse functions are coordinated by numerous associated proteins. Actin filaments and myosins mediate the transport of secretory vehicles to the growing tip and precisely control the delivery of cell wall material. In addition, both cell types use a very efficient actomyosin-based system to control and correct the position of their statoliths and to direct sedimenting statoliths to confined graviperception sites at the plasma membrane. The studies presented in this paper provide evidence for the essential role of actin in plant gravity sensing and the gravitropic responses. A unique actin-organizing center exists in the tip of characean rhizoids and protonemata which is associated with and dynamically regulated by a specific set of actin-dynamizing proteins. It is concluded that this highly dynamic apical actin array is an essential prerequisite for gravity sensing and gravity-oriented tip growth.

Vaginal Speculum For Photodynamic Therapy And Method Of Using The Same

DOEpatents

Tadir, Yona; Berns, Michael W.; Monk, Brad J.; Profeta, Glen; Tromberg, Bruce J.

1995-10-17

An improved vaginal speculum for photodynamic therapy of intraepithelial tissue and in particular vaginal, cervical and vulvar neoplasia utilizes a precisely and accurately positionable optic fiber through which a predetermined dose of light in the range of 620 to 700 nanometers is delivered over a controlled area which has been previously treated with photodynamic therapeutic substances. In particular, the neoplastic area has been treated with hematoporphyrin derivatives and other photosensitizers which are selectively taken into the cancerous tissue. Exposure to the appropriate wavelength laser light photoactivates the absorbed hematoporphyrins causing the release of singlet oxygen which internally oxidizes and ultimately causes cell death. The fiber optic tip from which the laser light is transmitted is precisely positioned within the body cavity at a predetermined distance from the intraepithelial neoplasia in order to obtain the appropriate spot size and location to minimize damage to healthy tissue and maximize damage to the selectively impregnated cancerous tissue.

Aluminium and hydrogen ions inhibit a mechanosensory calcium-selective cation channel

NASA Technical Reports Server (NTRS)

Ding, J. P.; Pickard, B. G.

1993-01-01

The tension-dependent activity of mechanosensory calcium-selective cation channels in excised plasmalemmal patches from onion bulb scale epidermis is modulated by pH in the physiologically meaningful range between 4.5 and 7.2. It is rapidly lowered by lowering pH and rapidly raised by raising pH. Channel activity is effectively inhibited by low levels of aluminium ions and activity can be partially restored by washing for a few minutes. We suggest that under normal conditions the sensitivity of the mechanosensory channels to pH of the wall free space plays important roles in regulation of plant activities such as growth. We further suggest that, when levels of acid and aluminium ions in the soil solution are high, they might inhibit similar sensory channels in cells of the root tip, thus contributing critically to the acid soil syndrome.

Dynamics of cell wall elasticity pattern shapes the cell during yeast mating morphogenesis.

PubMed

Goldenbogen, Björn; Giese, Wolfgang; Hemmen, Marie; Uhlendorf, Jannis; Herrmann, Andreas; Klipp, Edda

2016-09-01

The cell wall defines cell shape and maintains integrity of fungi and plants. When exposed to mating pheromone, Saccharomyces cerevisiae grows a mating projection and alters in morphology from spherical to shmoo form. Although structural and compositional alterations of the cell wall accompany shape transitions, their impact on cell wall elasticity is unknown. In a combined theoretical and experimental approach using finite-element modelling and atomic force microscopy (AFM), we investigated the influence of spatially and temporally varying material properties on mating morphogenesis. Time-resolved elasticity maps of shmooing yeast acquired with AFM in vivo revealed distinct patterns, with soft material at the emerging mating projection and stiff material at the tip. The observed cell wall softening in the protrusion region is necessary for the formation of the characteristic shmoo shape, and results in wider and longer mating projections. The approach is generally applicable to tip-growing fungi and plants cells. © 2016 The Authors.

Root gravitropism

NASA Technical Reports Server (NTRS)

Masson, P. H.

1995-01-01

When a plant root is reoriented within the gravity field, it responds by initiating a curvature which eventually results in vertical growth. Gravity sensing occurs primarily in the root tip. It may involve amyloplast sedimentation in the columella cells of the root cap, or the detection of forces exerted by the mass of the protoplast on opposite sides of its cell wall. Gravisensing activates a signal transduction cascade which results in the asymmetric redistribution of auxin and apoplastic Ca2+ across the root tip, with accumulation at the bottom side. The resulting lateral asymmetry in Ca2+ and auxin concentration is probably transmitted to the elongation zone where differential cellular elongation occurs until the tip resumes vertical growth. The Cholodny-Went theory proposes that gravity-induced auxin redistribution across a gravistimulated plant organ is responsible for the gravitropic response. However, recent data indicate that the gravity-induced reorientation is more complex, involving both auxin gradient-dependent and auxin gradient-independent events.

Transition from a planar interface to cellular and dendritic structures during rapid solidification processing

NASA Technical Reports Server (NTRS)

Laxmanan, V.

1986-01-01

The development of theoretical models which characterize the planar-cellular and cell-dendrite transitions is described. The transitions are analyzed in terms of the Chalmers number, the solute Peclet number, and the tip stability parameter, which correlate microstructural features and processing conditions. The planar-cellular transition is examined using the constitutional supercooling theory of Chalmers et al., (1953) and it is observed that the Chalmers number is between 0 and 1 during dendritic and cellular growth. Analysis of cell-dendrite transition data reveal that the transition occurs when the solute Peclet number goes through a minimum, the primary arm spacings go through a maximum, and the Chalmers number is equal to 1/2. The relation between the tip stability parameter and the solute Peclet number is investigated and it is noted that the tip stability parameter is useful for studying dendritic growth in alloys.

Neuropilin-2 mediates VEGF-C–induced lymphatic sprouting together with VEGFR3

PubMed Central

Xu, Yunling; Yuan, Li; Mak, Judy; Pardanaud, Luc; Caunt, Maresa; Kasman, Ian; Larrivée, Bruno; del Toro, Raquel; Suchting, Steven; Medvinsky, Alexander; Silva, Jillian; Yang, Jian; Thomas, Jean-Léon; Koch, Alexander W.; Alitalo, Kari

2010-01-01

Vascular sprouting is a key process-driving development of the vascular system. In this study, we show that neuropilin-2 (Nrp2), a transmembrane receptor for the lymphangiogenic vascular endothelial growth factor C (VEGF-C), plays an important role in lymphatic vessel sprouting. Blocking VEGF-C binding to Nrp2 using antibodies specifically inhibits sprouting of developing lymphatic endothelial tip cells in vivo. In vitro analyses show that Nrp2 modulates lymphatic endothelial tip cell extension and prevents tip cell stalling and retraction during vascular sprout formation. Genetic deletion of Nrp2 reproduces the sprouting defects seen after antibody treatment. To investigate whether this defect depends on Nrp2 interaction with VEGF receptor 2 (VEGFR2) and/or 3, we intercrossed heterozygous mice lacking one allele of these receptors. Double-heterozygous nrp2vegfr2 mice develop normally without detectable lymphatic sprouting defects. In contrast, double-heterozygote nrp2vegfr3 mice show a reduction of lymphatic vessel sprouting and decreased lymph vessel branching in adult organs. Thus, interaction between Nrp2 and VEGFR3 mediates proper lymphatic vessel sprouting in response to VEGF-C. PMID:20065093

Cytotoxicity of aluminum oxide nanoparticles on Allium cepa root tip--effects of oxidative stress generation and biouptake.

PubMed

Rajeshwari, A; Kavitha, S; Alex, Sruthi Ann; Kumar, Deepak; Mukherjee, Anita; Chandrasekaran, Natarajan; Mukherjee, Amitava

2015-07-01

The commercial usage of Al2O3 nanoparticles (Al2O3 NPs) has gone up significantly in the recent times, enhancing the risk of environmental contamination with these agents and their consequent adverse effects on living systems. The current study has been designed to evaluate the cytogenetic potential of Al2O3 NPs in Allium cepa (root tip cells) at a range of exposure concentrations (0.01, 0.1, 1, 10, and 100 μg/mL), their uptake/internalization profile, and the oxidative stress generated. We noted a dose-dependent decrease in the mitotic index (42 to 28 %) and an increase in the number of chromosomal aberrations. Various chromosomal aberrations, e.g. sticky, multipolar and laggard chromosomes, chromosomal breaks, and the formation of binucleate cells, were observed by optical, fluorescence, and confocal laser scanning microscopy. FT-IR analysis demonstrated the surface chemical interaction between the nanoparticles and root tip cells. The biouptake of Al2O3 in particulate form led to reactive oxygen species generation, which in turn probably contributed to the induction of chromosomal aberrations.

Emergent behavior of cells on microfabricated soft polymeric substrates

NASA Astrophysics Data System (ADS)

Anand, Sandeep Venkit

In recent years, cell based bio-actuators like cardiomyocytes and skeletal muscle cells have emerged as popular choices for powering biological machines consisting of soft polymeric scaffolds at the micro and macro scales. This is owing to their unique ability to generate spontaneous, synchronous contractions either autonomously or under externally applied fields. Most of the biological machine designs reported in literature use single cells or cell clusters conjugated with biocompatible soft polymers like polydimethylsiloxane (PDMS) and hydrogels to produce some form of locomotion by converting chemical energy of the cells to mechanical energy. The mode of locomotion may vary, but the fundamental mechanism that these biological machines exploit to achieve locomotion stems from cell substrate interactions leading to large deformations of the substrates (relative to the cell size). However, the effect of such large scale, dynamic deformation of the substrates on the cellular and cluster level organization of the cells remains elusive. This dissertation tries to explore the emergent behavior of cells on different types of micro-scale deformable, soft polymeric substrates. In the first part of the dissertation, contractile dynamics of primary cardiomyocyte clusters is studied by culturing them on deformable thin polymeric films. The cell clusters beat and generate sufficient forces to deform the substrates out of plane. Over time, the clusters reorient their force dipoles along the direction of maximum compliance. This suggests that the cells are capable of sensing substrate deformations through a mechanosensitive feedback mechanism and dynamically reorganizing themselves. Results are further validated through finite element analysis. The development, characterization and quantification of a novel 1D/2D like polymeric platform for cell culture is presented in the second part. The platform consists of a 2D surface anchoring a long (few millimeters) narrow filament (1D) with a single cell scale (micro scale) cross section. We plate C2C12 cells on the platform and characterize their migration, proliferation, and differentiation patterns in contrast to 2D culture. We find that the cells land on the 2D surface, and then migrate to the filament only when the 2D surface has become nearly confluent. Individual and isolated cells randomly approaching the filament always retract away towards the 2D surface. Once on the filament, their differentiation to myotubes is expedited compared to that on 2D substrate. The myotubes generate periodic twitching forces that deform the filament producing more than 17 um displacement at the tip. Such flagellar motion can be used to develop autonomous micro scale bio-bots. Finally, the design and fabrication of a polymeric micro-pillar based force sensor capable of measuring cellular focal-adhesion forces under externally applied stretch is discussed. The force sensor consists of arrays of uniformly spaced PDMS micro-pillars of 1-2 um diameter and 2-3 um spacing on a macroscale PDMS substrate. The tips of the micro-pillars are selectively patterned with fluorescently labeled ECM proteins using micro-contact printing to promote cell adhesion while simultaneously acting as markers for strain measurements. Cells adhere and spread on top of the pillars causing them to deform. When stretched, the cells reorganize their internal structure and modulate their traction forces in response to the applied stretch. The dynamically varying cellular forces in response to the stretch are computed by measuring the cell induced displacements estimated by isolating the displacements caused by the applied stretch from the net displacements of the tips.

Conductance maps of quantum rings due to a local potential perturbation.

PubMed

Petrović, M D; Peeters, F M; Chaves, A; Farias, G A

2013-12-11

We performed a numerical simulation of the dynamics of a Gaussian shaped wavepacket inside a small sized quantum ring, smoothly connected to two leads and exposed to a perturbing potential of a biased atomic force microscope tip. Using the Landauer formalism, we calculated conductance maps of this system in the case of single and two subband transport. We explain the main features in the conductance maps as due to the AFM tip influence on the wavepacket phase and amplitude. In the presence of an external magnetic field, the tip modifies the ϕ0 periodic Aharonov-Bohm oscillation pattern into a ϕ0/2 periodic Al'tshuler-Aronov-Spivak oscillation pattern. Our results in the case of multiband transport suggest tip selectivity to higher subbands, making them more observable in the total conductance map.

CFD simulations of a wind turbine for analysis of tip vortex breakdown

NASA Astrophysics Data System (ADS)

Kimura, K.; Tanabe, Y.; Aoyama, T.; Matsuo, Y.; Arakawa, C.; Iida, M.

2016-09-01

This paper discusses about the wake structure of wind turbine via the use of URANS and Quasi-DNS, focussing on the tip vortex breakdown. The moving overlapped structured grids CFD Solver based on a fourth-order reconstruction and an all-speed scheme, rFlow3D is used for capturing the characteristics of tip vortices. The results from the Model Experiments in Controlled Conditions project (MEXICO) was accordingly selected for executing wake simulations through the variation of tip speed ratio (TSR); in an operational wind turbine, TSR often changes in value. Therefore, it is important to assess the potential effects of TSR on wake characteristics. The results obtained by changing TSR show the variations of the position of wake breakdown and wake expansion. The correspondence between vortices and radial/rotational flow is also confirmed.

Nanofabrication of insulated scanning probes for electromechanical imaging in liquid solutions

PubMed Central

Noh, Joo Hyon; Nikiforov, Maxim; Kalinin, Sergei V.; Vertegel, Alexey A.; Rack, Philip D.

2011-01-01

In this paper, the fabrication and electrical and electromechanical characterization of insulated scanning probes have been demonstrated in liquid solutions. The silicon cantilevers were sequentially coated with chromium and silicon dioxide, and the silicon dioxide was selectively etched at tip apex using focused electron beam induced etching (FEBIE) with XeF2 The chromium layer acted not only as the conductive path from the tip, but also as an etch resistant layer. This insulated scanning probe fabrication process is compatible with any commercial AFM tip and can be used to easily tailor the scanning probe tip properties because FEBIE does not require lithography. The suitability of the fabricated probes is demonstrated by imaging of standard topographical calibration grid as well as piezoresponse force microscopy (PFM) and electrical measurements in ambient and liquid environments. PMID:20702930

System and method for improved rotor tip performance

NASA Technical Reports Server (NTRS)

Zientek, Thomas A. (Inventor); Bussom, Richard (Inventor); McVeigh, Michael A. (Inventor); Narducci, Robert P. (Inventor)

2007-01-01

The present invention discloses systems and methods for the performance enhancement of rotary wing aircraft through reduced torque, noise and vibration. In one embodiment, a system includes a sail having an aerodynamic shape positioned proximate to a tip of the rotor blade. An actuator may be configured to rotate the sail relative to the blade tip. a A control system receives information from a rotorcraft system and commands the actuator to rotate the sail to a predetermined favorable rotor blade operating condition. In another embodiment, a method includes configuring the rotorcraft in a selected flight condition, communicating input signals to a control system operable to position sails coupled to tips of blades of a rotor assembly, processing the input signals according to a constraint condition to generate sail positional information, and transferring the sail positional information to the sail.

Tips for Teachers Selecting Toys to Facilitate Social Interaction

ERIC Educational Resources Information Center

Vail, Cynthia O.; Elmore, Shannon Renee

2011-01-01

Toy selection is an important role for early childhood teachers. This research-to-practice article describes what research tells us about how toys can affect the social interactions and communication of young children including those with developmental delays.

Nanostructure and force spectroscopy analysis of human peripheral blood CD4+ T cells using atomic force microscopy.

PubMed

Hu, Mingqian; Wang, Jiongkun; Cai, Jiye; Wu, Yangzhe; Wang, Xiaoping

2008-09-12

To date, nanoscale imaging of the morphological changes and adhesion force of CD4(+) T cells during in vitro activation remains largely unreported. In this study, we used atomic force microscopy (AFM) to study the morphological changes and specific binding forces in resting and activated human peripheral blood CD4(+) T cells. The AFM images revealed that the volume of activated CD4(+) T cells increased and the ultrastructure of these cells also became complex. Using a functionalized AFM tip, the strength of the specific binding force of the CD4 antigen-antibody interaction was found to be approximately three times that of the unspecific force. The adhesion forces were not randomly distributed over the surface of a single activated CD4(+) T cell, indicated that the CD4 molecules concentrated into nanodomains. The magnitude of the adhesion force of the CD4 antigen-antibody interaction did not change markedly with the activation time. Multiple bonds involved in the CD4 antigen-antibody interaction were measured at different activation times. These results suggest that the adhesion force involved in the CD4 antigen-antibody interaction is highly selective and of high affinity.

Human embryonic lung epithelial tips are multipotent progenitors that can be expanded in vitro as long-term self-renewing organoids

PubMed Central

Nikolić, Marko Z; Caritg, Oriol; Jeng, Quitz; Johnson, Jo-Anne; Sun, Dawei; Howell, Kate J; Brady, Jane L; Laresgoiti, Usua; Allen, George; Butler, Richard; Zilbauer, Matthias; Giangreco, Adam; Rawlins, Emma L

2017-01-01

The embryonic mouse lung is a widely used substitute for human lung development. For example, attempts to differentiate human pluripotent stem cells to lung epithelium rely on passing through progenitor states that have only been described in mouse. The tip epithelium of the branching mouse lung is a multipotent progenitor pool that self-renews and produces differentiating descendants. We hypothesized that the human distal tip epithelium is an analogous progenitor population and tested this by examining morphology, gene expression and in vitro self-renewal and differentiation capacity of human tips. These experiments confirm that human and mouse tips are analogous and identify signalling pathways that are sufficient for long-term self-renewal of human tips as differentiation-competent organoids. Moreover, we identify mouse-human differences, including markers that define progenitor states and signalling requirements for long-term self-renewal. Our organoid system provides a genetically-tractable tool that will allow these human-specific features of lung development to be investigated. DOI: http://dx.doi.org/10.7554/eLife.26575.001 PMID:28665271

Thermo-solutal and kinetic modes of stable dendritic growth with different symmetries of crystalline anisotropy in the presence of convection

NASA Astrophysics Data System (ADS)

Alexandrov, Dmitri V.; Galenko, Peter K.; Toropova, Lyubov V.

2018-01-01

Motivated by important applications in materials science and geophysics, we consider the steady-state growth of anisotropic needle-like dendrites in undercooled binary mixtures with a forced convective flow. We analyse the stable mode of dendritic evolution in the case of small anisotropies of growth kinetics and surface energy for arbitrary Péclet numbers and n-fold symmetry of dendritic crystals. On the basis of solvability and stability theories, we formulate a selection criterion giving a stable combination between dendrite tip diameter and tip velocity. A set of nonlinear equations consisting of the solvability criterion and undercooling balance is solved analytically for the tip velocity V and tip diameter ρ of dendrites with n-fold symmetry in the absence of convective flow. The case of convective heat and mass transfer mechanisms in a binary mixture occurring as a result of intensive flows in the liquid phase is detailed. A selection criterion that describes such solidification conditions is derived. The theory under consideration comprises previously considered theoretical approaches and results as limiting cases. This article is part of the theme issue `From atomistic interfaces to dendritic patterns'. This article is part of the theme issue `From atomistic interfaces to dendritic patterns'.

Estimation of Nasal Tip Support Using Computer-Aided Design and 3-Dimensional Printed Models

PubMed Central

Gray, Eric; Maducdoc, Marlon; Manuel, Cyrus; Wong, Brian J. F.

2016-01-01

IMPORTANCE Palpation of the nasal tip is an essential component of the preoperative rhinoplasty examination. Measuring tip support is challenging, and the forces that correspond to ideal tip support are unknown. OBJECTIVE To identify the integrated reaction force and the minimum and ideal mechanical properties associated with nasal tip support. DESIGN, SETTING, AND PARTICIPANTS Three-dimensional (3-D) printed anatomic silicone nasal models were created using a computed tomographic scan and computer-aided design software. From this model, 3-D printing and casting methods were used to create 5 anatomically correct nasal models of varying constitutive Young moduli (0.042, 0.086, 0.098, 0.252, and 0.302 MPa) from silicone. Thirty rhinoplasty surgeons who attended a regional rhinoplasty course evaluated the reaction force (nasal tip recoil) of each model by palpation and selected the model that satisfied their requirements for minimum and ideal tip support. Data were collected from May 3 to 4, 2014. RESULTS Of the 30 respondents, 4 surgeons had been in practice for 1 to 5 years; 9 surgeons, 6 to 15 years; 7 surgeons, 16 to 25 years; and 10 surgeons, 26 or more years. Seventeen surgeons considered themselves in the advanced to expert skill competency levels. Logistic regression estimated the minimum threshold for the Young moduli for adequate and ideal tip support to be 0.096 and 0.154 MPa, respectively. Logistic regression estimated the thresholds for the reaction force associated with the absolute minimum and ideal requirements for good tip recoil to be 0.26 to 4.74 N and 0.37 to 7.19 N during 1- to 8-mm displacement, respectively. CONCLUSIONS AND RELEVANCE This study presents a method to estimate clinically relevant nasal tip reaction forces, which serve as a proxy for nasal tip support. This information will become increasingly important in computational modeling of nasal tip mechanics and ultimately will enhance surgical planning for rhinoplasty. LEVEL OF EVIDENCE NA. PMID:27124818

The Microstructural Evolution of Fatigue Cracks in FCC Metals

NASA Astrophysics Data System (ADS)

Gross, David William

The microstructural evolution during fatigue crack propagation was investigated in a variety of planar and wavy slip FCC metals. The planar materials included Haynes 230, Nitronic 40, and 316 stainless steel, and the wavy materials included pure nickel and pure copper. Three different sets of experiments were performed to fully characterize the microstructural evolution. The first, performed on Haynes 230, mapped the strain field ahead a crack tip using digital image correlation and electron backscatter diffraction techniques. Focused ion beam (FIB) lift-out techniques were then utilized to extract transmission electron microscopy (TEM) samples at specific distances from the crack tip. TEM investigations compared the measured strain to the microstructure. Overall, the strain measured via DIC and EBSD was only weakly correlated to the density of planar slip bands in the microstructure. The second set of experiments concerned the dislocation structure around crack tips. This set of experiments was performed on all the materials. The microstructure at arrested fatigue cracks on the free surface was compared to the microstructure found beneath striations on the fracture surfaces by utilizing FIB micromachining to create site-specific TEM samples. The evolved microstructure depended on the slip type. Strong agreement was found between the crack tip microstructure at the free surface and the fracture surface. In the planar materials, the microstructure in the plastic zone consisted of bands of dislocations or deformation twins, before transitioning to a refined sub-grain microstructure near the crack flank. The sub-grain structure extended 300-500 nm away from the crack flank in all the planar slip materials studied. In contrast, the bulk structure in the wavy slip material consisted of dislocation cells and did not transition to a different microstructure as the crack tip was approached. The strain in wavy slip was highest near the crack tip, as the misorientations between the dislocation cells increased and the cell size decreased as the crack flank was approached. The final set of experiments involved reloading the arrested crack tips in monotonic tension. This was performed on both the Haynes 230 and 316 stainless steel. This technique exposed the fracture surface and location of the arrested crack tip away from the free surface, allowing for a sample to be extracted via FIB micromachining and TEM evaluation of the microstructure. This permitted the crack tip microstructure to be investigated without exposing the microstructure to crack closure or free surface effects. These experiments confirmed what was inferred from the earlier experiments, namely that the banded structure was a product of the crack tip plastic zone and the refined structure was a product of the strain associated with crack advance. Overall the microstructural complexity presented in this work was much higher than would be predicted by current models of fatigue crack propagation. It is recommended that future models attempt to simulate interactions between the dislocations emitted during fatigue crack growth and the pre-existing microstructure to more accurately simulate the processes occurring at the crack tip during crack growth.

Dark tip-tilt sensing

NASA Astrophysics Data System (ADS)

Arcidiacono, Carmelo; Ragazzoni, Roberto; Viotto, Valentina; Bergomi, Maria; Farinato, Jacopo; Magrin, Demetrio; Dima, Marco; Gullieuszik, Marco; Marafatto, Luca

2016-07-01

Dark wavefront sensing in its simplest and more crude form is a quad-cell with a round spot of dark ink acting as occulting disk at the center. This sensor exhibits fainter limiting magnitude than a conventional quad-cell, providing that the size of the occulting disk is slightly smaller than the size of the spot and smaller than the residual jitter movement in closed loop. We present simulations focusing a generic Adaptive Optics system using Natural Guide Stars to provide the tip-tilt signal. We consider a jitter spectrum of the residual correction including amplitudes exceeding the dark disk size.

Buder revisited: cell and organ polarity during phototropism.

PubMed

Nick, P; Furuya, M

1996-10-01

The induction of a radial polarity by environmental stimuli was studied at the cellular and organ levels, with phototropism chosen as a model. The light gradient acting on the whole coleoptile was opposed to the light direction acting upon individual cells in the classical Buder experiment, irradiating from the inside out. Alternatively, the stimulus was administered to the coleoptile tip with a microbeam-irradiation device. Tropistic curvature was assayed as a marker for the response of the whole organ, whereas cell elongation and the orientation of cortical microtubules were taken as markers for the responses of individual cells. Upon tip irradiation, signals much faster than basipetal auxin transport migrate towards the base. The data are discussed in terms of an organ polarity that is the primary result of the asymmetric light signal and affects, in a second step, an endogenous radial polarity of epidermal cells.

Survival rate of eukaryotic cells following electrophoretic nanoinjection

PubMed Central

Simonis, Matthias; Hübner, Wolfgang; Wilking, Alice; Huser, Thomas; Hennig, Simon

2017-01-01

Insertion of foreign molecules such as functionalized fluorescent probes, antibodies, or plasmid DNA to living cells requires overcoming the plasma membrane barrier without harming the cell during the staining process. Many techniques such as electroporation, lipofection or microinjection have been developed to overcome the cellular plasma membrane, but they all result in reduced cell viability. A novel approach is the injection of cells with a nanopipette and using electrophoretic forces for the delivery of molecules. The tip size of these pipettes is approximately ten times smaller than typical microinjection pipettes and rather than pressure pulses as delivery method, moderate DC electric fields are used to drive charged molecules out of the tip. Here, we show that this approach leads to a significantly higher survival rate of nanoinjected cells and that injection with nanopipettes has a significantly lower impact on the proliferation behavior of injected cells. Thus, we propose that injection with nanopipettes using electrophoretic delivery is an excellent alternative when working with valuable and rare living cells, such as primary cells or stem cells. PMID:28120926

Dielectrophoresis-based particle sensor using nanoelectrode arrays

NASA Technical Reports Server (NTRS)

Arumugam, Prabhu U. (Inventor); Li, Jun (Inventor); Cassell, Alan M. (Inventor)

2009-01-01

A method for concentrating or partly separating particles of a selected species from a liquid or fluid containing these particles and flowing in a channel, and for determining if the selected species particle is present in the liquid or fluid. A time varying electrical field E, having a root-mean-square intensity E.sup.2.sub.rms with a non-zero gradient in a direction transverse to the liquid or fluid flow direction, is produced by a nanostructure electrode array, with a very high magnitude gradient near exposed electrode tips. A dielectrophoresis force causes the selected particles to accumulate near the electrode tips, if the medium and selected particles have substantially different dielectric constants. An insulating material surrounds most of each of the nanostructure electrodes, and a region of the insulating material surface is functionalized to promote attachment of the selected species particles to the surface. An electrical property value Z(meas) is measured at the functionalized surface and is compared with a reference value Z(ref) to determine if the selected species particles are attached to the functionalized surface.

Effects of aluminum on nucleoli in root tip cells and selected physiological and biochemical characters in Allium cepa var. agrogarum L

PubMed Central

2010-01-01

Background Increased Al concentration causes reduction of mitotic activity, induction of nucleolar alteration, increase of the production of ROS and alteration of several antioxidant enzyme activities in plant cells. Allium cepa is an excellent plant and a useful biomarker for environmental monitoring. Limited information is available about the effects of Al on nucleoli, antioxidant enzyme system, contents of MDA and soluble protein in A. cepa. Therefore, we carried out the investigation in order to better understand the effects of Al on the growth, nucleoli in root tip cells and selected physiological and biochemical characters. Results The results showed that the root growth exposed to 50 μM Al was inhibited significantly. 50 μM Al could induce some particles of argyrophilic proteins scattered in the nuclei and extruded from the nucleoli into the cytoplasm. The nucleolus did not disaggregate normally and still remained its characteristic structure during metaphase. Nucleolar reconstruction was inhibited. 50 μM Al induced high activities of SOD and POD in leaves and roots significantly (P < 0.05) when compared with control, whereas the level of CAT was low significantly (P < 0.05). At 50 μM Al the content of MDA in leaves was high significantly (P < 0.05) at 9th day and in roots increased (P < 0.05) with prolonging the treatment time during 6-12 days. The soluble protein content in leaves treated with 50 μM Al was high significantly (P < 0.05) at 6th day and increased with prolonging the treatment time. Conclusions We suggest that variations in nucleoli and the alterations of antioxidant enzyme activities, MDA and soluble protein contents in Allium cepa can serve as useful biomarkers, which can provide valuable information for monitoring and forecasting effects of exposure to Al in real scenarios conditions. Among the antioxidant enzymes SOD and POD appear to play a key role in the antioxidant defense mechanism under Al toxicity condition. Data from MDA concentration show that Al indirectly produces superoxide radicals, resulting in increased lipid peroxidative products and oxidative stress. PMID:20964828

Effects of aluminum on nucleoli in root tip cells and selected physiological and biochemical characters in Allium cepa var. agrogarum L.

PubMed

Qin, Rong; Jiao, Yunqiu; Zhang, Shanshan; Jiang, Wusheng; Liu, Donghua

2010-10-21

Increased Al concentration causes reduction of mitotic activity, induction of nucleolar alteration, increase of the production of ROS and alteration of several antioxidant enzyme activities in plant cells. Allium cepa is an excellent plant and a useful biomarker for environmental monitoring. Limited information is available about the effects of Al on nucleoli, antioxidant enzyme system, contents of MDA and soluble protein in A. cepa. Therefore, we carried out the investigation in order to better understand the effects of Al on the growth, nucleoli in root tip cells and selected physiological and biochemical characters. The results showed that the root growth exposed to 50 μM Al was inhibited significantly. 50 μM Al could induce some particles of argyrophilic proteins scattered in the nuclei and extruded from the nucleoli into the cytoplasm. The nucleolus did not disaggregate normally and still remained its characteristic structure during metaphase. Nucleolar reconstruction was inhibited. 50 μM Al induced high activities of SOD and POD in leaves and roots significantly (P < 0.05) when compared with control, whereas the level of CAT was low significantly (P < 0.05). At 50 μM Al the content of MDA in leaves was high significantly (P < 0.05) at 9(th) day and in roots increased (P < 0.05) with prolonging the treatment time during 6-12 days. The soluble protein content in leaves treated with 50 μM Al was high significantly (P < 0.05) at 6(th) day and increased with prolonging the treatment time. We suggest that variations in nucleoli and the alterations of antioxidant enzyme activities, MDA and soluble protein contents in Allium cepa can serve as useful biomarkers, which can provide valuable information for monitoring and forecasting effects of exposure to Al in real scenarios conditions. Among the antioxidant enzymes SOD and POD appear to play a key role in the antioxidant defense mechanism under Al toxicity condition. Data from MDA concentration show that Al indirectly produces superoxide radicals, resulting in increased lipid peroxidative products and oxidative stress.

IRSp53 is colocalised with WAVE2 at the tips of protruding lamellipodia and filopodia independently of Mena.

PubMed

Nakagawa, Hiroyuki; Miki, Hiroaki; Nozumi, Motohiro; Takenawa, Tadaomi; Miyamoto, Shigeaki; Wehland, Jürgen; Small, J Victor

2003-06-15

The insulin receptor tyrosine kinase substrate p53 (IRSp53) links Rac and WAVE2 and has been implicated in lamellipodia protrusion. Recently, however, IRSp53 has been reported to bind to both Cdc42 and Mena to induce filopodia. To shed independent light on IRSp53 function we determined the localisations and dynamics of IRSp53 and WAVE2 in B16 melanoma cells. In cells spread well on a laminin substrate, IRSp53 was localised by antibody labelling at the tips of both lamellipodia and filopodia. The same localisation was observed in living cells with IRSp53 tagged with enhanced green florescence protein (EGFP-IRSp53), but only during protrusion. From the transfection of deletion mutants the N-terminal region of IRSp53, which binds active Rac, was shown to be responsible for its localisation. Although IRSp53 has been reported to regulate filopodia formation with Mena, EGFP-IRSp53 showed the same localisation in MVD7 Ena/VASP (vasodilator stimulated phosphoprotein) family deficient cells. WAVE2 tagged with DsRed1 colocalised with EGFP-IRSp53 at the tips of protruding lamellipodia and filopodia and, in double-transfected cells, the IRSp53 signal in filopodia decreased before that of WAVE2 during retraction. These results suggest an alternative modulatory role for IRSp53 in the extension of both filopodia and lamellipodia, through WAVE2.

Note: Design and fabrication of a simple versatile microelectrochemical cell and its accessories

NASA Astrophysics Data System (ADS)

Rajan, Viswanathan; Neelakantan, Lakshman

2015-09-01

A microelectrochemical cell housed in an optical microscope and custom-made accessories have been designed and fabricated, which allows performing spatially resolved corrosion measurements. The cell assembly was designed to directly integrate the reference electrode close to the capillary tip to avoid air bubbles. A hard disk along with an old optical microscope was re-engineered into a microgrinder, which made the vertical grinding of glass capillary tips very easy. A stepper motor was customized into a microsyringe pump to dispense a controlled volume of electrolyte through the capillary. A force sensitive resistor was used to achieve constant wetting area. The functionality of the developed instrument is demonstrated by studying μ-electrochemical behavior of worn surface on AA2014-T6 alloy.

Numerical investigation of the unsteady tip leakage flow and rotating stall inception in a transonic compressor

NASA Astrophysics Data System (ADS)

Zhang, Yanfeng; Lu, Xingen; Chu, Wuli; Zhu, Junqiang

2010-08-01

It is well known that tip leakage flow has a strong effect on the compressor performance and stability. This paper reports on a numerical investigation of detailed flow structures in an isolated transonic compressor rotor-NASA Rotor 37 at near stall and stalled conditions aimed at improving understanding of changes in 3D tip leakage flow structures with rotating stall inception. Steady and unsteady 3D Navier-Stokes analyses were conducted to investigate flow structures in the same rotor. For steady analysis, the predicted results agree well with the experimental data for the estimation of compressor rotor global performance. For unsteady flow analysis, the unsteady flow nature caused by the breakdown of the tip leakage vortex in blade tip region in the transonic compressor rotor at near stall condition has been captured with a single blade passage. On the other hand, the time-accurate unsteady computations of multi-blade passage at near stall condition indicate that the unsteady breakdown of the tip leakage vortex triggered the short length-scale — spike type rotating stall inception at blade tip region. It was the forward spillage of the tip leakage flow at blade leading edge resulting in the spike stall inception. As the mass flow ratio is decreased, the rotating stall cell was further developed in the blade passage.

Antisense expression of an Arabidopsis ran binding protein renders transgenic roots hypersensitive to auxin and alters auxin-induced root growth and development by arresting mitotic progress

NASA Technical Reports Server (NTRS)

Kim, S. H.; Arnold, D.; Lloyd, A.; Roux, S. J.

2001-01-01

We cloned a cDNA encoding an Arabidopsis Ran binding protein, AtRanBP1c, and generated transgenic Arabidopsis expressing the antisense strand of the AtRanBP1c gene to understand the in vivo functions of the Ran/RanBP signal pathway. The transgenic plants showed enhanced primary root growth but suppressed growth of lateral roots. Auxin significantly increased lateral root initiation and inhibited primary root growth in the transformants at 10 pM, several orders of magnitude lower than required to induce these responses in wild-type roots. This induction was followed by a blockage of mitosis in both newly emerged lateral roots and in the primary root, ultimately resulting in the selective death of cells in the tips of both lateral and primary roots. Given the established role of Ran binding proteins in the transport of proteins into the nucleus, these findings are consistent with a model in which AtRanBP1c plays a key role in the nuclear delivery of proteins that suppress auxin action and that regulate mitotic progress in root tips.

Trans-Golgi network localized small GTPase RabA1d is involved in cell plate formation and oscillatory root hair growth.

PubMed

Berson, Tobias; von Wangenheim, Daniel; Takáč, Tomáš; Šamajová, Olga; Rosero, Amparo; Ovečka, Miroslav; Komis, George; Stelzer, Ernst H K; Šamaj, Jozef

2014-09-27

Small Rab GTPases are important regulators of vesicular trafficking in plants. AtRabA1d, a member of the RabA1 subfamily of small GTPases, was previously found in the vesicle-rich apical dome of growing root hairs suggesting a role during tip growth; however, its specific intracellular localization and role in plants has not been well described. The transient expression of 35S::GFP:RabA1d construct in Allium porrum and Nicotiana benthamiana revealed vesicular structures, which were further corroborated in stable transformed Arabidopsis thaliana plants. GFP-RabA1d colocalized with the trans-Golgi network marker mCherry-VTI12 and with early FM4-64-labeled endosomal compartments. Late endosomes and endoplasmic reticulum labeled with FYVE-DsRed and ER-DsRed, respectively, were devoid of GFP-RabA1d. The accumulation of GFP-RabA1d in the core of brefeldin A (BFA)-induced-compartments and the quantitative upregulation of RabA1d protein levels after BFA treatment confirmed the association of RabA1d with early endosomes/TGN and its role in vesicle trafficking. Light-sheet microscopy revealed involvement of RabA1d in root development. In root cells, GFP-RabA1d followed cell plate expansion consistently with cytokinesis-related vesicular trafficking and membrane recycling. GFP-RabA1d accumulated in disc-like structures of nascent cell plates, which progressively evolved to marginal ring-like structures of the growing cell plates. During root hair growth and development, GFP-RabA1d was enriched at root hair bulges and at the apical dome of vigorously elongating root hairs. Importantly, GFP-RabA1d signal intensity exhibited an oscillatory behavior in-phase with tip growth. Progressively, this tip localization dissapeared in mature root hairs suggesting a link between tip localization of RabA1d and root hair elongation. Our results support a RabA1d role in events that require vigorous membrane trafficking. RabA1d is located in early endosomes/TGN and is involved in vesicle trafficking. RabA1d participates in both cell plate formation and root hair oscillatory tip growth. The specific GFP-RabA1d subcellular localization confirms a correlation between its specific spatio-temporal accumulation and local vesicle trafficking requirements during cell plate and root hair formation.

Human iPSC-Derived Endothelial Cell Sprouting Assay in ...

EPA Pesticide Factsheets

Activation of vascular endothelial cells (ECs) by growth factors initiates a cascade of events in vivo consisting of EC tip cell selection, sprout formation, EC stalk cell proliferation, and ultimately vascular stabilization by support cells. Although EC functional assays can recapitulate one or more aspects of angiogenesis in vitro, they are often limited by a lack of definition to the substratum and lack of dependence on key angiogenic signaling axes. Here, we designed and characterized a chemically-defined model of endothelial sprouting behavior in vitro using human induced pluripotent stem cell-derived endothelial cells (iPSC-ECs). Thiol-ene photopolymerization was used to rapidly encapsulate iPSC-ECs at high density in poly(ethylene glycol) (PEG) hydrogel spheres and subsequently to rapidly encapsulate iPSC-EC-containing hydrogel spheres in a cell-free over-layer. The hydrogel sprouting array here maintained pro-angiogenic phenotype of iPSC-ECs and supported growth factor-dependent proliferation and sprouting behavior. The sprouting model responded appropriately to several reference pharmacological angiogenesis inhibitors, which suggests the functional role of vascular endothelial growth factor, NF-κB, matrix metalloproteinase-2/9, protein kinase activity, and β-tubulin in endothelial sprouting. A blinded screen of 38 putative vascular disrupting compounds (pVDCs) from the US Environmental Protection Agency’s ToxCast library identified five compounds th

Conformational Changes in IpaD from Shigella flexneri Upon Binding Bile Salts Provide Insight into the Second Step of Type III Secretion†

PubMed Central

Dickenson, Nicholas E.; Zhang, Lingling; Epler, Chelsea R.; Adam, Philip R.; Picking, Wendy L.; Picking, William D.

2011-01-01

Shigella flexneri uses its type III secretion apparatus (TTSA) to inject host-altering proteins into targeted eukaryotic cells. The TTSA is composed of a basal body and an exposed needle with invasion plasmid antigen D (IpaD) forming a tip complex that controls secretion. The bile salt deoxycholate (DOC) stimulates recruitment of the translocator protein IpaB into the maturing TTSA needle tip complex. This process appears to be triggered by a direct interaction between DOC and IpaD. Fluorescence spectroscopy and NMR spectroscopy are used here to confirm the DOC-IpaD interaction and to reveal that IpaD conformational changes upon DOC binding trigger the appearance of IpaB at the needle tip. Förster resonance energy transfer between specific sites on IpaD was used here to identify changes in distances between IpaD domains as a result of DOC binding. To further explore the effects of DOC binding on IpaD structure, NMR chemical shift mapping was employed. The environments of residues within the proposed DOC binding site and additional residues within the “distal” globular domain were perturbed upon DOC binding, further indicating that conformational changes occur within IpaD upon DOC binding. These events are proposed to be responsible for the recruitment of IpaB at the TTSA needle tip. Mutation analyses combined with additional spectroscopic analyses confirms that conformational changes in IpaD induced by DOC binding contribute to the recruitment of IpaB to the S. flexneri TTSA needle tip. These findings lay the foundation for determining how environmental factors promote TTSA needle tip maturation prior to host cell contact. PMID:21126091

Gravitropism of Arabidopsis thaliana roots requires the polarization of PIN2 toward the root tip in meristematic cortical cells.

PubMed

Rahman, Abidur; Takahashi, Maho; Shibasaki, Kyohei; Wu, Shuang; Inaba, Takehito; Tsurumi, Seiji; Baskin, Tobias I

2010-06-01

In the root, the transport of auxin from the tip to the elongation zone, referred to here as shootward, governs gravitropic bending. Shootward polar auxin transport, and hence gravitropism, depends on the polar deployment of the PIN-FORMED auxin efflux carrier PIN2. In Arabidopsis thaliana, PIN2 has the expected shootward localization in epidermis and lateral root cap; however, this carrier is localized toward the root tip (rootward) in cortical cells of the meristem, a deployment whose function is enigmatic. We use pharmacological and genetic tools to cause a shootward relocation of PIN2 in meristematic cortical cells without detectably altering PIN2 polarization in other cell types or PIN1 polarization. This relocation of cortical PIN2 was negatively regulated by the membrane trafficking factor GNOM and by the regulatory A1 subunit of type 2-A protein phosphatase (PP2AA1) but did not require the PINOID protein kinase. When GNOM was inhibited, PINOID abundance increased and PP2AA1 was partially immobilized, indicating both proteins are subject to GNOM-dependent regulation. Shootward PIN2 specifically in the cortex was accompanied by enhanced shootward polar auxin transport and by diminished gravitropism. These results demonstrate that auxin flow in the root cortex is important for optimal gravitropic response.

Dynamic gradients of an intermediate filament-like cytoskeleton are recruited by a polarity landmark during apical growth.

PubMed

Fuchino, Katsuya; Bagchi, Sonchita; Cantlay, Stuart; Sandblad, Linda; Wu, Di; Bergman, Jessica; Kamali-Moghaddam, Masood; Flärdh, Klas; Ausmees, Nora

2013-05-21

Intermediate filament (IF)-like cytoskeleton emerges as a versatile tool for cellular organization in all kingdoms of life, underscoring the importance of mechanistically understanding its diverse manifestations. We showed previously that, in Streptomyces (a bacterium with a mycelial lifestyle similar to that of filamentous fungi, including extreme cell and growth polarity), the IF protein FilP confers rigidity to the hyphae by an unknown mechanism. Here, we provide a possible explanation for the IF-like function of FilP by demonstrating its ability to self-assemble into a cis-interconnected regular network in vitro and its localization into structures consistent with a cytoskeletal network in vivo. Furthermore, we reveal that a spatially restricted interaction between FilP and DivIVA, the main component of the Streptomyces polarisome complex, leads to formation of apical gradients of FilP in hyphae undergoing active tip extension. We propose that the coupling between the mechanism driving polar growth and the assembly of an IF cytoskeleton provides each new hypha with an additional stress-bearing structure at its tip, where the nascent cell wall is inevitably more flexible and compliant while it is being assembled and matured. Our data suggest that recruitment of cytoskeleton around a cell polarity landmark is a broadly conserved strategy in tip-growing cells.

Cyclic AMP Response Element Binding Protein Mediates Pathological Retinal Neovascularization via Modulating DLL4-NOTCH1 Signaling

PubMed Central

Singh, Nikhlesh K.; Kotla, Sivareddy; Kumar, Raj; Rao, Gadiparthi N.

2015-01-01

Retinal neovascularization is the most common cause of moderate to severe vision loss in all age groups. Despite the use of anti-VEGFA therapies, this complication continues to cause blindness, suggesting a role for additional molecules in retinal neovascularization. Besides VEGFA and VEGFB, hypoxia induced VEGFC expression robustly. Based on this finding, we tested the role of VEGFC in pathological retinal angiogenesis. VEGFC induced proliferation, migration, sprouting and tube formation of human retinal microvascular endothelial cells (HRMVECs) and these responses require CREB-mediated DLL4 expression and NOTCH1 activation. Furthermore, down regulation of VEGFC levels substantially reduced tip cell formation and retinal neovascularization in vivo. In addition, we observed that CREB via modulating the DLL4-NOTCH1 signaling mediates VEGFC-induced tip cell formation and retinal neovascularization. In regard to upstream mechanism, we found that down regulation of p38β levels inhibited hypoxia-induced CREB-DLL4-NOTCH1 activation, tip cell formation, sprouting and retinal neovascularization. Based on these findings, it may be suggested that VEGFC besides its role in the regulation of lymphangiogenesis also plays a role in pathological retinal angiogenesis and this effect depends on p38β and CREB-mediated activation of DLL4-NOTCH1 signaling. PMID:26870802

Microtubules are reversibly depolymerized in response to changing gaseous microenvironments within Aspergillus nidulans biofilms

PubMed Central

Shukla, Nandini; Osmani, Aysha H.; Osmani, Stephen A.

2017-01-01

How microtubules (MTs) are regulated during fungal biofilm formation is unknown. By tracking MT +end–binding proteins (+TIPS) in Aspergillus nidulans, we find that MTs are regulated to depolymerize within forming fungal biofilms. During this process, EB1, dynein, and ClipA form transient fibrous and then bar-like structures, novel configurations for +TIPS. Cells also respond in an autonomous manner, with cells separated by a septum able to maintain different MT dynamics. Surprisingly, all cells with depolymerized MTs rapidly repolymerize their MTs after air exchange above the static culture medium of biofilms. Although the specific gasotransmitter for this biofilm response is not known, we find that addition of hydrogen sulfide gas to growing cells recapitulates all aspects of reversible MT depolymerization and transient formation of +TIPs bars. However, as biofilms mature, physical removal of part of the biofilm is required to promote MT repolymerization, which occurs at the new biofilm edge. We further show MT depolymerization within biofilms is regulated by the SrbA hypoxic transcription factor and that without SrbA, MTs are maintained as biofilms form. This reveals a new mode of MT regulation in response to changing gaseous biofilm microenvironments, which could contribute to the unique characteristics of fungal biofilms in medical and industrial settings. PMID:28057761

Assessment of arsenic toxicity using Allium/Vicia root tip micronucleus assays.

PubMed

Wu, Lihua; Yi, Huilan; Yi, Min

2010-04-15

Arsenic is ubiquitous in the environment and is a potential human carcinogen. Its carcinogenicity has been demonstrated in several models. In this study, broad bean (Vicia faba L.) and common onion (Allium cepa L.), two plant species which are commonly used for detecting the genotoxic effects of environmental pollutants, were used to measure possible genotoxic effect of arsenite (0.3-30 mg/l). Present results showed that arsenite (As(III)) induced micronuclei (MN) formation in both Allium and Vicia root tips. MN frequency significantly increased in Vicia root cells exposed to 0.3-10 mg/l arsenite and in Allium root cells exposed to 1-30 mg/l arsenite, which indicated that Vicia root tip cells are more sensitive to arsenite than Allium. Mitotic index (MI) decreased in a concentration-dependent manner and showed significant differences in Vicia/Allium roots among treatments and the control, after exposure to 1-30 mg/l arsenite for at least 4 h. In the present study, MN frequency was positively associated with lipid peroxidation, which indicated that arsenite exposure can induce oxidative stress, cytotoxicity and genotoxicity in plant cells. The results also suggested that Vicia/Allium root micronucleus (MN) assays are simple, efficient and reproducible methods for the genotoxicity monitoring of arsenic water contamination. 2009 Elsevier B.V. All rights reserved.

Positive and negative tropic curvature induced by microbeam irradiation of protonemal tip cells of the moss Ceratodon purpureus.

PubMed

Lamparter, T; Kagawa, T; Brücker, G; Wada, M

2004-01-01

The photoreceptor phytochrome mediates tropic responses in protonemata of the moss Ceratodon purpureus. Under standard conditions the tip cells grow towards unilateral red light, or perpendicular to the electrical vector of polarized light. In this study the response of tip cells to partial irradiation of the apical region was analysed using a microbeam apparatus. The fluence response curve gave an unexpected pattern: whereas a 15-min microbeam with light intensities around 3 micro mol m (-2) s (-1) induced a growth curvature towards the irradiated side, higher light intensities around 100 micro mol m (-2) s (-1) caused a negative response, the cells grew away from the irradiated side. This avoidance response is explained by two effects: the light intensity is high enough to induce photoconversion into the active Pfr form of phytochrome, not only on the irradiated but also on the non-irradiated side by stray light. At the same time, the strong light on the irradiated side acts antagonistically to Pfr. As a result of this inhibition, the growth direction is moved to the light-avoiding side. Such a Pfr-independent mechanism might be important for the phototropic response to distinguish between the light-directed and light-avoiding side under unilateral light.

Interaction of intact porcine spermatozoa with epithelial cells and neutrophilic granulocytes during uterine passage.

PubMed

Taylor, U; Rath, D; Zerbe, H; Schuberth, H J

2008-04-01

New insemination techniques allow a tremendous sperm reduction for successful artificial insemination (AI) if highly diluted semen is deposited in the tip of the uterine horn and close to the utero-tubal junction. High sperm losses are known to occur during uterine passage and it was the general question whether specific binding mechanisms are involved. Upon arrival in the uterus, spermatozoa are confronted with mainly two different cell types: uterine epithelial cells (UEC) and neutrophilic granulocytes (polymorphonuclear neutrophil, PMN). As cell-sperm interactions can hardly be observed in vivo, an ex vivo system was established to study the interaction between spermatozoa and the UEC. Uterine segments (10 cm) from freshly slaughtered synchronized juvenile gilts were inseminated for 60 min at 38 degrees C. Thereafter spermatozoa were recovered, counted flow cytometrically and examined for changes in viability and mitochondrial membrane potential (MMP). Significantly less spermatozoa with a functioning MMP and intact plasma membranes could be retrieved (55 +/- 7%), while the number of damaged spermatozoa hardly changed (93 +/- 12%), indicating retention of viable sperm cells in the uterine lumen. The interactions between porcine PMN and spermatozoa (motile, immotile, membrane-damaged) were studied in coincubation assays in vitro. The binding of membrane-damaged sperm cells to PMN was virtually non-existent (3 +/- 2%). Viable and motile spermatozoa attached to PMN without being phagocytosed within 60 min (45 +/- 3%), whereas binding to sodium fluoride (NaF)-immobilized spermatozoa was reduced to 20 +/- 2%. The binding of viable sperm to PMN is most likely not lectin-dependent; although both viable cell types were shown to express a broad range of different lectin-binding sugar residues, none of the lectins tested was able to selectively block PMN-sperm binding significantly. The results of the study suggest that viable spermatozoa are already subject to selective processes within the uterus before further selection is initiated at the utero-tubal junction and in the oviductal isthmus.

Observation of Biological Tissues Using Common Path Optical Coherence Tomography with Gold Coated Conical Tip Lens Fiber

NASA Astrophysics Data System (ADS)

Taguchi, K.; Sugiyama, J.; Totsuka, M.; Imanaka, S.

2012-03-01

In this paper, we proposed a high lateral resolution common-path Fourier domain optical coherence tomography(OCT) system with the use of a chemically etched single mode fiber. In our experiments, single mode optical fiber for 1310nm was used for preparing the tapered tips. Our system used a conical microlens that was chemically etched by selective chemical etching technique using an etching solution of buffered hydrofluoric acid (BHF). From experimental results, we verified that our proposed optical coherence tomography system could operate as a common-path Fourier domain OCT system and conical tip lens fiber was very useful for a high lateral resolution common-path Fourier domain OCT system. Furthermore, we could observe a surface of paramecium bursaria and symbiotic chlorella in the paramecium bursaria using gold coated conical-tip fiber in the water.

Nanomachining by rubbing at ultrasonic frequency under controlled shear force.

PubMed

Muraoka, Mikio

2011-03-01

This study proposes a new method of proximal-probe machining that uses a rubbing process by introducing concentrated-mass (CM) cantilevers. At the second resonance of the CM cantilever vibration, the tip site of the cantilever becomes a node of the standing deflection wave because of the sufficient inertia of the attached concentrated mass. The tip makes a cyclic motion that is tangential to the sample surface, not vertical to it, as in a tapping motion. This lateral tip motion that is selectively excited by CM cantilevers was effective for the material modification of a sample due to the friction between the tip and the sample. Imaging and nanomachining under controlled shear force were demonstrated by means of the modified cantilever and a normal atomic force microscope. We were able to write a micron-sized letter "Z" having a line width of 30-100 nm on a resin surface.

Thermal study of bare tips with various system parameters and incision sizes.

PubMed

Osher, Robert H; Injev, Valentine P

2006-05-01

To identify major and minor surgeon-controlled parameters that affect incision temperature when performing microincision lens removal using the Alcon Infiniti Vision System. In vitro research and development laboratory, Alcon Research, Irvine, California, USA. Phacoemulsification was performed in eye-bank cadaver eyes and the following parameters evaluated: incision, duty cycle, ultrasound (US) power, aspiration flow rate (AFR), vacuum, pulse, bottle height and balanced salt solution temperature, and tip design/size. Each parameter was varied while the others remained constant. The resulting temperature of the incision and US tip was measured using a thermal camera. Major contributors to elevated incision temperature included incision size, US power, duty cycle, AFR, vacuum setting, tip design, and presence of an ophthalmic viscosurgical device (OVD). Minor contributors included pulse frequency, bottle height, and temperature of the infusate. Microincision lens removal can be performed at safe temperatures with the knowledgeable selection of surgeon-controlled parameters.

Intrauterine device for laser light diffusion and method of using the same

DOEpatents

Tadir, Yona; Berns, Michael W.; Svaasand, Lars O.; Tromberg, Bruce J.

1995-01-01

An improved device for delivery of photoenergy from a light source, such as a laser, into a uterine cavity for photodynamic therapy is comprised of a plurality of optic fibers, which are bundled together and inserted into the uterine cavity by means of a uterine cannula. The cannula is positioned within the uterine cavity at a preferred location and then withdrawn thereby allowing the plurality of optic fibers to splay or diverge one from the other within the cavity. Different portions of the distal tip of the optic fiber is provided with a light diffusing tip, the remainder being provided with a nondiffusing tip portion. The fiber optic shape, as well as the segment which is permitted to actively diffuse light through the tip, is selected in order to provide a more uniform exposure intensity of the photo energy or at least sufficient radiation directed to each segment of the uterine walls.

Intrauterine device for laser light diffusion and method of using the same

DOEpatents

Tadir, Y.; Berns, M.W.; Svaasand, L.O.; Tromberg, B.J.

1995-12-26

An improved device for delivery of photoenergy from a light source, such as a laser, into a uterine cavity for photodynamic therapy is comprised of a plurality of optic fibers, which are bundled together and inserted into the uterine cavity by means of a uterine cannula. The cannula is positioned within the uterine cavity at a preferred location and then withdrawn thereby allowing the plurality of optic fibers to splay or diverge one from the other within the cavity. Different portions of the distal tip of the optic fiber is provided with a light diffusing tip, the remainder being provided with a nondiffusing tip portion. The fiber optic shape, as well as the segment which is permitted to actively diffuse light through the tip, is selected in order to provide a more uniform exposure intensity of the photo energy or at least sufficient radiation directed to each segment of the uterine walls. 5 figs.

Spatiotemporal changes in the distribution of LHFPL5 in mice cochlear hair bundles during development and in the absence of PCDH15

PubMed Central

Mahendrasingam, Shanthini; Fettiplace, Robert; Alagramam, Kumar N.; Cross, Ellen

2017-01-01

Mechanosensory transduction by vertebrate hair cells depends on a protein complex at the tips of shorter stereocilia associated with mechanoelectrical transduction channels activated by tip links in the hair bundle. In mammalian hair cells, this complex includes transmembrane channel-like protein subunit 1 (TMC1), lipoma HMGIC fusion partner-like 5 protein (LHFPL5) and protocadherin 15 (PCDH15), a lower-end component of the tip link. TMC1 interacts with LHFPL5 and PCDH15 but how the complex develops to maturity, and the relationships between these proteins, remains uncertain. Here we evaluate the spatiotemporal development of LHFPL5 distributions in mouse cochlear hair bundles by immunofluorescence and immunogold transmission electron microscopy, from postnatal day 0 (P0) through P21 in wild type and PCDH15-deficient mice. At P0, hair bundles contain many short microvilli-like processes which we term unranked stereocilia, and a subset of lengthening rows, adjacent to a kinocilium. LHFPL5 is distributed throughout the bundle, including on stereocilia tips and the kinocilium. At P3, 4-to-6 rows of ranked stereocilia are evident, total LHFPL5 expression peaks, and LHFPL5 is localised to ranked stereocilia tips of all rows and to lower shaft/ankle links. By P12, the bundle has a mature pattern with 3 ranked rows but virtually no unranked stereocilia or kinocilium; LHFPL5 expression has declined and become restricted to the tips of shorter stereocilia. Throughout development from P0, expression of LHFPL5 is greater overall on apical than basal bundles, but there is, on average, an equal amount of labelling per labelled tip. In P3 mice lacking PCDH15, LHFPL5 labelling is not at the tips but is primarily on unranked stereocilia and lower lateral links. These data show that LHFPL5 is already present in the MET apparatus at P0 but requires PCDH15 at P3 to remain there. Shaft/ankle link localisation suggests it interacts with link proteins other than PCDH15. PMID:29069081

Propidium iodide competes with Ca(2+) to label pectin in pollen tubes and Arabidopsis root hairs.

PubMed

Rounds, Caleb M; Lubeck, Eric; Hepler, Peter K; Winship, Lawrence J

2011-09-01

We have used propidium iodide (PI) to investigate the dynamic properties of the primary cell wall at the apex of Arabidopsis (Arabidopsis thaliana) root hairs and pollen tubes and in lily (Lilium formosanum) pollen tubes. Our results show that in root hairs, as in pollen tubes, oscillatory peaks in PI fluorescence precede growth rate oscillations. Pectin forms the primary component of the cell wall at the tip of both root hairs and pollen tubes. Given the electronic structure of PI, we investigated whether PI binds to pectins in a manner analogous to Ca(2+) binding. We first show that Ca(2+) is able to abrogate PI growth inhibition in a dose-dependent manner. PI fluorescence itself also relies directly on the amount of Ca(2+) in the growth solution. Exogenous pectin methyl esterase treatment of pollen tubes, which demethoxylates pectins, freeing more Ca(2+)-binding sites, leads to a dramatic increase in PI fluorescence. Treatment with pectinase leads to a corresponding decrease in fluorescence. These results are consistent with the hypothesis that PI binds to demethoxylated pectins. Unlike other pectin stains, PI at low yet useful concentration is vital and specifically does not alter the tip-focused Ca(2+) gradient or growth oscillations. These data suggest that pectin secretion at the apex of tip-growing plant cells plays a critical role in regulating growth, and PI represents an excellent tool for examining the role of pectin and of Ca(2+) in tip growth.

Nitric oxide is involved in the oxytetracycline-induced suppression of root growth through inhibiting hydrogen peroxide accumulation in the root meristem

NASA Astrophysics Data System (ADS)

Yu, Qing-Xiang; Ahammed, Golam Jalal; Zhou, Yan-Hong; Shi, Kai; Zhou, Jie; Yu, Yunlong; Yu, Jing-Quan; Xia, Xiao-Jian

2017-02-01

Use of antibiotic-contaminated manure in crop production poses a severe threat to soil and plant health. However, few studies have studied the mechanism by which plant development is affected by antibiotics. Here, we used microscopy, flow cytometry, gene expression analysis and fluorescent dyes to study the effects of oxytetracycline (OTC), a widely used antibiotic in agriculture, on root meristem activity and the accumulation of hydrogen peroxide (H2O2) and nitric oxide (NO) in the root tips of tomato seedlings. We found that OTC caused cell cycle arrest, decreased the size of root meristem and inhibited root growth. Interestingly, the inhibition of root growth by OTC was associated with a decline in H2O2 levels but an increase in NO levels in the root tips. Diphenyliodonium (DPI), an inhibitor of H2O2 production, showed similar effects on root growth as those of OTC. However, exogenous H2O2 partially reversed the effects on the cell cycle, meristem size and root growth. Importantly, cPTIO (the NO scavenger) and tungstate (an inhibitor of nitrate reductase) significantly increased H2O2 levels in the root tips and reversed the inhibition of root growth by OTC. Out results suggest that OTC-induced NO production inhibits H2O2 accumulation in the root tips, thus leading to cell cycle arrest and suppression of root growth.

Biochemical and Immunocytological Characterizations of Arabidopsis Pollen Tube Cell Wall1[C][W][OA

PubMed Central

Dardelle, Flavien; Lehner, Arnaud; Ramdani, Yasmina; Bardor, Muriel; Lerouge, Patrice; Driouich, Azeddine; Mollet, Jean-Claude

2010-01-01

During plant sexual reproduction, pollen germination and tube growth require development under tight spatial and temporal control for the proper delivery of the sperm cells to the ovules. Pollen tubes are fast growing tip-polarized cells able to perceive multiple guiding signals emitted by the female organ. Adhesion of pollen tubes via cell wall molecules may be part of the battery of signals. In order to study these processes, we investigated the cell wall characteristics of in vitro-grown Arabidopsis (Arabidopsis thaliana) pollen tubes using a combination of immunocytochemical and biochemical techniques. Results showed a well-defined localization of cell wall epitopes. Low esterified homogalacturonan epitopes were found mostly in the pollen tube wall back from the tip. Xyloglucan and arabinan from rhamnogalacturonan I epitopes were detected along the entire tube within the two wall layers and the outer wall layer, respectively. In contrast, highly esterified homogalacturonan and arabinogalactan protein epitopes were found associated predominantly with the tip region. Chemical analysis of the pollen tube cell wall revealed an important content of arabinosyl residues (43%) originating mostly from (1→5)-α-l-arabinan, the side chains of rhamnogalacturonan I. Finally, matrix-assisted laser desorption ionization time-of-flight mass spectrometry analysis of endo-glucanase-sensitive xyloglucan showed mass spectra with two dominant oligosaccharides (XLXG/XXLG and XXFG), both being mono O-acetylated, and accounting for over 68% of the total ion signals. These findings demonstrate that the Arabidopsis pollen tube wall has its own characteristics compared with other cell types in the Arabidopsis sporophyte. These structural features are discussed in terms of pollen tube cell wall biosynthesis and growth dynamics. PMID:20547702

Biochemical and immunocytological characterizations of Arabidopsis pollen tube cell wall.

PubMed

Dardelle, Flavien; Lehner, Arnaud; Ramdani, Yasmina; Bardor, Muriel; Lerouge, Patrice; Driouich, Azeddine; Mollet, Jean-Claude

2010-08-01

During plant sexual reproduction, pollen germination and tube growth require development under tight spatial and temporal control for the proper delivery of the sperm cells to the ovules. Pollen tubes are fast growing tip-polarized cells able to perceive multiple guiding signals emitted by the female organ. Adhesion of pollen tubes via cell wall molecules may be part of the battery of signals. In order to study these processes, we investigated the cell wall characteristics of in vitro-grown Arabidopsis (Arabidopsis thaliana) pollen tubes using a combination of immunocytochemical and biochemical techniques. Results showed a well-defined localization of cell wall epitopes. Low esterified homogalacturonan epitopes were found mostly in the pollen tube wall back from the tip. Xyloglucan and arabinan from rhamnogalacturonan I epitopes were detected along the entire tube within the two wall layers and the outer wall layer, respectively. In contrast, highly esterified homogalacturonan and arabinogalactan protein epitopes were found associated predominantly with the tip region. Chemical analysis of the pollen tube cell wall revealed an important content of arabinosyl residues (43%) originating mostly from (1-->5)-alpha-L-arabinan, the side chains of rhamnogalacturonan I. Finally, matrix-assisted laser desorption ionization time-of-flight mass spectrometry analysis of endo-glucanase-sensitive xyloglucan showed mass spectra with two dominant oligosaccharides (XLXG/XXLG and XXFG), both being mono O-acetylated, and accounting for over 68% of the total ion signals. These findings demonstrate that the Arabidopsis pollen tube wall has its own characteristics compared with other cell types in the Arabidopsis sporophyte. These structural features are discussed in terms of pollen tube cell wall biosynthesis and growth dynamics.

PeakForce Tapping resolves individual microvilli on living cells.

PubMed

Schillers, Hermann; Medalsy, Izhar; Hu, Shuiqing; Slade, Andrea L; Shaw, James E

2016-02-01

Microvilli are a common structure found on epithelial cells that increase the apical surface thus enhancing the transmembrane transport capacity and also serve as one of the cell's mechanosensors. These structures are composed of microfilaments and cytoplasm, covered by plasma membrane. Epithelial cell function is usually coupled to the density of microvilli and its individual size illustrated by diseases, in which microvilli degradation causes malabsorption and diarrhea. Atomic force microscopy (AFM) has been widely used to study the topography and morphology of living cells. Visualizing soft and flexible structures such as microvilli on the apical surface of a live cell has been very challenging because the native microvilli structures are displaced and deformed by the interaction with the probe. PeakForce Tapping® is an AFM imaging mode, which allows reducing tip-sample interactions in time (microseconds) and controlling force in the low pico-Newton range. Data acquisition of this mode was optimized by using a newly developed PeakForce QNM-Live Cell probe, having a short cantilever with a 17-µm-long tip that minimizes hydrodynamic effects between the cantilever and the sample surface. In this paper, we have demonstrated for the first time the visualization of the microvilli on living kidney cells with AFM using PeakForce Tapping. The structures observed display a force dependence representing either the whole microvilli or just the tips of the microvilli layer. Together, PeakForce Tapping allows force control in the low pico-Newton range and enables the visualization of very soft and flexible structures on living cells under physiological conditions. © 2015 The Authors Journal of Molecular Recognition Published by John Wiley & Sons Ltd.

Atomizing nozzle and method

DOEpatents

Ting, Jason; Anderson, Iver E.; Terpstra, Robert L.

2000-03-16

A high pressure close-coupled gas atomizing nozzle includes multiple discrete gas jet discharge orifices having aerodynamically designed convergent-divergent geometry with an first converging section communicated to a gas supply manifold and to a diverging section by a constricted throat section to increase atomizing gas velocity. The gas jet orifices are oriented at gas jet apex angle selected relative to the melt supply tip apex angle to establish a melt aspiration condition at the melt supply tip.

Sunlight decreased genotoxicity of azadirachtin on root tip cells of Allium cepa and Eucrosia bicolor.

PubMed

Kwankua, W; Sengsai, S; Kuleung, C; Euawong, N

2010-07-01

Utilization of neem plant (Azadirachta indica A. Juss) extract for pest control in agriculture has raised concerns over contamination by the residues to the environment. Such residues, particularly azadirachtin (Aza), may cause deleterious effect to non-target organisms. This investigation was conducted to find out if Aza could be inactivated through exposures to sunlight. Activity of Aza was assessed as its ability to cause cytotoxic and genotoxic effects in the forms of nuclei abnormality and chromosome aberration as measured by mitotic index (MI) and mitotic aberration (MA). Varying concentrations of Aza were tested on Allium cepa and Eucrosia bicolor. It was found that the MI of all root tip meristematic cells of A. cepa and E. bicolor treated with 0.00005%, 0.00010%, 0.00015%, and 0.00020% (w/v) Aza-containing neem extract for 24h, were significantly lower than the controls. Complementary to the lower levels of MI, the Aza-treated groups showed higher MA levels in all cases investigated. Furthermore, the decreasing levels of MI and the increasing levels of MA related well with the increasing concentration of Aza. Microscopic examination of root tip meristematic cells revealed that the anomaly found most often were mitotic disturbances and chromosomal bridges. Exposures of 0.00020% (w/v) Aza to sunlight for 3 days and 7 days decreased Aza ability to induce cytotoxicity and genotoxicity, both in terms of MI and MA, to root tip meristematic cells in A. cepa and E. bicolor. Photodegradation of Aza upon exposure to direct sunlight was confirmed by HPLC. The study implicates that Aza would unlikely cause long term deleterious effects to the environment since it would be inactivated by sunlight. Copyright (c) 2010 Elsevier Inc. All rights reserved.

Inhibition of the MRP1-mediated transport of the menadione-glutathione conjugate (thiodione) in HeLa cells as studied by SECM

PubMed Central

Koley, Dipankar; Bard, Allen J.

2012-01-01

Oxidative stress induced in live HeLa cells by menadione (2-methyl-1,4-napthaquinone) was studied in real time by scanning electrochemical microscopy (SECM). The hydrophobic molecule menadione diffuses through a living cell membrane where it is toxic to the cell. However, in the cell it is conjugated with glutathione to form thiodione. Thiodione is then recognized and transported across the cell membrane via the ATP-driven MRP1 pump. In the extracellular environment, thiodione was detected by the SECM tip at levels of 140, 70, and 35 µM upon exposure of the cells to menadione concentrations of 500, 250, and 125 µM, respectively. With the aid of finite element modeling, the kinetics of thiodione transport was determined to be 1.6 × 10-7 m/s, about 10 times faster than menadione uptake. Selective inhibition of these MRP1 pumps inside live HeLa cells by MK571 produced a lower thiodione concentration of 50 µM in presence of 500 µM menadione and 50 µM MK571. A similar reduced (50% drop) thiodione efflux was observed in the presence of monoclonal antibody QCRL-4, a selective blocking agent of the MRP1 pumps. The reduced thiodione flux confirmed that thiodione was transported by MRP1, and that glutathione is an essential substrate for MRP1-mediated transport. This finding demonstrates the usefulness of SECM in quantitative studies of MRP1 inhibitors and suggests that monoclonal antibodies can be a useful tool in inhibiting the transport of these MDR pumps, and thereby aiding in overcoming multidrug resistance. PMID:22679290

Inhibition of the MRP1-mediated transport of the menadione-glutathione conjugate (thiodione) in HeLa cells as studied by SECM.

PubMed

Koley, Dipankar; Bard, Allen J

2012-07-17

Oxidative stress induced in live HeLa cells by menadione (2-methyl-1,4-napthaquinone) was studied in real time by scanning electrochemical microscopy (SECM). The hydrophobic molecule menadione diffuses through a living cell membrane where it is toxic to the cell. However, in the cell it is conjugated with glutathione to form thiodione. Thiodione is then recognized and transported across the cell membrane via the ATP-driven MRP1 pump. In the extracellular environment, thiodione was detected by the SECM tip at levels of 140, 70, and 35 µM upon exposure of the cells to menadione concentrations of 500, 250, and 125 µM, respectively. With the aid of finite element modeling, the kinetics of thiodione transport was determined to be 1.6 10(-7) m/s, about 10 times faster than menadione uptake. Selective inhibition of these MRP1 pumps inside live HeLa cells by MK571 produced a lower thiodione concentration of 50 µM in presence of 500 µM menadione and 50 µM MK571. A similar reduced (50% drop) thiodione efflux was observed in the presence of monoclonal antibody QCRL-4, a selective blocking agent of the MRP1 pumps. The reduced thiodione flux confirmed that thiodione was transported by MRP1, and that glutathione is an essential substrate for MRP1-mediated transport. This finding demonstrates the usefulness of SECM in quantitative studies of MRP1 inhibitors and suggests that monoclonal antibodies can be a useful tool in inhibiting the transport of these MDR pumps, and thereby aiding in overcoming multidrug resistance.

Acoustic measurements of a full-scale rotor with four tip shapes. Volume 2: Appendices C, D, E and F

NASA Technical Reports Server (NTRS)

Mosher, M.

1984-01-01

A full scale helicopter with four different blade tip geometries is tested in a 40- by 80-foot wind tunnel. Performance, loads, and noise are measured. The four tip shapes tested were rectangular, tapered, swept, and swept/tapered. The noise data include measurements of the sound pressure levels in decibels dB, decibels adjusted dBA, and tone-corrected PNdB, for all of the conditions tested. Also included are the detailed measurements, 1/3 octave spectra and time histories for some selected data, and plots of dBA as function of test condition. Some performance measurements are included to aid interpretation of the noise data.

Fundamental aspects of electric double layer force-distance measurements at liquid-solid interfaces using atomic force microscopy

PubMed Central

Black, Jennifer M.; Zhu, Mengyang; Zhang, Pengfei; Unocic, Raymond R.; Guo, Daqiang; Okatan, M. Baris; Dai, Sheng; Cummings, Peter T.; Kalinin, Sergei V.; Feng, Guang; Balke, Nina

2016-01-01

Atomic force microscopy (AFM) force-distance measurements are used to investigate the layered ion structure of Ionic Liquids (ILs) at the mica surface. The effects of various tip properties on the measured force profiles are examined and reveal that the measured ion position is independent of tip properties, while the tip radius affects the forces required to break through the ion layers as well as the adhesion force. Force data is collected for different ILs and directly compared with interfacial ion density profiles predicted by molecular dynamics. Through this comparison it is concluded that AFM force measurements are sensitive to the position of the ion with the larger volume and mass, suggesting that ion selectivity in force-distance measurements are related to excluded volume effects and not to electrostatic or chemical interactions between ions and AFM tip. The comparison also revealed that at distances greater than 1 nm the system maintains overall electroneutrality between the AFM tip and sample, while at smaller distances other forces (e.g., van der waals interactions) dominate and electroneutrality is no longer maintained. PMID:27587276

The Mechanism Forming the Cell Surface of Tip-Growing Rooting Cells Is Conserved among Land Plants.

PubMed

Honkanen, Suvi; Jones, Victor A S; Morieri, Giulia; Champion, Clement; Hetherington, Alexander J; Kelly, Steve; Proust, Hélène; Saint-Marcoux, Denis; Prescott, Helen; Dolan, Liam

2016-12-05

To discover mechanisms that controlled the growth of the rooting system in the earliest land plants, we identified genes that control the development of rhizoids in the liverwort Marchantia polymorpha. 336,000 T-DNA transformed lines were screened for mutants with defects in rhizoid growth, and a de novo genome assembly was generated to identify the mutant genes. We report the identification of 33 genes required for rhizoid growth, of which 6 had not previously been functionally characterized in green plants. We demonstrate that members of the same orthogroup are active in cell wall synthesis, cell wall integrity sensing, and vesicle trafficking during M. polymorpha rhizoid and Arabidopsis thaliana root hair growth. This indicates that the mechanism for constructing the cell surface of tip-growing rooting cells is conserved among land plants and was active in the earliest land plants that existed sometime more than 470 million years ago [1, 2]. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

Semicircular Canals Circumvent Brownian Motion Overload of Mechanoreceptor Hair Cells.

PubMed

Muller, Mees; Heeck, Kier; Elemans, Coen P H

2016-01-01

Vertebrate semicircular canals (SCC) first appeared in the vertebrates (i.e. ancestral fish) over 600 million years ago. In SCC the principal mechanoreceptors are hair cells, which as compared to cochlear hair cells are distinctly longer (70 vs. 7 μm), 10 times more compliant to bending (44 vs. 500 nN/m), and have a 100-fold higher tip displacement threshold (< 10 μm vs. <400 nm). We have developed biomechanical models of vertebrate hair cells where the bundle is approximated as a stiff, cylindrical elastic rod subject to friction and thermal agitation. Our models suggest that the above differences aid SCC hair cells in circumventing the masking effects of Brownian motion noise of about 70 nm, and thereby permit transduction of very low frequency (<10 Hz) signals. We observe that very low frequency mechanoreception requires increased stimulus amplitude, and argue that this is adaptive to circumvent Brownian motion overload at the hair bundles. We suggest that the selective advantage of detecting such low frequency stimuli may have favoured the evolution of large guiding structures such as semicircular canals and otoliths to overcome Brownian Motion noise at the level of the mechanoreceptors of the SCC.

Analytical solution of reaction-diffusion equations for calcium wave propagation in a starburst amacrine cell.

PubMed

Poznanski, R R

2010-09-01

A reaction-diffusion model is presented to encapsulate calcium-induced calcium release (CICR) as a potential mechanism for somatofugal bias of dendritic calcium movement in starburst amacrine cells. Calcium dynamics involves a simple calcium extrusion (pump) and a buffering mechanism of calcium binding proteins homogeneously distributed over the plasma membrane of the endoplasmic reticulum within starburst amacrine cells. The system of reaction-diffusion equations in the excess buffer (or low calcium concentration) approximation are reformulated as a nonlinear Volterra integral equation which is solved analytically via a regular perturbation series expansion in response to calcium feedback from a continuously and uniformly distributed calcium sources. Calculation of luminal calcium diffusion in the absence of buffering enables a wave to travel at distances of 120 μm from the soma to distal tips of a starburst amacrine cell dendrite in 100 msec, yet in the presence of discretely distributed calcium-binding proteins it is unknown whether the propagating calcium wave-front in the somatofugal direction is further impeded by endogenous buffers. If so, this would indicate CICR to be an unlikely mechanism of retinal direction selectivity in starburst amacrine cells.

Semicircular Canals Circumvent Brownian Motion Overload of Mechanoreceptor Hair Cells

PubMed Central

Muller, Mees; Heeck, Kier

2016-01-01

Vertebrate semicircular canals (SCC) first appeared in the vertebrates (i.e. ancestral fish) over 600 million years ago. In SCC the principal mechanoreceptors are hair cells, which as compared to cochlear hair cells are distinctly longer (70 vs. 7 μm), 10 times more compliant to bending (44 vs. 500 nN/m), and have a 100-fold higher tip displacement threshold (< 10 μm vs. <400 nm). We have developed biomechanical models of vertebrate hair cells where the bundle is approximated as a stiff, cylindrical elastic rod subject to friction and thermal agitation. Our models suggest that the above differences aid SCC hair cells in circumventing the masking effects of Brownian motion noise of about 70 nm, and thereby permit transduction of very low frequency (<10 Hz) signals. We observe that very low frequency mechanoreception requires increased stimulus amplitude, and argue that this is adaptive to circumvent Brownian motion overload at the hair bundles. We suggest that the selective advantage of detecting such low frequency stimuli may have favoured the evolution of large guiding structures such as semicircular canals and otoliths to overcome Brownian Motion noise at the level of the mechanoreceptors of the SCC. PMID:27448330

Three-dimensional Architecture of Hair-bundle Linkages Revealed by Electron-microscopic Tomography

PubMed Central

Auer, Manfred; Koster, Abrahram J.; Ziese, Ulrike; Bajaj, Chandrajit; Volkmann, Niels; Wang, Da Neng

2008-01-01

The senses of hearing and balance rest upon mechanoelectrical transduction by the hair bundles of hair cells in the inner ear. Located at the apical cellular surface, each hair bundle comprises several tens of stereocilia and a single kinocilium that are interconnected by extracellular proteinaceous links. Using electron-microscopic tomography of bullfrog saccular sensory epithelia, we examined the three-dimensional structures of basal links, kinociliary links, and tip links. We observed significant differences in the appearances and dimensions of these three structures and found two distinct populations of tip links suggestive of the involvement of different proteins, splice variants, or protein–protein interactions. We noted auxiliary links connecting the upper portions of tip links to the taller stereocilia. Tip links and auxiliary links show a tendency to adopt a globular conformation when disconnected from the membrane surface. PMID:18421501

NMR Model of PrgI-SipD Interaction and its Implications in the Needle-Tip Assembly of the Salmonella Type III Secretion System

PubMed Central

Rathinavelan, Thenmalarchelvi; Lara-Tejero, Maria; Lefebre, Matthew; Chatterjee, Srirupa; McShan, Andrew C.; Guo, Da-Chuan; Tang, Chun; Galan, Jorge E.; De Guzman, Roberto N.

2014-01-01

Salmonella and other pathogenic bacteria use the type III secretion system (T3SS) to inject virulence proteins into human cells to initiate infections. The structural component of the T3SS contains a needle and a needle tip. The needle is assembled from PrgI needle protomers and the needle tip is capped with several copies of the SipD tip protein. How a tip protein docks on the needle is unclear. A crystal structure of a PrgI-SipD fusion protein docked on the PrgI needle results in steric clash of SipD at the needle tip when modeled on the recent atomic structure of the needle. Thus, there is currently no good model of how SipD is docked on the PrgI needle tip. Previously, we showed by NMR paramagnetic relaxation enhancement (PRE) methods that a specific region in the SipD coiled-coil is the binding site for PrgI. Others have hypothesized that a domain of the tip protein – the N-terminal α-helical hairpin, has to swing away during the assembly of the needle apparatus. Here, we show by PRE methods that a truncated form of SipD lacking the α-helical hairpin domain binds more tightly to PrgI. Further, PRE-based structure calculations revealed multiple PrgI binding sites on the SipD coiled-coil. Our PRE results together with the recent NMR-derived atomic structure of the Salmonella needle suggest a possible model of how SipD might dock at the PrgI needle tip. SipD and PrgI are conserved in other bacterial T3SSs, thus our results have wider implication in understanding other needle-tip complexes. PMID:24951833

Epigenetic control of learning and memory in Drosophila by Tip60 HAT action.

PubMed

Xu, Songjun; Wilf, Rona; Menon, Trisha; Panikker, Priyalakshmi; Sarthi, Jessica; Elefant, Felice

2014-12-01

Disruption of epigenetic gene control mechanisms in the brain causes significant cognitive impairment that is a debilitating hallmark of most neurodegenerative disorders, including Alzheimer's disease (AD). Histone acetylation is one of the best characterized of these epigenetic mechanisms that is critical for regulating learning- and memory- associated gene expression profiles, yet the specific histone acetyltransferases (HATs) that mediate these effects have yet to be fully characterized. Here, we investigate an epigenetic role for the HAT Tip60 in learning and memory formation using the Drosophila CNS mushroom body (MB) as a well-characterized cognition model. We show that Tip60 is endogenously expressed in the Kenyon cells, the intrinsic neurons of the MB, and in the MB axonal lobes. Targeted loss of Tip60 HAT activity in the MB causes thinner and shorter axonal lobes while increasing Tip60 HAT levels cause no morphological defects. Functional consequences of both loss and gain of Tip60 HAT levels in the MB are evidenced by defects in immediate-recall memory. Our ChIP-Seq analysis reveals that Tip60 target genes are enriched for functions in cognitive processes, and, accordingly, key genes representing these pathways are misregulated in the Tip60 HAT mutant fly brain. Remarkably, we find that both learning and immediate-recall memory deficits that occur under AD-associated, amyloid precursor protein (APP)-induced neurodegenerative conditions can be effectively rescued by increasing Tip60 HAT levels specifically in the MB. Together, our findings uncover an epigenetic transcriptional regulatory role for Tip60 in cognitive function and highlight the potential of HAT activators as a therapeutic option for neurodegenerative disorders. Copyright © 2014 by the Genetics Society of America.

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

Merkle, J.G.

In order to study effects of constraint on fracture toughness, it is important to select the right location within the crack-tip field for investigation. In 1950 Hill postulated that close to a circular notch tip the principal stress directions would be radial and circumferential, so that the plastic slip lines (maximum shear stress trajectories) would be logarithmic spirals. The resulting equation for stress normal to the notch symmetry plane, neglecting strain hardening, was identical to that for the circumferential stress near the bore of an ideally plastic thick-walled hollow cylinder under external radial tension, because the relevant geometries are identical.more » In 1969, Rice and Johnson developed a near crack-tip, plane strain, large-strain rigid-plastic analysis considering strain hardening and assuming an infinitely sharp initial crack tip. Shortly afterwards, Merkle, following Hill's suggestion, proposed an approximate analysis of the stresses and strains ahead of a blunted crack tip on the plane of symmetry, based on a circular blunted crack tip. The analysis amounted to a hollow cylinder analogy, including the effects of strain hardening. The original hollow cylinder analogy was based on small strain theory, and the calculated strain distributions did not agree well with the Rice and Johnson results very near the blunted crack tip. Therefore, the hollow cylinder analogy equations have been rederived, based on large strain theory, and the agreement with the Rice and Johnson results and other more recent numerical results is good. Calculations illustrate the effects of transverse strain on the principal stresses very close to a blunting crack tip and show that, theoretically, a singularity still exists at the tip of a blunting crack. 10 refs., 9 figs.« less

Temporal and non-permanent division of labor during sorocarp formation in the social amoeba Acytostelium subglobosum.

PubMed

Mohri, Kurato; Kiyota, Yu; Kuwayama, Hidekazu; Urushihara, Hideko

2013-03-15

Somatic cell differentiation is crucial for the development of multicellular organisms. While the development of a fruiting body in Dictyostelium discoideum represents a simple model of this process with separation of stalk cells from the spore lineage, that of Acytostelium subglobosum is not accompanied by cell type separation. This species produces acellular stalks and, seemingly, all aggregated amoebae become spores; however, it possesses homologs for the stalk-cell marker genes of D. discoideum. In this study, we analyzed the spatio-temporal expression of A. subglobosum orthologs for D. discoideum stalk- or spore-lineage markers to clarify the developmental process of A. subglobosum. We first found that the prespore vesicles, which contained spore coat proteins, started to accumulate in the tip region and were observed in the entire sorogen throughout later development, confirming that all A. subglobosum cells became spores. The expression of a stalk-lineage gene ortholog, As-ecmA, started at the mound stage and was prominent in the protruding sorogen. Although two spore-lineage gene orthologs, As-cotD1 and -cotD2, were likewise detected shortly after cell aggregation and increased in intensity until tip formation, their expression diminished in the protruding sorogen. Double-fluorescence staining of these prestalk and prespore marker genes revealed that the expression of these marker genes was mutually exclusive and that expression switching occurred in the early tip. Our results indicate that A. subglobosum cells become committed to the spore lineage first, and then, while keeping this commitment intact, participate in stalk formation. Instead of the permanent division of labor observed in D. discoideum, A. subglobosum produces fruiting bodies by all cells contributing to the formation of the stalk as well as forming spores. Copyright © 2013 Elsevier Inc. All rights reserved.

[A comparison of three different needles used for spinal anesthesia in terms of squamous epithelial cell transport risk].

PubMed

Çiğdem, Ünal Kantekin; Sevinç, Şahin; Esef, Bolat; Süreyya, Öztürk; Muzaffer, Gencer; Akif, Demirel

To investigate the differences in the number of squamous epithelial cells carried to the spinal canal by three different types of spinal needle tip of the same size. Patients were allocated into three groups (Group I, Group II, Group III). Spinal anesthesia was administered to Group I (n=50) using a 25G Quincke needle, to Group II (n=50) using a 25G pencil point spinal needle, and to Group III (n=50) using a non-cutting atraumatic needle with special bending. The first and third drops of cerebral spinal fluid (CSF) samples were taken from each patient and each drop was placed on a slide for cytological examination. Nucleated and non-nucleated squamous epithelial cells on the smear preparations were counted. There was statistically significant difference between the groups in respect to the number of squamous epithelial cells in the first drop (p<0.05). Group III had lower number of squamous epithelial cells in the first drop compared to that of Group I and Group II. Mean while Group I had higher number of squamous epithelial cells in the third drop compared to the other groups. The number of squamous epithelial cells in the first and third drops was statistically similar in each group respectively (p>0.05 for each group). In this study of different needle tips, it was seen that with atraumatic needle with special bending a significantly smaller number of cells were transported when compared to the Quincke tip needles, and with pencil point needles. Copyright © 2016 Sociedade Brasileira de Anestesiologia. Publicado por Elsevier Editora Ltda. All rights reserved.

The tip link protein Cadherin-23: From Hearing Loss to Cancer.

PubMed

Vanniya S, Paridhy; Srisailapathy, C R Srikumari; Kunka Mohanram, Ramkumar

2018-04-01

Cadherin-23 is an atypical member of the cadherin superfamily, with a distinctly long extracellular domain. It has been known to be a part of the tip links of the inner ear mechanosensory hair cells. Several studies have been carried out to understand the role of Cadherin-23 in the hearing mechanism and defects in the CDH23 have been associated with hearing impairment resulting from defective or absence of tip links. Recent studies have highlighted the role of Cadherin-23 in several pathological conditions, including cancer, suggesting the presence of several unknown functions. Initially, it was proposed that Cadherin-23 represents a yet unspecified subtype of Cadherins; however, no other proteins with similar characteristics have been identified, till date. It has a unique cytoplasmic domain that does not bear a β-catenin binding region, but has been demonstrated to mediate cell-cell adhesions. Several protein interacting partners have been identified for Cadherin-23 and the roles of their interactions in various cellular mechanisms are yet to be explored. This review summarizes the characteristics of Cadherin-23 and its roles in several pathologies including cancer. Copyright © 2018 Elsevier Ltd. All rights reserved.

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

Altieri, Amanda S.; Ladner, Jane E.; Li, Zhuo

Proliferating cell nuclear antigen (PCNA) forms a trimeric ring that encircles duplex DNA and acts as an anchor for a number of proteins involved in DNA metabolic processes. PCNA has two structurally similar domains (I and II) linked by a long loop (inter-domain connector loop, IDCL) on the outside of each monomer of the trimeric structure that makes up the DNA clamp. All proteins that bind to PCNA do so via a PCNA-interacting peptide (PIP) motif that binds near the IDCL. A small protein, called TIP, binds to PCNA and inhibits PCNA-dependent activities although it does not contain a canonicalmore » PIP motif. The X-ray crystal structure of TIP bound to PCNA reveals that TIP binds to the canonical PIP interaction site, but also extends beyond it through a helix that relocates the IDCL. TIP alters the relationship between domains I and II within the PCNA monomer such that the trimeric ring structure is broken, while the individual domains largely retain their native structure. Small angle X-ray scattering (SAXS) confirms the disruption of the PCNA trimer upon addition of the TIP protein in solution and together with the X-ray crystal data, provides a structural basis for the mechanism of PCNA inhibition by TIP.« less

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

Altieri, Amanda S.; Ladner, Jane E.; Li, Zhuo

Here, proliferating cell nuclear antigen (PCNA) forms a trimeric ring that encircles duplex DNA and acts as an anchor for a number of proteins involved in DNA metabolic processes. PCNA has two structurally similar domains (I and II) linked by a long loop (inter-domain connector loop, IDCL) on the outside of each monomer of the trimeric structure that makes up the DNA clamp. All proteins that bind to PCNA do so via a PCNA-interacting peptide (PIP) motif that binds near the IDCL. A small protein, called TIP, binds to PCNA and inhibits PCNA-dependent activities although it does not contain amore » canonical PIP motif. The X-ray crystal structure of TIP bound to PCNA reveals that TIP binds to the canonical PIP interaction site, but also extends beyond it through a helix that relocates the IDCL. TIP alters the relationship between domains I and II within the PCNA monomer such that the trimeric ring structure is broken, while the individual domains largely retain their native structure. Small angle X-ray scattering (SAXS) confirms the disruption of the PCNA trimer upon addition of the TIP protein in solution and together with the X-ray crystal data, provides a structural basis for the mechanism of PCNA inhibition by TIP.« less

Pore structure and function of synthetic nanopores with fixed charges: tip shape and rectification properties

NASA Astrophysics Data System (ADS)

Ramírez, Patricio; Apel, Pavel Yu; Cervera, Javier; Mafé, Salvador

2008-08-01

We present a complete theoretical study of the relationship between the structure (tip shape and dimensions) and function (selectivity and rectification) of asymmetric nanopores on the basis of previous experimental studies. The theoretical model uses a continuum approach based on the Nernst-Planck equations. According to our results, the nanopore transport properties, such as current-voltage (I-V) characteristics, conductance, rectification ratio, and selectivity, are dictated mainly by the shape of the pore tip (we have distinguished bullet-like, conical, trumpet-like, and hybrid shapes) and the concentration of pore surface charges. As a consequence, the nanopore performance in practical applications will depend not only on the base and tip openings but also on the pore shape. In particular, we show that the pore opening dimensions estimated from the pore conductance can be very different, depending on the pore shape assumed. The results obtained can also be of practical relevance for the design of nanopores, nanopipettes, and nanoelectrodes, where the electrical interactions between the charges attached to the nanostructure and the mobile charges confined in the reduced volume of the inside solution dictate the device performance in practical applications. Because single tracks are the elementary building blocks for nanoporous membranes, the understanding and control of their individual properties should also be crucial in protein separation, water desalination, and bio-molecule detection using arrays of identical nanopores.

Pore structure and function of synthetic nanopores with fixed charges: tip shape and rectification properties.

PubMed

Ramírez, Patricio; Apel, Pavel Yu; Cervera, Javier; Mafé, Salvador

2008-08-06

We present a complete theoretical study of the relationship between the structure (tip shape and dimensions) and function (selectivity and rectification) of asymmetric nanopores on the basis of previous experimental studies. The theoretical model uses a continuum approach based on the Nernst-Planck equations. According to our results, the nanopore transport properties, such as current-voltage (I-V) characteristics, conductance, rectification ratio, and selectivity, are dictated mainly by the shape of the pore tip (we have distinguished bullet-like, conical, trumpet-like, and hybrid shapes) and the concentration of pore surface charges. As a consequence, the nanopore performance in practical applications will depend not only on the base and tip openings but also on the pore shape. In particular, we show that the pore opening dimensions estimated from the pore conductance can be very different, depending on the pore shape assumed. The results obtained can also be of practical relevance for the design of nanopores, nanopipettes, and nanoelectrodes, where the electrical interactions between the charges attached to the nanostructure and the mobile charges confined in the reduced volume of the inside solution dictate the device performance in practical applications. Because single tracks are the elementary building blocks for nanoporous membranes, the understanding and control of their individual properties should also be crucial in protein separation, water desalination, and bio-molecule detection using arrays of identical nanopores.

PhenoTips: patient phenotyping software for clinical and research use.

PubMed

Girdea, Marta; Dumitriu, Sergiu; Fiume, Marc; Bowdin, Sarah; Boycott, Kym M; Chénier, Sébastien; Chitayat, David; Faghfoury, Hanna; Meyn, M Stephen; Ray, Peter N; So, Joyce; Stavropoulos, Dimitri J; Brudno, Michael

2013-08-01

We have developed PhenoTips: open source software for collecting and analyzing phenotypic information for patients with genetic disorders. Our software combines an easy-to-use interface, compatible with any device that runs a Web browser, with a standardized database back end. The PhenoTips' user interface closely mirrors clinician workflows so as to facilitate the recording of observations made during the patient encounter. Collected data include demographics, medical history, family history, physical and laboratory measurements, physical findings, and additional notes. Phenotypic information is represented using the Human Phenotype Ontology; however, the complexity of the ontology is hidden behind a user interface, which combines simple selection of common phenotypes with error-tolerant, predictive search of the entire ontology. PhenoTips supports accurate diagnosis by analyzing the entered data, then suggesting additional clinical investigations and providing Online Mendelian Inheritance in Man (OMIM) links to likely disorders. By collecting, classifying, and analyzing phenotypic information during the patient encounter, PhenoTips allows for streamlining of clinic workflow, efficient data entry, improved diagnosis, standardization of collected patient phenotypes, and sharing of anonymized patient phenotype data for the study of rare disorders. Our source code and a demo version of PhenoTips are available at http://phenotips.org. © 2013 WILEY PERIODICALS, INC.

Cell Internal Treatable Microplasma Jets in Cancer Therapies

NASA Astrophysics Data System (ADS)

Kim, Jae Young; Wei, Yanzhang; Li, Jinhua; Kim, Sung-O.

2011-10-01

We developed a 15- μm-sized, single-cellular-level, and cell-manipulatable microplasma jet device with a microcapillary glass tip and described its potential in physical cancer therapies. The microcapillary tip is a funnel shaped glass tube and its nozzle has an inner diameter of 15 μm and an outer diameter of 20 μm with 20 capillary angle. The electrical and optical properties of this plasma jet and apoptosis results of cultured murine B16F0 melanoma tumor cells and CL.7 fibroblast cells treated with the plasma jets were described. In spite of the small inner diameter and the low gas flow rate of the microplasma jet device, the generated plasma jets are stable enough to treat tumor cells. The microplasma jet was observed inducing apoptosis in cultured murine melanoma tumor cells in a dose-dependent manner. Furthermore, the percentage of apoptotic cells of murine melanoma tumor cells induced by this plasma device was approximately 2.5 times bigger than that of murine fibroblast cells as indicated by an Annex V-FITC method. This highly precise plasma medicine, which enables new directed cancer therapies, can be combined with current cell manipulation and cell culturing technologies without much difficulty.

Formation of a PKCζ/β-catenin complex in endothelial cells promotes angiopoietin-1–induced collective directional migration and angiogenic sprouting

PubMed Central

Oubaha, Malika; Lin, Michelle I.; Margaron, Yoran; Filion, Dominic; Price, Emily N.; Zon, Leonard I.; Côté, Jean-François

2012-01-01

Angiogenic sprouting requires that cell-cell contacts be maintained during migration of endothelial cells. Angiopoietin-1 (Ang-1) and vascular endothelial growth factor act oppositely on endothelial cell junctions. We found that Ang-1 promotes collective and directional migration and, in contrast to VEGF, induces the formation of a complex formed of atypical protein kinase C (PKC)-ζ and β-catenin at cell-cell junctions and at the leading edge of migrating endothelial cells. This complex brings Par3, Par6, and adherens junction proteins at the front of migrating cells to locally activate Rac1 in response to Ang-1. The colocalization of PKCζ and β-catenin at leading edge along with PKCζ-dependent stabilization of cell-cell contacts promotes directed and collective endothelial cell migration. Consistent with these results, down-regulation of PKCζ in endothelial cells alters Ang-1–induced sprouting in vitro and knockdown in developing zebrafish results in intersegmental vessel defects caused by a perturbed directionality of tip cells and by loss of cell contacts between tip and stalk cells. These results reveal that PKCζ and β-catenin function in a complex at adherens junctions and at the leading edge of migrating endothelial cells to modulate collective and directional migration during angiogenesis. PMID:22936663

Characteristics of nanosecond pulse needle-to-plane discharges at high pressure: a particle-in-cell Monte Carlo collision simulation

NASA Astrophysics Data System (ADS)

Sang, Chaofeng; Sun, Jizhong; Ren, Chunsheng; Wang, Dezhen

2009-02-01

A model of one dimensional in position and three dimensional in velocity space self-consistent particle in cell with Monte Carlo collision technique was employed to simulate the argon discharge between the needle and plane electrodes at high pressure, in which a nanosecond rectangular pulse was applied to the needle electrode. The work focused on the investigation of the spatiotemporal evolution of the discharge versus the needle tip size and working gas pressure. The simulation results showed that the discharge occurred mainly in the region near the needle tip at atmospheric pressure, and that the small radius of the needle tip led to easy discharge. Reducing the gas pressure gave rise to a transition from a corona discharge to a glowlike discharge along the needle-to-plane direction. The microscopic mechanism for the transition can arguably be attributed to the peak of high-energy electrons occurring before the breakdown; the magnitude of the number of these electrons determined whether the breakdown can take place.

Robust syntaxin-4 immunoreactivity in mammalian horizontal cell processes

PubMed Central

HIRANO, ARLENE A.; BRANDSTÄTTER, JOHANN HELMUT; VILA, ALEJANDRO; BRECHA, NICHOLAS C.

2009-01-01

Horizontal cells mediate inhibitory feed-forward and feedback communication in the outer retina; however, mechanisms that underlie transmitter release from mammalian horizontal cells are poorly understood. Toward determining whether the molecular machinery for exocytosis is present in horizontal cells, we investigated the localization of syntaxin-4, a SNARE protein involved in targeting vesicles to the plasma membrane, in mouse, rat, and rabbit retinae using immunocytochemistry. We report robust expression of syntaxin-4 in the outer plexiform layer of all three species. Syntaxin-4 occurred in processes and tips of horizontal cells, with regularly spaced, thicker sandwich-like structures along the processes. Double labeling with syntaxin-4 and calbindin antibodies, a horizontal cell marker, demonstrated syntaxin-4 localization to horizontal cell processes; whereas, double labeling with PKC antibodies, a rod bipolar cell (RBC) marker, showed a lack of co-localization, with syntaxin-4 immunolabeling occurring just distal to RBC dendritic tips. Syntaxin-4 immunolabeling occurred within VGLUT-1-immunoreactive photoreceptor terminals and underneath synaptic ribbons, labeled by CtBP2/RIBEYE antibodies, consistent with localization in invaginating horizontal cell tips at photoreceptor triad synapses. Vertical sections of retina immunostained for syntaxin-4 and peanut agglutinin (PNA) established that the prominent patches of syntaxin-4 immunoreactivity were adjacent to the base of cone pedicles. Horizontal sections through the OPL indicate a one-to-one co-localization of syntaxin-4 densities at likely all cone pedicles, with syntaxin-4 immunoreactivity interdigitating with PNA labeling. Pre-embedding immuno-electron microscopy confirmed the subcellular localization of syntaxin-4 labeling to lateral elements at both rod and cone triad synapses. Finally, co-localization with SNAP-25, a possible binding partner of syntaxin-4, indicated co-expression of these SNARE proteins in the same subcellular compartment of the horizontal cell. Taken together, the strong expression of these two SNARE proteins in the processes and endings of horizontal cells at rod and cone terminals suggests that horizontal cell axons and dendrites are likely sites of exocytotic activity. PMID:17640443

Space-saving tips to lower stress

NASA Astrophysics Data System (ADS)

Gordon, Vernita D.

2018-03-01

Understanding how some single cells evolved into multicellular life means figuring out how they overcome the stresses associated with crowding as they multiply. New insights from yeast suggest that changes in the shape of cells may provide an answer.

Technical tips during implantation of selective upper airway stimulation.

PubMed

Heiser, Clemens; Thaler, Erica; Soose, Ryan J; Woodson, B Tucker; Boon, Maurits

2018-03-01

Selective upper airway stimulation is now well-established in the United States and in several European countries, with more than 1,000 patients implanted since U.S. Food and Drug Administration approval in April 2014. The authors herein, all head and neck surgeons, account for approximately one of every five implants completed to date. Several of the authors also provide comprehensive longitudinal care of their patients as dual-specialty sleep medicine physicians. Multi-center, retrospective clinical analysis. More than 300 implants have been evaluated and reviewed in five different implant centers (Germany, United States). This analysis shares tips and techniques from the collective experiences with more than 300 implants, which can help newer implanters learn vicariously both for standard practices in executing routine implants through activation and, importantly, for working through more challenging encounters with anatomy, special patient phenotypes, system testing, and troubleshooting. These tips should help new implanters handle most of the situations arising during implantation and avoid common pitfalls. Laryngoscope, 128:756-762, 2018. © 2017 The American Laryngological, Rhinological and Otological Society, Inc.

Theoretical and practical investigation into sustainable metal joining process for the automotive industry

NASA Astrophysics Data System (ADS)

Al-Jader, M. A.; Cullen, J. D.; Shaw, Andy; Al-Shamma'a, A. I.

2011-08-01

Currently there are about 4300 weld points on the average steel vehicle. Errors and problems due to tip damage and wear can cause great losses due to production line downtime. Current industrial monitoring systems check the quality of the nugget after processing 15 cars average once every two weeks. The nuggets are examined off line using a destructive process, which takes approximately 10 days to complete causing a long delay in the production process. In this paper a simulation results using software package, SORPAS, will be presented to determined the sustainability factors in spot welding process including Voltage, Current, Force, Water cooling rates, Material thicknesses and usage. The experimental results of various spot welding processes will be investigated and reported. The correlation of experimental results shows that SORPAS simulations can be used as an off line measurement to reduce factory energy usage. This paper also provides an overview of electrode current selection and its variance over the lifetime of the electrode tip, and describes the proposed analysis system for the selection of welding parameters for the spot welding process, as the electrode tip wears.

Are central venous catheter tip cultures reliable after 6-day refrigeration?

PubMed

Bouza, Emilio; Guembe, Maria; Gómez, Haydee; Martín-Rabadán, Pablo; Rivera, Marisa; Alcalá, Luis

2009-07-01

Present guidelines recommend culturing only central venous catheter (CVC) tips from patients with suspected catheter-related bloodstream infection (CR-BSI). However, a high proportion of these suspicions are not confirmed. Moreover, CVC tip culture increases laboratory workload, and reports of colonization may be meaningless or misleading for the clinician. Our working hypothesis was that CVC tips should be refrigerated and cultured only in patients with positive blood cultures. We evaluated the effect of 6-day refrigeration of 215 CVC tips. We selected all the catheters with a significant count according to the Maki's roll-plate technique and randomly assigned them to 2 groups. In group A, the catheters were recultured after 24 h of refrigeration, and in group B, the catheters were recultured after 6 days more of refrigeration, so that the refrigeration time evaluated would be of 6 days. The yield of refrigerated CVC tips that grow significant colony counts of primary culture in group B was compared with the yield of refrigerated catheter tips in group A. The difference showed that 6-day refrigeration reduced the number of significant CVCs by 15.2%. Only 61 CVCs were obtained from patients with CR-BSI, and in most of them, blood cultures were already positive before CVC culture, so only 0.91% of the CR-BSI episodes would have been misdiagnosed as culture negative after refrigeration. Refrigeration of CVC tips sent for culture and culturing only those from patients with positive blood cultures reduce the workload in the microbiology laboratory without misdiagnosing CR-BSI.

Cytogenetical and ultrastructural effects of copper on root meristem cells of Allium sativum L.

PubMed

Liu, Donghua; Jiang, Wusheng; Meng, Qingmin; Zou, Jin; Gu, Jiegang; Zeng, Muai

2009-04-01

Different copper concentrations, as well as different exposure times, were applied to investigate both cytogenetical and ultrastructural alterations in garlic (Allium sativum L.) meristem cells. Results showed that the mitotic index decreased progressively when either copper concentration or exposure time increased. C-mitosis, anaphase bridges, chromosome stickiness and broken nuclei were observed in the copper treated root tip cells. Some particulates containing the argyrophilic NOR-associated proteins were distributed in the nucleus of the root-tip cells and the amount of this particulate material progressively increased with increasing exposure time. Finally, the nucleolar material was extruded from the nucleus into the cytoplasm. Also, increased dictyosome vesicles in number, formation of cytoplasmic vesicles containing electron dense granules, altered mitochondrial shape, disruption of nuclear membranes, condensation of chromatin material, disintegration of organelles were observed. The mechanisms of detoxification and tolerance of copper are briefly discussed.

Hypoxia-driven angiogenesis: role of tip cells and extracellular matrix scaffolding.

PubMed

Germain, Stéphane; Monnot, Catherine; Muller, Laurent; Eichmann, Anne

2010-05-01

Angiogenesis is a highly coordinated tissue remodeling process leading to blood vessel formation. Hypoxia triggers angiogenesis via induction of expression of growth factors such as vascular endothelial growth factor (VEGF). VEGF instructs endothelial cells to form tip cells, which lead outgrowing capillary sprouts, whereas Notch signaling inhibits sprout formation. Basement membrane deposition and mechanical cues from the extracellular matrix (ECM) induced by hypoxia may participate to coordinated vessel sprouting in conjunction with the VEGF and Notch signaling pathways. Hypoxia regulates ECM composition, deposition, posttranslational modifications and rearrangement. In particular, hypoxia-driven vascular remodeling is dynamically regulated through modulation of ECM-modifying enzyme activities that eventually affect both matricellular proteins and growth factor availability. Better understanding of the complex interplay between endothelial cells and soluble growth factors and mechanical factors from the ECM will certainly have significant implications for understanding the regulation of developmental and pathological angiogenesis driven by hypoxia.

The Cdc42 GTPase-associated proteins Gic1 and Gic2 are required for polarized cell growth in Saccharomyces cerevisiae

PubMed Central

Chen, Guang-Chao; Kim, Yung-Jin; Chan, Clarence S.M.

1997-01-01

BEM2 of Saccharomyces cerevisiae encodes a Rho-type GTPase-activating protein that is required for proper bud site selection at 26°C and for bud emergence at elevated temperatures. We show here that the temperature-sensitive growth phenotype of bem2 mutant cells can be suppressed by increased dosage of the GIC1 gene. The Gic1 protein, together with its structural homolog Gic2, are required for cell size and shape control, bud site selection, bud emergence, actin cytoskeletal organization, mitotic spindle orientation/positioning, and mating projection formation in response to mating pheromone. Each protein contains a CRIB (Cdc42/Rac-interactive binding) motif and each interacts in the two-hybrid assay with the GTP-bound form of the Rho-type Cdc42 GTPase, a key regulator of polarized growth in yeast. The CRIB motif of Gic1 and the effector domain of Cdc42 are required for this association. Genetic experiments indicate that Gic1 and Gic2 play positive roles in the Cdc42 signal transduction pathway, probably as effectors of Cdc42. Subcellular localization studies with a functional green fluorescent protein–Gic1 fusion protein indicate that this protein is concentrated at the incipient bud site of unbudded cells, at the bud tip and mother-bud neck of budded cells, and at cortical sites on large-budded cells that may delimit future bud sites in the two progeny cells. The ability of Gic1 to associate with Cdc42 is important for its function but is apparently not essential for its subcellular localization. PMID:9367979

Microtubule distribution in gravitropic protonemata of the moss Ceratodon

NASA Technical Reports Server (NTRS)

Schwuchow, J.; Sack, F. D.; Hartmann, E.

1990-01-01

Tip cells of dark-grown protonemata of the moss Ceratodon purpureus are negatively gravitropic (grow upward). They possess a unique longitudinal zonation: (1) a tip group of amylochloroplasts in the apical dome, (2) a plastid-free zone, (3) a zone of significant plastid sedimentation, and (4) a zone of mostly non-sedimenting plastids. Immunofluorescence of vertical cells showed microtubules distributed throughout the cytoplasm in a mostly axial orientation extending through all zones. Optical sectioning revealed a close spatial association between microtubules and plastids. A majority (two thirds) of protonemata gravistimulated for > 20 min had a higher density of microtubules near the lower flank compared to the upper flank in the plastid-free zone. This apparent enrichment of microtubules occurred just proximal to sedimented plastids and near the part of the tip that presumably elongates more to produce curvature. Fewer than 5% of gravistimulated protonemata had an enrichment in microtubules near the upper flank, whereas 14% of vertical protonemata were enriched near one of the side walls. Oryzalin and amiprophos-methyl (APM) disrupted microtubules, gravitropism, and normal tip growth and zonation, but did not prevent plastid sedimentation. We hypothesize that a microtubule redistribution plays a role in gravitropism in this protonema. This appears to be the first report of an effect of gravity on microtubule distribution in plants.

An evolutionarily conserved NIMA-related kinase directs rhizoid tip growth in the basal land plant Marchantia polymorpha.

PubMed

Otani, Kento; Ishizaki, Kimitsune; Nishihama, Ryuichi; Takatani, Shogo; Kohchi, Takayuki; Takahashi, Taku; Motose, Hiroyasu

2018-03-01

Tip growth is driven by turgor pressure and mediated by the polarized accumulation of cellular materials. How a single polarized growth site is established and maintained is unclear. Here, we analyzed the function of NIMA-related protein kinase 1 (MpNEK1) in the liverwort Marchantia polymorpha In the wild type, rhizoid cells differentiate from the ventral epidermis and elongate through tip growth to form hair-like protrusions. In Mp nek1 knockout mutants, rhizoids underwent frequent changes in growth direction, resulting in a twisted and/or spiral morphology. The functional MpNEK1-Citrine protein fusion localized to microtubule foci in the apical growing region of rhizoids. Mp nek1 knockouts exhibited increases in both microtubule density and bundling in the apical dome of rhizoids. Treatment with the microtubule-stabilizing drug taxol phenocopied the Mp nek1 knockout. These results suggest that MpNEK1 directs tip growth in rhizoids through microtubule organization. Furthermore, MpNEK1 expression rescued ectopic outgrowth of epidermal cells in the Arabidopsis thaliana nek6 mutant, strongly supporting an evolutionarily conserved NEK-dependent mechanism of directional growth. It is possible that such a mechanism contributed to the evolution of the early rooting system in land plants. © 2018. Published by The Company of Biologists Ltd.

Impact of tip-gap size and periodicity on turbulent transition

NASA Astrophysics Data System (ADS)

Pogorelov, Alexej; Meinke, Matthias; Schroeder, Wolfgang

2015-11-01

Large-Eddy Simulations of the flow field in an axial fan are performed at a Reynolds number of 936.000 based on the diameter and the rotational speed of the casing wall. A finite-volume flow solver based on a conservative Cartesian cut-cell method is used to solve the unsteady compressible Navier-Stokes equations. Computations are performed at a flow rate coefficient of 0.165 and a tip-gap size of s/D =0.01, for a 72 degrees fan section resolving only one out of five blades and a full fan resolving all five blades to investigate the impact of the periodic boundary condition. Furthermore, a grid convergence study is performed using four computational grids. Results of the flow field are analyzed for the computational grid with 1 billion cells. An interaction of the turbulent wake, generated by the tip-gap vortex, with the downstream blade, is observed, which leads to a cyclic transition with high pressure fluctuations on the suction side of the blade. Two dominant frequencies are identified which perfectly match with the characteristic frequencies in the experimental sound power level such that their physical origin is explained. A variation of the tip-gap size alters the transition on the suction side, i.e., no cyclic transition is observed.

SERS investigations and electrical recording of neuronal networks with three-dimensional plasmonic nanoantennas (Conference Presentation)

NASA Astrophysics Data System (ADS)

De Angelis, Francesco

2017-06-01

SERS investigations and electrical recording of neuronal networks with three-dimensional plasmonic nanoantennas Michele Dipalo, Valeria Caprettini, Anbrea Barbaglia, Laura Lovato, Francesco De Angelis e-mail: francesco.deangelis@iit.it Istituto Italiano di Tecnologia, Via Morego 30, 16163, Genova Biological systems are analysed mainly by optical, chemical or electrical methods. Normally each of these techniques provides only partial information about the environment, while combined investigations could reveal new phenomena occurring in complex systems such as in-vitro neuronal networks. Aiming at the merging of optical and electrical investigations of biological samples, we introduced three-dimensional plasmonic nanoantennas on CMOS-based electrical sensors [1]. The overall device is then capable of enhanced Raman Analysis of cultured cells combined with electrical recording of neuronal activity. The Raman measurements show a much higher sensitivity when performed on the tip of the nanoantenna in respect to the flat substrate [2]; this effect is a combination of the high plasmonic field enhancement and of the tight adhesion of cells on the nanoantenna tip. Furthermore, when plasmonic opto-poration is exploited [3] the 3D nanoelectrodes are able to penetrate through the cell membrane thus accessing the intracellular environment. Our latest results (unpublished) show that the technique is completely non-invasive and solves many problems related to state-of-the-art intracellular recording approaches on large neuronal networks. This research received funding from ERC-IDEAS Program: "Neuro-Plasmonics" [Grant n. 616213]. References: [1] M. Dipalo, G. C. Messina, H. Amin, R. La Rocca, V. Shalabaeva, A. Simi, A. Maccione, P. Zilio, L. Berdondini, F. De Angelis, Nanoscale 2015, 7, 3703. [2] R. La Rocca, G. C. Messina, M. Dipalo, V. Shalabaeva, F. De Angelis, Small 2015, 11, 4632. [3] G. C. Messina et al., Spatially, Temporally, and Quantitatively Controlled Delivery of Broad Range of Molecules into Selected Cells through Plasmonic Nanotubes. Advanced Materials 2015.

Help Your Students Nail that College Audition!

ERIC Educational Resources Information Center

Keating, Andrea

2003-01-01

Focuses on how music teachers can assist high school students to prepare for a college audition and to select the right music program. Explains what audition committees look for in music students. Includes tips teachers can share with their students on program selection and the audition. (CMK)

Paclitaxel, Ifosfamide, and Cisplatin Efficacy for First-Line Treatment of Patients With Intermediate- or Poor-Risk Germ Cell Tumors

PubMed Central

Hu, James; Dorff, Tanya B.; Lim, Kristina; Patil, Sujata; Woo, Kaitlin M.; Carousso, Maryann; Hughes, Amanda; Sheinfeld, Joel; Bains, Manjit; Daneshmand, Siamak; Ketchens, Charlene; Bajorin, Dean F.; Bosl, George J.; Quinn, David I.; Motzer, Robert J.

2016-01-01

Purpose Paclitaxel, ifosfamide, and cisplatin (TIP) achieved complete responses (CRs) in two thirds of patients with advanced germ cell tumors (GCTs) who relapsed after first-line chemotherapy with cisplatin and etoposide with or without bleomycin. We tested the efficacy of first-line TIP in patients with intermediate- or poor-risk disease. Patients and Methods In this prospective, multicenter, single-arm phase II trial, previously untreated patients with International Germ Cell Cancer Collaborative Group poor-risk or modified intermediate-risk GCTs received four cycles of TIP (paclitaxel 240 mg/m2 over 2 days, ifosfamide 6 g/m2 over 5 days with mesna support, and cisplatin 100 mg/m2 over 5 days) once every 3 weeks with granulocyte colony-stimulating factor support. The primary end point was the CR rate. Results Of the first 41 evaluable patients, 28 (68%) achieved a CR, meeting the primary efficacy end point. After additional accrual on an extension phase, total enrollment was 60 patients, including 40 (67%) with poor risk and 20 (33%) with intermediate risk. Thirty-eight (68%) of 56 evaluable patients achieved a CR and seven (13%) achieved partial responses with negative markers (PR-negative) for a favorable response rate of 80%. Five of seven achieving PR-negative status had seminoma and therefore did not undergo postchemotherapy resection of residual masses. Estimated 3-year progression-free survival and overall survival rates were 72% (poor risk, 63%; intermediate risk, 90%) and 91% (poor risk, 87%; intermediate risk, 100%), respectively. Grade 3 to 4 toxicities consisted primarily of reversible hematologic or electrolyte abnormalities, including neutropenic fever in 18%. Conclusion TIP demonstrated efficacy as first-line therapy for intermediate- and poor-risk GCTs with an acceptable safety profile. Given higher rates of favorable response, progression-free survival, and overall survival compared with prior first-line studies, TIP warrants further study in this population. PMID:27185842

Pragmatic setup for bioparticle responses by dielectrophoresis for resource limited environment application

NASA Astrophysics Data System (ADS)

Ali, Mohd Anuar Md; Yeop Majlis, Burhanuddin; Kayani, Aminuddin Ahmad

2017-12-01

Various dielectrophoretic responses of bioparticles, including cell-chain, spinning, rotation and clustering, are of high interest in the field due to their benefit into application for biomedical and clinical implementation potential. Numerous attempts using sophisticated equipment setup have been studied to perform those dielectrophoretic responses, however, for development into resource limited environment application, such as portable, sustainable and environmental friendly diagnostic tools, establishment of pragmatic setup using standard, non-sophisticated and low-cost equipment is of important task. Here we show the advantages in the judicious design optimization of tip microelectrode, also with selection of suspending medium and optimization of electric signal configuration in establishing setup that can promote the aforementioned dielectrophoretic responses within standard equipments, i.e. pragmatic setup.

Shape selection criterion for cellular array during constrained growth of binary alloys - Need for low gravity experiment

NASA Technical Reports Server (NTRS)

Tewari, Surendra N.; Trivedi, Rohit

1991-01-01

Development of steady-state periodic cellular array is one of the critical problems in the study of nonlinear pattern formation during directional solidification of binary alloys. The criterion which establishes the values of cell tip radius and spacing under given growth condition is not known. Theoretical models, such as marginal stability and microscopic solvability, have been developed for purely diffusive regime. However, the experimental conditions where cellular structures are stable are precisely the ones where the convection effects are predominant. Thus, the critical data for meaningful evaluation of cellular array growth models can only be obtained by partial directional solidification and quenching experiments carried out in the low gravity environment of space.

Solid surface wetting and the deployment of drops in microgravity

NASA Technical Reports Server (NTRS)

Trinh, E. H.; Depew, J.

1994-01-01

The complete or partial deployment of liquid samples in low gravity is primarily influenced by the interfacial properties of the specific liquid and solid materials used because the overwhelming bias of the Earth gravitational acceleration is removed. This study addresses the engineering aspects of injecting and deploying drops of prescribed volume into an acoustic positioning chamber in microgravity. The specific problems of interest are the design, testing, and implementation of injector tips to be used in a simultaneously retracting dual-injector system in the Drop Physics Module microgravity experiment facility. Prior to release, the liquid to be deployed must be retained within a restricted area at the very end of the injectors under dynamic stimuli from the continuous injection flow as well as from the stepped motion of the injectors. The final released drop must have a well determined volume and negligible residual linear or angular momentum. The outcome of Earth-based short-duration low gravity experiments had been the selection of two types of injector tips which were flown as back-up parts. They were successfully utilized during the USML-1 Spacelab mission as the primary tips. The combination of a larger contact surface, liquid pinning with a sharp edge, and selective coating of strategic tip surfaces with a non-wetting compound has allowed a significant increase in the success rate of deployment of simple and compound drops of aqueous solutions of glycerol and silicone oil. The diameter of the samples studied in the Drop Physics Module range between 0.3 and 2.7 cm. The tests conducted on-orbit with a manually operated small device have allowed the calibration of the volume deployed for a few drop sizes. The design for improved tips to be used during the next USML flight is based on these results.

Phytotoxic cyanamide affects maize (Zea mays) root growth and root tip function: from structure to gene expression.

PubMed

Soltys, Dorota; Rudzińska-Langwald, Anna; Kurek, Wojciech; Szajko, Katarzyna; Sliwinska, Elwira; Bogatek, Renata; Gniazdowska, Agnieszka

2014-05-01

Cyanamide (CA) is a phytotoxic compound produced by four Fabaceae species: hairy vetch, bird vetch, purple vetch and black locust. Its toxicity is due to complex activity that involves the modification of both cellular structures and physiological processes. To date, CA has been investigated mainly in dicot plants. The goal of this study was to investigate the effects of CA in the restriction of the root growth of maize (Zea mays), representing the monocot species. CA (3mM) reduced the number of border cells in the root tips of maize seedlings and degraded their protoplasts. However, CA did not induce any significant changes in the organelle structure of other root cells, apart from increased vacuolization. CA toxicity was also demonstrated by its effect on cell cycle activity, endoreduplication intensity, and modifications of cyclins CycA2, CycD2, and histone HisH3 gene expression. In contrast, the arrangement of microtubules was not altered by CA. Treatment of maize seedlings with CA did not completely arrest mitotic activity, although the frequency of dividing cells was reduced. Furthermore, prolonged CA treatment increased the proportion of endopolyploid cells in the root tip. Cytological malformations were accompanied by an induction of oxidative stress in root cells, which manifested as enhanced accumulation of H2O2. Exposure of maize seedlings to CA resulted in an increased concentration of auxin and stimulated ethylene emission. Taken together, these findings suggested that the inhibition of root growth by CA may be a consequence of stress-induced morphogenic responses. Copyright © 2014. Published by Elsevier GmbH.

Mammalian Cell Encapsulation in Alginate Beads Using a Simple Stirred Vessel.

PubMed

Hoesli, Corinne A; Kiang, Roger L J; Raghuram, Kamini; Pedroza, René G; Markwick, Karen E; Colantuoni, Antonio M R; Piret, James M

2017-06-29

Cell encapsulation in alginate beads has been used for immobilized cell culture in vitro as well as for immunoisolation in vivo. Pancreatic islet encapsulation has been studied extensively as a means to increase islet survival in allogeneic or xenogeneic transplants. Alginate encapsulation is commonly achieved by nozzle extrusion and external gelation. Using this method, cell-containing alginate droplets formed at the tip of nozzles fall into a solution containing divalent cations that cause ionotropic alginate gelation as they diffuse into the droplets. The requirement for droplet formation at the nozzle tip limits the volumetric throughput and alginate concentration that can be achieved. This video describes a scalable emulsification method to encapsulate mammalian cells in 0.5% to 10% alginate with 70% to 90% cell survival. By this alternative method, alginate droplets containing cells and calcium carbonate are emulsified in mineral oil, followed by a decrease in pH leading to internal calcium release and ionotropic alginate gelation. The current method allows the production of alginate beads within 20 min of emulsification. The equipment required for the encapsulation step consists in simple stirred vessels available to most laboratories.

Lanthanum resulted in unbalance of nutrient elements and disturbance of cell proliferation cycles in V. faba L. seedlings.

PubMed

Wang, Chengrun; Lu, Xianwen; Tian, Yuan; Cheng, Tao; Hu, Lingling; Chen, Fenfen; Jiang, Chuanjun; Wang, Xiaorong

2011-11-01

Effects of lanthanum (La) on mineral nutrients, cell cycles, and root lengthening have been little reported. The present work investigated these physiological responses in roots of Vicia faba seedlings cultivated in La3+-contained solutions for 15 days. The results showed that the increasing contents of La in the roots and leaves contributed to disbalances of contents of Ca, Fe, Cu, Zn, Mg, Mn, P, and K elements, and potential redistributions of some elements in the roots and leaves. These disbalances might be involved in the subsequent alteration of cell cycle phases in the root tips. Low-dose promotion and high-dose inhibition (Hormetic effects) were demonstrated as the dose responses of G0/G1-, S- or G2/M-phase ratios. The cell cycles were most probably arrested at G1/S interphase by La3+ in the root tips. The fact that the root lengths were not consistent with the changes of cell cycle phases suggested that the cell proliferation activities might be masked by other factors (e.g., cell expansion) under long-time exposure to La3+.

Experimental and theoretical investigation of HT-S/PMR-PI composites for application to advanced aircraft engines. [High-Tip-Speed/Polymerization of Monomeric Reactant

NASA Technical Reports Server (NTRS)

Hanson, M. P.; Chamis, C. C.

1974-01-01

A combined experimental and theoretical investigation was performed in order to: (1) demonstrate that high quality angleplied laminates can be made from HT-S/PMR-PI (PMR in situ polymerization of monomeric reactants), (2) characterize the PMR-PI material and to determine the HT-S unidirectional composite properties required for composite micro and macromechanics and laminate analyses, (3) select HT-S/PMR laminate configurations to meet the general design requirements for high-tip-speed compressor blades. The results of the investigation showed that: HT-S/PMR laminate configurations can be fabricated which satisfy the high-tip-speed compressor blade design requirements when operating within the temperature capability of the polymide matrix.

Alar setback technique: a controlled method of nasal tip deprojection.

PubMed

Foda, H M

2001-11-01

To describe an alar cartilage-modifying technique aimed at decreasing nasal tip projection in cases with overdeveloped alar cartilages and to compare it with other deprojection techniques used to correct such deformity. Selected case series. University and private practice settings in Alexandria, Egypt. Twenty patients presenting for rhinoplasty who had overprojected nasal tips primarily due to overdeveloped alar cartilages. All cases were primary cases except for one patient, who had undergone 2 previous rhinoplasties. An external rhinoplasty approach was used to set back the alar cartilages by shortening their medial and lateral crura. The choice of performing a high or low setback depended on the preexisting lobule-to-columella ratio. Following the setback, the alar cartilages were reconstructed in a fashion that increased the strength and stability of the tip complex. Subjective evaluation included clinical examination, analysis of preoperative and postoperative photographs, and patient satisfaction. Objective evaluation of nasal tip projection, using the Goode ratio and the nasofacial angle, was performed preoperatively and repeated at least 6 months postoperatively. A low setback was performed in 16 cases (80%) and a high setback in 4 (20%). The mean follow-up period was 18 months (range, 6-36 months). The technique effectively deprojected the nasal tip as evidenced by the considerable postoperative decrease in values of the Goode ratio and the nasofacial angle. No complications were encountered and no revision surgical procedures were required. The alar setback technique has many advantages; it results in precise predictable amounts of deprojection, controls the degree of tip rotation, preserves the natural contour of the nasal tip, respects the tip support mechanisms, increases the strength and stability of nasal tip complex, preserves or restores the normal lobule-to-columella proportion, and does not lead to alar flaring. However, the technique requires an external rhinoplasty approach and fine technical precision.

Sarcopenia Is Risk Factor for Development of Hepatic Encephalopathy After Transjugular Intrahepatic Portosystemic Shunt Placement.

PubMed

Nardelli, Silvia; Lattanzi, Barbara; Torrisi, Sabrina; Greco, Francesca; Farcomeni, Alessio; Gioia, Stefania; Merli, Manuela; Riggio, Oliviero

2017-06-01

Hepatic encephalopathy (HE) is an important complication in patients with cirrhosis who received transjugular intrahepatic portosystemic shunts (TIPS). We investigated whether a decrease in muscle mass was associated independently with the occurrence of HE after TIPS. We performed a prospective study of 46 consecutive patients with cirrhosis (mean age, 58.6 ± 9.1 y; mean model for end-stage liver disease score, 11.3 ± 3.3; mean Child-Pugh score, 7.6 ± 1.5) who received TIPS from January 2013 through December 2014 at a tertiary center in Rome, Italy. All patients underwent computed tomography analysis at the level of the third lumbar vertebrae to determine the skeletal muscle index; sarcopenia was defined by sex-specific cut-off values. We estimated the incidence of the first episode of HE after TIPS, taking into account the competing risk nature of the data (death or liver transplantation). Twenty-six patients (57%) were found to have sarcopenia. Twenty-one patients (46%) developed overt HE in the 7 ± 9 months after TIPS placement; all of these patients were sarcopenic, according to the skeletal muscle index. Of the 25 patients without HE after TIPS, only 5 had sarcopenia. In multivariate analysis, model for end-stage liver disease score (subdistribution hazard ratio, 1.16; 95% confidence interval, 1.01-1.34; P = .043) and sarcopenia (subdistribution hazard ratio, 31.3; 95% confidence interval, 4.5-218.07; P < .001) were associated independently with the development of HE after TIPS placement. In a prospective study of 46 patients with cirrhosis, we found muscle wasting, probably owing to reduced processing of ammonia, to be associated with the development of HE after TIPS placement. Sarcopenia should be considered in selecting patients for TIPS therapy. Nutritional status should be evaluated in patients with sarcopenia before TIPS placement, which might reduce the incidence of HE. Copyright © 2017 AGA Institute. Published by Elsevier Inc. All rights reserved.

Propidium Iodide Competes with Ca2+ to Label Pectin in Pollen Tubes and Arabidopsis Root Hairs1[W][OA

PubMed Central

Rounds, Caleb M.; Lubeck, Eric; Hepler, Peter K.; Winship, Lawrence J.

2011-01-01

We have used propidium iodide (PI) to investigate the dynamic properties of the primary cell wall at the apex of Arabidopsis (Arabidopsis thaliana) root hairs and pollen tubes and in lily (Lilium formosanum) pollen tubes. Our results show that in root hairs, as in pollen tubes, oscillatory peaks in PI fluorescence precede growth rate oscillations. Pectin forms the primary component of the cell wall at the tip of both root hairs and pollen tubes. Given the electronic structure of PI, we investigated whether PI binds to pectins in a manner analogous to Ca2+ binding. We first show that Ca2+ is able to abrogate PI growth inhibition in a dose-dependent manner. PI fluorescence itself also relies directly on the amount of Ca2+ in the growth solution. Exogenous pectin methyl esterase treatment of pollen tubes, which demethoxylates pectins, freeing more Ca2+-binding sites, leads to a dramatic increase in PI fluorescence. Treatment with pectinase leads to a corresponding decrease in fluorescence. These results are consistent with the hypothesis that PI binds to demethoxylated pectins. Unlike other pectin stains, PI at low yet useful concentration is vital and specifically does not alter the tip-focused Ca2+ gradient or growth oscillations. These data suggest that pectin secretion at the apex of tip-growing plant cells plays a critical role in regulating growth, and PI represents an excellent tool for examining the role of pectin and of Ca2+ in tip growth. PMID:21768649

Simulated Tip Rub Testing of Low-Density Metal Foam

NASA Technical Reports Server (NTRS)

Bowman, Cheryl L.; Jones, Michael G.

2009-01-01

Preliminary acoustic studies have indicated that low-density, open-cell, metal foams may be suitable acoustic liner material for noise suppression in high by-pass engines. Metal foam response under simulated tip rub conditions was studied to assess whether its durability would be sufficient for the foam to serve both as a rub strip above the rotor as well as an acoustic treatment. Samples represented four metal alloys, nominal cell dimensions ranging from 60 to 120 cells per inch (cpi), and relative densities ranging from 3.4 to 10 percent. The resulting rubbed surfaces were relatively smooth and the open cell structure of the foam was not adversely affected. Sample relative density appeared to have significant influence on the forces induced by the rub event. Acoustic responses of various surface preparations were measured using a normal incidence tube. The results of this study indicate that the foam s open-cell structure was retained after rubbing and that the acoustic absorption spectra variation was minimal.

High-speed AFM for scanning the architecture of living cells

NASA Astrophysics Data System (ADS)

Li, Jing; Deng, Zhifeng; Chen, Daixie; Ao, Zhuo; Sun, Quanmei; Feng, Jiantao; Yin, Bohua; Han, Li; Han, Dong

2013-08-01

We address the modelling of tip-cell membrane interactions under high speed atomic force microscopy. Using a home-made device with a scanning area of 100 × 100 μm2, in situ imaging of living cells is successfully performed under loading rates from 1 to 50 Hz, intending to enable detailed descriptions of physiological processes in living samples.We address the modelling of tip-cell membrane interactions under high speed atomic force microscopy. Using a home-made device with a scanning area of 100 × 100 μm2, in situ imaging of living cells is successfully performed under loading rates from 1 to 50 Hz, intending to enable detailed descriptions of physiological processes in living samples. Electronic supplementary information (ESI) available: Movie of the real-time change of inner surface within fresh blood vessel. The movie was captured at a speed of 30 Hz in the range of 80 μm × 80 μm. See DOI: 10.1039/c3nr01464a

Nonlinear unstable viscous fingers in Hele--Shaw flows. I. Experiments

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

Kopf-Sill, A.R.; Homsy, G.M.

1988-02-01

Post-instability viscous fingering in rectilinear flow in a Hele--Shaw cell has been studied experimentally. Of particular interest was the characterization of the range of length scales associated with tip splitting, over a reasonably wide range of parameters. A digital imaging system was used to record the patterns as a function of time, which allowed properties such as the tip velocity, finger width, perimeter, and area to be studied as functions of time and capillary number. The tip velocity was observed to be approximately constant regardless of the occurrence of splitting events, and the average finger width decreased as the degreemore » of supercriticality increased. Quantitative measures of the fact that there is a limit to the complexity of viscous fingers are provided, and that over the range of parameters studied, no evidence for fractal fingering exists. A discussion of the dynamics of tip splitting explains why this is so.« less

A dendrite-autonomous mechanism for direction selectivity in retinal starburst amacrine cells.

PubMed

Hausselt, Susanne E; Euler, Thomas; Detwiler, Peter B; Denk, Winfried

2007-07-01

Detection of image motion direction begins in the retina, with starburst amacrine cells (SACs) playing a major role. SACs generate larger dendritic Ca(2+) signals when motion is from their somata towards their dendritic tips than for motion in the opposite direction. To study the mechanisms underlying the computation of direction selectivity (DS) in SAC dendrites, electrical responses to expanding and contracting circular wave visual stimuli were measured via somatic whole-cell recordings and quantified using Fourier analysis. Fundamental and, especially, harmonic frequency components were larger for expanding stimuli. This DS persists in the presence of GABA and glycine receptor antagonists, suggesting that inhibitory network interactions are not essential. The presence of harmonics indicates nonlinearity, which, as the relationship between harmonic amplitudes and holding potential indicates, is likely due to the activation of voltage-gated channels. [Ca(2+)] changes in SAC dendrites evoked by voltage steps and monitored by two-photon microscopy suggest that the distal dendrite is tonically depolarized relative to the soma, due in part to resting currents mediated by tonic glutamatergic synaptic input, and that high-voltage-activated Ca(2+) channels are active at rest. Supported by compartmental modeling, we conclude that dendritic DS in SACs can be computed by the dendrites themselves, relying on voltage-gated channels and a dendritic voltage gradient, which provides the spatial asymmetry necessary for direction discrimination.

Root Tip Shape Governs Root Elongation Rate under Increased Soil Strength1[OPEN

PubMed Central

Kirchgessner, Norbert; Walter, Achim

2017-01-01

Increased soil strength due to soil compaction or soil drying is a major limitation to root growth and crop productivity. Roots need to exert higher penetration force, resulting in increased penetration stress when elongating in soils of greater strength. This study aimed to quantify how the genotypic diversity of root tip geometry and root diameter influences root elongation under different levels of soil strength and to determine the extent to which roots adjust to increased soil strength. Fourteen wheat (Triticum aestivum) varieties were grown in soil columns packed to three bulk densities representing low, moderate, and high soil strength. Under moderate and high soil strength, smaller root tip radius-to-length ratio was correlated with higher genotypic root elongation rate, whereas root diameter was not related to genotypic root elongation. Based on cavity expansion theory, it was found that smaller root tip radius-to-length ratio reduced penetration stress, thus enabling higher root elongation rates in soils with greater strength. Furthermore, it was observed that roots could only partially adjust to increased soil strength. Root thickening was bounded by a maximum diameter, and root tips did not become more acute in response to increased soil strength. The obtained results demonstrated that root tip geometry is a pivotal trait governing root penetration stress and root elongation rate in soils of greater strength. Hence, root tip shape needs to be taken into account when selecting for crop varieties that may tolerate high soil strength. PMID:28600344

Transjugular portal vein recanalization with creation of intrahepatic portosystemic shunt (PVR-TIPS) in patients with chronic non-cirrhotic, non-malignant portal vein thrombosis.

PubMed

Klinger, Christoph; Riecken, Bettina; Schmidt, Arthur; De Gottardi, Andrea; Meier, Benjamin; Bosch, Jaime; Caca, Karel

2018-03-01

To determine safety and efficacy of transjugular portal vein recanalization with creation of intrahepatic portosystemic shunt (PVR-TIPS) in patients with chronic non-cirrhotic, non-malignant portal vein thrombosis (PVT). This retrospective study includes 17 consecutive patients with chronic non-cirrhotic PVT (cavernous transformation n = 15). PVR-TIPS was indicated because of variceal bleeding (n = 13), refractory ascites (n = 2), portal biliopathy with recurrent cholangitis (n = 1), or abdominal pain (n = 1). Treatment consisted of a combination of transjugular balloon angioplasty, mechanical thrombectomy, and-depending on extent of residual thrombosis-transjugular intrahepatic portosystemic shunt and additional stenting of the portal venous system. Recanalization was successful in 76.5 % of patients despite cavernous transformation in 88.2 %. Both 1- and 2-year secondary PV and TIPS patency rates were 69.5 %. Procedure-related bleeding complications occurred in 2 patients (intraperitoneal bleeding due to capsule perforation, n = 1; liver hematoma, n = 1) and resolved spontaneously. However, 1 patient died due to subsequent nosocomial pneumonia. During follow-up, 3 patients with TIPS occlusion and PVT recurrence experienced portal hypertensive complications. PVR-TIPS is safe and effective in selected patients with chronic non-cirrhotic PVT. Due to technical complexity and possible complications, it should be performed only in specialized centers with high experience in TIPS procedures. © Georg Thieme Verlag KG Stuttgart · New York.

Fabrication and characterization of a nanometer-sized optical fiber electrode based on selective chemical etching for scanning electrochemical/optical microscopy.

PubMed

Maruyama, Kenichi; Ohkawa, Hiroyuki; Ogawa, Sho; Ueda, Akio; Niwa, Osamu; Suzuki, Koji

2006-03-15

We have already reported a method for fabricating ultramicroelectrodes (Suzuki, K. JP Patent, 2004-45394, 2004). This method is based on the selective chemical etching of optical fibers. In this work, we undertake a detailed investigation involving a combination of etched optical fibers with various types of tapered tip (protruding-shape, double- (or pencil-) shape and triple-tapered electrode) and insulation with electrophoretic paint. Our goal is to establish a method for fabricating nanometer-sized optical fiber electrodes with high reproducibility. As a result, we realized pencil-shaped and triple-tapered electrodes that had radii in the nanometer range with high reproducibility. These nanometer-sized electrodes showed well-defined sigmoidal curves and stable diffusion-limited responses with cyclic voltammetry. The pencil-shaped optical fiber, which has a conical tip with a cone angle of 20 degrees , was effective for controlling the electrode radius. The pencil-shaped electrodes had higher reproducibility and smaller electrode radii (r(app) < 1.0 nm) than those of other etched optical fiber electrodes. By using a pencil-shaped electrode with a 105-nm radius as a probe, we obtained simultaneous electrochemical and optical images of an implantable interdigitated array electrode. We achieved nanometer-scale resolution with a combination of scanning electrochemical microscopy SECM and optical microscopy. The resolution of the electrochemical and optical images indicated sizes of 300 and 930 nm, respectively. The neurites of living PC12 cells were also successfully imaged on a 1.6-microm scale by using the negative feedback mode of an SECM.

Gravitropism of Arabidopsis thaliana Roots Requires the Polarization of PIN2 toward the Root Tip in Meristematic Cortical Cells[C][W

PubMed Central

Rahman, Abidur; Takahashi, Maho; Shibasaki, Kyohei; Wu, Shuang; Inaba, Takehito; Tsurumi, Seiji; Baskin, Tobias I.

2010-01-01

In the root, the transport of auxin from the tip to the elongation zone, referred to here as shootward, governs gravitropic bending. Shootward polar auxin transport, and hence gravitropism, depends on the polar deployment of the PIN-FORMED auxin efflux carrier PIN2. In Arabidopsis thaliana, PIN2 has the expected shootward localization in epidermis and lateral root cap; however, this carrier is localized toward the root tip (rootward) in cortical cells of the meristem, a deployment whose function is enigmatic. We use pharmacological and genetic tools to cause a shootward relocation of PIN2 in meristematic cortical cells without detectably altering PIN2 polarization in other cell types or PIN1 polarization. This relocation of cortical PIN2 was negatively regulated by the membrane trafficking factor GNOM and by the regulatory A1 subunit of type 2-A protein phosphatase (PP2AA1) but did not require the PINOID protein kinase. When GNOM was inhibited, PINOID abundance increased and PP2AA1 was partially immobilized, indicating both proteins are subject to GNOM-dependent regulation. Shootward PIN2 specifically in the cortex was accompanied by enhanced shootward polar auxin transport and by diminished gravitropism. These results demonstrate that auxin flow in the root cortex is important for optimal gravitropic response. PMID:20562236

Reliability of plant root comet assay in comparison with human leukocyte comet assay for assessment environmental genotoxic agents.

PubMed

Reis, Gabriela Barreto Dos; Andrade-Vieira, Larissa Fonseca; Moraes, Isabella de Campos; César, Pedro Henrique Souza; Marcussi, Silvana; Davide, Lisete Chamma

2017-08-01

Comet assay is an efficient test to detect genotoxic compounds based on observation of DNA damage. The aim of this work was to compare the results obtained from the comet assay in two different type of cells extracted from the root tips from Lactuca sativa L. and human blood. For this, Spent Pot Liner (SPL), and its components (aluminum and fluoride) were applied as toxic agents. SPL is a solid waste generated in industry from the aluminum mining and processing with known toxicity. Three concentrations of all tested solutions were applied and the damages observed were compared to negative and positive controls. It was observed an increase in the frequency of DNA damage for human leukocytes and plant cells, in all treatments. On human leukocytes, SPL induced the highest percentage of damage, with an average of 87.68%. For root tips cells of L. sativa the highest percentage of damage was detected for aluminum (93.89%). Considering the arbitrary units (AU), the average of nuclei with high levels of DNA fragmentation was significant for both cells type evaluated. The tested cells demonstrated equal effectiveness for detection of the genotoxicity induced by the SPL and its chemical components, aluminum and fluoride. Further, using a unique method, the comet assay, we proved that cells from root tips of Lactuca sativa represent a reliable model to detect DNA damage induced by genotoxic pollutants is in agreement of those observed in human leukocytes as model. So far, plant cells may be suggested as important system to assess the toxicological risk of environmental agents. Copyright © 2017 Elsevier Inc. All rights reserved.

Mapping HA-tagged protein at the surface of living cells by atomic force microscopy.

PubMed

Formosa, C; Lachaize, V; Galés, C; Rols, M P; Martin-Yken, H; François, J M; Duval, R E; Dague, E

2015-01-01

Single-molecule force spectroscopy using atomic force microscopy (AFM) is more and more used to detect and map receptors, enzymes, adhesins, or any other molecules at the surface of living cells. To be specific, this technique requires antibodies or ligands covalently attached to the AFM tip that can specifically interact with the protein of interest. Unfortunately, specific antibodies are usually lacking (low affinity and specificity) or are expensive to produce (monoclonal antibodies). An alternative strategy is to tag the protein of interest with a peptide that can be recognized with high specificity and affinity with commercially available antibodies. In this context, we chose to work with the human influenza hemagglutinin (HA) tag (YPYDVPDYA) and labeled two proteins: covalently linked cell wall protein 12 (Ccw12) involved in cell wall remodeling in the yeast Saccharomyces cerevisiae and the β2-adrenergic receptor (β2-AR), a G protein-coupled receptor (GPCR) in higher eukaryotes. We first described the interaction between HA antibodies, immobilized on AFM tips, and HA epitopes, immobilized on epoxy glass slides. Using our system, we then investigated the distribution of Ccw12 proteins over the cell surface of the yeast S. cerevisiae. We were able to find the tagged protein on the surface of mating yeasts, at the tip of the mating projections. Finally, we could unfold multimers of β2-AR from the membrane of living transfected chinese hamster ovary cells. This result is in agreement with GPCR oligomerization in living cell membranes and opens the door to the study of the influence of GPCR ligands on the oligomerization process. Copyright © 2014 John Wiley & Sons, Ltd.

Nanostructure and force spectroscopy analysis of human peripheral blood CD4{sup +} T cells using atomic force microscopy

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

Hu Mingqian; Wang Jiongkun; Cai Jiye

2008-09-12

To date, nanoscale imaging of the morphological changes and adhesion force of CD4{sup +} T cells during in vitro activation remains largely unreported. In this study, we used atomic force microscopy (AFM) to study the morphological changes and specific binding forces in resting and activated human peripheral blood CD4{sup +} T cells. The AFM images revealed that the volume of activated CD4{sup +} T cells increased and the ultrastructure of these cells also became complex. Using a functionalized AFM tip, the strength of the specific binding force of the CD4 antigen-antibody interaction was found to be approximately three times thatmore » of the unspecific force. The adhesion forces were not randomly distributed over the surface of a single activated CD4{sup +} T cell, indicated that the CD4 molecules concentrated into nanodomains. The magnitude of the adhesion force of the CD4 antigen-antibody interaction did not change markedly with the activation time. Multiple bonds involved in the CD4 antigen-antibody interaction were measured at different activation times. These results suggest that the adhesion force involved in the CD4 antigen-antibody interaction is highly selective and of high affinity.« less

Personnel Recruitment/Selection/Induction in Small/Rural Schools.

ERIC Educational Resources Information Center

Neely, Steve

This paper provides tips and resources for the recruitment, selection, and induction of teachers and other school personnel. These materials were developed with rural and small schools in mind; some resources focus on Kansas. Means of recruitment are discussed, including internal search, referrals, college and university placement services,…

Percutaneous Pulmonary Embolism Thrombectomy and Thrombolysis: Technical Tips and Tricks.

PubMed

Devcic, Zlatko; Kuo, William T

2018-06-01

Catheter-directed therapy (CDT) is now acknowledged as a treatment option for select patients with acute massive or submassive pulmonary embolism (PE), and more patients are being considered for CDT if there is available expertise. Therefore, interventionalists should be aware of the variety of catheter-based treatment options, specific pitfalls to avoid during therapy, and the appropriate treatment endpoints. This article reviews currently available techniques and protocols for treating acute massive and submassive PE, with tips to safely and successfully perform percutaneous PE interventions.

Oligomeric Neuronal Protein Aggregates as Biomarkers for Traumatic Brain Injury (TBI) and Alzheimer Disease (AD)

DTIC Science & Technology

2013-10-01

displayed at the tip of the bacteriophage. The M13 hyperphage system can produce phage with multiple copies of the scFv expressed at the tip. Using C6T...antibody is one of the morphology specific nanobodies and the detection antibody is a phage displayed version of the capture nanobody. The phage ...selected for future ELISAs development. To ensure that the phage - displayed scFvs can still bind to their antigens, the different protein targets

Final Technical Report: Electrohydrodynamic Tip Streaming

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

Basaran, Osman

2016-01-06

When subjected to strong electric fields, liquid drops and films form conical tips and emit thin jets from their tips. Such electrodydrodynamic (EDH) tip streaming or cone-jetting phenomena, which are sometimes referred to as electrospraying, occur widely in nature, e.g., in ejection of streams of small charged drops from pointed tips of raindrops in thunderclouds, and technology, e.g., in electrospray mass spectrometry or electric field-driven solvent extraction. More recently, EHD cone-jetting has emerged as a powerful technique for direct printing of solar cells, micro- and nano- particle production, and microencapsulation for controlled release. In many of the aforementioned situations, ofmore » equal importance to the processes by which one drop disintegrates to form several drops are those by which (a) two drops come together and coalesce and (b) two drops are coupled to form a double droplet system (DDS) or a capillary switch (CS). the main objective of this research program is to advance through simulation, theory, and experiment the breakup, coalescence, and oscillatory dynamics of single and pairs of charged as well as uncharged drops.« less

Aerodynamic effect of a honeycomb rotor tip shroud on a 50.8-centimeter-tip-diameter core turbine

NASA Technical Reports Server (NTRS)

Moffitt, T. P.; Whitney, W. J.

1983-01-01

A 50.8-cm-tip-diameter turbine equipped with a rotor tip shroud of hexagonal cell (or honeycomb) cross section has been tested in warm air (416 K) for a range of shroud coolant to primary flow rates. Test results were also obtained for the same turbine operated with a solid shroud for comparison. The results showed that the combined effect of the honeycomb shroud and the coolant flow was to cause a reduction of 2.8 points in efficiency at design speed, pressure ratio, and coolant flow rate. With the coolant system inactivated, the honeycomb shroud caused a decrease in efficiency of 2.3 points. These results and those obtained from a small reference turbine indicate that the dominant factor governing honeycomb tip shroud loss is the ratio of honeycomb depth to blade span. The loss results of the two shrouds could be correlated on this basis. The same honeycomb and coolant effects are expected to occur for the hot (2200 K) version of this turbine.

Time Dependence of Tip Morphology during Cellular/Dendritic Arrayed Growth

NASA Technical Reports Server (NTRS)

Song, H.; Tewari, S. N.

1996-01-01

Succinonitrile-1.9 wt pct acetone has been directionally solidified in 0.7 X 0.7-cm-square cross section pyrex ampoules in order to observe the cell/dendrite tip morphologies, not influenced by the 'wall effects', which are present during growth in the generally used thin (about 200 gm) crucibles. The tips do not maintain a steady-state shape, as is generally assumed. Instead, they fluctuate within a shape envelope. The extent of fluctuation increases with decreasing growth speed, as the micro structure changes from the dendritic to cellular. The influence of natural convection has been examined by comparing these morphologies with those grown, without convection, in the thin ampoules.

Evaluation of Hydroprocessed Renewable Diesel (HRD) Fuel in a Caterpillar Engine Using the 210 Hour TWV Cycle

DTIC Science & Technology

2014-05-01

TERMS Hydroprocessed Renewable Diesel , Reference Diesel Fuel, C7, emissions, power, performance, deposition, ambient, desert, synthetic fuel injector ...the engine run-in, the engine was disassembled to determine injector nozzle tip deposits, and the piston crowns and engine combustion chamber deposits...removed from the test cell and disassembled to determine injector nozzle tip and piston crown and engine combustion chamber deposits. Post- test

Nanoengineering Testbed for Nanosolar Cell and Piezoelectric Compounds

DTIC Science & Technology

2012-02-29

element mesh. The third model was a 3D finite element mesh that included complete geometric representation of Berkovich tip. This model allows for a...height of the specimen. These simulations suggest the proper specimen size to approximate a body of semi-infinite extent for a given indentation depth...tip nanoindentation model was the third and final finite element mesh created for analysis and comparison. The material model and the finite element

Micromorphological effect of calcium phosphate coating on compatibility of magnesium alloy with osteoblast

PubMed Central

Hiromoto, Sachiko; Yamazaki, Tomohiko

2017-01-01

Abstract Octacalcium phosphate (OCP) and hydroxyapatite (HAp) coatings were developed to control the degradation speed and to improve the biocompatibility of biodegradable magnesium alloys. Osteoblast MG-63 was cultured directly on OCP- and HAp-coated Mg-3Al-1Zn (wt%, AZ31) alloy (OCP- and HAp-AZ31) to evaluate cell compatibility. Cell proliferation was remarkably improved with OCP and HAp coatings which reduced the corrosion and prevented the H2O2 generation on Mg alloy substrate. OCP-AZ31 showed sparse distribution of living cell colonies and dead cells. HAp-AZ31 showed dense and homogeneous distribution of living cells, with dead cells localized over and around corrosion pits, some of which were formed underneath the coating. These results demonstrated that cells were dead due to changes in the local environment, and it is necessary to evaluate the local biocompatibility of magnesium alloys. Cell density on HAp-AZ31 was higher than that on OCP-AZ31 although there was not a significant difference in the amount of Mg ions released in medium between OCP- and HAp-AZ31. The outer layer of OCP and HAp coatings consisted of plate-like crystal with a thickness of around 0.1 μm and rod-like crystals with a diameter of around 0.1 μm, respectively, which grew from a continuous inner layer. Osteoblasts formed focal contacts on the tips of plate-like OCP and rod-like HAp crystals, with heights of 2–5 μm. The spacing between OCP tips of 0.8–1.1 μm was wider than that between HAp tips of 0.2–0.3 μm. These results demonstrated that cell proliferation depended on the micromorphology of the coatings which governed spacing of focal contacts. Consequently, HAp coating is suitable for improving cell compatibility and bone-forming ability of the Mg alloy. PMID:28179963

Micromorphological effect of calcium phosphate coating on compatibility of magnesium alloy with osteoblast

NASA Astrophysics Data System (ADS)

Hiromoto, Sachiko; Yamazaki, Tomohiko

2017-12-01

Octacalcium phosphate (OCP) and hydroxyapatite (HAp) coatings were developed to control the degradation speed and to improve the biocompatibility of biodegradable magnesium alloys. Osteoblast MG-63 was cultured directly on OCP- and HAp-coated Mg-3Al-1Zn (wt%, AZ31) alloy (OCP- and HAp-AZ31) to evaluate cell compatibility. Cell proliferation was remarkably improved with OCP and HAp coatings which reduced the corrosion and prevented the H2O2 generation on Mg alloy substrate. OCP-AZ31 showed sparse distribution of living cell colonies and dead cells. HAp-AZ31 showed dense and homogeneous distribution of living cells, with dead cells localized over and around corrosion pits, some of which were formed underneath the coating. These results demonstrated that cells were dead due to changes in the local environment, and it is necessary to evaluate the local biocompatibility of magnesium alloys. Cell density on HAp-AZ31 was higher than that on OCP-AZ31 although there was not a significant difference in the amount of Mg ions released in medium between OCP- and HAp-AZ31. The outer layer of OCP and HAp coatings consisted of plate-like crystal with a thickness of around 0.1 μm and rod-like crystals with a diameter of around 0.1 μm, respectively, which grew from a continuous inner layer. Osteoblasts formed focal contacts on the tips of plate-like OCP and rod-like HAp crystals, with heights of 2-5 μm. The spacing between OCP tips of 0.8-1.1 μm was wider than that between HAp tips of 0.2-0.3 μm. These results demonstrated that cell proliferation depended on the micromorphology of the coatings which governed spacing of focal contacts. Consequently, HAp coating is suitable for improving cell compatibility and bone-forming ability of the Mg alloy.

Observations on the Feeding and Symptomatology of Xiphinema and Longidorus on Selected Host Roots

PubMed Central

Cohn, E.

1970-01-01

In vitro feeding of Xiphinema brevicolle, X. index and Longidorus africanus on roots of host seedlings is described. Both Xiphinema spp. fed mainly along roots rather than at tips and up to several days at a single site. Feeding of L. africanus was confined to root tips and lasted up to 15 min. No visible short term reaction of roots parasitized by the Xiphinema spp. could be discerned, but both swelling and cessation of growth of root tips were observed within 20 hr after feeding by L. africanus. Long-term (12-month) symptoms on roots of several host plants caused by cultured populations of X. brevicolle, X. index, X. italiae, L. africanus and L. brevicaudatus are described. All the Xiphinema spp. caused a thinning and distinct darkening of root systems and, at some sites, a breakdown of the cortex. Both species of Longidorus caused stubby and swollen root tips. Root symptom severity was in proportion to nematode population levels. PMID:19322291

Colloidal Random Terpolymers: Controlling Reactivity Ratios of Colloidal Comonomers via Metal Tipping

DOE PAGES

Pavlopoulos, Nicholas G.; Dubose, Jeffrey T.; Hartnett, Erin D.; ...

2016-07-26

We report on a versatile synthetic m-shell nanoparticles (NPs) in the backbone, along with semiconductor CdSe@CdS nanorod (NR), or tetrapod (TP) side chain groups. A seven-step colloidal total synthesis enabled the synthesis of well-defined colloidal comonomers composed of a dipolar Au@CoNP attached to a single CdSe@CdS NR, or TP, where magnetic dipolar associations between Au@CoNP units promoted the formation of colloidal co- or terpolymers. The key step in this synthesis was the ability to photodeposit a single AuNP tip onto CdSe@CdS NR or TP that enables selective seeding of a dipolar CoNP onto the AuNP seed. In conclusion, we showmore » that the variation of the AuNP size directly controlled the size and dipolar character of the CoNP tip, where the size modulation of the Au and Au@CoNP tips is analogous to control of comonomer reactivity ratios in classical copolymerization processes.« less

Piezoelectric cantilever sensors

NASA Technical Reports Server (NTRS)

Shih, Wan Y. (Inventor); Shih, Wei-Heng (Inventor); Shen, Zuyan (Inventor)

2008-01-01

A piezoelectric cantilever with a non-piezoelectric, or piezoelectric tip useful as mass and viscosity sensors. The change in the cantilever mass can be accurately quantified by monitoring a resonance frequency shift of the cantilever. For bio-detection, antibodies or other specific receptors of target antigens may be immobilized on the cantilever surface, preferably on the non-piezoelectric tip. For chemical detection, high surface-area selective absorbent materials are coated on the cantilever tip. Binding of the target antigens or analytes to the cantilever surface increases the cantilever mass. Detection of target antigens or analytes is achieved by monitoring the cantilever's resonance frequency and determining the resonance frequency shift that is due to the mass of the adsorbed target antigens on the cantilever surface. The use of a piezoelectric unimorph cantilever allows both electrical actuation and electrical sensing. Incorporating a non-piezoelectric tip (14) enhances the sensitivity of the sensor. In addition, the piezoelectric cantilever can withstand damping in highly viscous liquids and can be used as a viscosity sensor in wide viscosity range.

Sickle Cell Disease

MedlinePlus

... message, please visit this page: About CDC.gov . Learn Tips for Receiving Better Care in the Emergency Department in Our Fact ... related care in the United States. Read Supplement » VIDEO Sickle Cell Disease: When to Transfuse Learn about indications for blood transfusion in patients with ...

Stroboscobic near-field scanning optical microscopy for 3D mapping of mode profiles of plasmonic nanostructures (Conference Presentation)

NASA Astrophysics Data System (ADS)

Dana, Aykutlu; Ozgur, Erol; Torunoglu, Gamze

2016-09-01

We present a dynamic approach to scanning near field optical microscopy that extends the measurement technique to the third dimension, by strobing the illumination in sync with the cantilever oscillation. Nitrogen vacancy (NV) centers in nanodiamonds placed on cantilever tips are used as stable emitters for emission enhancement. Local field enhancement and modulation of optical density states are mapped in three dimensions based on fluorescence intensity and spectrum changes as the tip is scanned over plasmonic nanostructures. The excitation of NV centers is done using a total internal reflection setup. Using a digital phase locked loop to pulse the excitation in various tip sample separations, 2D slices of fluorescence enhancement can be recorded. Alternatively, a conventional SNOM tip can be used to selectively couple wideband excitation to the collection path, with subdiffraction resolution of 60 nm in x and y and 10 nm in z directions. The approach solves the problem of tip-sample separation stabilization over extended periods of measurement time, required to collect data resolved in emission wavelength and three spatial dimensions. The method can provide a unique way of accessing the three dimensional field and mode profiles of nanophotonics structures.

Fundamental aspects of electric double layer force-distance measurements at liquid-solid interfaces using atomic force microscopy

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

Black, Jennifer M.; Zhu, Mengyang; Zhang, Pengfei

In this paper, atomic force microscopy (AFM) force-distance measurements are used to investigate the layered ion structure of Ionic Liquids (ILs) at the mica surface. The effects of various tip properties on the measured force profiles are examined and reveal that the measured ion position is independent of tip properties, while the tip radius affects the forces required to break through the ion layers as well as the adhesion force. Force data is collected for different ILs and directly compared with interfacial ion density profiles predicted by molecular dynamics. Through this comparison it is concluded that AFM force measurements aremore » sensitive to the position of the ion with the larger volume and mass, suggesting that ion selectivity in force-distance measurements are related to excluded volume effects and not to electrostatic or chemical interactions between ions and AFM tip. Finally, the comparison also revealed that at distances greater than 1 nm the system maintains overall electroneutrality between the AFM tip and sample, while at smaller distances other forces (e.g., van der waals interactions) dominate and electroneutrality is no longer maintained.« less

Fundamental aspects of electric double layer force-distance measurements at liquid-solid interfaces using atomic force microscopy

DOE PAGES

Black, Jennifer M.; Zhu, Mengyang; Zhang, Pengfei; ...

2016-09-02

In this paper, atomic force microscopy (AFM) force-distance measurements are used to investigate the layered ion structure of Ionic Liquids (ILs) at the mica surface. The effects of various tip properties on the measured force profiles are examined and reveal that the measured ion position is independent of tip properties, while the tip radius affects the forces required to break through the ion layers as well as the adhesion force. Force data is collected for different ILs and directly compared with interfacial ion density profiles predicted by molecular dynamics. Through this comparison it is concluded that AFM force measurements aremore » sensitive to the position of the ion with the larger volume and mass, suggesting that ion selectivity in force-distance measurements are related to excluded volume effects and not to electrostatic or chemical interactions between ions and AFM tip. Finally, the comparison also revealed that at distances greater than 1 nm the system maintains overall electroneutrality between the AFM tip and sample, while at smaller distances other forces (e.g., van der waals interactions) dominate and electroneutrality is no longer maintained.« less

Experimental study of main rotor tip geometry and tail rotor interactions in hover. Volume 2: Run log and tabulated data

NASA Technical Reports Server (NTRS)

Balch, D. T.; Lombardi, J.

1985-01-01

A model scale hover test was conducted in the Sikorsky Aircraft Model Rotor hover Facility to identify and quantify the impact of the tail rotor on the demonstrated advantages of advanced geometry tip configurations. The existence of mutual interference between hovering main rotor and a tail rotor was acknowledged in the test. The test was conducted using the Basic Model Test Rig and two scaled main rotor systems, one representing a 1/5.727 scale UH-60A BLACK HAWK and the others a 1/4.71 scale S-76. Eight alternate rotor tip configurations were tested, 3 on the BLACK HAWK rotor and 6 on the S-76 rotor. Four of these tips were then selected for testing in close proximity to an operating tail rotor (operating in both tractor and pusher modes) to determine if the performance advantages that could be obtained from the use of advanced geometry tips in a main rotor only environment would still exist in the more complex flow field involving a tail rotor. This volume contains the test run log and tabulated data.

Tracking Blade Tip Vortices for Numerical Flow Simulations of Hovering Rotorcraft

NASA Technical Reports Server (NTRS)

Kao, David L.

2016-01-01

Blade tip vortices generated by a helicopter rotor blade are a major source of rotor noise and airframe vibration. This occurs when a vortex passes closely by, and interacts with, a rotor blade. The accurate prediction of Blade Vortex Interaction (BVI) continues to be a challenge for Computational Fluid Dynamics (CFD). Though considerable research has been devoted to BVI noise reduction and experimental techniques for measuring the blade tip vortices in a wind tunnel, there are only a handful of post-processing tools available for extracting vortex core lines from CFD simulation data. In order to calculate the vortex core radius, most of these tools require the user to manually select a vortex core to perform the calculation. Furthermore, none of them provide the capability to track the growth of a vortex core, which is a measure of how quickly the vortex diffuses over time. This paper introduces an automated approach for tracking the core growth of a blade tip vortex from CFD simulations of rotorcraft in hover. The proposed approach offers an effective method for the quantification and visualization of blade tip vortices in helicopter rotor wakes. Keywords: vortex core, feature extraction, CFD, numerical flow visualization

UWB Wind Turbine Blade Deflection Sensing for Wind Energy Cost Reduction.

PubMed

Zhang, Shuai; Jensen, Tobias Lindstrøm; Franek, Ondrej; Eggers, Patrick C F; Olesen, Kim; Byskov, Claus; Pedersen, Gert Frølund

2015-08-12

A new application of utilizing ultra-wideband (UWB) technology to sense wind turbine blade deflections is introduced in this paper for wind energy cost reduction. The lower UWB band of 3.1-5.3 GHz is applied. On each blade, there will be one UWB blade deflection sensing system, which consists of two UWB antennas at the blade root and one UWB antenna at the blade tip. The detailed topology and challenges of this deflection sensing system are addressed. Due to the complexity of the problem, this paper will first realize the on-blade UWB radio link in the simplest case, where the tip antenna is situated outside (and on the surface of) a blade tip. To investigate this case, full-blade time-domain measurements are designed and conducted under different deflections. The detailed measurement setups and results are provided. If the root and tip antenna locations are properly selected, the first pulse is always of sufficient quality for accurate estimations under different deflections. The measured results reveal that the blade tip-root distance and blade deflection can be accurately estimated in the complicated and lossy wireless channels around a wind turbine blade. Some future research topics on this application are listed finally.

New application system for laser and ultrasonic therapy in endoscopic surgery

NASA Astrophysics Data System (ADS)

Desinger, Kai; Helfmann, Juergen; Stein, Thomas; Mueller, Gerhard J.

1996-12-01

Flexible acoustic waveguides for selective tissue fragmentation are not yet commercially available. Experimental studies have shown the possibility of transmission of acoustical transients via optical silica glass fibers. The aim of this project is the development of a new endoscopic application system that would enable surgeons to use the laser and the ultrasound technique for therapy simultaneously. The concept of this application system is based on the transmission of laser radiation and ultrasound power via flexible silica glass fibers. Theoretical and experimental results on the feasibility of such an application system for an ultrasonic power delivery system are presented. Piezo-electric transducers are used to provide a high efficiency in generating the ultrasonic power. With reference to the CUSA-technique, a special flexible guiding system has been designed for providing aspiration at the tip and for protection of the fiber. The system transmits via an optical fiber up to 100 Watt Nd:YAG laser radiation. The axial oscillation of the fiber tip is +/- micrometers at a frequency of 27 kHz. First results of in vitro experiments are presented. The parenchymatous cells of liver can be fragmented without destruction of the collagenous matrix. The laser can be optionally used to coagulate bleedings or to cut collagenous tissues in contact. Applications for an acoustical and optical waveguide in ultrasonic surgery are demonstrated. This new approach in developing a first application system for the therapeutical use of laser radiation and power ultrasound in minimal invasive surgery via optical waveguides offers new possibilities in surgery. The laser ultrasonic surgical therapy (LUST) with its thin and flexible applicator provides new working fields especially for neuro or liver surgery. The tip can be bent and thus areas which could not be treated before have now been made accessible. Without changing the instrumentation, the surgeon can use the laser for tissue coagulation or cutting, next to the selective ultrasonic tissue fragmentation, where nerves or vessels will not be affected. Such a LUST-application system could be ready for clinical use in two to four years.

Smart helicopter rotor with active blade tips

NASA Astrophysics Data System (ADS)

Bernhard, Andreas Paul Friedrich

2000-10-01

The smart active blade tip (SABT) rotor is an on-blade rotor vibration reduction system, incorporating active blade tips that can be independently pitched with respect to the main blade. The active blade tip rotor development included an experimental test program culminating in a Mach scale hover test, and a parallel development of a coupled, elastic actuator and rotor blade analysis for preliminary design studies and hover performance prediction. The experimental testing focussed on a small scale rotor on a bearingless Bell-412 hub. The fabricated Mach-scale active-tip rotor has a diameter of 1.524 m, a blade chord of 76.2 mm and incorporated a 10% span active tip. The nominal operating speed is 2000 rpm, giving a tip Mach number of 0.47. The blade tips are driven by a novel piezo-induced bending-torsion coupled actuator beam, located spanwise in the hollow mid-cell of the main rotor blade. In hover at 2000 rpm, at 2 deg collective, and for an actuation of 125 Vrms, the measured blade tip deflection at the first four rotor harmonics is between +/-1.7 and +/-2.8 deg, increasing to +/-5.3 deg at 5/rev with resonant amplification. The corresponding oscillatory amplitude of the rotor thrust coefficient is between 0.7 · 10-3 and 1.3 · 10-1 at the first four rotor harmonics, increasing to 2.1 · 10-3 at 5/rev. In general, the experimental blade tip frequency response and corresponding rotor thrust response are well captured by the analysis. The flexbeam root flap bending moment is predicted in trend, but is significantly over-estimated. The blade tips did not deflect as expected at high collective settings, because of the blade tip shaft locking up in the bearing. This is caused by the high flap bending moment on the blade tip shaft. Redesign of the blade tip shaft assembly and bearing support is identified as the primary design improvement for future research. The active blade tip rotor was also used as a testbed for the evaluation of an adaptive neural-network based control algorithm. Effective background vibration reduction of an intentional 1/rev hover imbalance was demonstrated. The control algorithm also showed the capability to generate desired multi-frequency control loads on the hub, based on artificial signal injection into the vibration measurement. The research program demonstrates the technical feasibility of the active blade tip concept for vibration reduction and warrants further investigation in terms of closed loop forward flight tests in the windtunnel and full scale design studies.