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Sample records for nonplanar secretory iga2

  1. The Nonplanar Secretory IgA2 and Near Planar Secretory IgA1 Solution Structures Rationalize Their Different Mucosal Immune Responses*S⃞

    PubMed Central

    Bonner, Alexandra; Almogren, Adel; Furtado, Patricia B.; Kerr, Michael A.; Perkins, Stephen J.

    2009-01-01

    Secretory IgA (SIgA) is the most prevalent human antibody and is central to mucosal immunity. It exists as two subclasses, SIgA1 and SIgA2, where SIgA2 has a shorter hinge joining the Fab and Fc regions. Both forms of SIgA are predominantly dimeric and contain an additional protein called the secretory component (SC) that is attached during the secretory process and is believed to protect SIgA in harsh mucosal conditions. Here we locate the five SC domains relative to dimeric IgA2 within SIgA2 using constrained scattering modeling. The x-ray and sedimentation parameters showed that SIgA2 has an extended solution structure. The constrained modeling of SIgA2 was initiated using two IgA2 monomers that were positioned according to our best fit solution structure for dimeric IgA1. SC was best located along the convex edge of the Fc-Fc region. The best fit models showed that SIgA2 is significantly nonplanar in its structure, in distinction to our previous near planar SIgA1 structure. Both the shorter IgA2 hinges and the presence of SC appear to displace the four Fab regions out of the Fc plane in SIgA2. This may explain the noncovalent binding of SC in some SIgA2 molecules. This nonplanar structure is predicted to result in specific immune properties for SIgA2 and SIgA1. It may explain differences observed between the SIgA1 and SIgA2 subclasses in terms of their interactions with antigens, susceptibility to proteases, effects on receptors, and distribution in different tissues. The different structures account for the prevalence of both forms in mucosal secretions. PMID:19109255

  2. Highly nonplanar lifting systems

    NASA Technical Reports Server (NTRS)

    Kroo, Ilan; McMasters, John; Smith, Stephen C.

    1996-01-01

    This paper deals with nonplanar wing concepts -- their advantages and possible applications in a variety of aircraft designs. A brief review and assessment of several concepts from winglets to ring wings is followed by a more detailed look at two recent ideas: exploiting nonplanar wakes to reduce induced drag, and applying a 'C-wing' design to large commercial transports. Results suggest that potential efficiency gains may be significant, while several nonaerodynamic characteristics are particularly interesting.

  3. Non-planar chemical preconcentrator

    DOEpatents

    Manginell, Ronald P.; Adkins, Douglas R.; Sokolowski, Sara S.; Lewis, Patrick R.

    2006-10-10

    A non-planar chemical preconcentrator comprises a high-surface area, low mass, three-dimensional, flow-through sorption support structure that can be coated or packed with a sorptive material. The sorptive material can collect and concentrate a chemical analyte from a fluid stream and rapidly release it as a very narrow temporal plug for improved separations in a microanalytical system. The non-planar chemical preconcentrator retains most of the thermal and fabrication benefits of a planar preconcentrator, but has improved ruggedness and uptake, while reducing sorptive coating concerns and extending the range of collectible analytes.

  4. Evidence for a nonplanar amplituhedron

    SciTech Connect

    Bern, Zvi; Herrmann, Enrico; Litsey, Sean; Stankowicz, James; Trnka, Jaroslav

    2016-06-17

    The scattering amplitudes of planar N = 4 super-Yang-Mills exhibit a number of remarkable analytic structures, including dual conformal symmetry and logarithmic singularities of integrands. The amplituhedron is a geometric construction of the integrand that incorporates these structures. This geometric construction further implies the amplitude is fully specified by constraining it to vanish on spurious residues. By writing the amplitude in a dlog basis, we provide nontrivial evidence that these analytic properties and “zero conditions” carry over into the nonplanar sector. Finally, this suggests that the concept of the amplituhedron can be extended to the nonplanar sector of N = 4 super-Yang-Mills theory.

  5. Evidence for a nonplanar amplituhedron

    DOE PAGES

    Bern, Zvi; Herrmann, Enrico; Litsey, Sean; ...

    2016-06-17

    The scattering amplitudes of planar N = 4 super-Yang-Mills exhibit a number of remarkable analytic structures, including dual conformal symmetry and logarithmic singularities of integrands. The amplituhedron is a geometric construction of the integrand that incorporates these structures. This geometric construction further implies the amplitude is fully specified by constraining it to vanish on spurious residues. By writing the amplitude in a dlog basis, we provide nontrivial evidence that these analytic properties and “zero conditions” carry over into the nonplanar sector. Finally, this suggests that the concept of the amplituhedron can be extended to the nonplanar sector of N =more » 4 super-Yang-Mills theory.« less

  6. Monlithic nonplanar ring oscillator and method

    NASA Technical Reports Server (NTRS)

    Nilsson, Alan C. (Inventor); Byer, Robert L. (Inventor)

    1991-01-01

    A monolithic nonplanar ring oscillator having an optically isotropic solid-state laser body for propagating laser radiation about a nonplanar ring path internal to the laser body is disclosed. The monolithic laser body is configured to produce a 2N reflection nonplanar ring light path, where N is an integer greater than or equal to 2, comprising 2N-1 total internal reflections and one reflection at a coupler in a single round trip. Undirectional traveling wave oscillation of the laser is induced by the geometry of the nonplanar ring path together with the effect of an applied magnetic field and partial polarizer characteristics of the oblique reflection from the coupler. The 6-reflection nonplanar ring oscillator makes possible otpimal unidirectional oscillation (low loss for the oscillating direction of propagation and, simultaneously high loss for the nonoscillating direction of propagation) in monolithic NPROs using materials with index of refraction smaller than the square root of 3, for example, laser glass.

  7. Nonplanar electrostatic shock waves in dense plasmas

    SciTech Connect

    Masood, W.; Rizvi, H.

    2010-02-15

    Two-dimensional quantum ion acoustic shock waves (QIASWs) are studied in an unmagnetized plasma consisting of electrons and ions. In this regard, a nonplanar quantum Kadomtsev-Petviashvili-Burgers (QKPB) equation is derived using the small amplitude perturbation expansion method. Using the tangent hyperbolic method, an analytical solution of the planar QKPB equation is obtained and subsequently used as the initial profile to numerically solve the nonplanar QKPB equation. It is observed that the increasing number density (and correspondingly the quantum Bohm potential) and kinematic viscosity affect the propagation characteristics of the QIASW. The temporal evolution of the nonplanar QIASW is investigated both in Cartesian and polar planes and the results are discussed from the numerical stand point. The results of the present study may be applicable in the study of propagation of small amplitude localized electrostatic shock structures in dense astrophysical environments.

  8. Nonplanar waves with electronegative dusty plasma

    SciTech Connect

    Zobaer, M. S.; Mukta, K. N.; Nahar, L.; Mamun, A. A.; Roy, N.

    2013-04-15

    A rigorous theoretical investigation has been made of basic characteristics of the nonplanar dust-ion-acoustic shock and solitary waves in electronegative dusty plasma containing Boltzmann electrons, Boltzmann negative ions, inertial positive ions, and charge fluctuating (negatively charged) stationary dust. The Burgers' and Korteweg-de Vries (K-dV) equations, which is derived by reductive perturbation technique, is numerically solved to examine the effects of nonplanar geometry on the basic features of the DIA shock and solitary waves formed in the electronegative dusty plasma. The implications of the results (obtained from this investigation) in space and laboratory experiments are briefly discussed.

  9. Nonplanar solitons collision in ultracold neutral plasmas

    SciTech Connect

    El-Tantawy, S. A.; Moslem, W. M.; El-Metwally, M.; Sabry, R.; El-Labany, S. K.; Schlickeiser, R.

    2013-09-15

    Collisions between two nonplanar ion-acoustic solitons in strongly coupled ultracold neutral plasmas composed of ion fluid and non-Maxwellian (nonthermal or superthermal) electron distributions are investigated. The extended Poincare-Lighthill-Kuo method is used to obtain coupled nonplanar Kortweg-de Vries equations for describing the system. The nonplanar phase shifts after the interaction of the two solitons are calculated. It is found that the properties of the nonplanar colliding solitons and its corresponding phase shifts are different from those in the planar case. The polarity of the colliding solitons strongly depends on the type of the non-Maxwellian electron distributions. A critical nonthermality parameter β{sub c} is identified. For values of β ≤ β{sub c} solitons with double polarity exist, while this behavior cannot occur for superthermal plasmas. The phase shift for nonthermal plasmas increases below β{sub c} for a positive soliton, but it decreases for β > β{sub c} for a negative soliton. For superthermal plasmas, the phase shift enhances rapidly for low values of spectral index κ and higher values of ions effective temperature ratio σ{sub *}. For 2 ≲ κ<10, the phase shift decreases but does not change for κ > 10. The nonlinear structure, as reported here, is useful for controlling the solitons created in forthcoming ultracold neutral plasma experiments.

  10. Nonplanar Shock Waves in Dusty Plasmas

    SciTech Connect

    Mamun, A. A.; Shukla, P. K.

    2011-11-29

    Nonplanar (viz. cylindrical and spherical) electro-acoustic [dust-ion-acoustic (DIA) and dust-acoustic (DA)] shock waves have been investigated by employing the reductive perturbation method. The dust charge fluctuation (strong correlation among highly charged dust) is the source of dissipation, and is responsible for the formation of the DIA (DA) shock structures. The effects of cylindrical and spherical geometries on the time evolution of DIA and DA shock structures are examined and identified. The combined effects of vortex-like electron distribution and dust charge fluctuation (dust-correlation and effective dust-temperature) on the basic features of nonplanar DIA (DA) shock waves are pinpointed. The implications of our results in laboratory dusty plasma experiments are briefly discussed.

  11. Nonplanar Shock Waves in Dusty Plasmas

    NASA Astrophysics Data System (ADS)

    Mamun, A. A.; Shukla, P. K.

    2011-11-01

    Nonplanar (viz. cylindrical and spherical) electro-acoustic [dust-ion-acoustic (DIA) and dust-acoustic (DA)] shock waves have been investigated by employing the reductive perturbation method. The dust charge fluctuation (strong correlation among highly charged dust) is the source of dissipation, and is responsible for the formation of the DIA (DA) shock structures. The effects of cylindrical and spherical geometries on the time evolution of DIA and DA shock structures are examined and identified. The combined effects of vortex-like electron distribution and dust charge fluctuation (dust-correlation and effective dust-temperature) on the basic features of nonplanar DIA (DA) shock waves are pinpointed. The implications of our results in laboratory dusty plasma experiments are briefly discussed.

  12. Structural mechanisms of nonplanar hemes in proteins

    SciTech Connect

    Shelnutt, J.A.

    1997-05-01

    The objective is to assess the occurrence of nonplanar distortions of hemes and other tetrapyrroles in proteins and to determine the biological function of these distortions. Recently, these distortions were found by us to be conserved among proteins belonging to a functional class. Conservation of the conformation of the heme indicates a possible functional role. Researchers have suggested possible mechanisms by which heme distortions might influence biological properties; however, no heme distortion has yet been shown conclusively to participate in a structural mechanism of hemoprotein function. The specific aims of the proposed work are: (1) to characterize and quantify the distortions of the hemes in all of the more than 300 hemoprotein X-ray crystal structures in terms of displacements along the lowest-frequency normal coordinates, (2) to determine the structural features of the protein component that generate and control these nonplanar distortions by using spectroscopic studies and molecular-mechanics calculations for the native proteins, their mutants and heme-peptide fragments, and model porphyrins, (3) to determine spectroscopic markers for the various types of distortion, and, finally, (4) to discover the functional significance of the nonplanar distortions by correlating function with porphyrin conformation for proteins and model porphyrins.

  13. Non-planar microfabricated gas chromatography column

    DOEpatents

    Lewis, Patrick R.; Wheeler, David R.

    2007-09-25

    A non-planar microfabricated gas chromatography column comprises a planar substrate having a plurality of through holes, a top lid and a bottom lid bonded to opposite surfaces of the planar substrate, and inlet and outlet ports for injection of a sample gas and elution of separated analytes. A plurality of such planar substrates can be aligned and stacked to provide a longer column length having a small footprint. Furthermore, two or more separate channels can enable multi-channel or multi-dimensional gas chromatography. The through holes preferably have a circular cross section and can be coated with a stationary phase material or packed with a porous packing material. Importantly, uniform stationary phase coatings can be obtained and band broadening can be minimized with the circular channels. A heating or cooling element can be disposed on at least one of the lids to enable temperature programming of the column.

  14. Influence of nonplanarity and extended conjugation on porphyrin basicity.

    PubMed

    Finikova, Olga S; Cheprakov, Andrei V; Carroll, Patrick J; Dalosto, Sergio; Vinogradov, Sergei A

    2002-12-30

    Thermodynamic basicities of several new nonplanar water soluble tetraaryltetracyclohexano- (Ar(4)TCHP) and tetraaryltetrabenzoporphyrins (Ar(4)TBP) have been measured and correlated with their structural parameters. While the degrees of nonplanarity in these porphyrins are similar, Ar(4)TCHPs exhibit significantly higher basicities than Ar(4)TBPs and than planar tetraarylporphyrins. Low basicities of distorted Ar(4)TBPs are believed to be due to extended pi-conjugation, which causes delocalization of the core electron density in these porphyrins and reduces negative charges at the protonation site.

  15. Experimental investigation of non-planar sheared outboard wing planforms

    NASA Technical Reports Server (NTRS)

    Naik, D. A.; Ostowari, C.

    1988-01-01

    The outboard planforms of wings have been found to be of prime importance in studies of induced drag reduction. This conclusion is based on an experimental and theoretical study of the aerodynamic characteristics of planar and nonplanar outboard wing forms. Six different configurations; baseline rectangular, planar sheared, sheared with dihedral, sheared with anhedral, rising arc, and drooping arc were investigated for two different spans. Span efficiencies as much as 20 percent greater than baseline can be realized with nonplanar wing forms. Optimization studies show that this advantage can be achieved along with a bending moment benefit. Parasite drag and lateral stability estimations were not included in the analysis.

  16. Constrained Quantum Mechanics: Chaos in Non-Planar Billiards

    ERIC Educational Resources Information Center

    Salazar, R.; Tellez, G.

    2012-01-01

    We illustrate some of the techniques to identify chaos signatures at the quantum level using as guiding examples some systems where a particle is constrained to move on a radial symmetric, but non-planar, surface. In particular, two systems are studied: the case of a cone with an arbitrary contour or "dunce hat billiard" and the rectangular…

  17. Constrained quantum mechanics: chaos in non-planar billiards

    NASA Astrophysics Data System (ADS)

    Salazar, R.; Téllez, G.

    2012-07-01

    We illustrate some of the techniques to identify chaos signatures at the quantum level using as guiding examples some systems where a particle is constrained to move on a radial symmetric, but non-planar, surface. In particular, two systems are studied: the case of a cone with an arbitrary contour or dunce hat billiard and the rectangular billiard with an inner Gaussian surface.

  18. Mammary Analogue Secretory Carcinoma.

    PubMed

    Stevens, Todd M; Parekh, Vishwas

    2016-09-01

    Mammary analogue secretory carcinoma (MASC) is a recently described salivary gland tumor that shares the same histologic appearance and ETV6 gene (12p13) rearrangement as secretory carcinoma of the breast. Prior to its recognition, MASC cases were commonly labeled acinic cell carcinoma and adenocarcinoma, not otherwise specified. Despite distinctive histologic features, MASC may be difficult to distinguish from other salivary gland tumors, in particular zymogen-poor acinic cell carcinoma and low-grade salivary duct carcinoma. Although characteristic morphologic and immunohistochemical features form the basis of a diagnosis of MASC, the presence of an ETV6-NTRK3 gene fusion is confirmatory. Given its recent recognition the true prognostic import of MASC is not yet clearly defined.

  19. Study of nonplanarity of peptide bond using theoretical calculations.

    PubMed

    Selvarengan, P; Kolandaivel, P

    2005-08-01

    The conformational dependence of nonplanarity of the peptide bond of formylglycinamide has been studied using ab initio and density functional theory methods. Hartree-Fock self-consistent field theory (HF), Møller-Plesset perturbation theory (MP2) of ab initio and B3LYP level of theory of dft method have been used employing 6-31++G** basis set. The MP2 method predicts better results than HF and B3LYP levels of theory for conformational stability dependence of nonplanarity. Systematic dependence of planarity deviation has been observed in MP2 theory. The chemical hardness values successfully predict the conformational region, but fail to obey maximum hardness principle. It is concluded that the most reliable dft method could not successfully predict the planarity of peptide bond in comparison with electron correlated method of ab initio method.

  20. Defective non-planar surfaces of MgO

    NASA Astrophysics Data System (ADS)

    Colbourn, E. A.; Kendrick, J.; Mackrodt, W. C.

    1983-03-01

    Defect lattice and ab initio SCF calculations are reported for defective non-planar surfaces of MgO. It is suggested that the concentration of free vacancies and Ca 2+ will be much lower at non-planar irregularities than at planar surfaces and in the bulk, whereas the level of Li + and trapped holes will be higher. The binding of H atoms and CO to three-coordinated comer sites is found to be appreciably higher than to the five-coordinated {001} surface. Thus it is concluded that calcium doping of MgO will affect neither CO chemisorption nor the dissociative chemisorption of hydrogen whereas lithium doping will influence both.

  1. Application of Random Ferns for non-planar object detection

    NASA Astrophysics Data System (ADS)

    Mastov, Alexey; Konovalenko, Ivan; Grigoryev, Anton

    2015-12-01

    The real time object detection task is considered as a part of a project devoted to development of autonomous ground robot. This problem has been successfully solved with Random Ferns algorithm, which belongs to keypoint-based method and uses fast machine learning algorithms for keypoint matching step. As objects in the real world are not always planar, in this article we describe experiments of applying this algorithm for non-planar objects. Also we introduce a method for fast detection of a special class of non-planar objects | those which can be decomposed into planar parts (e.g. faces of a box). This decomposition needs one detector for each side, which may significantly affect speed of detection. Proposed approach copes with it by omitting repeated steps for each detector and organizing special queue of detectors. It makes the algorithm three times faster than naive one.

  2. Increased serum concentration of BAFF/APRIL and IgA2 subclass in patients with mixed connective tissue disease complicated by interstitial lung disease.

    PubMed

    Kaneko, Toshiyuki; Amano, Hirofumi; Kawano, Shinya; Minowa, Kentaro; Ando, Seiichiro; Watanabe, Takashi; Nakano, Soichiro; Suzuki, Jun; Morimoto, Shinji; Tokano, Yoshiaki; Takasaki, Yoshinari

    2014-03-01

    B cell activating factor (BAFF) and a proliferation-inducing ligand (APRIL) are known to be crucial for B cell maturation and survival, and increased expression of these factors in various autoimmune diseases has been reported. Human B cells produce two IgA subclasses: IgA1 and IgA2, the latter being abundant in the distal intestine, saliva, colostrum and bronchial fluid. We investigated these parameters in patients with mixed connective tissue disease (MCTD) complicated by interstitial lung disease (ILD+), and compared them with those in MCTD patients without ILD (ILD-). Sixty-three MCTD patients were divided into two groups: 21 ILD+ patients and 42 ILD- patients. In each patient group we analyzed soluble BAFF/APRIL using ELISA, and IgA1 and IgA2 using double immunodiffusion. Furthermore, we analyzed BAFF-APRIL receptors, BCMA, BAFF-R and TACI, using flow cytometry. The ILD+ patients had significantly higher levels of BAFF/APRIL than the ILD- patients. There were significant correlations between BAFF/APRIL, BAFF/KL-6 and APRIL/KL-6. Although there was no significant inter-group difference in the serum IgA1 level, ILD+ patients had a significantly elevated IgA2 level in comparison with ILD- patients. Moreover, although there were no significant inter-group differences in the expression of BCMA, BAFF-R and TACI on B cells, the expression of BAFF-R was significantly decreased in the ILD+ patients. In recent years, relationships between BAFF/APRIL and IgA subclass have been reported. Our results suggest that an elevated level of BAFF/APRIL drives the maturation of B cells, subsequently leading to IgA2 class switching, and possibly to the development of ILD in patients with MCTD.

  3. Solution structure determination of monomeric human IgA2 by X-ray and neutron scattering, analytical ultracentrifugation and constrained modelling: a comparison with monomeric human IgA1.

    PubMed

    Furtado, Patricia B; Whitty, Patrick W; Robertson, Alexis; Eaton, Julian T; Almogren, Adel; Kerr, Michael A; Woof, Jenny M; Perkins, Stephen J

    2004-05-14

    Immunoglobulin A (IgA), the most abundant human immunoglobulin, mediates immune protection at mucosal surfaces as well as in plasma. It exists as two subclasses IgA1 and IgA2, and IgA2 is found in at least two allotypic forms, IgA2m(1) or IgA2m(2). Compared to IgA1, IgA2 has a much shorter hinge region, which joins the two Fab and one Fc fragments. In order to assess its solution structure, monomeric recombinant IgA2m(1) was studied by X-ray and neutron scattering. Its Guinier X-ray radius of gyration R(G) is 5.18 nm and its neutron R(G) is 5.03 nm, both of which are significantly smaller than those for monomeric IgA1 at 6.1-6.2 nm. The distance distribution function P(r)for IgA2m(1) showed a broad peak with a subpeak and gave a maximum dimension of 17 nm, in contrast to the P(r) curve for IgA1, which showed two distinct peaks and a maximum dimension of 21 nm. The sedimentation coefficients of IgA1 and IgA2m(1) were 6.2S and 6.4S, respectively. These data show that the solution structure of IgA2m(1) is significantly more compact than IgA1. The complete monomeric IgA2m(1) structure was modelled using molecular dynamics to generate random IgA2 hinge structures, to which homology models for the Fab and Fc fragments were connected to generate 10,000 full models. A total of 104 compact best-fit IgA2m(1) models gave good curve fits. These best-fit models were modified by linking the two Fab light chains with a disulphide bridge that is found in IgA2m(1), and subjecting these to energy refinement to optimise this linkage. The averaged solution structure of the arrangement of the Fab and Fc fragments in IgA2m(1) was found to be predominantly T-shaped and flexible, but also included Y-shaped structures. The IgA2 models show full steric access to the two FcalphaRI-binding sites at the Calpha2-Calpha3 interdomain region in the Fc fragment. Since previous scattering modelling had shown that IgA1 also possessed a flexible T-shaped solution structure, such a T-shape may be common to both IgA1 and IgA2. The final models suggest that the combination of the more compact IgA2m(1) and the more extended IgA1 structures will enable human IgA to access a broader range of antigens than either acting alone. The hinges of both IgA subclasses appear to show reduced flexibility when compared to their equivalents in IgG, and this may be important for maintaining an extended IgA structure.

  4. Dynamics of rotationally fissioned asteroids: non-planar case

    NASA Astrophysics Data System (ADS)

    Boldrin, L. A. G.; Scheeres, D. J.; Winter, O. C.

    2016-10-01

    The rotational fission of asteroids has been studied previously with simplified models restricted to planar motion. However, the observed physical configuration of contact binaries leads one to conclude that most of them are not in a planar configuration and hence would not be restricted to planar motion once they undergo rotational fission. This motivated a study of the evolution of initially non-planar binaries created by fission. Using a two-ellipsoid model, we performed simulations taking only gravitational interactions between components into account. We simulate 91 different initial inclinations of the equator of the secondary body for 19 different mass ratios. After disruption, the binary system dynamics are chaotic, as predicted from theory. Starting the system in a non-planar configuration leads to a larger energy and enhanced coupling between the rotation state of the smaller fissioned body and the evolving orbital system, and enables re-impact to occur. This leads to differences with previous planar studies, with collisions and secondary spin fission occurring for all mass ratios with inclinations θ0 ≥ 40o, and mimics a Lidov-Kozai mechanism. Out of 1729 studied cases, we found that ˜14 per cent result in secondary fission, ˜25 per cent result in collisions and ˜6 per cent have lifetimes longer than 200 yr. In Jacobson & Scheeres stable binaries only formed in cases with mass ratios q < 0.20. Our results indicate that it should be possible to obtain a stable binary with the same mechanisms for cases with mass ratios larger than this limit, but that the system should start in a non-planar configuration.

  5. Spin Vortex Resonance in Non-planar Ferromagnetic Dots

    PubMed Central

    Ding, Junjia; Lapa, Pavel; Jain, Shikha; Khaire, Trupti; Lendinez, Sergi; Zhang, Wei; Jungfleisch, Matthias B.; Posada, Christian M.; Yefremenko, Volodymyr G.; Pearson, John E.; Hoffmann, Axel; Novosad, Valentine

    2016-01-01

    In planar structures, the vortex resonance frequency changes little as a function of an in-plane magnetic field as long as the vortex state persists. Altering the topography of the element leads to a vastly different dynamic response that arises due to the local vortex core confinement effect. In this work, we studied the magnetic excitations in non-planar ferromagnetic dots using a broadband microwave spectroscopy technique. Two distinct regimes of vortex gyration were detected depending on the vortex core position. The experimental results are in qualitative agreement with micromagnetic simulations. PMID:27143405

  6. Tunneling properties of nonplanar molecules in a gas medium

    SciTech Connect

    Bahrami, Mohammad; Bassi, Angelo

    2011-12-15

    We propose a simple, general, and accurate formula for analyzing the tunneling between classical configurations of a nonplanar molecule in a gas medium, as a function of the thermodynamic parameters of the gas. We apply it to two interesting cases: (i) the shift to zero frequency of the inversion line of ammonia, upon an increase in the pressure of the gas; and (ii) the destruction of the coherent tunneling of D{sub 2}S{sub 2} molecules in a He gas. In both cases, we compare our analysis with previous theoretical and experimental results.

  7. Secretory Carcinoma of the Breast

    PubMed Central

    Aktepe, Fatma; Sarsenov, Dauren; Özmen, Vahit

    2016-01-01

    Secretory carcinoma is a very rare subtype of breast carcinoma. These tumors are generally associated with a favorable prognosis, although having triple-negative phenotype (estrogen receptor (ER), progesterone receptor (PR) negative and c-erbB2 (HER2) negative). In this presentation, a rare secretory carcinoma of the breast in a woman aged 24 years is discussed and the literature is reviewed. PMID:28331758

  8. Probabilistic Prognosis of Non-Planar Fatigue Crack Growth

    NASA Technical Reports Server (NTRS)

    Leser, Patrick E.; Newman, John A.; Warner, James E.; Leser, William P.; Hochhalter, Jacob D.; Yuan, Fuh-Gwo

    2016-01-01

    Quantifying the uncertainty in model parameters for the purpose of damage prognosis can be accomplished utilizing Bayesian inference and damage diagnosis data from sources such as non-destructive evaluation or structural health monitoring. The number of samples required to solve the Bayesian inverse problem through common sampling techniques (e.g., Markov chain Monte Carlo) renders high-fidelity finite element-based damage growth models unusable due to prohibitive computation times. However, these types of models are often the only option when attempting to model complex damage growth in real-world structures. Here, a recently developed high-fidelity crack growth model is used which, when compared to finite element-based modeling, has demonstrated reductions in computation times of three orders of magnitude through the use of surrogate models and machine learning. The model is flexible in that only the expensive computation of the crack driving forces is replaced by the surrogate models, leaving the remaining parameters accessible for uncertainty quantification. A probabilistic prognosis framework incorporating this model is developed and demonstrated for non-planar crack growth in a modified, edge-notched, aluminum tensile specimen. Predictions of remaining useful life are made over time for five updates of the damage diagnosis data, and prognostic metrics are utilized to evaluate the performance of the prognostic framework. Challenges specific to the probabilistic prognosis of non-planar fatigue crack growth are highlighted and discussed in the context of the experimental results.

  9. Planning maximally smooth hand movements constrained to nonplanar workspaces.

    PubMed

    Liebermann, Dario G; Krasovsky, Tal; Berman, Sigal

    2008-11-01

    The article characterizes hand paths and speed profiles for movements performed in a nonplanar, 2-dimensional workspace (a hemisphere of constant curvature). The authors assessed endpoint kinematics (i.e., paths and speeds) under the minimum-jerk model assumptions and calculated minimal amplitude paths (geodesics) and the corresponding speed profiles. The authors also calculated hand speeds using the 2/3 power law. They then compared modeled results with the empirical observations. In all, 10 participants moved their hands forward and backward from a common starting position toward 3 targets located within a hemispheric workspace of small or large curvature. Comparisons of modeled observed differences using 2-way RM-ANOVAs showed that movement direction had no clear influence on hand kinetics (p < .05). Workspace curvature affected the hand paths, which seldom followed geodesic lines. Constraining the paths to different curvatures did not affect the hand speed profiles. Minimum-jerk speed profiles closely matched the observations and were superior to those predicted by 2/3 power law (p < .001). The authors conclude that speed and path cannot be unambiguously linked under the minimum-jerk assumption when individuals move the hand in a nonplanar 2-dimensional workspace. In such a case, the hands do not follow geodesic paths, but they preserve the speed profile, regardless of the geometric features of the workspace.

  10. Nonplanar push-pull chromophores for opto-electronic applications.

    PubMed

    Breiten, Benjamin; Biaggio, Ivan; Diederich, François

    2010-01-01

    Donor-substituted cyanoethynylethenes (CEEs) are planar push-pull chromophores featuring intense intramolecular charge-transfer (CT) interactions and high third-order optical nonlinearities. Their thermal stability allows for the formation of crystalline thin films by vapor-phase deposition. On the other hand, high-quality amorphous thin films are preferred for opto-electronic applications and such films can be prepared using nonplanar push-pull chromophores with a less pronounced propensity to crystallize. By taking advantage of a versatile, atom-economic 'click-chemistry'-type transformation, involving a formal [2 + 2] cycloaddition of tetracyanoethene (TCNE) to electron-rich alkynes, followed by cycloreversion, stable donor-substituted 1,1,4,4-tetracyanobuta-1,3-dienes (TCBDs) are obtained in high yield and large quantities. These nonplanar push-pull chromophores also feature intense intramolecular CT and, in many cases, high third-order optical nonlinearities. Some of these compounds form high-optical-quality amorphous thin films by vapor-phase deposition, and first applications in next-generation opto-electronic devices have already been demonstrated. Chiral derivatives display high helical twisting power and are efficient dopants to translate molecular into macroscopic chirality, by switching nematic into cholesteric liquid crystalline phases.

  11. Photoinduced axial ligation and deligation dynamics of nonplanar nickel dodecaarylporphyrins.

    PubMed

    Retsek, Jennifer L; Drain, Charles Michael; Kirmaier, Christine; Nurco, Daniel J; Medforth, Craig J; Smith, Kevin M; Sazanovich, Igor V; Chirvony, Vladimir S; Fajer, Jack; Holten, Dewey

    2003-08-13

    The ground- and excited-state metal-ligand dynamics of nonplanar nickel(II) 2,3,5,7,8,10,12,13,15,17,18,20-dodecaphenylporphyrin (NiDPP) and two fluorinated analogues (NiF(20)DPP and NiF(28)DPP) have been investigated using static and time-resolved absorption spectroscopy in toluene and in ligating media that differ in basicity, aromaticity, and steric encumbrance. Because of the electronic and steric consequences of nonplanarity, NiDPP does not bind axial ligands in the ground state, but metal coordination does occur after photoexcitation with multistep dynamics that depend on the properties of the ligand. Following the structural relaxations that occur in all nickel porphyrins within approximately 10 ps, ligand binding to photoexcited NiDPP is progressively longer in pyridine, piperidine, and 3,5-lutidine (25-100 ps) but does not occur at all in 2,6-lutidine in which the ligating nitrogen is sterically encumbered. The transient intermediate that is formed, which nominally could be either a five- or six-coordinate species, also has a ligand-dependent lifetime (200-550 ps). Decay of this intermediate occurs partially via ligand release to re-form the uncoordinated species, in competition with binding of the second axial ligand and/or conformational/electronic relaxations (of a six-coordinate intermediate) to give the ground state of the bis-ligated photoproduct. The finding that the photoproduct channel principally depends on ligand characteristics along with the time-evolving spectra suggests that the transient intermediate may involve a five-coordinate species. In contrast to NiDPP, the fluorinated analogues NiF(20)DPP and NiF(28)DPP do coordinate axial ligands in the ground state but eject them after photoexcitation. Collectively, these results demonstrate the sensitivity with which the electronic and structural characteristics of the macrocycle, substituents, and solvent (ligands) can govern the photophysical and photochemical properties of nonplanar porphyrins

  12. Cell projection of meshes with non-planar faces

    SciTech Connect

    Max, N; Williams, P; Silva, C

    2000-11-27

    Volume rendering converts a scalar function on a 3D volume into varying colors and opacities, and creates an image by integrating the color and opacity effects along viewing rays through each pixel [1]. For data specified on a regular grid, the ray tracing is straightforward [2,3,4,5], and similar effects can be obtained with 3D textures [6]. For curvilinear or irregular grids, these methods are only applicable after the data has been resampled. An alternative, which works directly on these more general grids, is cell projection [7,8,9]. The cells composited onto the image in back to front sorted order. The projections of the edges of a single cell divide the image plane into polygons, which can be scan converted and composited by standard graphics hardware. In references [9,10,11], we assumed that the cells were polyhedra with planar faces. A curvilinear grid maps a rectangular grid onto a curved volume, for example to fit next to an airplane wing or ship hull, and quadrilateral faces may map to non-planar surfaces. Irregular grids are fitted to complex geometries, for example mechanical parts, and even initially flat faces may become non-planar as the grid elements deform, for example, in a car crash simulation. Non-planar faces cause problems in the sorting and compositing when a viewing ray crosses the same face twice. We call such a face a ''problem face''. For example, the ray may leave cell A through face F, enter cell B , and then enter cell A again through the same face F. If a viewing ray intersects a cell like A in two disjoint segments, we call the cell a ''problem cell''. This makes it impossible to sort cells A and B in back-to-front compositing order. Our solution is to divide problem cells into tetrahedra, which have planar faces. A single hexahedron can be projected and composited more quickly than the five or six tetrahedra into which it is subdivided, so we subdivide only the problem cells. In the example above, cell B might not turn out to be a

  13. Strain and the optoelectronic properties of nonplanar phosphorene monolayers

    PubMed Central

    Mehboudi, Mehrshad; Utt, Kainen; Terrones, Humberto; Harriss, Edmund O.; Pacheco SanJuan, Alejandro A.; Barraza-Lopez, Salvador

    2015-01-01

    Lattice kirigami, ultralight metamaterials, polydisperse aggregates, ceramic nanolattices, and 2D atomic materials share an inherent structural discreteness, and their material properties evolve with their shape. To exemplify the intimate relation among material properties and the local geometry, we explore the properties of phosphorene––a new 2D atomic material––in a conical structure, and document a decrease of the semiconducting gap that is directly linked to its nonplanar shape. This geometrical effect occurs regardless of phosphorene allotrope considered, and it provides a unique optical vehicle to single out local structural defects on this 2D material. We also classify other 2D atomic materials in terms of their crystalline unit cells, and propose means to obtain the local geometry directly from their diverse 2D structures while bypassing common descriptions of shape that are based from a parametric continuum. PMID:25918413

  14. Strain and the optoelectronic properties of nonplanar phosphorene monolayers.

    PubMed

    Mehboudi, Mehrshad; Utt, Kainen; Terrones, Humberto; Harriss, Edmund O; Pacheco SanJuan, Alejandro A; Barraza-Lopez, Salvador

    2015-05-12

    Lattice kirigami, ultralight metamaterials, polydisperse aggregates, ceramic nanolattices, and 2D atomic materials share an inherent structural discreteness, and their material properties evolve with their shape. To exemplify the intimate relation among material properties and the local geometry, we explore the properties of phosphorene--a new 2D atomic material--in a conical structure, and document a decrease of the semiconducting gap that is directly linked to its nonplanar shape. This geometrical effect occurs regardless of phosphorene allotrope considered, and it provides a unique optical vehicle to single out local structural defects on this 2D material. We also classify other 2D atomic materials in terms of their crystalline unit cells, and propose means to obtain the local geometry directly from their diverse 2D structures while bypassing common descriptions of shape that are based from a parametric continuum.

  15. The Secretory System of Arabidopsis

    PubMed Central

    Bassham, Diane C.; Brandizzi, Federica; Otegui, Marisa S.; Sanderfoot, Anton A.

    2008-01-01

    Over the past few years, a vast amount of research has illuminated the workings of the secretory system of eukaryotic cells. The bulk of this work has been focused on the yeast Saccharomyces cerevisiae, or on mammalian cells. At a superficial level, plants are typical eukaryotes with respect to the operation of the secretory system; however, important differences emerge in the function and appearance of endomembrane organelles. In particular, the plant secretory system has specialized in several ways to support the synthesis of many components of the complex cell wall, and specialized kinds of vacuole have taken on a protein storage role—a role that is intended to support the growing seedling, but has been co-opted to support human life in the seeds of many crop plants. In the past, most research on the plant secretory system has been guided by results in mammalian or fungal systems but recently plants have begun to stand on their own as models for understanding complex trafficking events within the eukaryotic endomembrane system. PMID:22303241

  16. Hybrid orbital deformation (HOD) effect and spectral red-shift property of nonplanar porphyrin.

    PubMed

    Zhou, Zaichun; Cao, Chenzhong; Liu, Qiuhua; Jiang, Rongqing

    2010-04-16

    A series of 5,15-meso,meso-strapped nonplanar porphyrins with different degrees of ruffling distortion, as a model system, have been synthesized and characterized. The spectral red-shift of the nonplanar porphyrins was experimentally demonstrated to mainly originate from the hybrid orbital deformation (HOD) effect due to the distortion in the tetrapyrrole macrocycle, which confirmed previous explanations to the red-shift phenomenon.

  17. A nonplanar porphyrin-based receptor molecule for chiral amine ligands

    SciTech Connect

    MUZZI,CINZIA M.; MEDFORTH,CRAIG J.; SMITH,KEVIN M.; JIA,SONG-LING; SHELNUTT,JOHN A.

    2000-03-06

    A novel porphyrin-based receptor molecule for chiral amine ligands is described in which nonplanarity of the porphyrin macrocycle is used to orient the ligand and to enhance porphyrin-ligand interactions. The porphyrin macrocycle provides a versatile platform upon which to build elaborate superstructures, and this feature coupled with a rich and well-developed synthetic chemistry has led to the synthesis of many elegant models of heme protein active sites and numerous porphyrin-based receptor molecules. One design feature which is not usually considered in the design of porphyrin-based receptor molecules is nonplanarity of the porphyrin ring, although there are a few systems such as the pyridine sensitive Venus Flytrap and the chirality-memory molecule which illustrate that nonplanar porphyrin-based receptors can display unique and interesting behavior. Given the novel properties of these receptors and the continuing interest in the effects of nonplanarity on the properties of porphyrins the authors decided to investigate in more detail the potential applications of nonplanarity in the design of porphyrin-based receptors. Herein, they describe the design, synthesis, and characterization of a new kind of nonplanar porphyrin-based receptor molecule for chiral amines.

  18. Formation of secretory granules by chromogranins.

    PubMed

    Inomoto, Chie; Osamura, Robert Yoshiyuki

    2009-12-01

    This review article covers the molecular mechanisms of secretory granule formation by chromogranin transfection. Recently, a few investigators have reported that the transfection of chromogranin A and B produces the structures of secretory granules. We used the GFP-chromogranin A transfection method to nonendocrine cells, COS-7 cells, which are not equipped with secretory granules. Despite the absence of endogenous secretory granules in nontransfected COS-7 cells, COS-7 cells transfected with chromogranin A contained granule-like structures in electron micrographs. The granules were composed of an outer limiting membrane with core structures that were interpreted as secretory granules. Human chromogranin A (CgA) labeled with 5-nm gold particles was present in several dense-core granules in our previous electron microscopy study. This review depicts the role of chromogranin A in the formation of secretory granules. It emphasizes the application of recently developed new technologies and the genesis of secretory granules.

  19. Constraint of fault parameters inferred from nonplanar fault modeling

    NASA Astrophysics Data System (ADS)

    Aochi, Hideo; Madariaga, Raul; Fukuyama, Eiichi

    2003-02-01

    We study the distribution of initial stress and frictional parameters for the 28 June 1992 Landers, California, earthquake through dynamic rupture simulation along a nonplanar fault system. We find that observational evidence of large slip distribution near the ground surface requires large nonzero cohesive forces in the depth-dependent friction law. This is the only way that stress can accumulate and be released at shallow depths. We then study the variation of frictional parameters along the strike of the fault. For this purpose we mapped into our segmented fault model the initial stress heterogeneity inverted by Peyrat et al. [2001] using a planar fault model. Simulations with this initial stress field improved the overall fit of the rupture process to that inferred from kinematic inversions, and also improved the fit to the ground motion observed in Southern California. In order to obtain this fit, we had to introduce an additional variations of frictional parameters along the fault. The most important is a weak Kickapoo fault and a strong Johnson Valley fault.

  20. Adsorption Behavior of Nonplanar Phthalocyanines: Competition of Different Adsorption Conformations

    PubMed Central

    2016-01-01

    Using density functional theory augmented with state-of-the-art van der Waals corrections, we studied the geometric and electronic properties of nonplanar chlorogallium-phthalocyanine GaClPc molecules adsorbed on Cu(111). Comparing these results with published experimental data for adsorption heights, we found indications for breaking of the metal–halogen bond when the molecule is heated during or after the deposition process. Interestingly, the work-function change induced by this dissociated geometry is the same as that computed for an intact adsorbate layer in the “Cl-down” configuration, with both agreeing well with the experimental photoemission data. This is unexpected, as the chemical natures of the adsorbates and the adsorption distances are markedly different in the two cases. The observation is explained as a consequence of Fermi-level pinning due to fractional charge transfer at the interface. Our results show that rationalizing the adsorption configurations on the basis of electronic interface properties alone can be ambiguous and that additional insight from dispersion-corrected DFT simulations is desirable. PMID:27066160

  1. Method for brazing together planar and nonplanar metal members

    DOEpatents

    Hammersand, Fred G.; Witkowski, Anthony J.

    1985-01-01

    The invention relates to a method and apparatus for brazing two metal members together, at least one of which is nonplanar, in a brazing furnace using a substantially pure brazing material. The method comprises the steps of utilizing a brazing fixture to hold the two metal members in tangential relation to one another along a portion of each member so that a cavity is formed adjacent to the contacting portions. A braze material is then positioned within the cavity. The braze fixture, the metal members, and the braze material are then placed in a brazing furnace. A heat shield is then placed over the braze fixture, the metal members, and the braze material to shield the braze material from direct furnace radiation. The furnace temperature is linearly increased at a rate of about 180.degree. C. per hour until a temperature of 350.degree. C. is achieved. Heat is transferred by conduction from the metal members to the braze material to cause the braze material to melt. Some material from the metal members slowly diffuses into the braze material forming a braze joint. The furnace is rapidly cooled to room temperature using nitrogen gas. The brazed assemblies made according to this method are superior to assemblies formed by heliarc welding.

  2. Assumptions and ambiguities in nonplanar acoustic soliton theory

    SciTech Connect

    Verheest, Frank; Hellberg, Manfred A.

    2014-02-15

    There have been many recent theoretical investigations of the nonlinear evolution of electrostatic modes with cylindrical or spherical symmetry. Through a reductive perturbation analysis based on a quasiplanar stretching, a modified form of the Korteweg-de Vries or related equation is derived, containing an additional term which is linear in the electrostatic potential and singular at time t = 0. Unfortunately, these analyses contain several restrictive assumptions and ambiguities which are normally neither properly explained nor discussed, and severely limit the applicability of the technique. Most glaring are the use of plane-wave stretchings, the assumption that shape-preserving cylindrical modes can exist and that, although time is homogeneous, the origin of time (which can be chosen arbitrarily) needs to be avoided. Hence, only in the domain where the nonlinear modes are quasiplanar, far from the axis of cylindrical or from the origin of spherical symmetry can acceptable but unexciting results be obtained. Nonplanar nonlinear modes are clearly an interesting topic of research, as some of these phenomena have been observed in experiments. However, it is argued that a proper study of such modes needs numerical simulations rather than ill-suited analytical approximations.

  3. Electronic circular dichroism of the chiral rigid tricyclic dilactam with nonplanar tertiary amide groups.

    PubMed

    Pazderková, Markéta; Profant, Václav; Seidlerová, Beata; Dlouhá, Helena; Hodačová, Jana; Jávorfi, Tamás; Siligardi, Giuliano; Baumruk, Vladimír; Bednárová, Lucie; Maloň, Petr

    2014-09-25

    Electronic circular dichroism (ECD) of the spirocyclic dilactam 5,8-diazatricyclo[6,3,0,0(1,5)]undecane-4,9-dione has been measured in the extended wavelength range (170-260 nm) utilizing far-UV CD instrumentation including synchrotron radiation light source. The data of this model of two nonplanar tertiary amide groups interacting within the rigid chiral environment provided new information particularly about the shorter wavelength π-π* transition region below 190 nm. The interpretation using TDDFT calculations confirmed that effects of amide nonplanarity follow our previous observations on monolactams as far as amide n-π* transitions are concerned. ECD band in the n-π* transition region of the nonplanar diamide exhibits an identical bathochromic shift and its sign remains tied to the sense of nonplanar deformation in the same way. As far as n-π* transitions are concerned amide nonplanarity acts as a local phenomenon independently reflecting sum properties of single amide groups. On the other hand, CD bands associated with π-π* transitions (found between ∼170 to 210 nm) form an exciton-like couplet with the sign pattern determined by mutual orientation of the associated electric transition moments. This sign pattern follows predictions pertaining to a coupled oscillator. The influence of amide nonplanarity on π-π* transitions is only minor and concentrates into the shorter wavelength lobe of the π-π* couplet. The detailed analysis of experimental ECD with the aid of TDDFT calculations shows that there is only little interaction between effects of inherent chirality caused by nonplanarity of amide groups and amide-amide coupling. Consequently these two effects can be studied nearly independently using ECD. In addition, the calculations indicate that participation of other type of transitions (n-σ*, π-σ* or Rydberg type transitions) is only minor and is concentrated below 180 nm.

  4. Nonplanar tertiary amides in rigid chiral tricyclic dilactams. Peptide group distortions and vibrational optical activity.

    PubMed

    Pazderková, Markéta; Profant, Václav; Hodačová, Jana; Sebestík, Jaroslav; Pazderka, Tomáš; Novotná, Pavlína; Urbanová, Marie; Safařík, Martin; Buděšínský, Miloš; Tichý, Miloš; Bednárová, Lucie; Baumruk, Vladimír; Maloň, Petr

    2013-08-22

    We investigate amide nonplanarity in vibrational optical activity (VOA) spectra of tricyclic spirodilactams 5,8-diazatricyclo[6,3,0,0(1,5)]undecan-4,9-dione (I) and its 6,6',7,7'-tetradeuterio derivative (II). These rigid molecules constrain amide groups to nonplanar geometries with twisted pyramidal arrangements of bonds to amide nitrogen atoms. We have collected a full range vibrational circular dichroism (VCD) and Raman optical activity (ROA) spectra including signals of C-H and C-D stretching vibrations. We report normal-mode analysis and a comparison of calculated to experimental VCD and ROA. The data provide band-to-band assignment and offer a possibility to evaluate roles of constrained nonplanar tertiary amide groups and rigid chiral skeletons. Nonplanarity shows as single-signed VCD and ROA amide I signals, prevailing the couplets expected to arise from the amide-amide interaction. Amide-amide coupling dominates amide II (mainly C'-N stretching, modified in tertiary amides by the absence of a N-H bond) transitions (strong couplet in VCD, no significant ROA) probably due to the close proximity of amide nitrogen atoms. At lower wavenumbers, ROA spectra exhibit another likely manifestation of amide nonplanarity, showing signals of amide V (δ(oop)(N-C) at ~570 cm(-1)) and amide VI (δ(oop)(C'═O) at ~700 cm(-1) and ~650 cm(-1)) vibrations.

  5. Realization of autofocusing system for laser direct writing on non-planar surfaces.

    PubMed

    Luo, Jianbo; Liang, Yiyong; Yang, Guoguang

    2012-05-01

    This paper presents an autofocusing system for laser direct writing on non-planar surfaces, including focus error signal detection and focusing control. The focus error signal detection is based on modified confocal techniques, which features easy implementation, independence of the tilt angles of non-planar surfaces, and excellent suppression of common-mode noise or variable system factors. We also present a macro/micro dual-drive mechanism and its synchronous operation for focusing control on non-planar surfaces. Finally, a performance evaluation of the autofocusing system is presented. The uniform line width of 2.1 μm for a pattern on a convex spherical substrate with a curvature radius of 100 mm shows the autofocusing system performs well.

  6. Fracturing from highly deviated and horizontal wells: Numerical analysis of non-planar fracture propagation

    SciTech Connect

    Olson, J.E.

    1995-12-31

    Fracturing from horizontal and highly deviated wells can often result in complex, non-planar fracture geometry. A two-dimensional model was developed to analyze the effects of non-planar fracture propagation for different in situ boundary conditions and hydraulic fracturing parameters. Numerical simulations show that curving fracture geometry reduces created fracture length compared to a planar fracture and causes a fracture width restriction at the wellbore. Reduction in fracture length can reduce expected well stimulation effects and jeopardize well economics. Near-wellbore width restrictions increase fracture treating pressure and may cause wellbore screen-out during the proppant stages of a fracturing treatment. The negative impact of non-planar geometry can be mitigated with short perforated intervals, high viscosity fracturing fluids, proper wellbore alignment and pre-pad proppant slugs for near-wellbore erosion.

  7. Nonplanar dust acoustic solitary waves in a strongly coupled dusty plasma with superthermal ions

    SciTech Connect

    El-Labany, S. K. Zedan, N. A.; El-Taibany, W. F. E-mail: eltaibany@du.edu.eg; El-Shamy, E. F.

    2014-12-15

    The nonplanar amplitude modulation of dust acoustic (DA) envelope solitary waves in a strongly coupled dusty plasma (SCDP) has been investigated. By using a reductive perturbation technique, a modified nonlinear Schrödinger equation (NLSE) including the effects of geometry, polarization, and ion superthermality is derived. The modulational instability (MI) of the nonlinear DA wave envelopes is investigated in both planar and nonplanar geometries. There are two stable regions for the DA wave propagation strongly affected by polarization and ion superthermality. Moreover, it is found that the nonlinear DA waves in spherical geometry are the more structurally stable. The larger growth rate of the nonlinear DA MI is observed in the cylindrical geometry. The salient characteristics of the MI in the nonplanar geometries cannot be found in the planar one. The DA wave propagation and the NLSE solutions are investigated both analytically and numerically.

  8. Evolution of apicomplexan secretory organelles

    PubMed Central

    Gubbels, Marc-Jan; Duraisingh, Manoj T.

    2013-01-01

    The alveolate superphylum includes many free-living and parasitic organisms, which are united by the presence of alveolar sacs lying proximal to the plasma membrane, providing cell structure. All species comprising the apicomplexan group of alveolates are parasites and have adapted to the unique requirements of the parasitic lifestyle. Here the evolution of apicomplexan secretory organelles that are involved in the critical process of egress from one cell and invasion of another is explored. The variations within the Apicomplexa and how these relate to species-specific biology will be discussed. In addition, recent studies have identified specific calcium-sensitive molecules that coordinate the various events and regulate the release of these secretory organelles within apicomplexan parasites. Some aspects of this machinery are conserved outside the Apicomplexa, and are beginning to elucidate the conserved nature of the machinery. Briefly, the relationship of this secretion machinery within the Apicomplexa will be discussed, compared with free-living and predatory alveolates, and how these might have evolved from a common ancestor. PMID:23068912

  9. Fabrication of small-scale structures with non-planar features

    DOEpatents

    Burckel, David B.; Ten Eyck, Gregory A.

    2015-11-19

    The fabrication of small-scale structures is disclosed. A unit-cell of a small-scale structure with non-planar features is fabricated by forming a membrane on a suitable material. A pattern is formed in the membrane and a portion of the substrate underneath the membrane is removed to form a cavity. Resonators are then directionally deposited on the wall or sides of the cavity. The cavity may be rotated during deposition to form closed-loop resonators. The resonators may be non-planar. The unit-cells can be formed in a layer that includes an array of unit-cells.

  10. Fabrication method for small-scale structures with non-planar features

    DOEpatents

    Burckel, David Bruce; Ten Eyck, Gregory A.

    2016-09-20

    The fabrication of small-scale structures is disclosed. A unit-cell of a small-scale structure with non-planar features is fabricated by forming a membrane on a suitable material. A pattern is formed in the membrane and a portion of the substrate underneath the membrane is removed to form a cavity. Resonators are then directionally deposited on the wall or sides of the cavity. The cavity may be rotated during deposition to form closed-loop resonators. The resonators may be non-planar. The unit-cells can be formed in a layer that includes an array of unit-cells.

  11. The interaction of two nonplanar solitary waves in electron-positron-ion plasmas: An application in active galactic nuclei

    SciTech Connect

    EL-Labany, S. K.; Khedr, D. M.; El-Shamy, E. F.; Sabry, R.

    2013-01-15

    In the present research paper, the effect of bounded nonplanar (cylindrical and spherical) geometry on the interaction between two nonplanar electrostatic solitary waves (NESWs) in electron-positron-ion plasmas has been studied. The extended Poincare-Lighthill-Kuo method is used to obtain nonplanar phase shifts after the interaction of the two NESWs. This study is a first attempt to investigate nonplanar phase shifts and trajectories for NESWs in a two-fluid plasma (a pair-plasma) consisting of electrons and positrons, as well as immobile background positive ions in nonplanar geometry. The change of phase shifts and trajectories for NESWs due to the effect of cylindrical geometry, spherical geometry, the physical processes (either isothermal or adiabatic), and the positions of two NESWs are discussed. The present investigation may be beneficial to understand the interaction between two NESWs that may occur in active galactic nuclei.

  12. Cloning and structural analysis of two highly divergent IgA isotypes, IgA1 and IgA2 from the duck billed platypus, Ornithorhynchus anatinus.

    PubMed

    Vernersson, M; Belov, K; Aveskogh, M; Hellman, L

    2010-01-01

    To trace the emergence of modern IgA isotypes during vertebrate evolution we have studied the immunoglobulin repertoire of a model monotreme, the platypus. Two highly divergent IgA-like isotypes (IgA1 and IgA2) were identified and their primary structures were determined from full-length cDNAs. A comparative analysis of the amino acid sequences for IgA from various animal species showed that the two platypus IgA isotypes form a branch clearly separated from their eutherian (placental) counterparts. However, they still conform to the general structure of eutherian IgA, with a hinge region and three constant domains. This indicates that the deletion of the second domain and the formation of a hinge region in IgA did occur very early during mammalian evolution, more than 166 million years ago. The two IgA isotypes in platypus differ in primary structure and appear to have arisen from a very early gene duplication, possibly preceding the metatherian eutherian split. Interestingly, one of these isotypes, IgA1, appears to be expressed in only the platypus, but is present in the echidna based on Southern blot analysis. The platypus may require a more effective mucosal immunity, with two highly divergent IgA forms, than the terrestrial echidna, due to its lifestyle, where it is exposed to pathogens both on land and in the water.

  13. Muscle as a secretory organ.

    PubMed

    Pedersen, Bente K

    2013-07-01

    Skeletal muscle is the largest organ in the body. Skeletal muscles are primarily characterized by their mechanical activity required for posture, movement, and breathing, which depends on muscle fiber contractions. However, skeletal muscle is not just a component in our locomotor system. Recent evidence has identified skeletal muscle as a secretory organ. We have suggested that cytokines and other peptides that are produced, expressed, and released by muscle fibers and exert either autocrine, paracrine, or endocrine effects should be classified as "myokines." The muscle secretome consists of several hundred secreted peptides. This finding provides a conceptual basis and a whole new paradigm for understanding how muscles communicate with other organs such as adipose tissue, liver, pancreas, bones, and brain. In addition, several myokines exert their effects within the muscle itself. Many proteins produced by skeletal muscle are dependent upon contraction. Therefore, it is likely that myokines may contribute in the mediation of the health benefits of exercise.

  14. An experimental study of the aerodynamic characteristics of planar and non-planar outboard wing planforms

    NASA Technical Reports Server (NTRS)

    Naik, D. A.; Ostowari, C.

    1987-01-01

    A series of wind tunnel experiments have been conducted to investigate the aerodynamic characteristics of several planar and nonplanar wingtip planforms. Seven different configurations: base-line rectangular, elliptical, swept and tapered, swept and tapered with dihedral, swept and tapered with anhedral, rising arc, and drooping arc, were investigated for two different spans. The data are available in terms of coefficient plots of force data, flow visualization photographs, and velocity and pressure flowfield surveys. All planforms, particularly the nonplanar, have some advantages over the baseline rectangular planform. Span efficiencies up to 20-percent greater than baseline are a possibility. However, it is suggested that the span efficiency concept might need refinement for nonplanar wings. Flow survey data show the change in effective span with vortex roll-up. The flow visualization shows the occurrence of mushroom-cell-separation flow patterns at angles of attack corresponding to stall. These grow with an increase in post-stall angle of attack. For the larger aspect ratios, the cells are observed to split into sub-cells at the higher angles of attack. For all angles of attack, some amount of secondary vortex flow is observed for the planar and nonplanar out-board planforms with sweep and taper.

  15. 2D and 3D Non-planar Dynamic Rupture by a Finite Volume Method

    NASA Astrophysics Data System (ADS)

    Benjemaa, M.; Glinsky-Olivier, N.; Cruz-Atienza, V. M.; Virieux, J.; Piperno, S.; Lanteri, S.

    2006-12-01

    Understanding the physics of the rupture process requires very sophisticated and accurate tools in which both the geometry of the fault surface and realistic frictional behaviours could interact during rupture propagation. New formulations have been recently proposed for modelling the dynamic shear rupture of non-planar faults (Ando et al., 2004; Cruz-Atienza &Virieux, 2004; Huang &Costanzo, 2004) providing highly accurate field estimates nearby the crack edges at the expanse of a simple medium description or high computational cost. We propose a new method based on the finite volume formulation to model the dynamic rupture propagation of non-planar faults. After proper transformations of the velocity-stress elastodynamic system of partial differential equations following an explicit conservative law, we construct an unstructured time-domain numerical formulation of the crack problem. As a result, arbitrary non-planar faults can be explicitly represented without extra computational cost. The analysis of the total discrete energy through the fault surface leads us to the specification of dynamic rupture boundary conditions which insure the correct discrete energy time variation and, therefore, the system stability. These boundary conditions are set on stress fluxes and not on stress values, which makes the fracture to have no thickness. Different shapes of cracks are analysed. We present an example of a bidimensional non-planar spontaneous fault growth in heterogeneous media as well as preliminary results of a highly efficient extension to the three dimensional rupture model based on the standard MPI.

  16. Non-planar Feynman diagrams and Mellin-Barnes representations with AMBRE 3.0

    NASA Astrophysics Data System (ADS)

    Dubovyk, Ievgen; Gluza, Janusz; Riemann, Tord

    2015-05-01

    We introduce the Mellin-Barnes representation of general Feynman integrals and discuss their evaluation. The Mathematica package AMBRE has been recently extended in order to cover consistently non-planar Feynman integrals with two loops. Prospects for the near future are outlined. This write-up is an introduction to new results which have also been presented elsewhere.

  17. Analysis and design of planar and non-planar wings for induced drag minimization

    NASA Technical Reports Server (NTRS)

    Mortara, K.; Straussfogel, Dennis M.; Maughmer, Mark D.

    1991-01-01

    The goal of the work was to develop and validate computational tools to be used for the design of planar and non-planar wing geometries for minimum induced drag. Because of the iterative nature of the design problem, it is important that, in addition to being sufficiently accurate for the problem at hand, they are reasonably fast and computationally efficient. Toward this end, a method of predicting induced drag in the presence of a non-rigid wake is coupled with a panel method. The induced drag prediction technique is based on the Kutta-Joukowski law applied at the trailing edge. Until recently, the use of this method has not been fully explored and pressure integration and Trefftz-plane calculations favored. As is shown in this report, however, the Kutta-Joukowski method is able to give better results for a given amount of effort than the more common techniques, particularly when relaxed wakes and non-planar wing geometries are considered. Using these tools, a workable design method is in place which takes into account relaxed wakes and non-planar wing geometries. It is recommended that this method be used to design a wind-tunnel experiment to verify the predicted aerodynamic benefits of non-planar wing geometries.

  18. Electroimmunodiffusion Studies of alpha Chain, Secretory Piece and Secretory IgA.

    DTIC Science & Technology

    1977-03-15

    riflIy b, bloeti nw~.b.r) V • V V VCommercially available antisera to secretory piece, colostrum , and a chain ‘ V were evaluated for use in...secretorj IgA Is laborious and expensive.. Problems in using currently available V commercial antisera to colostrum and free secretory piece arise because...V Coninercially available antisera to secretory piece, colostrum , V and ~t chain were evaluated for use in electroimmunodiffusj on of

  19. Secretory diarrhoea: mechanisms and emerging therapies.

    PubMed

    Thiagarajah, Jay R; Donowitz, Mark; Verkman, Alan S

    2015-08-01

    Diarrhoeal disease remains a major health burden worldwide. Secretory diarrhoeas are caused by certain bacterial and viral infections, inflammatory processes, drugs and genetic disorders. Fluid secretion across the intestinal epithelium in secretory diarrhoeas involves multiple ion and solute transporters, as well as activation of cyclic nucleotide and Ca(2+) signalling pathways. In many secretory diarrhoeas, activation of Cl(-) channels in the apical membrane of enterocytes, including the cystic fibrosis transmembrane conductance regulator and Ca(2+)-activated Cl(-) channels, increases fluid secretion, while inhibition of Na(+) transport reduces fluid absorption. Current treatment of diarrhoea includes replacement of fluid and electrolyte losses using oral rehydration solutions, and drugs targeting intestinal motility or fluid secretion. Therapeutics in the development pipeline target intestinal ion channels and transporters, regulatory proteins and cell surface receptors. This Review describes pathogenic mechanisms of secretory diarrhoea, current and emerging therapeutics, and the challenges in developing antidiarrhoeal therapeutics.

  20. Secretory diarrhoea: mechanisms and emerging therapies

    PubMed Central

    Thiagarajah, Jay R.; Donowitz, Mark; Verkman, Alan S.

    2016-01-01

    Diarrhoeal disease remains a major health burden worldwide. Secretory diarrhoeas are caused by certain bacterial and viral infections, inflammatory processes, drugs and genetic disorders. Fluid secretion across the intestinal epithelium in secretory diarrhoeas involves multiple ion and solute transporters, as well as activation of cyclic nucleotide and Ca2+ signalling pathways. In many secretory diarrhoeas, activation of Cl− channels in the apical membrane of enterocytes, including the cystic fibrosis transmembrane conductance regulator and Ca2+-activated Cl− channels, increases fluid secretion, while inhibition of Na+ transport reduces fluid absorption. Current treatment of diarrhoea includes replacement of fluid and electrolyte losses using oral rehydration solutions, and drugs targeting intestinal motility or fluid secretion. Therapeutics in the development pipeline target intestinal ion channels and transporters, regulatory proteins and cell surface receptors. This Review describes pathogenic mechanisms of secretory diarrhoea, current and emerging therapeutics, and the challenges in developing antidiarrhoeal therapeutics. PMID:26122478

  1. [Overweight and secretory male infertility].

    PubMed

    Oshakbaev, K P; Abylaĭuly, Zh; Dukenbaeva, B A

    2009-01-01

    We have performed a trial with participation of 60 males aged 23-52. Of them, 30 had secretory male iufertility (SMI) and obesity. The control 30 patients were healthy volunteers. The protocol was performed by two stages. Stage 1 included: investigation of a clinico-laboratory status, of correlation between a sorption function of erythrocytes, endogenic metabolic intoxication (EMI) and spermogram parameters, concentration of serum testosterone in SMI patients. Stage 2 consisted in treatment of the intoxication by reducing body mass. All the infertile men were obese; 30% of them had low glucose tolerance, 46.7% had stage 2 hypertension, 23.3%--seasonal allergic symptoms. The level of organic substances on the surface of erythrocytes in infertile men was higher than in the controls (p < 0.01). A negative correlation was seen between spermogram parameters and organic substances content on erythrocytic surface (p < 0/05), concentration of serum testosterone and the above substances (p < 0.01). The loss of fat tissue by 7-14 kg by infertile men resulted in a positive trend in spermogram parameters and the level of serum testosterone (p < 0.01).

  2. Histochemical Analysis of Plant Secretory Structures.

    PubMed

    Demarco, Diego

    2017-01-01

    Histochemical analysis is essential for the study of plant secretory structures whose classification is based, at least partially, on the composition of their secretion. As each gland may produce one or more types of substances, a correct analysis of its secretion should be done using various histochemical tests to detect metabolites of different chemical classes. Here I describe some of the most used methods to detect carbohydrates, proteins, lipids, phenolic compounds, and alkaloids in the secretory structures.

  3. Secretory granule biogenesis: rafting to the SNARE.

    PubMed

    Tooze, S A; Martens, G J; Huttner, W B

    2001-03-01

    Regulated secretion of hormones occurs when a cell receives an external stimulus, triggering the secretory granules to undergo fusion with the plasma membrane and release their content into the extracellular milieu. The formation of a mature secretory granule (MSG) involves a series of discrete and unique events such as protein sorting, formation of immature secretory granules (ISGs), prohormone processing and vesicle fusion. Regulated secretory proteins (RSPs), the proteins stored and secreted from MSGs, contain signals or domains to direct them into the regulated secretory pathway. Recent data on the role of specific domains in RSPs involved in sorting and aggregation suggest that the cell-type-specific composition of RSPs in the trans-Golgi network (TGN) has an important role in determining how the RSPs get into ISGs. The realization that lipid rafts are implicated in sorting RSPs in the TGN and the identification of SNARE molecules represent further major advances in our understanding of how MSGs are formed. At the heart of these findings is the elucidation of molecular mechanisms driving protein--lipid and protein--protein interactions specific for secretory granule biogenesis.

  4. Non-planar vibrations of a string in the presence of a boundary obstacle

    NASA Astrophysics Data System (ADS)

    Singh, Harkirat; Wahi, Pankaj

    2017-02-01

    We analyze planar and non-planar motions of a string vibrating against a unilateral curved obstacle. Our model incorporates the change in tension due to stretching of the string, which introduces nonlinear coupling between motions in mutually perpendicular directions, as well as the wrapping nonlinearity due to the presence of the obstacle. The system of equations has been discretized by assuming functional form of the displacements which satisfies all the geometrical boundary conditions. This discretized system is then used to investigate the various motions possible both in the absence as well as the presence of the obstacle. In the absence of the obstacle, there are infinitely many planar and two non-planar motions viz. a circular trajectory and a precessing elliptical trajectory for a fixed magnitude of the disturbance. In contrast, the string has only one planar motion when the obstacle is present and two non-planar motions, either an oscillating orbit or a whirling orbit depending on the magnitude of the initial disturbance. To obtain the transition from oscillating to whirling orbits, we perform a stability analysis of the planar motion using Floquet theory. This analysis reveals that there exists a critical amplitude below which the planar motion is neutrally stable and the typical trajectories are ellipses with major and minor radii changing both in magnitude and direction. Beyond the critical amplitude, the planar motion is unstable and we get whirling trajectories which are precessing ellipses again with varying major and minor radii. We further study the effect of changing obstacle parameters on the critical amplitude, and obtain the stability boundaries in the space spanned by the obstacle parameters and the amplitude of the planar vibration. We obtain some interesting values of the obstacle parameters for which small and large amplitude planar motions are stable resulting in oscillating ellipses while motions with intermediate amplitudes are unstable

  5. Porosome: The Universal Secretory Portal in Cells

    NASA Astrophysics Data System (ADS)

    Jena, Bhanu

    2012-10-01

    In the past 50 years it was believed that during cell secretion, membrane-bound secretory vesicles completely merge at the cell plasma membrane resulting in the diffusion of intra-vesicular contents to the cell exterior and the compensatory retrieval of the excess membrane by endocytosis. This explanation made no sense or logic, since following cell secretion partially empty vesicles accumulate as demonstrated in electron micrographs. Furthermore, with the ``all or none'' mechanism of cell secretion by complete merger of secretory vesicle membrane at the cell plasma membrane, the cell is left with little regulation and control of the amount of content release. Moreover, it makes no sense for mammalian cells to possess such `all or none' mechanism of cell secretion, when even single-cell organisms have developed specialized and sophisticated secretory machinery, such as the secretion apparatus of Toxoplasma gondii, the contractile vacuoles in paramecium, or the various types of secretory structures in bacteria. Therefore, in 1993 in a News and Views article in Nature, E. Neher wrote ``It seems terribly wasteful that, during the release of hormones and neurotransmitters from a cell, the membrane of a vesicle should merge with the plasma membrane to be retrieved for recycling only seconds or minutes later.'' This conundrum in the molecular mechanism of cell secretion was finally resolved in 1997 following discovery of the ``Porosome,'' the universal secretory machinery in cells. Porosomes are supramolecular lipoprotein structures at the cell plasma membrane, where membrane-bound secretory vesicles transiently dock and fuse to release inravesicular contents to the outside during cell secretion. In the past decade, the composition of the porosome, its structure and dynamics at nm resolution and in real time, and its functional reconstitution into artificial lipid membrane, have all been elucidated. Since porosomes in exocrine and neuroendocrine cells measure 100-180 nm

  6. Inversion and rotation processes involving non-planar aromatic compounds catalyzed by extended polycyclic aromatic hydrocarbons

    NASA Astrophysics Data System (ADS)

    Karton, Amir

    2014-10-01

    Using accurate quantum chemical calculations, we show that extended planar polycyclic aromatic hydrocarbons (PAHs) can efficiently catalyze a range of chemical processes involving non-planar aromatic systems. These include (i) bowl-to-bowl inversion of curved PAHs (e.g. corannulene and sumanene), (ii) 'flip-flop' inversion of helicenes (e.g. benzo[c]phenanthrene), and (iii) rotation about the Phsbnd Ph bond in biphenyls. Non-covalent π-π interactions between the planar catalyst and the substrate stabilize the planar transition structures to a greater extent than they stabilize the non-planar reactants. These result in surprisingly large catalytic enhancements (namely, the reaction barrier heights are reduced by 21-63% of the uncatalyzed reaction barriers).

  7. Demonstration of the stabilization technique for nonplanar optical resonant cavities utilizing polarization

    SciTech Connect

    Akagi, T.; Araki, S.; Funahashi, Y.; Honda, Y.; Okugi, T.; Omori, T.; Shimizu, H.; Terunuma, N.; Urakawa, J.; Miyoshi, S.; Takahashi, T. Tanaka, R.; Uesugi, Y.; Yoshitama, H.; Sakaue, K.; Washio, M.

    2015-04-15

    Based on our previously developed scheme to stabilize nonplanar optical resonant cavities utilizing polarization caused by a geometric phase in electromagnetic waves traveling along a twisted path, we report an application of the technique for a cavity installed in the Accelerator Test Facility, a 1.3-GeV electron beam accelerator at KEK, in which photons are generated by laser-Compton scattering. We successfully achieved a power enhancement of 1200 with 1.4% fluctuation, which means that the optical path length of the cavity has been controlled with a precision of 14 pm under an accelerator environment. In addition, polarization switching utilizing a geometric phase of the nonplanar cavity was demonstrated.

  8. Analysis of Nonplanar Wing-tip-mounted Lifting Surfaces on Low-speed Airplanes

    NASA Technical Reports Server (NTRS)

    Vandam, C. P.; Roskam, J.

    1983-01-01

    Nonplanar wing tip mounted lifting surfaces reduce lift induced drag substantially. Winglets, which are small, nearly vertical, winglike surfaces, are an example of these devices. To achieve reduction in lift induced drag, winglets produce significant side forces. Consequently, these surfaces can seriously affect airplane lateral directional aerodynamic characteristics. Therefore, the effects of nonplanar wing tip mounted surfaces on the lateral directional stability and control of low speed general aviation airplanes were studied. The study consists of a theoretical and an experimental, in flight investigation. The experimental investigation involves flight tests of winglets on an agricultural airplane. Results of these tests demonstrate the significant influence of winglets on airplane lateral directional aerodynamic characteristics. It is shown that good correlations exist between experimental data and theoretically predicted results. In addition, a lifting surface method was used to perform a parametric study of the effects of various winglet parameters on lateral directional stability derivatives of general aviation type wings.

  9. Uniform aluminum deposits on large nonplanar and planar polyimide substrates by physical vapor deposition

    SciTech Connect

    Hale, G.J.

    1980-07-01

    The capability to deposit 10- to 12-..mu..m-thick aluminum films with less than 2 percent thickness uniformity (defined as standard deviation/average thickness) on large nonplanar and planar polyimide substrates has been developed using physical vapor deposition (PVD). A technique called shadow masking was developed to provide this consistent deposit uniformity over the entire substrate surfaces (greater than 2000 cm/sup 2/). The specific dimensions of a shadow mask for each substrate are generated from a computer-reduced plot of beta-backscatter thickness measurements of a previous deposit. With this developed method, initial thickness uniformities of 9.7 and 24.7 percent obtained for the nonplanar and planar deposits were improved to 1.07 and 1.80 percent, respectively.

  10. Planar and nonplanar ion acoustic shock waves in relativistic degenerate astrophysical electron-positron-ion plasmas

    SciTech Connect

    Ata-ur-Rahman,; Qamar, A.; Ali, S.; Mirza, Arshad M.

    2013-04-15

    We have studied the propagation of ion acoustic shock waves involving planar and non-planar geometries in an unmagnetized plasma, whose constituents are non-degenerate ultra-cold ions, relativistically degenerate electrons, and positrons. By using the reductive perturbation technique, Korteweg-deVries Burger and modified Korteweg-deVries Burger equations are derived. It is shown that only compressive shock waves can propagate in such a plasma system. The effects of geometry, the ion kinematic viscosity, and the positron concentration are examined on the ion acoustic shock potential and electric field profiles. It is found that the properties of ion acoustic shock waves in a non-planar geometry significantly differ from those in planar geometry. The present study has relevance to the dense plasmas, produced in laboratory (e.g., super-intense laser-dense matter experiments) and in dense astrophysical objects.

  11. A Vector Study of Linearized Supersonic Flow Applications to Nonplanar Problems

    NASA Technical Reports Server (NTRS)

    Martin, John C

    1953-01-01

    A vector study of the partial-differential equation of steady linearized supersonic flow is presented. General expressions which relate the velocity potential in the stream to the conditions on the disturbing surfaces, are derived. In connection with these general expressions the concept of the finite part of an integral is discussed. A discussion of problems dealing with planar bodies is given and the conditions for the solution to be unique are investigated. Problems concerning nonplanar systems are investigated, and methods are derived for the solution of some simple nonplanar bodies. The surface pressure distribution and the damping in roll are found for rolling tails consisting of four, six, and eight rectangular fins for the Mach number range where the region of interference between adjacent fins does not affect the fin tips.

  12. Manifestations of non-planar adsorption geometries of lead pyrenocyanine at the liquid-solid interface.

    PubMed

    Mali, Kunal S; Zöphel, Lukas; Ivasenko, Oleksandr; Müllen, Klaus; De Feyter, Steven

    2013-10-01

    In this work, we provide evidence for multiple non-planar adsorption geometries of a novel pyrenocyanine derivative at the liquid-solid interface under ambient conditions. When adsorbed at the organic liquid-solid interface, lead pyrenocyanine forms well-ordered monolayers that exhibit peculiar non-periodic contrast variation. The different contrast of the adsorbed molecules is attributed to dissimilar adsorption geometries which arise from the non-planar conformation of the molecules. The non-planarity of the molecular backbone in turn arises due to a combination of the angularly extended pyrene subunits and the presence of the large lead ion, which is too big to fit inside the central cavity and thus is located out of the aromatic plane. The two possible locations of the lead atom, namely below and above the aromatic plane, could be identified as depression and protrusion in the central cavity, respectively. The manifestation of such multiple adsorption geometries on the structure of the resultant monolayer is discussed in detail. The packing density of these 2D arrays of molecules could be tuned by heating of the sample wherein the molecular packing changes from a low-density, pseudo six-fold symmetric to a high-density, two-fold symmetric arrangement. Finally, a well-ordered two-component system could be constructed by incorporating C60 molecules in the adlayer of lead pyrenocyanine at the liquid-solid interface.

  13. Analysis and design of planar and non-planar wings for induced drag minimization

    NASA Technical Reports Server (NTRS)

    Mortara, Karl W.; Straussfogel, Dennis M.; Maughmer, Mark D.

    1992-01-01

    The goal of the work reported herein is to develop and validate computational tools to be used for the design of planar and non-planar wing geometries for minimum induced drag. Because of the iterative nature of the design problem, it is important that, in addition to being sufficiently accurate for the problem at hand, these tools need to be reasonably fast and computationally efficient. Toward this end, a method of predicting induced drag in the presence of a free wake has been coupled with a panel method. The induced drag prediction technique is based on the application of the Kutta-Joukowski law at the trailing edge. Until now, the use of this method has not been fully explored and pressure integration and Trefftz-plane calculations favored. As is shown in this report, however, the Kutta-Joukowski method is able to give better results for a given amount of effort than the more commonly used techniques, particularly when relaxed wakes and non-planar wing geometries are considered. Using these methods, it is demonstrated that a reduction in induced drag can be achieved through non-planar wing geometries. It remains to determine what overall drag reductions are possible when the induced drag reduction is traded-off against increased wetted area. With the design methodology that is described herein, such trade studies can be performed in which the non-linear effects of the free wake are taken into account.

  14. Toxoplasma secretory granules: one population or more?

    PubMed

    Mercier, Corinne; Cesbron-Delauw, Marie-France

    2015-02-01

    In Toxoplasma gondii, dense granules are known as the storage secretory organelles of the so-called GRA proteins (for dense granule proteins), which are destined to the parasitophorous vacuole (PV) and the PV-derived cyst wall. Recently, newly annotated GRA proteins targeted to the host cell nucleus have enlarged this view. Here we provide an update on the latest developments on the Toxoplasma secreted proteins, which to date have been mainly studied at both the tachyzoite and bradyzoite stages, and we point out that recent discoveries could open the issue of a possible, yet uncharacterized, distinct secretory pathway in Toxoplasma.

  15. Quality control in the secretory assembly line.

    PubMed Central

    Helenius, A

    2001-01-01

    As a rule, only proteins that have reached a native, folded and assembled structure are transported to their target organelles and compartments within the cell. In the secretory pathway of eukaryotic cells, this type of sorting is particularly important. A variety of molecular mechanisms are involved that distinguish between folded and unfolded proteins, modulate their intracellular transport, and induce degradation if they fail to fold. This phenomenon, called quality control, occurs at several levels and involves different types of folding sensors. The quality control system provides a stringent and versatile molecular sorting system that guaranties fidelity of protein expression in the secretory pathway. PMID:11260794

  16. [Secretory immunoglobulin A in amniotic fluid].

    PubMed

    Briese, V; Straube, W; Brock, J; Lorenz, U

    1983-01-01

    Secretory immunoglobulin A (S-IgA) was estimated in amniotic fluid samples by means of the single radial immunodiffusion according to Mancini. A monospecific antiserum against human secretory component was used. 163 amniotic fluid samples from normal pregnancies and risk pregnancies respectively were investigated. Within the 3rd trimenon the S-IgA content in amniotic fluid increased significantly. With respect to literature and examinations performed previously a connection between S-IgA content in amniotic fluid and fetal lung maturity seems to be possible.

  17. Synaptic Control of Secretory Trafficking in Dendrites

    PubMed Central

    Hanus, Cyril; Kochen, Lisa; Dieck, Susanne tom; Racine, Victor; Sibarita, Jean-Baptiste; Schuman, Erin M.; Ehlers, Michael D.

    2016-01-01

    Summary Localized signaling in neuronal dendrites requires tight spatial control of membrane composition. Upon initial synthesis, nascent secretory cargo in dendrites exits the endoplasmic reticulum (ER) from local zones of ER complexity that are spatially coupled to post-ER compartments. Although newly synthesized membrane proteins can be processed locally, the mechanisms that control the spatial range of secretory cargo transport in dendritic segments are unknown. Here, we monitored the dynamics of nascent membrane proteins in dendritic post-ER compartments under regimes of low or increased neuronal activity. In response to activity blockade, post-ER carriers are highly mobile and are transported over long distances. Conversely, increasing synaptic activity dramatically restricts the spatial scale of post-ER trafficking along dendrites. This activity-induced confinement of secretory cargo requires site-specific phosphorylation of the kinesin motor KIF17 by Ca2+/calmodulin-dependent protein kinases (CaMK). Thus, the length scales of early secretory trafficking in dendrites are tuned by activity-dependent regulation of microtubule-dependent transport. PMID:24931613

  18. Ursodeoxycholic acid attenuates colonic epithelial secretory function

    PubMed Central

    Kelly, Orlaith B; Mroz, Magdalena S; Ward, Joseph B J; Colliva, Carolina; Scharl, Michael; Pellicciari, Roberto; Gilmer, John F; Fallon, Padraic G; Hofmann, Alan F; Roda, Aldo; Murray, Frank E; Keely, Stephen J

    2013-01-01

    Dihydroxy bile acids, such as chenodeoxycholic acid (CDCA), are well known to promote colonic fluid and electrolyte secretion, thereby causing diarrhoea associated with bile acid malabsorption. However, CDCA is rapidly metabolised by colonic bacteria to ursodeoxycholic acid (UDCA), the effects of which on epithelial transport are poorly characterised. Here, we investigated the role of UDCA in the regulation of colonic epithelial secretion. Cl− secretion was measured across voltage-clamped monolayers of T84 cells and muscle-stripped sections of mouse or human colon. Cell surface biotinylation was used to assess abundance/surface expression of transport proteins. Acute (15 min) treatment of T84 cells with bilateral UDCA attenuated Cl− secretory responses to the Ca2+ and cAMP-dependent secretagogues carbachol (CCh) and forskolin (FSK) to 14.0 ± 3.8 and 40.2 ± 7.4% of controls, respectively (n= 18, P < 0.001). Investigation of the molecular targets involved revealed that UDCA acts by inhibiting Na+/K+-ATPase activity and basolateral K+ channel currents, without altering their cell surface expression. In contrast, intraperitoneal administration of UDCA (25 mg kg−1) to mice enhanced agonist-induced colonic secretory responses, an effect we hypothesised to be due to bacterial metabolism of UDCA to lithocholic acid (LCA). Accordingly, LCA (50–200 μm) enhanced agonist-induced secretory responses in vitro and a metabolically stable UDCA analogue, 6α-methyl-UDCA, exerted anti-secretory actions in vitro and in vivo. In conclusion, UDCA exerts direct anti-secretory actions on colonic epithelial cells and metabolically stable derivatives of the bile acid may offer a new approach for treating intestinal diseases associated with diarrhoea. PMID:23507881

  19. Polarizability matrix retrieval of a non-planar chiral particle through scattering parameters

    NASA Astrophysics Data System (ADS)

    Karamanos, Theodosios; Kantartzis, Nikolaos

    2016-04-01

    An efficient technique for the polarizability matrix extraction of non-planar chiral particles is introduced in this paper. Assuming electrically small sizes, the particles are modeled via electric and magnetic dipoles, whose induced moments are derived from the surface S-parameter response of three normal wave incidences. Then, these moments are inserted in the initial convention and polarizabilities are acquired through a linear system formed by the measured or simulated S-parameters. To validate the novel method, the retrieved polarizabilities are involved in RCS calculations for diverse incidences and compared with simulated ones.

  20. A Compact Non-Planar Coil Design for the SFLM Hybrid

    NASA Astrophysics Data System (ADS)

    Hagnestål, A.; Ågren, O.; Moiseenko, V. E.

    2012-08-01

    A non-planar single layer semiconductor coil set for a version of the Straight Field Line Mirror Hybrid concept with reduced magnetic field has been computed. The coil set consists of 30 coils that are somewhat similar to baseball coils with skewed sides. The coil set has been modeled with filamentary current distributions and basic scaling assumptions have been made regarding the coil widths. This coil set is expected to be considerably cheaper than a previous computed coil set. The coils can probably be produced with technologies known today.

  1. The interaction between two planar and nonplanar quantum electron acoustic solitary waves in dense electron-ion plasmas

    SciTech Connect

    EL-Labany, S. K.; El-Mahgoub, M. G.; EL-Shamy, E. F.

    2012-06-15

    The interaction between two planar and nonplanar (cylindrical and spherical) quantum electron acoustic solitary waves (QEASWs) in quantum dense electron-ion plasmas has been studied. The extended Poincare-Lighthill-Kuo method is used to obtain planar and nonplanar phase shifts after the interaction of the two QEASWs. The change of phase shifts and trajectories for QEASWs due to the effect of the different geometries, the quantum corrections of diffraction, and the cold electron-to-hot electron number density ratio are discussed. It is shown that the interaction of the QEASWs in planar geometry, cylindrical geometry, and spherical geometry are different. The present investigation may be beneficial to understand the interaction between two planar and nonplanar QEASWs that may occur in the quantum plasmas found in laser-produced plasmas as well as in astrophysical plasmas.

  2. Three-Dimensional Microstructural Characterization of GaN Nonplanar Substrate Laterally Epitaxially Overgrown by Metalorganic Chemical Vapor Deposition

    DTIC Science & Technology

    2005-07-20

    at 120 kV or an Akashi 002B operating at 200 kV. Some of the images are collaged from several photographs so as to gain better knowledge of the defect...views of the microstructures inside the GaN nonplanar substrate, with different TEM view directions indicated in Fig. 1(a). Fig. 1(b) is a collaged ...TEM photo of its typical ð1 1̄ 0 0Þ transversal cross-section under the ½1 1̄ 0 0 zone axis. The LEO nonplanar GaN substrate has a trapezoidal cross

  3. Effect of an applied electric field on a weakly anchored non-planar Nematic Liquid Crystal (NLC) layer

    NASA Astrophysics Data System (ADS)

    Mema, Ensela; Cummings, Linda J.; Kondic, Lou

    We consider a mathematical model that consists of a NLC layer sandwiched between two parallel bounding plates, across which an external field is applied. We investigate its effect on the director orientation by considering the dielectric and flexoelectric contributions and varying parameters that represent the anchoring conditions and the electric field strength. In particular, we investigate possible director configurations that occur in weakly anchored and non-planar systems. We observe that non-planar anchoring angles destroy any hysteresis seen in a planar system by eliminating the fully vertical director configuration and the ''saturation threshold'' seen in weakly anchored planar Freedericksz cells. Supported by NSF Grant No. DMS-1211713.

  4. Conformal growth of anodic nanotubes for dye-sensitized solar cells: part II. Nonplanar electrode.

    PubMed

    Sun, Lidong; Zhang, Sam; Wang, Qing

    2014-02-01

    Anodic titania nanotube array features highly ordered alignment as well as porous nature, and exhibits intriguing properties when employed in a variety of applications. All these profit from the continuous efforts on controlling the nanotube configurations. Recently, nonplanar electrodes have also been used to grow the nanotubes besides the conventional planar counterparts. As such, it is of great interest and significance to complete a picture to link the nanotubes grown on planar and various nonplanar electrodes for a comprehensive understanding of nanotube growing manners, in an attempt to boost their future applications. In the first part of this review, planar electrodes are focused with regard to nanotube growth and application in dye-sensitized solar cells. In this part, the nanotubes grown on patterned or curved surfaces are discussed first with reference to a similar structure of alumina nanopores, which are subsequently used to mirror the growth of nanotubes on cylindrical electrodes (i.e., titanium wires or meshes). The last section focuses on titanium tubular electrodes which are attractive for thermal fluids in view of the drastically reduced thermal conductivity in the presence of anodic nanotubes. As a recent hot topic, wire-shaped dye-sensitized solar cells are deliberated in terms of cell structure, efficiency calculation, merits, challenges and outlook.

  5. π-Conjugated Discrete Oligomers Containing Planar and Nonplanar Aromatic Motifs.

    PubMed

    Li, Ji; Terec, Anamaria; Wang, Yue; Joshi, Hrishikesh; Lu, Yunpeng; Sun, Handong; Stuparu, Mihaiela C

    2017-03-01

    A new family of π-conjugated oligomers featuring a nonplanar polycyclic aromatic hydrocarbon, corannulene, and a planar aromatic unit, thiophene, is synthesized through an iterative metal-catalyzed coupling protocol. The two structural motifs are connected through an acetylene linkage. In the shorter oligomers, a thiophene unit is attached to one or two corannulenes. In the higher analogues, two, three, and four thiophene units are placed in an alternating fashion with three, four, and five corannulene units, respectively. Photophysical studies reveal extended π-effects that initially increase and then attenuate as a function of the oligomer length. Notably, longer oligomers are found to be highly active for nonlinear absorption and emission properties. The oligomer with three corannulene and two thiophene units exhibits a two-photon absorption cross section of 600 GM and two-photon-excited intense green luminescence. This work, therefore, introduces the concept of combining planar and nonplanar aromatic motifs in the design of π-conjugated discrete oligomers, establishes synthetic feasibility of such hybrid materials, reports on their photophysical properties that is anticipated to have significant implications for future research targets, and features the discovery that corannulene derivatives can exhibit excellent nonlinear optical activity when extended through π-bridges.

  6. Rupture Dynamics Simulation for Non-Planar fault by a Curved Grid Finite Difference Method

    NASA Astrophysics Data System (ADS)

    Zhang, Z.; Zhu, G.; Chen, X.

    2011-12-01

    We first implement the non-staggered finite difference method to solve the dynamic rupture problem, with split-node, for non-planar fault. Split-node method for dynamic simulation has been used widely, because of that it's more precise to represent the fault plane than other methods, for example, thick fault, stress glut and so on. The finite difference method is also a popular numeric method to solve kinematic and dynamic problem in seismology. However, previous works focus most of theirs eyes on the staggered-grid method, because of its simplicity and computational efficiency. However this method has its own disadvantage comparing to non-staggered finite difference method at some fact for example describing the boundary condition, especially the irregular boundary, or non-planar fault. Zhang and Chen (2006) proposed the MacCormack high order non-staggered finite difference method based on curved grids to precisely solve irregular boundary problem. Based upon on this non-staggered grid method, we make success of simulating the spontaneous rupture problem. The fault plane is a kind of boundary condition, which could be irregular of course. So it's convinced that we could simulate rupture process in the case of any kind of bending fault plane. We will prove this method is valid in the case of Cartesian coordinate first. In the case of bending fault, the curvilinear grids will be used.

  7. Stringent limitations on reductive perturbation studies of nonplanar acoustic solitons in plasmas

    NASA Astrophysics Data System (ADS)

    Verheest, Frank; Hellberg, Manfred A.

    2016-06-01

    More than fifty years ago, the Korteweg-de Vries equation was shown to describe not only solitary surface waves on shallow water, but also nonlinear ion-acoustic waves. Because of the algorithmic ease of using reductive perturbation theory, intensive research followed on a wide range of wave types. Soon, the formalism was extended to nonplanar modes by introducing a stretching designed to accommodate spherically and cylindrically symmetric ion-acoustic waves. Over the last two decades many authors followed this approach, but almost all have ignored the severe restrictions in parameter space imposed by the Ansatz. In addition, for other steps in the formalism, the justification is often not spelled out, leading to effects that are physically undesirable or ambiguous. Hence, there is a need to critically assess this approach to nonplanar modes and to use it with the utmost care, respecting the restrictions on its validity. Only inward propagation may be meaningfully studied and respect for weak nonlinearities of at most 1/10 implies that one cannot get closer to the axis or centre of symmetry than about 30 Debye lengths. Thus, one is in a regime where the modes are quasi-planar and not particularly interesting. Most papers disregard these constraints and hence reach questionable conclusions.

  8. Elementary framework for cold field emission from quantum-confined, non-planar emitters

    SciTech Connect

    Patterson, A. A. Akinwande, A. I.

    2015-05-07

    For suitably small field emitters, the effects of quantum confinement at the emitter tip may have a significant impact on the emitter performance and total emitted current density (ECD). Since the geometry of a quantum system uniquely determines the magnitude and distribution of its energy levels, a framework for deriving ECD equations from cold field electron emitters of arbitrary geometry and dimensionality is developed. In the interest of obtaining semi-analytical ECD equations, the framework is recast in terms of plane wave solutions to the Schrödinger equation via the use of the Jeffreys-Wentzel-Kramers-Brillouin approximation. To demonstrate the framework's consistency with our previous work and its capabilities in treating emitters with non-planar geometries, ECD equations were derived for the normally unconfined cylindrical nanowire (CNW) and normally confined (NC) CNW emitter geometries. As a function of the emitter radius, the NC CNW emitter ECD profile displayed a strong dependence on the Fermi energy and had an average ECD that exceeded the Fowler-Nordheim equation for typical values of the Fermi energy due to closely spaced, singly degenerate energy levels (excluding electron spin), comparatively large electron supply values, and the lack of a transverse, zero-point energy. Such characteristics suggest that emitters with non-planar geometries may be ideal for emission from both an electron supply and electrostatics perspective.

  9. Nonplanar dust-acoustic Gardner solitons in a four-component dusty plasma

    NASA Astrophysics Data System (ADS)

    Mannan, A.; Mamun, A. A.

    2011-08-01

    The nonlinear propagation of Gardner solitons (GSs) in a nonplanar (cylindrical and spherical) four-component dusty plasma (composed of inertial positively and negatively dust, Boltzmann electrons, and ions) is studied by the reductive perturbation method. The modified Gardner equation is derived and numerically solved. It has been found that the basic characteristics of the dust-acoustic (DA) GSs, which are shown to exist for μ around its critical value μc [where μ=Zdpmdn/Zdnmdp, Zdn (Zdp) is the number of electrons (protons) residing on a negative (positive) dust, mdp (mdn) is the mass of the positive (negative) dust, μc is the value of μ corresponding to the vanishing of the nonlinear coefficient of the Korteweg-de Vries (KdV) equation, e.g., μc≃0.174 for μe=ne0/Zdnndn0=0.2, μi=ni0/Zdnndn0=0.4, and σ=Ti/Te=0.1, ne0, ni0, and ndn0 are, respectively, electron, ion, and dust number densities, and Ti (Te) is the ion (electron) temperature], are different from those of the KdV solitons, which do not exist for μ around μc. It has been also found that the propagation characteristics of nonplanar DA GSs significantly differ from those of planar ones.

  10. Docking of Secretory Vesicles Is Syntaxin Dependent

    PubMed Central

    de Wit, Heidi; Cornelisse, L. Niels; Toonen, Ruud F.G.; Verhage, Matthijs

    2006-01-01

    Secretory vesicles dock at the plasma membrane before they undergo fusion. Molecular docking mechanisms are poorly defined but believed to be independent of SNARE proteins. Here, we challenged this hypothesis by acute deletion of the target SNARE, syntaxin, in vertebrate neurons and neuroendocrine cells. Deletion resulted in fusion arrest in both systems. No docking defects were observed in synapses, in line with previous observations. However, a drastic reduction in morphologically docked secretory vesicles was observed in chromaffin cells. Syntaxin-deficient chromaffin cells showed a small reduction in total and plasma membrane staining for the docking factor Munc18-1, which appears insufficient to explain the drastic reduction in docking. The sub-membrane cortical actin network was unaffected by syntaxin deletion. These observations expose a docking role for syntaxin in the neuroendocrine system. Additional layers of regulation may have evolved to make syntaxin redundant for docking in highly specialized systems like synaptic active zones. PMID:17205130

  11. Parotid salivary secretory pattern in bulimia nervosa.

    PubMed

    Riad, M; Barton, J R; Wilson, J A; Freeman, C P; Maran, A G

    1991-01-01

    Parotid gland enlargement occurs in about 25% of patients with the binge eating syndrome of bulimia nervosa. The parotid salivary secretory patterns in 28 bulimics were determined in order to investigate the functional abnormality in the glands. Bulimia patients had a reduced resting flow rate. Bulimics who developed sialadenosis (4 patients) had reduced resting and stimulated flow rates. The salivary amylase activity was increased in both the resting and stimulated states in bulimics and the sialadenosis group. The resting total protein levels were greater in the bulimics. The electrolyte and immunoglobulin levels were within normal limits. The possibility of protein and enzymatic secretory disturbances due to autonomic nerve disorders as an explanation for the development of sialadenosis in bulimia nervosa is discussed.

  12. RFP tags for labeling secretory pathway proteins

    SciTech Connect

    Han, Liyang; Zhao, Yanhua; Xu, Pingyong; Huan, Shuangyan

    2014-05-09

    Highlights: • Membrane protein Orai1 can be used to report the fusion properties of RFPs. • Artificial puncta are affected by dissociation constant as well as pKa of RFPs. • Among tested RFPs mOrange2 is the best choice for secretory protein labeling. - Abstract: Red fluorescent proteins (RFPs) are useful tools for live cell and multi-color imaging in biological studies. However, when labeling proteins in secretory pathway, many RFPs are prone to form artificial puncta, which may severely impede their further uses. Here we report a fast and easy method to evaluate RFPs fusion properties by attaching RFPs to an environment sensitive membrane protein Orai1. In addition, we revealed that intracellular artificial puncta are actually colocalized with lysosome, thus besides monomeric properties, pKa value of RFPs is also a key factor for forming intracellular artificial puncta. In summary, our current study provides a useful guide for choosing appropriate RFP for labeling secretory membrane proteins. Among RFPs tested, mOrange2 is highly recommended based on excellent monomeric property, appropriate pKa and high brightness.

  13. Nonplanar fiber-optic sensing head for the compensation of bending-induced birefringence in Faraday current sensors.

    PubMed

    Perciante, César D; Aparicio, Sofía; Illa, Ricardo; Ferrari, José A

    2015-06-20

    We demonstrate the compensation of bending-induced linear birefringence in single-mode fibers coiled in a nonplanar path by alternating orthogonal bending planes. This effect can be applied for the construction of birefringence-free fiber coils in Faraday sensor heads (e.g., in current sensors) to improve their sensitivity. Validation experiments are presented.

  14. Beam position controlling method for 3D optical system and its application in non-planar ring resonators.

    PubMed

    Yuan, Jie; Chen, Meixiong; Long, Xingwu; Tan, Yanyang; Kang, Zhenglong; Li, Yingying

    2012-08-13

    A novel theoretical beam position controlling method for 3D optical system has been proposed in this paper. Non-planar ring resonator, which is a typical 3D optical system, has been chosen as an example to show its application. To the best of our knowledge, the generalized ray matrices, augmented 5 × 5 ray matrices for paraxial dielectric interface transmission and paraxial optical-wedge transmission, and their detailed deducing process have been proposed in this paper for the first time. By utilizing the novel coordinate system for Gaussian beam reflection and the generalized ray matrix of paraxial optical-wedge transmission, the rules and some novel results of the optical-axis perturbations of non-planar ring resonators have been obtained. Wedge angle-induced mismatching errors of non-planar ring resonators have been found out and two experimental beam position controlling methods to effectively eliminate the wedge angle-induced mismatching errors have been proposed. All those results have been confirmed by related alignment experiments and the experimental results have been described with diagrammatic representation. These findings are important to the beam control, cavity design, and cavity alignment of high precision non-planar ring laser gyroscopes. Those generalized ray matrices and their deducing methods are valuable for ray analysis of various kinds of paraxial optical-elements and resonators. This novel theoretical beam position controlling method for 3D optical system is valuable for the controlling of various kinds of 3D optical systems.

  15. Highly non-planar dendritic porphyrin for pH sensing: observation of porphyrin monocation.

    PubMed

    Thyagarajan, Sujatha; Leiding, Thom; Arsköld, Sindra Peterson; Cheprakov, Andrei V; Vinogradov, Sergei A

    2010-11-01

    Metal-free porphyrin-dendrimers provide a convenient platform for the construction of membrane-impermeable ratiometric probes for pH measurements in compartmentalized biological systems. In all previously reported molecules, electrostatic stabilization (shielding) of the core porphyrin by peripheral negative charges (carboxylates) was required to shift the intrinsically low porphyrin protonation pK(a)'s into the physiological pH range (pH 6-8). However, binding of metal cations (e.g., K(+), Na(+), Ca(2+), Mg(2+)) by the carboxylate groups on the dendrimer could affect the protonation behavior of such probes in biological environments. Here we present a dendritic pH nanoprobe based on a highly non-planar tetraaryltetracyclohexenoporphyrin (Ar(4)TCHP), whose intrinsic protonation pK(a)'s are significantly higher than those of regular tetraarylporphyrins, thereby eliminating the need for electrostatic core shielding. The porphyrin was modified with eight Newkome-type dendrons and PEGylated at the periphery, rendering a neutral water-soluble probe (TCHpH), suitable for measurements in the physiological pH range. The protonation of TCHpH could be followed by absorption (e.g., ε(Soret)(dication)∼270,000 M(-1) cm(-1)) or by fluorescence. Unlike most tetraarylporphyrins, TCHpH is protonated in two distinct steps (pK(a)'s 7.8 and 6.0). In the region between the pK(a)'s, an intermediate species with a well-defined spectroscopic signature, presumably a TCHpH monocation, could be observed in the mixture. The performance of TCHpH was evaluated by pH gradient measurements in large unilamellar vesicles. The probe was retained inside the vesicles and did not pass through and/or interact with vesicle membranes, proving useful for quantification of proton transport across phospholipid bilayers. To interpret the protonation behavior of TCHpH we developed a model relating structural changes on the porphyrin macrocycle upon protonation to its basicity. The model was validated by

  16. Influence of experimental hypokinesia on gastric secretory function

    NASA Technical Reports Server (NTRS)

    Markova, O. O.; Vavryshchuk, V. I.; Rozvodovskyy, V. I.; Proshcheruk, V. A.

    1980-01-01

    The gastric secretory function of rats was studied in 4, 8, 16 and 30 day hypokinesia. Inhibition of both the gastric juice secretory and acid producing functions was found. The greatest inhibition was observed on day 8 of limited mobility. By days 16 and 30 of the experiment, a tendency of the gastric secretory activity to return to normal was observed, although it remained reduced.

  17. Numerical investigation of the non-Newtonian pulsatile blood flow in a bifurcation model with a non-planar branch.

    PubMed

    Chen, Jie; Lu, Xi-Yun

    2006-01-01

    The pulsatile flow of non-Newtonian fluid in a bifurcation model with a non-planar daughter branch is investigated numerically by using the Carreau-Yasuda model to take into account the shear thinning behavior of the analog blood fluid. The objective of this study is to deal with the influence of the non-Newtonian property of fluid and of out-of-plane curvature in the non-planar daughter vessel on wall shear stress (WSS), oscillatory shear index (OSI), and flow phenomena during the pulse cycle. The non-Newtonian property in the daughter vessels induces a flattened axial velocity profile due to its shear thinning behavior. The non-planarity deflects flow from the inner wall of the vessel to the outer wall and changes the distribution of WSS along the vessel, in particular in systole phase. Downstream of the bifurcation, the velocity profiles are shifted toward the flow divider, and low WSS and high shear stress temporal oscillations characterized by OSI occur on the outer wall region of the daughter vessels close to the bifurcation. Secondary motions become stronger with the addition of the out-of-plane curvature induced by the bending of the vessel, and the secondary flow patterns swirl along the non-planar daughter vessel. A significant difference between the non-Newtonian and the Newtonian pulsatile flow is revealed during the pulse cycle; however, reasonable agreement between the non-Newtonian and the rescaled Newtonian flow is found. Calculated results for the pulsatile flow support the view that the non-planarity of blood vessels and the non-Newtonian properties of blood are an important factor in hemodynamics and may play a significant role in vascular biology and pathophysiology.

  18. A 5 meter range non-planar CMUT array for Automotive Collision Avoidance

    NASA Astrophysics Data System (ADS)

    Hernandez Aguirre, Jonathan

    A discretized hyperbolic paraboloid geometry capacitive micromachined ultrasonic transducer (CMUT) array has been designed and fabricated for automotive collision avoidance. The array is designed to operate at 40 kHz, beamwidth of 40° with a maximum sidelobe intensity of -10dB. An SOI based fabrication technology has been used for the 5x5 array with 5 sensing surfaces along each x and y axis and 7 elevation levels. An assembly and packaging technique has been developed to realize the non-planar geometry in a PGA-68 package. A highly accurate mathematical method has been presented for analytical characterization of capacitive micromachined ultrasonic transducers (CMUTs) built with square diaphragms. The method uses a new two-dimensional polynomial function to more accurately predict the deflection curve of a multilayer square diaphragm subject to both mechanical and electrostatic pressure and a new capacitance model that takes into account the contribution of the fringing field capacitances.

  19. Aerodynamic sensitivities from subsonic, sonic and supersonic unsteady, nonplanar lifting-surface theory

    NASA Technical Reports Server (NTRS)

    Yates, E. Carson, Jr.

    1987-01-01

    The technique of implicit differentiation has been used in combination with linearized lifting-surface theory to derive analytical expressions for aerodynamic sensitivities (i.e., rates of change of lifting pressures with respect to general changes in aircraft geometry, including planform variations) for steady or oscillating planar or nonplanar lifting surfaces in subsonic, sonic, or supersonic flow. The geometric perturbation is defined in terms of a single variable, and the user need only provide simple expressions or similar means for defining the continuous or discontinuous global or local perturbation of interest. Example expressions are given for perturbations of the sweep, taper, and aspect ratio of a wing with trapezoidal semispan planform. In addition to direct computational use, the analytical method presented here should provide benchmark criteria for assessing the accuracy of aerodynamic sensitivities obtained by approximate methods such as finite geometry perturbation and differencing. The present process appears to be readily adaptable to more general surface-panel methods.

  20. The Bouguer-Lambert-Beer Absorption Law and Non-Planar Geometries

    NASA Astrophysics Data System (ADS)

    Sinko, John E.; Oh, Benjamin I.

    2011-11-01

    The familiar Bouguer-Lambert-Beer absorption law, often called Beer's law, is an essential component of many laser ablation propulsion models. However, its treatment in non-planar conditions requires a consideration of irradiation geometry. Forms of the absorption law are derived for cylindrical and spherical normal incidence geometries, and for conical nozzles with flat and cylindrical targets. The results indicate that use of a concentrating nozzle optic with a transparent target could provide increased impulse generation for laser propulsion. This improvement would be accomplished using a combination of chosen optics and a transparent target material to generate highly confined ablation in-volume. The surface fluence and ablation depth on a cylindrical target in a parabolic optical nozzle is also derived, and the results are compared to literature raytracing model results and profilometry data, respectively.

  1. Dynamics of Nonplanar Quantized Vortex Rings Before Reconnection at Finite Temperatures

    NASA Astrophysics Data System (ADS)

    Andryushchenko, V. A.; Kondaurova, L. P.; Nemirovskii, S. K.

    2017-03-01

    The paper presents a numerical study of the dynamics of nonplanar quantized vortices at finite temperatures on their route to reconnection. We perform numerical simulations using the vortex filament method, solving the full Biot-Savart equation at a wide range of temperatures and initial conditions. We consider the dynamics of the two rings, lying initially in different planes and at different distances. The angles between planes are taken as equal to 30°, 45°, 60°, and 90°. It is observed that the temperature and the initial position of the vortices strongly affect the dynamics of the vortices on their route to reconnection. However, when the distances between the vertices of the vortices become smaller than the distances satisfying the Schwarz reconnection criterion, the dynamics of the system change drastically, and this trend is universal. The universality is expressed in the shapes and velocities of the vertices of the vortices.

  2. Calcium mediation of the pig jejunal secretory response.

    PubMed Central

    Forsyth, G W; Wong, P H; Maenz, D D

    1985-01-01

    The involvement of Ca++ ions as secretory mediators in pig jejunal epithelia has been investigated with an in vitro system. Omission of Ca++ from the Ringer-HCO3 bathing media on both sides of the tissue had minor effects on the basal electrical activity of pig jejunal mucosa. There were only slight decreases in transepithelial potential difference and increases in conductance with Ca++ free media. Low EGTA concentrations which reversibly blocked potential difference responses to secretory agents also had minimal effects on basal electrical activity. The in vitro secretory responses to A23187, to theophylline, and to Escherichia coli heat-stable enterotoxin were all eliminated by Ca++ depletion and restored by replacing normal Ca++ concentrations in the bathing media. Dantrolene prevented the secretory response but not the potential difference increases caused by heat-stable enterotoxin and A23187, suggesting that intracellular Ca++ stores may be reservoirs of secretory signal agent. Verapamil only blocked the secretory response to heat-stable enterotoxin. Chlorpromazine had negligible effects on basal conditions, but totally blocked both the secretory response and the Ca++-dependent effects of A23187 and heat-stable enterotoxin on potential difference. The response to theophylline was only partially inhibited by chlorpromazine, implying some involvement of both cAMP and Ca++ as secretory signals for theophylline. Cytoplasmic Ca++ concentrations appear to be at least as important as cyclic nucleotides in regulating the secretory effects of pig jejunum. PMID:2410089

  3. Nonplanar dust-acoustic Gardner solitons in a four-component dusty plasma

    SciTech Connect

    Mannan, A.; Mamun, A. A.

    2011-08-15

    The nonlinear propagation of Gardner solitons (GSs) in a nonplanar (cylindrical and spherical) four-component dusty plasma (composed of inertial positively and negatively dust, Boltzmann electrons, and ions) is studied by the reductive perturbation method. The modified Gardner equation is derived and numerically solved. It has been found that the basic characteristics of the dust-acoustic (DA) GSs, which are shown to exist for {mu} around its critical value {mu}{sub c}[where {mu}=Z{sub dp}m{sub dn}/Z{sub dn}m{sub dp}, Z{sub dn} (Z{sub dp}) is the number of electrons (protons) residing on a negative (positive) dust, m{sub dp} (m{sub dn}) is the mass of the positive (negative) dust, {mu}{sub c} is the value of {mu} corresponding to the vanishing of the nonlinear coefficient of the Korteweg-de Vries (KdV) equation, e.g., {mu}{sub c}{approx_equal}0.174 for {mu}{sub e}=n{sub e0}/Z{sub dn}n{sub dn0}=0.2, {mu}{sub i}=n{sub i0}/Z{sub dn}n{sub dn0}=0.4, and {sigma}=T{sub i}/T{sub e}=0.1, n{sub e0}, n{sub i0}, and n{sub dn0} are, respectively, electron, ion, and dust number densities, and T{sub i} (T{sub e}) is the ion (electron) temperature], are different from those of the KdV solitons, which do not exist for {mu} around {mu}{sub c}. It has been also found that the propagation characteristics of nonplanar DA GSs significantly differ from those of planar ones.

  4. Multi-dimensional modeling of atmospheric copper-sulfidation corrosion on non-planar substrates.

    SciTech Connect

    Chen, Ken Shuang

    2004-11-01

    This report documents the author's efforts in the deterministic modeling of copper-sulfidation corrosion on non-planar substrates such as diodes and electrical connectors. A new framework based on Goma was developed for multi-dimensional modeling of atmospheric copper-sulfidation corrosion on non-planar substrates. In this framework, the moving sulfidation front is explicitly tracked by treating the finite-element mesh as a pseudo solid with an arbitrary Lagrangian-Eulerian formulation and repeatedly performing re-meshing using CUBIT and re-mapping using MAPVAR. Three one-dimensional studies were performed for verifying the framework in asymptotic regimes. Limited model validation was also carried out by comparing computed copper-sulfide thickness with experimental data. The framework was first demonstrated in modeling one-dimensional copper sulfidation with charge separation. It was found that both the thickness of the space-charge layers and the electrical potential at the sulfidation surface decrease rapidly as the Cu{sub 2}S layer thickens initially but eventually reach equilibrium values as Cu{sub 2}S layer becomes sufficiently thick; it was also found that electroneutrality is a reasonable approximation and that the electro-migration flux may be estimated by using the equilibrium potential difference between the sulfidation and annihilation surfaces when the Cu{sub 2}S layer is sufficiently thick. The framework was then employed to model copper sulfidation in the solid-state-diffusion controlled regime (i.e. stage II sulfidation) on a prototypical diode until a continuous Cu{sub 2}S film was formed on the diode surface. The framework was also applied to model copper sulfidation on an intermittent electrical contact between a gold-plated copper pin and gold-plated copper pad; the presence of Cu{sub 2}S was found to raise the effective electrical resistance drastically. Lastly, future research needs in modeling atmospheric copper sulfidation are discussed.

  5. An evolving paradigm for the secretory pathway?

    PubMed Central

    Lippincott-Schwartz, Jennifer

    2011-01-01

    The paradigm that the secretory pathway consists of a stable endoplasmic reticulum and Golgi apparatus, using discrete transport vesicles to exchange their contents, gained important support from groundbreaking biochemical and genetic studies during the 1980s. However, the subsequent development of new imaging technologies with green fluorescent protein introduced data on dynamic processes not fully accounted for by the paradigm. As a result, we may be seeing an example of how a paradigm is evolving to account for the results of new technologies and their new ways of describing cellular processes. PMID:22039065

  6. Spiperone: evidence for uptake into secretory granules.

    PubMed Central

    Dannies, P S; Rudnick, M S; Fishkes, H; Rudnick, G

    1984-01-01

    Spiperone, a dopamine antagonist widely used as a specific ligand for dopamine and serotonin receptors, is actively accumulated into the F4C1 strain of rat pituitary tumor cells. The accumulation of 10 nM [3H]spiperone was linear for 3 min and reached a steady state after 10 min. Spiperone accumulation was reduced 50% by preincubation with 5 microM reserpine, an inhibitor of biogenic amine transport into secretory granules, and was also blocked by monensin and ammonium chloride, both of which increase the pH of intracellular storage organelles. Uptake was not affected by replacing sodium in the buffer with lithium at equimolar concentrations. Spiperone at 1 microM inhibited by over 50% serotonin transport into membrane vesicles isolated from platelet dense granules; this concentration inhibited the Na+-dependent plasma membrane transport system less than 10%. The data indicate spiperone specifically interacts with the secretory granule amine transport system and suggest that this transport system is found in the F4C1 pituitary cell strain as well as in platelets and neurons. The data also suggest that experiments utilizing spiperone to measure dopamine and serotonin receptors be interpreted with caution. PMID:6584920

  7. Brown adipose tissue as a secretory organ.

    PubMed

    Villarroya, Francesc; Cereijo, Rubén; Villarroya, Joan; Giralt, Marta

    2017-01-01

    Brown adipose tissue (BAT) is the main site of adaptive thermogenesis and experimental studies have associated BAT activity with protection against obesity and metabolic diseases, such as type 2 diabetes mellitus and dyslipidaemia. Active BAT is present in adult humans and its activity is impaired in patients with obesity. The ability of BAT to protect against chronic metabolic disease has traditionally been attributed to its capacity to utilize glucose and lipids for thermogenesis. However, BAT might also have a secretory role, which could contribute to the systemic consequences of BAT activity. Several BAT-derived molecules that act in a paracrine or autocrine manner have been identified. Most of these factors promote hypertrophy and hyperplasia of BAT, vascularization, innervation and blood flow, processes that are all associated with BAT recruitment when thermogenic activity is enhanced. Additionally, BAT can release regulatory molecules that act on other tissues and organs. This secretory capacity of BAT is thought to be involved in the beneficial effects of BAT transplantation in rodents. Fibroblast growth factor 21, IL-6 and neuregulin 4 are among the first BAT-derived endocrine factors to be identified. In this Review, we discuss the current understanding of the regulatory molecules (the so-called brown adipokines or batokines) that are released by BAT that influence systemic metabolism and convey the beneficial metabolic effects of BAT activation. The identification of such adipokines might also direct drug discovery approaches for managing obesity and its associated chronic metabolic diseases.

  8. ATP: The crucial component of secretory vesicles.

    PubMed

    Estévez-Herrera, Judith; Domínguez, Natalia; Pardo, Marta R; González-Santana, Ayoze; Westhead, Edward W; Borges, Ricardo; Machado, José David

    2016-07-12

    The colligative properties of ATP and catecholamines demonstrated in vitro are thought to be responsible for the extraordinary accumulation of solutes inside chromaffin cell secretory vesicles, although this has yet to be demonstrated in living cells. Because functional cells cannot be deprived of ATP, we have knocked down the expression of the vesicular nucleotide carrier, the VNUT, to show that a reduction in vesicular ATP is accompanied by a drastic fall in the quantal release of catecholamines. This phenomenon is particularly evident in newly synthesized vesicles, which we show are the first to be released. Surprisingly, we find that inhibiting VNUT expression also reduces the frequency of exocytosis, whereas the overexpression of VNUT drastically increases the quantal size of exocytotic events. To our knowledge, our data provide the first demonstration that ATP, in addition to serving as an energy source and purinergic transmitter, is an essential element in the concentration of catecholamines in secretory vesicles. In this way, cells can use ATP to accumulate neurotransmitters and other secreted substances at high concentrations, supporting quantal transmission.

  9. ATP: The crucial component of secretory vesicles

    PubMed Central

    Estévez-Herrera, Judith; Domínguez, Natalia; Pardo, Marta R.; González-Santana, Ayoze; Westhead, Edward W.; Borges, Ricardo; Machado, José David

    2016-01-01

    The colligative properties of ATP and catecholamines demonstrated in vitro are thought to be responsible for the extraordinary accumulation of solutes inside chromaffin cell secretory vesicles, although this has yet to be demonstrated in living cells. Because functional cells cannot be deprived of ATP, we have knocked down the expression of the vesicular nucleotide carrier, the VNUT, to show that a reduction in vesicular ATP is accompanied by a drastic fall in the quantal release of catecholamines. This phenomenon is particularly evident in newly synthesized vesicles, which we show are the first to be released. Surprisingly, we find that inhibiting VNUT expression also reduces the frequency of exocytosis, whereas the overexpression of VNUT drastically increases the quantal size of exocytotic events. To our knowledge, our data provide the first demonstration that ATP, in addition to serving as an energy source and purinergic transmitter, is an essential element in the concentration of catecholamines in secretory vesicles. In this way, cells can use ATP to accumulate neurotransmitters and other secreted substances at high concentrations, supporting quantal transmission. PMID:27342860

  10. Secretory antibody following oral influenza immunization.

    PubMed

    Waldman, R H; Stone, J; Bergmann, K C; Khakoo, R; Lazzell, V; Jacknowitz, A; Waldman, E R; Howard, S

    1986-12-01

    Secretory IgA antibody may be important in protection against respiratory viral infections, and the concept of a common mucosal immune system offers the theoretical basis for the convenient stimulation of this antibody. Therefore, the oral route was compared with intramuscular injection in a double-blind, placebo-controlled study in young healthy volunteers. A killed influenza vaccine, given in enteric-coated capsules (total of 98 ug hemagglutinin of A/Bangkok) led to significant salivary and nasal IgA antibody rises in a 4-week period. The preimmunization titers in secretions were inversely correlated with the antibody rise after immunization. The orally administered vaccine was associated with no more side effects than placebo, in contradistinction to reactions following the intramuscular route. The latter route also was without significant effect in regard to a stimulation of secretory antibodies. The observed simultaneous induction of antibodies in saliva and nasal secretions following oral administration of killed vaccine gives further evidence of a common mucosal immune system and its possible clinical use.

  11. 21 CFR 866.5380 - Free secretory component immuno-logical test system.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... immunochemical techniques free secretory component (normally a portion of the secretory IgA antibody molecule) in... repetitive lung infections and other hypogammaglobulinemic conditions (low antibody levels)....

  12. 21 CFR 866.5380 - Free secretory component immuno-logical test system.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... immunochemical techniques free secretory component (normally a portion of the secretory IgA antibody molecule) in... repetitive lung infections and other hypogammaglobulinemic conditions (low antibody levels)....

  13. 21 CFR 866.5380 - Free secretory component immuno-logical test system.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... immunochemical techniques free secretory component (normally a portion of the secretory IgA antibody molecule) in... repetitive lung infections and other hypogammaglobulinemic conditions (low antibody levels)....

  14. Numerical optimization techniques for bound circulation distribution for minimum induced drag of Nonplanar wings: Computer program documentation

    NASA Technical Reports Server (NTRS)

    Kuhlman, J. M.; Ku, T. J.

    1981-01-01

    A two dimensional advanced panel far-field potential flow model of the undistorted, interacting wakes of multiple lifting surfaces was developed which allows the determination of the spanwise bound circulation distribution required for minimum induced drag. This model was implemented in a FORTRAN computer program, the use of which is documented in this report. The nonplanar wakes are broken up into variable sized, flat panels, as chosen by the user. The wake vortex sheet strength is assumed to vary linearly over each of these panels, resulting in a quadratic variation of bound circulation. Panels are infinite in the streamwise direction. The theory is briefly summarized herein; sample results are given for multiple, nonplanar, lifting surfaces, and the use of the computer program is detailed in the appendixes.

  15. Some features of secretory systems in plants.

    PubMed

    Juniper, B E; Gilchrist, A J; Robins, R J

    1977-09-01

    Recent work on secretion in plants is reviewed, with emphasis on the anatomy and physiology of root cap cells in higher plants, the stalked glands of Drosera capensis, and the secretory mechanism of Dionaea muscipula. Cells of the root cap of higher plants switch from a geo-perceptive role to one of mucilage secretion at maturation. Features of this process, the role of the Golgi and the pathway for mucilage distribution are reviewed. In contrast, the stalked glands of the leaves of Drosera capensis are much longer lived and have a complex anatomy. The mechanisms for mucilage secretion, protein absorption and the role of the cell membranes in the internal secretion of the protein are described, using data from X-ray microscopv. The secretion of fluid and protein by Dionaea is stimulated by various nitrogen-containing compounds. Uric acid, often excreted by captured insects, is particularly effective in this respect.

  16. Secretion from Myeloid Cells: Secretory Lysosomes.

    PubMed

    Griffiths, Gillian M

    2016-08-01

    Many cells of the myeloid lineage use an unusual secretory organelle to deliver their effector mechanisms. In these cells, the lysosomal compartment is often modified not only to fulfill the degradative functions of a lysosome but also as a mechanism for secreting additional proteins that are found in the lysosomes of each specialized cell type. These extra proteins vary from one cell type to another according to the specialized function of the cell. For example, mast cells package histamine; cytotoxic T cells express perforin; azurophilic granules in neutrophils express antimicrobial peptides, and platelets von Willebrand factor. Upon release, these very different proteins can trigger inflammation, cell lysis, microbial death, and clotting, respectively, and hence deliver the very different effector mechanisms of these different myeloid cells.

  17. Numerical investigation of the non-Newtonian blood flow in a bifurcation model with a non-planar branch.

    PubMed

    Chen, Jie; Lu, Xi-Yun

    2004-12-01

    The non-Newtonian fluid flow in a bifurcation model with a non-planar daughter branch is investigated by using finite element method to solve the three-dimensional Navier-Stokes equations coupled with a non-Newtonian constitutive model, in which the shear thinning behavior of the blood fluid is incorporated by the Carreau-Yasuda model. The objective of this study is to investigate the influence of the non-Newtonian property of fluid as well as of curvature and out-of-plane geometry in the non-planar daughter vessel on wall shear stress (WSS) and flow phenomena. In the non-planar daughter vessel, the flows are typified by the skewing of the velocity profile towards the outer wall, creating a relatively low WSS at the inner wall. In the downstream of the bifurcation, the velocity profiles are shifted towards the flow divider. The low WSS is found at the inner walls of the curvature and the lateral walls of the bifurcation. Secondary flow patterns that swirl fluid from the inner wall of curvature to the outer wall in the middle of the vessel are also well documented for the curved and bifurcating vessels. The numerical results for the non-Newtonian fluid and the Newtonian fluid with original Reynolds number and the corresponding rescaled Reynolds number are presented. Significant difference between the non-Newtonian flow and the Newtonian flow is revealed; however, reasonable agreement between the non-Newtonian flow and the rescaled Newtonian flow is found. Results of this study support the view that the non-planarity of blood vessels and the non-Newtonian properties of blood are an important factor in hemodynamics and may play a significant role in vascular biology and pathophysiology.

  18. 30 Hz-linewidth, diode-laser-pumped, Nd:GGG nonplanar ring oscillators by active frequency stabilisation

    NASA Technical Reports Server (NTRS)

    Day, T.; Nilsson, A. C.; Fejer, M. M.; Farinas, A. D.; Gustafson, E. K.

    1989-01-01

    A heterodyne linewidth of less than 30 Hz for the beatnote between the outputs of two 282 THz Nd:GGG nonplanar ring oscillators (NPROs) is reported. The lasers were independently locked to adjacent axial modes of a high-finesse interferometer. The remnant frequency noise appears to be dominated by free spectral range fluctuations in the reference interferometer rather than by residual laser noise.

  19. Time evolution of nonplanar dust ion-acoustic solitary waves in a charge varying dusty plasma with superthermal electrons

    SciTech Connect

    Mayout, Saliha; Tribeche, Mouloud; Sahu, Biswajit

    2015-12-15

    A theoretical study on the nonlinear propagation of nonplanar (cylindrical and spherical) dust ion-acoustic solitary waves (DIASW) is carried out in a dusty plasma, whose constituents are inertial ions, superthermal electrons, and charge fluctuating stationary dust particles. Using the reductive perturbation theory, a modified Korteweg-de Vries equation is derived. It is shown that the propagation characteristics of the cylindrical and spherical DIA solitary waves significantly differ from those of their one-dimensional counterpart.

  20. AP-1A controls secretory granule biogenesis and trafficking of membrane secretory granule proteins.

    PubMed

    Bonnemaison, Mathilde; Bäck, Nils; Lin, Yimo; Bonifacino, Juan S; Mains, Richard; Eipper, Betty

    2014-10-01

    The adaptor protein 1A complex (AP-1A) transports cargo between the trans-Golgi network (TGN) and endosomes. In professional secretory cells, AP-1A also retrieves material from immature secretory granules (SGs). The role of AP-1A in SG biogenesis was explored using AtT-20 corticotrope tumor cells expressing reduced levels of the AP-1A μ1A subunit. A twofold reduction in μ1A resulted in a decrease in TGN cisternae and immature SGs and the appearance of regulated secretory pathway components in non-condensing SGs. Although basal secretion of endogenous SG proteins was unaffected, secretagogue-stimulated release was halved. The reduced μ1A levels interfered with the normal trafficking of carboxypeptidase D (CPD) and peptidylglycine α-amidating monooxygenase-1 (PAM-1), integral membrane enzymes that enter immature SGs. The non-condensing SGs contained POMC products and PAM-1, but not CPD. Based on metabolic labeling and secretion experiments, the cleavage of newly synthesized PAM-1 into PHM was unaltered, but PHM basal secretion was increased in sh-μ1A PAM-1 cells. Despite lacking a canonical AP-1A binding motif, yeast two-hybrid studies demonstrated an interaction between the PAM-1 cytosolic domain and AP-1A. Coimmunoprecipitation experiments with PAM-1 mutants revealed an influence of the luminal domains of PAM-1 on this interaction. Thus, AP-1A is crucial for normal SG biogenesis, function and composition.

  1. Scanning electron microscopy of the endometrium during the secretory phase.

    PubMed Central

    Motta, P M; Andrews, P M

    1976-01-01

    Scanning electron microscopy was used to study the surface morphology of the rabbit endometrium during the secretory phase of the oestrous cycle. The free surfaces of ciliated and of inactive active secretory cells are described. Changes in secretory cell surface morphology resulting from accumulation and secretion of material involve the apparent retraction of microvilli and the formation of one or more bulbous protrusions of the cell's apical surface. These protrusions may be relatively smooth surfaced or exhibit long slender micro-extensions. The protrusions grow in size and are eventually pinched off. Loss of the bulbous protrusions often leaves behind crater-like invaginations of the cell's surface. Secretory cells adjacent to the endometrial glands are the first to exhibit signs of mucin accumulation and secretion. The single cilium of a secretory cell is not apparently affected by the secretory process. Signs of ciliated and secretory cell degeneration, and possible sloughing, are also described. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5 Fig. 6 Fig. 7 PMID:1033932

  2. Magnetoresistance effects and phase coherent transport phenomena in a magnetic nonplanar two-dimensional hole system

    NASA Astrophysics Data System (ADS)

    Knott, Stefan; Hirschmann, Thomas Ch.; Wurstbauer, Ursula; Hansen, Wolfgang; Wegscheider, Werner

    2011-11-01

    Phase coherent transport phenomena are studied in a magnetic, two-dimensional hole system confined in a strained InAs quantum-well structure. At low magnetic fields we observe a well pronounced weak antilocalization signature, demonstrating that magnetic ions in proximity to the itinerant holes preserve phase coherent transport, although magnetic effects like the anomalous and planar Hall effects are present at low temperatures. From fits to the conductivity correction, spin and phase relaxation times are extracted in a large temperature range from 200 mK to 16 K. From the dependency of the spin relaxation time on temperature and in-plane magnetic fields, we conclude that the Dyakonov-Perel mechanism is the dominant spin relaxation mechanism. The metamorphic growth of the heterostructure causes a nonplanar quantum well that enables the formation of magnetic barriers in the presence of a parallel applied magnetic field. Transport through these magnetic barriers causes a positive magnetoresistance superimposed on the weak antilocalization signal.

  3. Infrared non-planar plasmonic perfect absorber for enhanced sensitive refractive index sensing

    NASA Astrophysics Data System (ADS)

    Cheng, Yongzhi; Mao, Xue Song; Wu, Chenjun; Wu, Lin; Gong, RongZhou

    2016-03-01

    We present a non-planar all-metal plasmonic perfect absorber (PA) with response polarization independent in infrared region, which can be served as a sensor for enhanced refractive index sensing. Distinct from previous designs, the proposed PA consisted of all metal structured film constructed with an assembly of four-tined rod resonators (FRRs). The PA with a high quality-factor (Q-factor) of 41.2 and an absorbance of 99.9% at 142.6 THz has been demonstrated numerically. The resonance behavior occurs in the space between the rods of the FRRs, which is remarkable different conventional sandwiched structural PAs. Based on equivalent LC circuit theory, the absorption peak can be finely tuned by varying the geometrical dimensions of the FRRs. Furthermore, the resonance frequency shows highly sensitive response to the change of refractive index in the surrounding medium. A careful design for refractive index sensor can yield a sensitivity of 1445 nm/refractive index unit (RIU) and a figure of merit (FOM) of 28.8. The demonstrated design of the plasmonic PA for sensing provides great potential application in enhancing refractive index sensors and the enhanced infrared spectroscopy.

  4. Asymptotic expansion for stellarator equilibria with a non-planar magnetic axis: Numerical results

    NASA Astrophysics Data System (ADS)

    Freidberg, Jeffrey; Cerfon, Antoine; Parra, Felix

    2012-10-01

    We have recently presented a new asymptotic expansion for stellarator equilibria that generalizes the classic Greene-Johnson expansion [1] to allow for 3D equilibria with a non-planar magnetic axis [2]. Our expansion achieves the two goals of reducing the complexity of the three-dimensional MHD equilibrium equations and of describing equilibria in modern stellarator experiments. The end result of our analysis is a set of two coupled partial differential equations for the plasma pressure and the toroidal vector potential which fully determine the stellarator equilibrium. Both equations are advection equations in which the toroidal angle plays the role of time. We show that the method of characteristics, following magnetic field lines, is a convenient way of solving these equations, avoiding the difficulties associated with the periodicity of the solution in the toroidal angle. By combining the method of characteristics with Green's function integrals for the evaluation of the magnetic field due to the plasma current, we obtain an efficient numerical solver for our expansion. Numerical equilibria thus calculated will be given.[4pt] [1] J.M. Greene and J.L. Johnson, Phys. Fluids 4, 875 (1961)[0pt] [2] A.J. Cerfon, J.P. Freidberg, and F.I. Parra, Bull. Am. Phys. Soc. 56, 16 GP9.00081 (2011)

  5. Asymptotic expansion for stellarator equilibria with a non-planar magnetic axis: Numerical progress

    NASA Astrophysics Data System (ADS)

    Cerfon, Antoine; Freidberg, Jeffrey; Parra, Felix

    2012-03-01

    We have recently presented a new asymptotic analysis [1], which reduces the complexity of the MHD equilibrium equations in stellarators and generalizes the asymptotic approach followed by Greene and Johnson in their classic paper [2]. As in [2], our expansion relies on the small ratio of the helical magnetic field to the vacuum toroidal field. However, our ordering relaxes the Greene and Johnson constraint which assumes a strong separation in length scales between the helical period and the major radius. In our expansion these two length scales are of comparable order, which provides a better match with modern stellarator experiments. Toroidal effects enter the analysis in the same order as helical effects, allowing the calculations of equilibria with multiple helicities and a non-planar magnetic axis. The end result of our analysis is a set of two coupled PDEs for the plasma pressure and the magnetic vector potential, which fully determine the stellarator equilibrium. We present simple analytic solutions to these equations, and discuss the numerical methods we are developping to calculate more general stellarator equilibria.[4pt] [1] A.J. Cerfon, J.P. Freidberg, and F.I. Parra, Bull. Am. Phys. Soc. 56, 16 GP9.00081[0pt] [2] J.M. Greene and J.L. Johnson, Phys. Fluids 4, 875 (1961)

  6. Elastic wave field computation in multilayered nonplanar solid structures: a mesh-free semianalytical approach.

    PubMed

    Banerjee, Sourav; Kundu, Tribikram

    2008-03-01

    Multilayered solid structures made of isotropic, transversely isotropic, or general anisotropic materials are frequently used in aerospace, mechanical, and civil structures. Ultrasonic fields developed in such structures by finite size transducers simulating actual experiments in laboratories or in the field have not been rigorously studied. Several attempts to compute the ultrasonic field inside solid media have been made based on approximate paraxial methods like the classical ray tracing and multi-Gaussian beam models. These approximate methods have several limitations. A new semianalytical method is adopted in this article to model elastic wave field in multilayered solid structures with planar or nonplanar interfaces generated by finite size transducers. A general formulation good for both isotropic and anisotropic solids is presented in this article. A variety of conditions have been incorporated in the formulation including irregularities at the interfaces. The method presented here requires frequency domain displacement and stress Green's functions. Due to the presence of different materials in the problem geometry various elastodynamic Green's functions for different materials are used in the formulation. Expressions of displacement and stress Green's functions for isotropic and anisotropic solids as well as for the fluid media are presented. Computed results are verified by checking the stress and displacement continuity conditions across the interface of two different solids of a bimetal plate and investigating if the results for a corrugated plate with very small corrugation match with the flat plate results.

  7. High-Reliability Pump Module for Non-Planar Ring Oscillator Laser

    NASA Technical Reports Server (NTRS)

    Liu, Duncan T.; Qiu, Yueming; Wilson, Daniel W.; Dubovitsky, Serge; Forouhar, Siamak

    2007-01-01

    We propose and have demonstrated a prototype high-reliability pump module for pumping a Non-Planar Ring Oscillator (NPRO) laser suitable for space missions. The pump module consists of multiple fiber-coupled single-mode laser diodes and a fiber array micro-lens array based fiber combiner. The reported Single-Mode laser diode combiner laser pump module (LPM) provides a higher normalized brightness at the combined beam than multimode laser diode based LPMs. A higher brightness from the pump source is essential for efficient NPRO laser pumping and leads to higher reliability because higher efficiency requires a lower operating power for the laser diodes, which in turn increases the reliability and lifetime of the laser diodes. Single-mode laser diodes with Fiber Bragg Grating (FBG) stabilized wavelength permit the pump module to be operated without a thermal electric cooler (TEC) and this further improves the overall reliability of the pump module. The single-mode laser diode LPM is scalable in terms of the number of pump diodes and is capable of combining hundreds of fiber-coupled laser diodes. In the proof-of-concept demonstration, an e-beam written diffractive micro lens array, a custom fiber array, commercial 808nm single mode laser diodes, and a custom NPRO laser head are used. The reliability of the proposed LPM is discussed.

  8. Research on the thermal relaxation effect of nonplanar four-frequency differential laser gyro.

    PubMed

    Wang, Guochen; Rao, Guyin; Han, Songlai; Zhang, Mei; Yuan, Baolun

    2013-07-01

    The nonplanar four-frequency differential laser gyro (NFFDLG, also called ZLG in the United States) has many advantages, such as the absence of mechanical noise, a whole solid state, high stability of scale factor, and no time delay. Because of its excellent performance, NFFDLG is currently the subject of research. In this paper, the error induced by the thermal relaxation effect is investigated both theoretically and experimentally, and some methods and conclusions that could restrain this error effectively are proposed. The experiment data confirm reliably that NFFDLG has a thermal relaxation effect error during overturning. At the same time, it is pointed out that the error can be minimized by reducing the stop interval. Furthermore, since the sum frequency could be affected more seriously by the overturning thermal relaxation effect than the difference frequency could, the former one can be used as one more sensitive condition in actual applications. As far as we know, the overturning thermal relaxation effect was first investigated in China, which is even not mentioned publicly.

  9. The secretory synapse: the secrets of a serial killer.

    PubMed

    Bossi, Giovanna; Trambas, Christina; Booth, Sarah; Clark, Richard; Stinchcombe, Jane; Griffiths, Gillian M

    2002-11-01

    Cytotoxic T lymphocytes (CTLs) destroy their targets by a process involving secretion of specialized granules. The interactions between CTLs and target can be very brief; nevertheless, adhesion and signaling proteins segregate into an immunological synapse. Secretion occurs in a specialized secretory domain. Use of live and fixed cell microscopy allows this secretory synapse to be visualized both temporally and spatially. The combined use of confocal and electron microscopy has produced some surprising findings, which suggest that the secretory synapse may be important both in delivering the lethal hit and in facilitating membrane transfer from target to CTL. Studies on the secretory synapse in wild-type and mutant CTLs have been used to identify proteins involved in secretion. Further clues as to the signals required for secretion are emerging from comparisons of inhibitory and activating synapses formed by natural killer cells.

  10. [Serum and secretory immunoglobulins in allergic diseases].

    PubMed

    Atovmian, O I; German, G P; Chernokhvostova, E V

    1985-07-01

    A total of 158 patients with pollinosis, bronchial asthma, urticaria and Quincke's edema were examined. The immunoglobulin and C3 levels in sera and the immunoglobulin and albumin levels in saliva were determined by the method of single radial immunodiffusion with the corresponding monospecific antisera. In all the groups of patients subjected to examination the presence of polyclonal hypergammaglobulinemia was detected, which was manifested by a rise in the levels of IgG, IgA and especially IgM; the level of IgD was low. A decrease in the level of C3 was detected in pollinosis patients in the absence of the exacerbation of the disease. No circulating immune complexes were detected. An essential increase in the level of IgG in saliva was revealed, which was due to the local synthesis of this immunoglobulin. In winter the level of salivary IgA in pollinosis patients was found to be essentially below normal, but at the period of exacerbation it increased twofold, probably in response to local stimulation with antigen-allergen. Patients with bronchial asthma and pollinosis were found to have a high level of free secretory component (SC); in pollinosis the level of free SC sharply increased during the stage of exacerbation, which was due to the increase of its synthesis and secretion by the epithelial cells of the mucous membranes. The importance of these data for the pathogenesis of allergic diseases are discussed.

  11. Mammary analogue secretory carcinoma mimicking salivary adenoma.

    PubMed

    Williams, Lindsay; Chiosea, Simion I

    2013-12-01

    Mammary analogue secretory carcinoma (MASC) is a recently described salivary gland tumor characterized by ETV6 translocation. It appears that prior studies have identified MASC by reviewing salivary gland carcinomas, such as acinic cell carcinoma and adenocarcinoma, not otherwise specified. To address the possibility of MASC mimicking benign salivary neoplasms we reviewed 12 salivary gland (cyst)adenomas diagnosed prior to the discovery of MASC. One encapsulated (cyst)adenoma of the parotid gland demonstrated features of MASC. The diagnosis was confirmed by fluorescence in situ hybridization with an ETV6 break-apart probe. An unusual complex pattern of ETV6 rearrangement with duplication of the telomeric/distal ETV6 probe was identified. This case illustrates that MASC may mimic salivary (cyst)adenomas. To more accurately assess true clinical and morphologic spectrum of MASC, future studies may have to include review of salivary (cyst)adenomas. The differential diagnosis of MASC may have to be expanded to include cases resembling salivary (cyst)adenomas.

  12. Episodic slow slip events in a non-planar subduction fault model for northern Cascadia

    NASA Astrophysics Data System (ADS)

    Li, D.; Liu, Y.; Matsuzawa, T.; Shibazaki, B.

    2014-12-01

    Episodic tremor and slow slip (ETS) events have been detected along the Cascadia margin, as well as many other subduction zones, by increasingly dense seismic and geodetic networks over the past decade. In northern Cascadia, ETS events arise on the thrust fault interface of 30~50 km depth, coincident with metamorphic dehydration of the subducting oceanic slab around temperatures of 350. Previous numerical simulations (e.g., Liu and Rice 2007) suggested that near-lithostatic pore pressure in the rate-state friction stability transition zone could give rise to slow slip events (SSE) down-dip of the seismogenic zone, which provides a plausible physical mechanism for these phenomena. Here we present a 3-D numerical simulation of inter-seismic SSEs based on the rate- and state- friction law, incorporating a non-planar, realistic northern Cascadia slab geometry compiled by McCrory et al. (2012) using triangular dislocation elements. Preliminary results show that the width and pore pressure level of the transition zone can remarkably affect the recurrence of SSEs. With effective normal stress of ~1-2 MPa and characteristic slip distance of ~1.4 mm, inter-seismic SSEs can arise about every year. The duration of each event is about 2~3 weeks, with the propagating speed along strike in the range of km/day. Furthermore, the slab bending beneath southern Vancouver Island and northern Washington State appears to accelerate the along-strike propagation of SSEs. Our next step is to constrain the rate-state frictional properties using geodetic inversion of SSE slip and inter-SSE plate coupling from the Plate Boundary Observatory (PBO) GPS measurements. Incorporating the realistic fault geometry into a physics model constrained by geodetic data will enable us to transition from a conceptual towards a quantitative and predictive understanding of SSEs mechanism.

  13. Dynamic earthquake rupture simulations on nonplanar faults embedded in 3D geometrically complex, heterogeneous elastic solids

    SciTech Connect

    Duru, Kenneth; Dunham, Eric M.

    2016-01-15

    Dynamic propagation of shear ruptures on a frictional interface in an elastic solid is a useful idealization of natural earthquakes. The conditions relating discontinuities in particle velocities across fault zones and tractions acting on the fault are often expressed as nonlinear friction laws. The corresponding initial boundary value problems are both numerically and computationally challenging. In addition, seismic waves generated by earthquake ruptures must be propagated for many wavelengths away from the fault. Therefore, reliable and efficient numerical simulations require both provably stable and high order accurate numerical methods. We present a high order accurate finite difference method for: a) enforcing nonlinear friction laws, in a consistent and provably stable manner, suitable for efficient explicit time integration; b) dynamic propagation of earthquake ruptures along nonplanar faults; and c) accurate propagation of seismic waves in heterogeneous media with free surface topography. We solve the first order form of the 3D elastic wave equation on a boundary-conforming curvilinear mesh, in terms of particle velocities and stresses that are collocated in space and time, using summation-by-parts (SBP) finite difference operators in space. Boundary and interface conditions are imposed weakly using penalties. By deriving semi-discrete energy estimates analogous to the continuous energy estimates we prove numerical stability. The finite difference stencils used in this paper are sixth order accurate in the interior and third order accurate close to the boundaries. However, the method is applicable to any spatial operator with a diagonal norm satisfying the SBP property. Time stepping is performed with a 4th order accurate explicit low storage Runge–Kutta scheme, thus yielding a globally fourth order accurate method in both space and time. We show numerical simulations on band limited self-similar fractal faults revealing the complexity of rupture

  14. Spatiotemporal evolution of slow slip events in a nonplanar fault model for northern Cascadia subduction zone

    NASA Astrophysics Data System (ADS)

    Li, Duo; Liu, Yajing

    2016-09-01

    Slow slip events (SSEs) are identified as the quasi-stable fault deformation in the deep transition zone from locked to continuous sliding in many subduction zones. In the well-instrumented Cascadia margin, a class of Mw6.0 slow slip events arise beneath Port Angeles every ˜14 months, as inferred from two decades of continuous geodetic monitoring. The along-strike bending of the incoming oceanic plate beneath north Washington is a unique geometric feature whose influence on slow slip processes is still unknown. Here we incorporate a realistic fault geometry of northern Cascadia in the framework of rate- and state-dependent friction law, to simulate the spatiotemporal evolution of slow slip events on a nonplanar subduction fault. The modeled SSEs capture the major characteristics revealed by GPS observations. The central 150 km long fault segment beneath Port Angeles acts as a repetitive slip patch, where SSEs appear every ˜1.5 years with a maximum slip of ˜2.5 cm. Two minor slip patches with smaller areas and cumulative slips straddle this central slip patch. The along-strike segmentation of slow slip is inversely related to the local fault dip and strike angles of the slow slip zone, suggesting strong geometrical control on the slow slip process. This correlation holds even after removing the effect of W/h∗, ratio between velocity-weakening SSE fault width and characteristic nucleation size. Besides the GPS-detectable fast-spreading phase, we find that each SSE cycle consists of deep pre-SSE preparation and post-SSE relaxation phases, which may be the driving mechanism for the deep tremor activity between major SSE episodes discovered in Cascadia.

  15. Numerical investigation of blood flow in a deformable coronary bifurcation and non-planar branch

    PubMed Central

    Razavi, Seyed Esmail; Omidi, Amir Ali; Saghafi Zanjani, Massoud

    2014-01-01

    Introduction: Among cardiovascular diseases, arterials stenosis is recognized more commonly than the others. Hemodynamic characteristics of blood play a key role in the incidence of stenosis. This paper numerically investigates the pulsatile blood flow in a coronary bifurcation with a non-planar branch. To create a more realistic analysis, the wall is assumed to be compliant. Furthermore, the flow is considered to be three-dimensional, incompressible, and laminar. Methods: The effects of non-Newtonian blood, compliant walls and different angles of bifurcation on hemodynamic characteristics of flow were evaluated. Shear thinning of blood was simulated with the Carreau-Yasuda model. The current research was mainly focused on the flow characteristics in bifurcations since atherosclerosis occurs mostly in bifurcations. Moreover, as the areas with low shear stresses are prone to stenosis, these areas were identified. Results: Our findings indicated that the compliant model of the wall, bifurcation’s angle, and other physical properties of flow have an impact on hemodynamics of blood flow. Lower wall shear stress was observed in the compliant wall than that in the rigid wall. The outer wall of bifurcation in all models had lower wall shear stress. In bifurcations with larger angles, wall shear stress was higher in outer walls, and lower in inner walls. Conclusion: The non-Newtonian blood vessels and different angles of bifurcation on hemodynamic characteristics of flow evaluation confirmed a lower wall shear stress in the compliant wall than that in the rigid wall, while the wall shear stress was higher in outer walls but lower in inner walls in the bifurcation regions with larger angles. PMID:25671176

  16. Progressive quality control of secretory proteins in the early secretory compartment by ERp44

    PubMed Central

    Sannino, Sara; Anelli, Tiziana; Cortini, Margherita; Masui, Shoji; Degano, Massimo; Fagioli, Claudio; Inaba, Kenji; Sitia, Roberto

    2014-01-01

    ERp44 is a pH-regulated chaperone of the secretory pathway. In the acidic milieu of the Golgi, its C-terminal tail changes conformation, simultaneously exposing the substrate-binding site for cargo capture and the RDEL motif for ER retrieval via interactions with cognate receptors. Protonation of cysteine 29 in the active site allows tail movements in vitro and in vivo. Here we show that also conserved histidines in the C-terminal tail regulate ERp44 in vivo. Mutants lacking these histidines are hyperactive in retaining substrates. Surprisingly, they are also O-glycosylated and partially secreted. Co-expression of client proteins prevents secretion of the histidine mutants, forcing tail opening and RDEL accessibility. Client-induced RDEL exposure allows retrieval of proteins from distinct stations along the secretory pathway, as indicated by the changes in O-glycosylation patterns upon over-expression of different partners. The ensuing gradients may help optimising folding and assembly of different cargoes. Endogenous ERp44 is O-glycosylated and secreted by human primary endometrial cells, suggesting possible pathophysiological roles of these processes. PMID:25097228

  17. Progressive quality control of secretory proteins in the early secretory compartment by ERp44.

    PubMed

    Sannino, Sara; Anelli, Tiziana; Cortini, Margherita; Masui, Shoji; Degano, Massimo; Fagioli, Claudio; Inaba, Kenji; Sitia, Roberto

    2014-10-01

    ERp44 is a pH-regulated chaperone of the secretory pathway. In the acidic milieu of the Golgi, its C-terminal tail changes conformation, simultaneously exposing the substrate-binding site for cargo capture and the RDEL motif for ER retrieval through interactions with cognate receptors. Protonation of cysteine 29 in the active site allows tail movements in vitro and in vivo. Here, we show that conserved histidine residues in the C-terminal tail also regulate ERp44 in vivo. Mutants lacking these histidine residues retain substrates more efficiently. Surprisingly, they are also O-glycosylated and partially secreted. Co-expression of client proteins prevents secretion of the histidine mutants, forcing tail opening and RDEL accessibility. Client-induced RDEL exposure allows retrieval of proteins from distinct stations along the secretory pathway, as indicated by the changes in O-glycosylation patterns upon overexpression of different partners. The ensuing gradients might help to optimize folding and assembly of different cargoes. Endogenous ERp44 is O-glycosylated and secreted by human primary endometrial cells, suggesting possible pathophysiological roles of these processes.

  18. Nonplanar positron-acoustic Gardner solitary waves in electron-positron-ion plasmas with superthermal electrons and positrons

    NASA Astrophysics Data System (ADS)

    Uddin, M. J.; Alam, M. S.; Mamun, A. A.

    2015-02-01

    Nonplanar (cylindrical and spherical) positron-acoustic (PA) Gardner solitary waves (SWs) in an unmagnetized plasma system consisting of immobile positive ions, mobile cold positrons, and superthermal (kappa distributed) hot positrons and electrons are investigated. The modified Gardner equation is derived by using the reductive perturbation technique. The effects of cylindrical and spherical geometries, superthermal parameter of hot positrons and electrons, relative temperature ratios, and relative number density ratios on the PA Gardner SWs are studied by using the numerical simulations. The implications of our results in various space and laboratory plasma environments are briefly discussed.

  19. Calculation of lateral-directional stability derivatives of wings by a nonplanar quasi-vortex-lattice method

    NASA Technical Reports Server (NTRS)

    Lan, C. E.

    1981-01-01

    The nonplanar quasi-vortex-lattice method is applied to the calculation of lateral-directional stability derivatives of wings with and without vortex-lift effect. Results for conventional configurations and those with winglets, V-tail, etc. are compared with available data. All rolling moment derivatives are found to be accurately predicted. The prediction of side force and yawing moment derivatives for some configurations is not as accurate. Causes of the discrepancy are discussed. A user's manual for the program and the program listing are also included.

  20. Nonplanar positron-acoustic Gardner solitary waves in electron-positron-ion plasmas with superthermal electrons and positrons

    SciTech Connect

    Uddin, M. J. Alam, M. S.; Mamun, A. A.

    2015-02-15

    Nonplanar (cylindrical and spherical) positron-acoustic (PA) Gardner solitary waves (SWs) in an unmagnetized plasma system consisting of immobile positive ions, mobile cold positrons, and superthermal (kappa distributed) hot positrons and electrons are investigated. The modified Gardner equation is derived by using the reductive perturbation technique. The effects of cylindrical and spherical geometries, superthermal parameter of hot positrons and electrons, relative temperature ratios, and relative number density ratios on the PA Gardner SWs are studied by using the numerical simulations. The implications of our results in various space and laboratory plasma environments are briefly discussed.

  1. Secretory granule biogenesis and neuropeptide sorting to the regulated secretory pathway in neuroendocrine cells.

    PubMed

    Loh, Y Peng; Kim, Taeyoon; Rodriguez, Yazmin M; Cawley, Niamh X

    2004-01-01

    Neuropeptide precursors synthesized at the rough endoplasmic reticulum are transported and sorted at the trans-Golgi network (TGN) to the granules of the regulated secretory pathway (RSP) of neuroendocrine cells. They are then processed into active peptides and stored in large dense-core granules (LDCGs) until secreted upon stimulation. We have studied the regulation of biogenesis of the LDCGs and the mechanism by which neuropeptide precursors, such as pro-opiomelanocortin (POMC), are sorted into these LDCGs of the RSP in neuroendocrine and endocrine cells. We provide evidence that chromogranin A (CgA), one of the most abundant acidic glycoproteins ubiquitously present in neuroendocrine/endocrine cells, plays an important role in the regulation of LDCG biogenesis. Specific depletion of CgA expression by antisense RNAs in PC12 cells led to a profound loss of secretory granule formation. Exogenously expressed POMC was neither stored nor secreted in a regulated manner in these CgA-deficient PC12 cells. Overexpression of CgA in a CgA- and LDCG-deficient endocrine cell line, 6T3, restored regulated secretion of transfected POMC and the presence of immunoreactive CgA at the tips of the processes of these cells. Unlike CgA, CgB, another granin protein, could not substitute for the role of CgA in regulating LDCG biogenesis. Thus, we conclude that CgA is a key player in the regulation of the biogenesis of LDCGs in neuroendocrine cells. To examine the mechanism of sorting POMC to the LDCGs, we carried out site-directed mutagenesis, transfected the POMC mutants into PC12 cells, and assayed for regulated secretion. Our previous molecular modeling studies predicted a three-dimensional sorting motif in POMC that can bind to a sorting receptor, membrane carboxypeptidase E (CPE). The sorting signal consists of four conserved residues at the N-terminal loop structure of POMC: two acidic residues and two hydrophobic residues. The two acidic residues were predicted to bind to a

  2. Finite Element Modeling of Dynamic Shear Rupture Experiments Along Non-Planar Faults

    NASA Astrophysics Data System (ADS)

    Templeton, E. L.; Baudet, A.; Bhat, H. S.; Rice, J. R.

    2004-12-01

    The study of dynamically propagating shear cracks along weak paths like faults is of great interest for the study of earthquakes. We adapted the ABAQUS/Explicit dynamic finite element program to analyze the nucleation and propagation of shear cracks along a non-planar, kinked, weak path corresponding to the one that was used in recent laboratory fracture studies by Rousseau and Rosakis [JGR, 2003]. Their experiments involved impact loading of thin plates of Homalite-100, a photoelastically sensitive brittle polymer, which had been cut along a kinked path and then weakly glued back together everywhere except along a starter notch near the impact site. Under different conditions, propagation speeds were observed in both the sub-Rayleigh and intersonic (supershear) regimes. Strain gage recordings and high speed photography of isochromatic lines (lines of constant difference between the in-plane principal strains) provided characterization of the transient deformation fields associated with the impact and fracture propagation. For the finite element analyses, we implemented a slip-weakening failure model through an option in the ABAQUS program allowing user defined constitutive relations. The analyses of impact loading and of rupture nucleation and propagation were then carried out in the 2D framework of plane stress. In a first set of studies of nucleation and propagation of rupture along a straight fault, we determined after some trial and error an appropriate CFL number, and examined different element types and layouts, finding that the most acceptable results were obtained for low order elements. We used constant strain triangles, arrayed in groups of four to effectively form four-sided elements with corner nodes and one internal node. The studies also showed that to obtain representations of slip rate and shear stress near the propagating rupture tip that were relatively free from numerical oscillations, it was necessary to have element side lengths of order Ro/50

  3. A New Physics-Based Modeling of Multiple Non-Planar Hydraulic Fractures Propagation

    SciTech Connect

    Zhou, Jing; Huang, Hai; Deo, Milind; Jiang, Shu

    2015-10-01

    Because of the low permeability in shale plays, closely spaced hydraulic fractures and multilateral horizontal wells are generally required to improve production. Therefore, understanding the potential fracture interaction and stress evolution is critical in optimizing fracture/well design and completion strategy in multi-stage horizontal wells. In this paper, a novel fully coupled reservoir flow and geomechanics model based on the dual-lattice system is developed to simulate multiple non-planar fractures propagation. The numerical model from Discrete Element Method (DEM) is used to simulate the mechanics of fracture propagations and interactions, while a conjugate irregular lattice network is generated to represent fluid flow in both fractures and formation. The fluid flow in the formation is controlled by Darcy’s law, but within fractures it is simulated by using cubic law for laminar flow through parallel plates. Initiation, growth and coalescence of the microcracks will lead to the generation of macroscopic fractures, which is explicitly mimicked by failure and removal of bonds between particles from the discrete element network. We investigate the fracture propagation path in both homogeneous and heterogeneous reservoirs using the simulator developed. Stress shadow caused by the transverse fracture will change the orientation of principal stress in the fracture neighborhood, which may inhibit or alter the growth direction of nearby fracture clusters. However, the initial in-situ stress anisotropy often helps overcome this phenomenon. Under large in-situ stress anisotropy, the hydraulic fractures are more likely to propagate in a direction that is perpendicular to the minimum horizontal stress. Under small in-situ stress anisotropy, there is a greater chance for fractures from nearby clusters to merge with each other. Then, we examine the differences in fracture geometry caused by fracturing in cemented or uncemented wellbore. Moreover, the impact of

  4. Secretory Granule Membrane Protein Recycles Through Multivesicular Bodies

    PubMed Central

    Bäck, Nils; Rajagopal, Chitra; Mains, Richard E.; Eipper, Betty A.

    2010-01-01

    The recycling of secretory granule membrane proteins that reach the plasma membrane following exocytosis is poorly understood. As a model, peptidylglycine α-amidating monooxygenase (PAM), a granule membrane protein that catalyzes a final step in peptide processing was examined. Ultrastructural analysis of antibody internalized by PAM and surface biotinylation demonstrated efficient return of plasma membrane PAM to secretory granules. Electron microscopy revealed the rapid movement of PAM from early endosomes to the limiting membranes of multivesicular bodies and then into intralumenal vesicles. Wheat germ agglutinin and PAM antibody internalized simultaneously were largely segregated when they reached multivesicular bodies. Mutation of basally phosphorylated residues (Thr946, Ser949) in the cytoplasmic domain of PAM to Asp (TS/DD) substantially slowed its entry into intralumenal vesicles. Mutation of the same sites to Ala (TS/AA) facilitated the entry of internalized PAM into intralumenal vesicles and its subsequent return to secretory granules. Entry of PAM into intralumenal vesicles is also associated with a juxtamembrane endoproteolytic cleavage that releases a 100 kDa soluble PAM fragment that can be returned to secretory granules. Controlled entry into the intralumenal vesicles of multivesicular bodies plays a key role in the recycling of secretory granule membrane proteins. PMID:20374556

  5. Stress modulates intestinal secretory immunoglobulin A

    PubMed Central

    Campos-Rodríguez, Rafael; Godínez-Victoria, Marycarmen; Abarca-Rojano, Edgar; Pacheco-Yépez, Judith; Reyna-Garfias, Humberto; Barbosa-Cabrera, Reyna Elizabeth; Drago-Serrano, Maria Elisa

    2013-01-01

    Stress is a response of the central nervous system to environmental stimuli perceived as a threat to homeostasis. The stress response triggers the generation of neurotransmitters and hormones from the hypothalamus pituitary adrenal axis, sympathetic axis and brain gut axis, and in this way modulates the intestinal immune system. The effects of psychological stress on intestinal immunity have been investigated mostly with the restraint/immobilization rodent model, resulting in an up or down modulation of SIgA levels depending on the intensity and time of exposure to stress. SIgA is a protein complex formed by dimeric (dIgA) or polymeric IgA (pIgA) and the secretory component (SC), a peptide derived from the polymeric immunoglobulin receptor (pIgR). The latter receptor is a transmembrane protein expressed on the basolateral side of gut epithelial cells, where it uptakes dIgA or pIgA released by plasma cells in the lamina propria. As a result, the IgA-pIgR complex is formed and transported by vesicles to the apical side of epithelial cells. pIgR is then cleaved to release SIgA into the luminal secretions of gut. Down modulation of SIgA associated with stress can have negative repercussions on intestinal function and integrity. This can take the form of increased adhesion of pathogenic agents to the intestinal epithelium and/or an altered balance of inflammation leading to greater intestinal permeability. Most studies on the molecular and biochemical mechanisms involved in the stress response have focused on systemic immunity. The present review analyzes the impact of stress (mostly by restraint/immobilization, but also with mention of other models) on the generation of SIgA, pIgR and other humoral and cellular components involved in the intestinal immune response. Insights into these mechanisms could lead to better therapies for protecting against pathogenic agents and avoiding epithelial tissue damage by modulating intestinal inflammation. PMID:24348350

  6. Effects of nonthermal distribution of electrons and polarity of net dust-charge number density on nonplanar dust-ion-acoustic solitary waves.

    PubMed

    Mamun, A A; Shukla, P K

    2009-09-01

    Effects of the nonthermal distribution of electrons as well as the polarity of the net dust-charge number density on nonplanar (viz. cylindrical and spherical) dust-ion-acoustic solitary waves (DIASWs) are investigated by employing the reductive perturbation method. It is found that the basic features of the DIASWs are significantly modified by the effects of nonthermal electron distribution, polarity of net dust-charge number density, and nonplanar geometry. The implications of our results in some space and laboratory dusty plasma environments are briefly discussed.

  7. A Comprehensive Model for Real Gas Transport in Shale Formations with Complex Non-planar Fracture Networks

    NASA Astrophysics Data System (ADS)

    Yang, Ruiyue; Huang, Zhongwei; Yu, Wei; Li, Gensheng; Ren, Wenxi; Zuo, Lihua; Tan, Xiaosi; Sepehrnoori, Kamy; Tian, Shouceng; Sheng, Mao

    2016-11-01

    A complex fracture network is generally generated during the hydraulic fracturing treatment in shale gas reservoirs. Numerous efforts have been made to model the flow behavior of such fracture networks. However, it is still challenging to predict the impacts of various gas transport mechanisms on well performance with arbitrary fracture geometry in a computationally efficient manner. We develop a robust and comprehensive model for real gas transport in shales with complex non-planar fracture network. Contributions of gas transport mechanisms and fracture complexity to well productivity and rate transient behavior are systematically analyzed. The major findings are: simple planar fracture can overestimate gas production than non-planar fracture due to less fracture interference. A “hump” that occurs in the transition period and formation linear flow with a slope less than 1/2 can infer the appearance of natural fractures. The sharpness of the “hump” can indicate the complexity and irregularity of the fracture networks. Gas flow mechanisms can extend the transition flow period. The gas desorption could make the “hump” more profound. The Knudsen diffusion and slippage effect play a dominant role in the later production time. Maximizing the fracture complexity through generating large connected networks is an effective way to increase shale gas production.

  8. A Comprehensive Model for Real Gas Transport in Shale Formations with Complex Non-planar Fracture Networks

    PubMed Central

    Yang, Ruiyue; Huang, Zhongwei; Yu, Wei; Li, Gensheng; Ren, Wenxi; Zuo, Lihua; Tan, Xiaosi; Sepehrnoori, Kamy; Tian, Shouceng; Sheng, Mao

    2016-01-01

    A complex fracture network is generally generated during the hydraulic fracturing treatment in shale gas reservoirs. Numerous efforts have been made to model the flow behavior of such fracture networks. However, it is still challenging to predict the impacts of various gas transport mechanisms on well performance with arbitrary fracture geometry in a computationally efficient manner. We develop a robust and comprehensive model for real gas transport in shales with complex non-planar fracture network. Contributions of gas transport mechanisms and fracture complexity to well productivity and rate transient behavior are systematically analyzed. The major findings are: simple planar fracture can overestimate gas production than non-planar fracture due to less fracture interference. A “hump” that occurs in the transition period and formation linear flow with a slope less than 1/2 can infer the appearance of natural fractures. The sharpness of the “hump” can indicate the complexity and irregularity of the fracture networks. Gas flow mechanisms can extend the transition flow period. The gas desorption could make the “hump” more profound. The Knudsen diffusion and slippage effect play a dominant role in the later production time. Maximizing the fracture complexity through generating large connected networks is an effective way to increase shale gas production. PMID:27819349

  9. Separation of rat pituitary secretory granules by continuous flow electrophoresis

    NASA Technical Reports Server (NTRS)

    Hayes, Daniel; Exton, Carrie; Salada, Thomas; Shellenberger, Kathy; Waddle, Jenny; Hymer, W. C.

    1990-01-01

    The separation of growth hormone-containing cytoplasmic secretory granules from the rat pituitary gland by continuous flow electrophoresis is described. The results are consistent with the hypothesis that granule subpopulations can be separated due to differences in surface charge; these, in turn, may be related to the oligomeric state of the hormone.

  10. Fine structure of the Caenorhabditis elegans secretory-excretory system.

    PubMed

    Nelson, F K; Albert, P S; Riddle, D L

    1983-02-01

    The secretory-excretory system of C. elegans, reconstructed from serial-section electron micrographs of larvae, is composed of four cells, the nuclei of which are located on the ventral side of the pharynx and adjacent intestine. (1) The pore cell encloses the terminal one-third of the excretory duct which leads to an excretory pore at the ventral midline. (2) The duct cell surrounds the excretory duct with a lamellar membrane from the origin of the duct at the excretory sinus to the pore cell boundary. (3) A large H-shaped excretory cell extends bilateral canals anteriorly and posteriorly nearly the entire length of the worm. The excretory sinus within the cell body joins the lumena of the canals with the origin of the duct. (4) A binucleate, A-shaped gland cell extends bilateral processes anteriorly from cell bodies located just behind the pharynx. These processes are fused at the anterior tip of the cell, where the cell enters the circumpharyngeal nerve ring. The processes are also joined at the anterior edge of the excretory cell body, where the excretory cell and gland are joined to the duct cell at the origin of the duct. Secretory granules may be concentrated in the gland near this secretory-excretory junction. Although the gland cells of all growing developmental stages stain positively with paraldehyde-fuchsin, the gland of the dauer larva stage (a developmentally arrested third-stage larva) does not stain, nor do glands of starved worms of other stages. Dauer larvae uniquely lack secretory granules, and the gland cytoplasm is displaced by a labyrinth of large, transparent spaces. Exit from the dauer stage results in the return of active secretory morphology in fourth-stage larvae.

  11. Resolving a Long-Standing Ambiguity: the Non-Planarity of gauche-1,3-BUTADIENE Revealed by Microwave Spectroscopy

    NASA Astrophysics Data System (ADS)

    Martin-Drumel, Marie-Aline; McCarthy, Michael C.; Patterson, David; Eibenberger, Sandra; Buckingham, Grant; Baraban, Joshua H.; Ellison, Barney; Stanton, John F.

    2016-06-01

    The preferred conformation of cis-1,3-butadiene (CH_2=CH-CH=CH_2) has been of long-standing importance in organic chemistry because of its role in Diels-Alder transition states. The molecule could adopt a planar s-cis conformation, in favor of conjugations in the carbon chain, or a non-planar gauche conformation, as a result of steric interactions between the terminal H atoms. To resolve this ambiguity, we have now measured the pure rotational spectrum of this isomer in the microwave region, unambiguously establishing a significant inertial defect, and therefore a gauche conformation. Experimental measurements of gauche-1,3-butadiene and several of its isotopologues using cavity Fourier-transform microwave (FTMW) spectroscopy in a supersonic expansion and chirped-pulse FTMW spectroscopy in a 4 K buffer gas cell will be summarized, as will new quantum chemical calculations.

  12. Comment on a spurious prediction of a non-planar geometry for benzene at the MP2 level of theory

    NASA Astrophysics Data System (ADS)

    Samala, Nagaprasad Reddy; Jordan, Kenneth D.

    2017-02-01

    MP2 calculations with the full aug-cc-pVTZ basis set give a non-planar structure for benzene. Although this non-physical result can be avoided by using the smaller aug-cc-pVDZ basis set or by scaling or deleting selected functions from the aug-cc-pVTZ basis set, such changes to the basis set can result in calculated values of the frequencies of the b2g out-of-plane vibrations that are considerably underestimated. The origin of this behavior is traced to linear dependency problems with the aug-cc-pVDZ and aug-cc-pVTZ basis sets when used for benzene.

  13. Analytical study of STOL Aircraft in ground effect. Part 1: Nonplanar, nonlinear wing/jet lifting surface method

    NASA Technical Reports Server (NTRS)

    Shollenberger, C. A.; Smyth, D. N.

    1978-01-01

    A nonlinear, nonplanar three dimensional jet flap analysis, applicable to the ground effect problem, is presented. Lifting surface methodology is developed for a wing with arbitrary planform operating in an inviscid and incompressible fluid. The classical, infintely thin jet flap model is employed to simulate power induced effects. An iterative solution procedure is applied within the analysis to successively approximate the jet shape until a converged solution is obtained which closely satisfies jet and wing boundary conditions. Solution characteristics of the method are discussed and example results are presented for unpowered, basic powered and complex powered configurations. Comparisons between predictions of the present method and experimental measurements indicate that the improvement of the jet with the ground plane is important in the analyses of powered lift systems operating in ground proximity. Further development of the method is suggested in the areas of improved solution convergence, more realistic modeling of jet impingement and calculation efficiency enhancements.

  14. The influence of non-planar geometry on the flow within a distal end-to-side anastomosis

    NASA Astrophysics Data System (ADS)

    Sherwin, S. J.; Doorly, D. J.; Peiro, J.; Caro, C. G.

    1998-11-01

    The pattern of the flow in arteries is strongly influenced by the three-dimensional shape of the geometry. Curvature and torsion of the wall geometry alters the axial velocity distribution, and introduces cross flow velocity components. In this investigation we have considered flow in a model geometry of a fully occluded 45^o distal end-to-side anastomosis. Previous investigations have typically focused on planar end-to-side anastomoses where the bypass and host vessels have a plane of symmetry. We have increased the complexity of the model by considering a non-planar geometry produced by deforming the bypass vessel out of the plane of symmetry. The flows have been numerically and experimentally investigated using a spectral/hp element algorithm and magnetic resonance imaging. The significant effect of the non-planar geometry is to introduce a bulk rotation of the two secondary flow cells present in flow within a planar geometry. A reduction in wall shear stress is observed at the bed of the anastomosis and a larger absolute flux of velocity is seen within the occluded region proximal to the anastomosis. Current investigations have considered the role of pulsatility in the form of a non-reversing sinusoidal oscillation. In this case a separation bubble, not present in the steady case, is seen at the toe of the anastomosis during the systolic part of the cycle. The role of geometry and pulsatility on particle motion has also been addressed with a view to determining the shear exposure on particle within these types of flows.

  15. Secretory function in subplate neurons during cortical development

    PubMed Central

    Kondo, Shinichi; Al-Hasani, Hannah; Hoerder-Suabedissen, Anna; Wang, Wei Zhi; Molnár, Zoltán

    2015-01-01

    Subplate cells are among the first generated neurons in the mammalian cerebral cortex and have been implicated in the establishment of cortical wiring. In rodents some subplate neurons persist into adulthood. Here we would like to highlight several converging findings which suggest a novel secretory function of subplate neurons during cortical development. Throughout the postnatal period in rodents, subplate neurons have highly developed rough endoplasmic reticulum (ER) and are under an ER stress condition. By comparing gene expression between subplate and layer 6, we found that several genes encoding secreted proteins are highly expressed in subplate neurons. One of these secreted proteins, neuroserpin, encoded by the serpini1 gene, is localized to the ER in subplate cells. We propose that subplate might influence cortical circuit formation through a transient secretory function. PMID:25859180

  16. [Mammary analog secretory carcinoma of the parotid gland].

    PubMed

    Guérin, Maxime; Diedhiou, Abdoulaye; Nallet, Emmanuel; Duflo, Suzy; Laé, Marick; Wassef, Michel

    2014-10-01

    Mammary analog secretory carcinoma (MASC) of the parotid gland is a rare and recently described lesion. We report the case of a 46-year-old man with a tumor of the parotid gland which was carried to the diagnosis of MASC. Diagnostic was confirmed by highlighting the ETV6-NTRK3 gene translocation. However, some morphologic and immunohistochemical features are suggestive of this entity. This carcinoma should be distinguished from its main differential diagnoses: acinic cell carcinoma and low grade cribriform cystadenocarcinoma.

  17. Neuroendocrine secretory protein 7B2: structure, expression and functions.

    PubMed Central

    Mbikay, M; Seidah, N G; Chrétien, M

    2001-01-01

    7B2 is an acidic protein residing in the secretory granules of neuroendocrine cells. Its sequence has been elucidated in many phyla and species. It shows high similarity among mammals. A Pro-Pro-Asn-Pro-Cys-Pro polyproline motif is its most conserved feature, being carried by both vertebrate and invertebrate sequences. It is biosynthesized as a precursor protein that is cleaved into an N-terminal fragment and a C-terminal peptide. In neuroendocrine cells, 7B2 functions as a specific chaperone for the proprotein convertase (PC) 2. Through the sequence around its Pro-Pro-Asn-Pro-Cys-Pro motif, it binds to an inactive proPC2 and facilitates its transport from the endoplasmic reticulum to later compartments of the secretory pathway where the zymogen is proteolytically matured and activated. Its C-terminal peptide can inhibit PC2 in vitro and may contribute to keep the enzyme transiently inactive in vivo. The PC2-7B2 model defines a new neuroendocrine paradigm whereby proteolytic activation of prohormones and proneuropeptides in the secretory pathway is spatially and temporally regulated by the dynamics of interactions between converting enzymes and their binding proteins. Interestingly, unlike PC2-null mice, which are viable, 7B2-null mutants die early in life from Cushing's disease due to corticotropin ('ACTH') hypersecretion by the neurointermediate lobe, suggesting a possible involvement of 7B2 in secretory granule formation and in secretion regulation. The mechanism of this regulation is yet to be elucidated. 7B2 has been shown to be a good marker of several neuroendocrine cell dysfunctions in humans. The possibility that anomalies in its structure and expression could be aetiological causes of some of these dysfunctions warrants investigation. PMID:11439082

  18. Secretory pattern and regulatory mechanism of growth hormone in cattle

    PubMed Central

    2016-01-01

    Abstract The ultradian rhythm of growth hormone (GH) secretion has been known in several animal species for years and has recently been observed in cattle. Although the physiological significance of the rhythm is not yet fully understood, it appears essential for normal growth. In this review, previous studies concerning the GH secretory pattern in cattle, including its ultradian rhythm, are introduced and the regulatory mechanism is discussed on the basis of recent findings. PMID:26260675

  19. Isolation of intact sub-dermal secretory cavities from Eucalyptus

    PubMed Central

    2010-01-01

    Background The biosynthesis of plant natural products in sub-dermal secretory cavities is poorly understood at the molecular level, largely due to the difficulty of physically isolating these structures for study. Our aim was to develop a protocol for isolating live and intact sub-dermal secretory cavities, and to do this, we used leaves from three species of Eucalyptus with cavities that are relatively large and rich in essential oils. Results Leaves were digested using a variety of commercially available enzymes. A pectinase from Aspergillus niger was found to allow isolation of intact cavities after a relatively short incubation (12 h), with no visible artifacts from digestion and no loss of cellular integrity or cavity contents. Several measurements indicated the potential of the isolated cavities for further functional studies. First, the cavities were found to consume oxygen at a rate that is comparable to that estimated from leaf respiratory rates. Second, mRNA was extracted from cavities, and it was used to amplify a cDNA fragment with high similarity to that of a monoterpene synthase. Third, the contents of the cavity lumen were extracted, showing an unexpectedly low abundance of volatile essential oils and a sizeable amount of non-volatile material, which is contrary to the widely accepted role of secretory cavities as predominantly essential oil repositories. Conclusions The protocol described herein is likely to be adaptable to a range of Eucalyptus species with sub-dermal secretory cavities, and should find wide application in studies of the developmental and functional biology of these structures, and the biosynthesis of the plant natural products they contain. PMID:20807444

  20. Altered secretory immunoglobulin A on skin surface after intensive exercise.

    PubMed

    Eda, Nobuhiko; Shimizu, Kazuhiro; Suzuki, Satomi; Tanabe, Yoko; Lee, Eunjae; Akama, Takao

    2013-09-01

    The aim of this study was to determine the effects of high-intensity endurance exercise on skin immunity by estimating secretory immunoglobulin A (SIgA) and staphylococci on skin surface. Seven healthy adult men (age, 22.3 ± 2.0 years) performed bicycle exercise at 75% HRmax for 60 minutes from 2030 to 2130 hours. Secretory immunoglobulin A was obtained from 1 ml extraction liquids stirred with the microtube homogenizer in the open end of a polypropylene tube for 60 seconds. Secretory immunoglobulin A concentrations were measured using enzyme-linked immunosorbent assay. Staphylococci were harvested by pressed agar-based media against skin surface. Skin surface samples were collected from the chest and the forearm on the first day at 2030 hours (before rest, A1), 2130 hours (after rest, A2), and 2230 hours (after showering, A3); the next morning at 0700 hours (A4); on the second day at 2030 hours (before exercise, B1), 2130 hours (after exercise, B2), and 2230 hours (after showering, B3); and the next morning at 0700 hours (B4). Secretory immunoglobulin A concentration on the forearm was significantly lower at B2 (p < 0.05) and B3 (p < 0.05) than that at B1 and that on the chest at B1 tended to be higher compared with B2 (p = 0.084) and B3 (p = 0.075). The number of staphylococci was significantly higher at B2 than that at B1 (p < 0.01) and B4 (p < 0.01) on the forearm. We conclude that high-intensity endurance exercise might depress immune function and enhance infectious risk on skin surface. Coaches should encourage their athletes to take a shower and change into clean clothes immediately after sports activities and athletes should maintain a clean skin surface to decrease the infectious risk on skin surface.

  1. Secretory pattern and regulatory mechanism of growth hormone in cattle.

    PubMed

    Kasuya, Etsuko

    2016-02-01

    The ultradian rhythm of growth hormone (GH) secretion has been known in several animal species for years and has recently been observed in cattle. Although the physiological significance of the rhythm is not yet fully understood, it appears essential for normal growth. In this review, previous studies concerning the GH secretory pattern in cattle, including its ultradian rhythm, are introduced and the regulatory mechanism is discussed on the basis of recent findings.

  2. RNAi knockdown of parafusin inhibits the secretory pathway.

    PubMed

    Liu, Li; Wyroba, Elzbieta; Satir, Birgit H

    2011-10-01

    Several glycolytic enzymes and their isoforms have been found to be important in cell signaling unrelated to glycolysis. The involvement of parafusin (PFUS), a member of the phosphoglucomutase (PGM) superfamily with no phosphoglucomutase activity, in Ca(2+)-dependent exocytosis has been controversial. This protein was first described in Paramecium tetraurelia, but is widely found. Earlier work showed that parafusin is a secretory vesicle scaffold component with unusual post-translational modifications (cyclic phosphorylation and phosphoglucosylation) coupled to stages in the exocytic process. Using RNAi, we demonstrate that parafusin synthesis can be reversibly blocked, with minor or no effect on other PGM isoforms. PFUS knockdown produces an inhibition of dense core secretory vesicle (DCSV) synthesis leading to an exo(-) phenotype. Although cell growth is unaffected, vesicle content is not packaged properly and no new DCSVs are formed. We conclude that PFUS and its orthologs are necessary for proper scaffold maturation. Because of this association, parafusin is an important signaling component for regulatory control of the secretory pathway.

  3. Observations of Calcium Dynamics in Cortical Secretory Vesicles

    PubMed Central

    Raveh, Adi; Valitsky, Michael; Shani, Liora; Coorssen, Jens R.; Blank, Paul S.; Zimmerberg, Joshua; Rahamimoff, Rami

    2012-01-01

    SUMMARY Calcium (Ca2+) dynamics were evaluated in fluorescently labeled sea urchin secretory vesicles using confocal microscopy. 71% of the vesicles examined exhibited one or more transient increases in the fluorescence signal that was damped in time. The detection of transient increases in signal was dependent upon the affinity of the fluorescence indicator; the free Ca2+ concentration in the secretory vesicles was estimated to be in the range of ~10 – 100 μM. Non-linear stochastic analysis revealed the presence of extra variance in the Ca2+ dependent fluorescence signal. This noise process increased linearly with the amplitude of the Ca2+ signal. Both the magnitude and spatial properties of this noise process were dependent upon the activity of vesicle p-type (Cav2.1) Ca2+ channels. Blocking the p-type Ca2+ channels with ω-agatoxin decreased signal variance, and altered the spatial noise pattern within the vesicle. These fluorescence signal properties are consistent with vesicle Ca2+ dynamics and not simply due to obvious physical properties such as gross movement artifacts or pH driven changes in Ca2+ indicator fluorescence. The results suggest that the free Ca2+ content of cortical secretory vesicles is dynamic; this property may modulate the exocytotic fusion process. PMID:22831912

  4. Unremitting Cell Proliferation in the Secretory Phase of Eutopic Endometriosis

    PubMed Central

    Franco-Murillo, Yanira; Miranda-Rodríguez, José Antonio; Rendón-Huerta, Erika; Montaño, Luis F.; Cornejo, Gerardo Velázquez; Gómez, Lucila Poblano; Valdez-Morales, Francisco Javier; Gonzalez-Sanchez, Ignacio

    2014-01-01

    Objective: Endometriosis is linked to altered cell proliferation and stem cell markers c-kit/stem cell factor (SCF) in ectopic endometrium. Our aim was to investigate whether c-kit/SCF also plays a role in eutopic endometrium. Design: Eutopic endometrium obtained from 35 women with endometriosis and 25 fertile eumenorrheic women was analyzed for in situ expression of SCF/c-kit, Ki67, RAC-alpha serine/threonine-protein kinase (Akt), phosphorylated RAC-alpha serine/threonin-protein kinase (pAkt), Glycogen synthase kinase 3 beta (GSK3β), and phosphorylated glycogen synthase kinase 3 beta (pGSK3β), throughout the menstrual cycle. Results: Expression of Ki67 and SCF was higher in endometriosis than in control tissue (P < .05) and greater in secretory rather than proliferative (P < .01) endometrium in endometriosis. Expression of c-kit was also higher in endometriosis although similar in both phases. Expression of Akt and GSK3β was identical in all samples and cycle phases, whereas pAkt and pGSK3β, opposed to control tissue, remained overexpressed in the secretory phase in endometriosis. Conclusion: Unceasing cell proliferation in the secretory phase of eutopic endometriosis is linked to deregulation of c-kit/SCF-associated signaling pathways. PMID:25194152

  5. Reversible condensation of mast cell secretory products in vitro.

    PubMed Central

    Fernandez, J M; Villalón, M; Verdugo, P

    1991-01-01

    We have investigated the mechanisms responsible for the condensation and decondensation of secretory products that occur in mast cell secretion. We show here that the hydrated matrix of an exocytosed secretory granule can be recondensed to its original volume by exposure to acidic solutions containing histamine at concentrations that mimic those found in vivo. Recondensation by acidic histamine began in the range of 1-10 mM with a dose response curve that was accurately predicted by a Hill type equation with four highly cooperative binding sites and a half maximum concentration of [Hi++] = 3.9 mM. Recondensation by histamine showed a sigmoidal dependency on pH (critical range pH 5.5-6.5) and was fully reversible. These experiments suggest that histamine, possibly by binding to anionic sites in the protein-heparin complex of the granule matrix, triggers a change in the polymeric structures of the granule matrix from an extended coil to a collapsed globular state. This may be a useful model for understanding the condensation of secretory products into dense core granules and their subsequent decondensation upon exocytosis. Images FIGURE 1 FIGURE 4 PMID:1868152

  6. Souffle/Spastizin controls secretory vesicle maturation during zebrafish oogenesis.

    PubMed

    Kanagaraj, Palsamy; Gautier-Stein, Amandine; Riedel, Dietmar; Schomburg, Christoph; Cerdà, Joan; Vollack, Nadine; Dosch, Roland

    2014-06-01

    During oogenesis, the egg prepares for fertilization and early embryogenesis. As a consequence, vesicle transport is very active during vitellogenesis, and oocytes are an outstanding system to study regulators of membrane trafficking. Here, we combine zebrafish genetics and the oocyte model to identify the molecular lesion underlying the zebrafish souffle (suf) mutation. We demonstrate that suf encodes the homolog of the Hereditary Spastic Paraplegia (HSP) gene SPASTIZIN (SPG15). We show that in zebrafish oocytes suf mutants accumulate Rab11b-positive vesicles, but trafficking of recycling endosomes is not affected. Instead, we detect Suf/Spastizin on cortical granules, which undergo regulated secretion. We demonstrate genetically that Suf is essential for granule maturation into secretion competent dense-core vesicles describing a novel role for Suf in vesicle maturation. Interestingly, in suf mutants immature, secretory precursors accumulate, because they fail to pinch-off Clathrin-coated buds. Moreover, pharmacological inhibition of the abscission regulator Dynamin leads to an accumulation of immature secretory granules and mimics the suf phenotype. Our results identify a novel regulator of secretory vesicle formation in the zebrafish oocyte. In addition, we describe an uncharacterized cellular mechanism for Suf/Spastizin activity during secretion, which raises the possibility of novel therapeutic avenues for HSP research.

  7. Souffle/Spastizin Controls Secretory Vesicle Maturation during Zebrafish Oogenesis

    PubMed Central

    Riedel, Dietmar; Schomburg, Christoph; Cerdà, Joan; Vollack, Nadine; Dosch, Roland

    2014-01-01

    During oogenesis, the egg prepares for fertilization and early embryogenesis. As a consequence, vesicle transport is very active during vitellogenesis, and oocytes are an outstanding system to study regulators of membrane trafficking. Here, we combine zebrafish genetics and the oocyte model to identify the molecular lesion underlying the zebrafish souffle (suf) mutation. We demonstrate that suf encodes the homolog of the Hereditary Spastic Paraplegia (HSP) gene SPASTIZIN (SPG15). We show that in zebrafish oocytes suf mutants accumulate Rab11b-positive vesicles, but trafficking of recycling endosomes is not affected. Instead, we detect Suf/Spastizin on cortical granules, which undergo regulated secretion. We demonstrate genetically that Suf is essential for granule maturation into secretion competent dense-core vesicles describing a novel role for Suf in vesicle maturation. Interestingly, in suf mutants immature, secretory precursors accumulate, because they fail to pinch-off Clathrin-coated buds. Moreover, pharmacological inhibition of the abscission regulator Dynamin leads to an accumulation of immature secretory granules and mimics the suf phenotype. Our results identify a novel regulator of secretory vesicle formation in the zebrafish oocyte. In addition, we describe an uncharacterized cellular mechanism for Suf/Spastizin activity during secretion, which raises the possibility of novel therapeutic avenues for HSP research. PMID:24967841

  8. In vitro aggregation of the regulated secretory protein chromogranin A.

    PubMed Central

    Jain, Renu K; Chang, Wen Tzu; Geetha, Chitta; Joyce, Paul B M; Gorr, Sven-Ulrik

    2002-01-01

    Aggregation chaperones, consisting of secretory proteins that contain a hexa-histidine epitope tag, enhance the calcium-induced aggregation of regulated secretory proteins and their sorting to secretory granules. The goal of this study was to gain a better understanding of this unusual aggregation mechanism. Hexa-histidine-epitope-tagged secreted alkaline phosphatase, an aggregation chaperone, enhanced the in vitro aggregation of chromogranin A in the presence of calcium, but not in the presence of magnesium or other divalent cations. As an exception, chromogranin was completely aggregated by zinc, even in the absence of the aggregation chaperone. In addition, fluorescence spectroscopy of the aggregation reaction mixture showed an increase in fluorescence intensity consistent with the formation of protein aggregates. The calcium-induced aggregation of chromogranin A was completely inhibited by 0.2% Triton X-100, suggesting that it involves hydrophobic interactions. In contrast, the detergent did not affect chaperone-enhanced aggregation, suggesting that this aggregation does not depend on hydrophobic interactions. EDTA-treated chaperone did not enhance chromogranin A aggregation, indicating that divalent cations are necessary for chaperone action. Although the structure of the aggregation chaperone was not important, the size of the chaperone was. Thus the free His-hexapeptide could not substitute for the aggregation chaperone. Based on these results, we propose that the hexa-histidine tag, in the context of a polypeptide, acts as a divalent cation-dependent nucleation site for chromogranin A aggregation. PMID:12175332

  9. Drug-induced secretory diarrhea: A role for CFTR.

    PubMed

    Moon, Changsuk; Zhang, Weiqiang; Sundaram, Nambirajan; Yarlagadda, Sunitha; Reddy, Vadde Sudhakar; Arora, Kavisha; Helmrath, Michael A; Naren, Anjaparavanda P

    2015-12-01

    Many medications induce diarrhea as a side effect, which can be a major obstacle to therapeutic efficacy and also a life-threatening condition. Secretory diarrhea can be caused by excessive fluid secretion in the intestine under pathological conditions. The cAMP/cGMP-regulated cystic fibrosis transmembrane conductance regulator (CFTR) is the primary chloride channel at the apical membrane of intestinal epithelial cells and plays a major role in intestinal fluid secretion and homeostasis. CFTR forms macromolecular complexes at discreet microdomains at the plasma membrane, and its chloride channel function is regulated spatiotemporally through protein-protein interactions and cAMP/cGMP-mediated signaling. Drugs that perturb CFTR-containing macromolecular complexes in the intestinal epithelium and upregulate intracellular cAMP and/or cGMP levels can hyperactivate the CFTR channel, causing excessive fluid secretion and secretory diarrhea. Inhibition of CFTR chloride-channel activity may represent a novel approach to the management of drug-induced secretory diarrhea.

  10. P-selectin targeting to secretory lysosomes of Rbl-2H3 cells.

    PubMed

    Kaur, Jasber; Cutler, Daniel F

    2002-03-22

    The biogenesis of secretory lysosomes, which combine characteristics of both lysosomes and secretory granules, is currently of high interest. In particular, it is not clear whether delivery of membrane proteins to the secretory lysosome requires lysosomal, secretory granule, or some novel targeting determinants. Heterologous expression of P-selectin has established that this membrane protein contains targeting signals for both secretory granules and lysosomes. P-selectin is therefore an ideal probe with which to determine the signals required for targeting to secretory lysosomes. We have exploited subcellular fractionation and immunofluorescence microscopy to monitor targeting of transiently expressed wild-type and mutant horseradish peroxidase (HRP)-P-selectin chimeras to secretory lysosomes of Rbl-2H3 cells. The exposure of the HRP chimeras to intracellular proteolysis was also determined as a third monitor of secretory lysosome targeting. Our data show that HRP-P-selectin accumulates in secretory lysosomes of Rbl-2H3 cells using those cytoplasmic sequences previously found to be sufficient for targeting to conventional lysosomes. This work highlights the similar sorting signals used for targeting of membrane proteins to conventional lysosomes and secretory lysosomes.

  11. BPIFB6 Regulates Secretory Pathway Trafficking and Enterovirus Replication

    PubMed Central

    Morosky, Stefanie; Lennemann, Nicholas J.

    2016-01-01

    ABSTRACT Bactericidal/permeability-increasing protein (BPI) fold-containing family B, member 3 (BPIFB3) is an endoplasmic reticulum (ER)-localized host factor that negatively regulates coxsackievirus B (CVB) replication through its control of the autophagic pathway. Here, we show that another member of the BPIFB family, BPIFB6, functions as a positive regulator of CVB, and other enterovirus, replication by controlling secretory pathway trafficking and Golgi complex morphology. We show that similar to BPIFB3, BPIFB6 localizes exclusively to the ER, where it associates with other members of the BPIFB family. However, in contrast to our findings that RNA interference (RNAi)-mediated silencing of BPIFB3 greatly enhances CVB replication, we show that silencing of BPIFB6 expression dramatically suppresses enterovirus replication in a pan-viral manner. Mechanistically, we show that loss of BPIFB6 expression induces pronounced alterations in retrograde and anterograde trafficking, which correlate with dramatic fragmentation of the Golgi complex. Taken together, these data implicate BPIFB6 as a key regulator of secretory pathway trafficking and viral replication and suggest that members of the BPIFB family participate in diverse host cell functions to regulate virus infections. IMPORTANCE Enterovirus infections are associated with a number of severe pathologies, such as aseptic meningitis, dilated cardiomyopathy, type I diabetes, paralysis, and even death. These viruses, which include coxsackievirus B (CVB), poliovirus (PV), and enterovirus 71 (EV71), co-opt the host cell secretory pathway, which controls the transport of proteins from the endoplasmic reticulum to the Golgi complex, to facilitate their replication. Here we report on the identification of a novel regulator of the secretory pathway, bactericidal/permeability-increasing protein (BPI) fold-containing family B, member 6 (BPIFB6), whose expression is required for enterovirus replication. We show that loss of

  12. A senescence secretory switch mediated by PI3K/AKT/mTOR activation controls chemoprotective endothelial secretory responses

    PubMed Central

    Bent, Eric H.; Gilbert, Luke A.; Hemann, Michael T.

    2016-01-01

    Cancer therapy targets malignant cells that are surrounded by a diverse complement of nonmalignant stromal cells. Therapy-induced damage of normal cells can alter the tumor microenvironment, causing cellular senescence and activating cancer-promoting inflammation. However, how these damage responses are regulated (both induced and resolved) to preserve tissue homeostasis and prevent chronic inflammation is poorly understood. Here, we detail an acute chemotherapy-induced secretory response that is self-limiting in vitro and in vivo despite the induction of cellular senescence. We used tissue-specific knockout mice to demonstrate that endothelial production of the proinflammatory cytokine IL-6 promotes chemoresistance and show that the chemotherapeutic doxorubicin induces acute IL-6 release through reactive oxygen species-mediated p38 activation in vitro. Doxorubicin causes endothelial senescence but, surprisingly, without a typical senescence secretory response. We found that endothelial cells repress senescence-associated inflammation through the down-regulation of PI3K/AKT/mTOR signaling and that reactivation of this pathway restores senescence-associated inflammation. Thus, we describe a mechanism by which damage-associated paracrine secretory responses are restrained to preserve tissue homeostasis and prevent chronic inflammation. PMID:27566778

  13. Synthesis and processing strategies to tune the film structure and optoelectronic properties of non-planar molecular semiconductors

    NASA Astrophysics Data System (ADS)

    Hiszpanski, Anna Maria

    Molecular semiconductors have generated significant interest for their potential use in lightweight and mechanically flexible electronic devices. Yet, predicting how new molecular semiconductors will perform in devices remains a challenge because devices are comprised of polycrystalline thin films of molecular semiconductors, and charge transport in these films depends greatly on the details of their microstructure whose heterogeneities can span multiple length scales. The microstructure typically evolves during deposition, and thus developing organic electronics not only hinges on the success of materials discovery, but also on the ability to fine-tune deposition and processing parameters to access the thin-film structure most conducive for charge transport. This thesis explores chemical modification of a non-planar organic semiconductor, contorted hexabenzocoronene, cHBC, to tune its optoelectronic properties and processing strategies to induce structural changes in thin films. We primarily explore fluorine- and chlorine-substitution at the peripheral aromatic rings of cHBC to lower its energy levels and optical bandgap, and we demonstrate such halogenated derivatives as electron acceptors in organic solar cells. Substitution with these larger atoms also increases cHBC's intramolecular steric hindrance, providing access to an alternative molecular conformation with an order of magnitude higher solubility and systematic shifts in absorption and emission characteristics. cHBC's non-planarity provides an added dimension of tunability as it frustrates crystallization during deposition, producing amorphous films that can be subsequently crystallized with post-deposition processing. Decoupling structural development from deposition allows us to fabricate transistors from differently treated cHBC films and elucidate the effects of changes in film structure on charge transport, as measured by the field-effect mobility. With different processing, the extent of c

  14. Room temperature single longitudinal mode laser output at 1645 nm from a laser-diode pumped Er:YAG nonplanar ring oscillator.

    PubMed

    Yao, B Q; Yu, X; Liu, X L; Duan, X M; Ju, Y L; Wang, Y Z

    2013-04-08

    We report on a monolithic 1645 nm Er:YAG nonplanar ring oscillator (NPRO) resonantly pumped by a fiber-coupled laser diode. In the experiment, an up to 550 mW single frequency laser output at 1645.2 nm was obtained, corresponding to a slope efficiency of 19.1% and an absolute efficiency of 6.0%. The beam quality M2 was measured to be 2.1 at the highest output power.

  15. HID-1 is required for homotypic fusion of immature secretory granules during maturation.

    PubMed

    Du, Wen; Zhou, Maoge; Zhao, Wei; Cheng, Dongwan; Wang, Lifen; Lu, Jingze; Song, Eli; Feng, Wei; Xue, Yanhong; Xu, Pingyong; Xu, Tao

    2016-10-18

    Secretory granules, also known as dense core vesicles, are generated at the trans-Golgi network and undergo several maturation steps, including homotypic fusion of immature secretory granules (ISGs) and processing of prehormones to yield active peptides. The molecular mechanisms governing secretory granule maturation are largely unknown. Here, we investigate a highly conserved protein named HID-1 in a mouse model. A conditional knockout of HID-1 in pancreatic β cells leads to glucose intolerance and a remarkable increase in the serum proinsulin/insulin ratio caused by defective proinsulin processing. Large volume three-dimensional electron microscopy and immunofluorescence imaging reveal that ISGs are much more abundant in the absence of HID-1. We further demonstrate that HID-1 deficiency prevented secretory granule maturation by blocking homotypic fusion of immature secretory granules. Our data identify a novel player during the early maturation of immature secretory granules.

  16. Structural stability, C--N internal rotations and vibrational spectral analysis of non-planar phenylurea and phenylthiourea.

    PubMed

    Badawi, Hassan M

    2009-04-01

    The structural stability and C-N internal rotations of phenylurea and phenylthiourea were investigated by DFT-B3LYP and ab initio MP2 and MP4//MP2 calculations with 6-311G** and/or 6-311+G** basis sets. The complex multirotor internal rotations in phenylurea and phenylthiourea were investigated at the B3LYP/6-311+G** level of theory from which several clear minima were predicted in the calculated potential energy scans of both molecules. For phenylurea two minima that correspond to non-planar- (CNCC dihedral angle of about 45 degrees ) cis (CNCO dihedral angle is near 0 degrees ) and trans (CNCO dihedral angle is near 180 degrees ) structures were predicted to have real frequency. For phenylthiourea only the non-planar-trans structure was predicted to be the low energy minimum for the molecule. The vibrational frequencies of the lowest energy non-planar-trans conformer of each of the two molecules were computed at the B3LYP level and tentative vibrational assignments were provided on the basis of normal coordinate analysis and experimental infrared and Raman data.

  17. Direct mapping of the electric permittivity of heterogeneous non-planar thin films at gigahertz frequencies by scanning microwave microscopy.

    PubMed

    Biagi, Maria Chiara; Badino, Giorgio; Fabregas, Rene; Gramse, Georg; Fumagalli, Laura; Gomila, Gabriel

    2017-02-01

    We obtained maps of electric permittivity at ∼19 GHz frequencies on non-planar thin film heterogeneous samples by means of combined atomic force-scanning microwave microscopy (AFM-SMM). We show that the electric permittivity maps can be obtained directly from the capacitance images acquired in contact mode, after removing the topographic cross-talk effects. This result demonstrates the possibility of identifying the electric permittivity of different materials in a thin film sample irrespectively of their thickness by just direct imaging and processing. We show, in addition, that quantitative maps of the electric permittivity can be obtained with no need for any theoretical calculation or complex quantification procedures when the electric permittivity of one of the materials is known. To achieve these results the use of contact mode imaging is a key factor. For non-contact imaging modes the effects of local sample thickness and of the imaging distance make the interpretation of the capacitance images in terms of the electric permittivity properties of the materials much more complex. The present results represent a substantial contribution to the field of nanoscale microwave dielectric characterization of thin film materials with important implications for the characterization of novel 3D electronic devices and 3D nanomaterials.

  18. Structural stability, C-N internal rotations and vibrational spectral analysis of non-planar phenylurea and phenylthiourea

    NASA Astrophysics Data System (ADS)

    Badawi, Hassan M.

    2009-04-01

    The structural stability and C-N internal rotations of phenylurea and phenylthiourea were investigated by DFT-B3LYP and ab initio MP2 and MP4//MP2 calculations with 6-311G** and/or 6-311+G** basis sets. The complex multirotor internal rotations in phenylurea and phenylthiourea were investigated at the B3LYP/6-311+G** level of theory from which several clear minima were predicted in the calculated potential energy scans of both molecules. For phenylurea two minima that correspond to non-planar- (CNCC dihedral angle of about 45°) cis (CNCO dihedral angle is near 0°) and trans (CNCO dihedral angle is near 180°) structures were predicted to have real frequency. For phenylthiourea only the non- planar- trans structure was predicted to be the low energy minimum for the molecule. The vibrational frequencies of the lowest energy non-planar-trans conformer of each of the two molecules were computed at the B3LYP level and tentative vibrational assignments were provided on the basis of normal coordinate analysis and experimental infrared and Raman data.

  19. Optical 3D Deformation Measurement Utilizing Non-planar Surface for the Development of an “Intelligent Tire”

    NASA Astrophysics Data System (ADS)

    Matsuzaki, Ryosuke; Hiraoka, Naoki; Todoroki, Akira; Mizutani, Yoshihiro

    Intelligent tires, also known as smart tires, are equipped with sensors to monitor the strain of the interior surface and the rolling radius of tire, and are expected to improve the reliability of tires and tire control systems such as anti-lock braking systems (ABS). However, the high stiffness of an attached sensor like a strain gauge causes sensors to debond from the tire rubber. In the present study, a novel optical method is used for the concurrent monitoring of in-plane strain and out-of-plane displacement (rolling radius) utilizing the non-planar surface of the monitoring object. The optical method enables noncontact measurement of strain distribution. The in-plane strain and out-of-plane displacement are calculated by using image processing with an image of the interior surface of a tire that is taken with a single CCD camera fixed on the wheel rim. This new monitoring system is applied to an aluminum beam and a commercially available radial tire. As a result, the monitoring system provides concurrent measurement of in-plane strain, out-of-plane displacement and tire pressure, and is shown to be an effective monitoring system for intelligent tires.

  20. Interaction of planar and nonplanar organic contaminants with coal fly ash: Effects of polar and nonpolar solvent solutions

    SciTech Connect

    Burgess, R.M.; Ryba, S.A.; Cantwell, M.G.; Gundersen, J.L.; Tien, R.; Perron, M.M.

    2006-08-15

    Coal fly ash has a very high sorption capacity for a variety of anthropogenic contaminants and has been used to cleanse wastewater of pollutants for approximately 40 years. Like other black carbons, the planar structure of the residual carbon in fly ash results in elevated affinities for planar organic contaminants, such as polycyclic aromatic hydrocarbons (PAHs) and some polychlorinated biphenyls (PCBs). The present study was performed to understand better the mechanisms affecting the strong interaction between planar contaminants and coal fly ash. The removal of 10 PCBs and 10 PAHs by several fly ashes and other sorbents was evaluated under different experimental conditions to highlight the intermolecular forces influencing adsorption. Varying fly ash concentration and solvent system composition indicated that dispersive interactions were most prevalent. For the PCBs, empirical results also were compared to molecular modeling estimates of the energy necessary for the PCB molecule to assume a planar conformation (PCe). The PCe levels ranged from 8 to 25 kcal/mol, depending on the degree of ortho-substituted chlorination of the PCB. A significant correlation between PCe and PCB removal from solution was observed for the fly ashes and activated carbon, whereas the nonplanar sorbent octadecyl (C{sub 18}) indicated no relationship. These findings demonstrate the strong interaction between black carbon fly ash and planar organic contaminants. Furthermore, as exemplified by the PCBs, these results show how this interaction is a function of a contaminant's ability to assume a planar conformation.

  1. [Estimation of secretory immunoglobulin A in serum of pregnant women (author's transl)].

    PubMed

    Briese, V; Lorenz, U; Brock, J; Straube, W

    1982-01-01

    Secretory immunoglobulin A (S-IgA) was measured by means of radial immunodiffusion, according to Mancini, with a self-made antiserum to the secretory component being used together with a S-IgA standard. S-IgA serum concentrations went up with significance in the course of pregnancy, which was probably attributable to increased production of secretory protein in the breasts in preparation for lactation.

  2. Secretory IgA: Designed for Anti-Microbial Defense.

    PubMed

    Brandtzaeg, Per

    2013-01-01

    Prevention of infections by vaccination remains a compelling goal to improve public health. Mucosal vaccines would make immunization procedures easier, be better suited for mass administration, and most efficiently induce immune exclusion - a term coined for non-inflammatory antibody shielding of internal body surfaces, mediated principally by secretory immunoglobulin A (SIgA). The exported antibodies are polymeric, mainly IgA dimers (pIgA), produced by local plasma cells (PCs) stimulated by antigens that target the mucose. SIgA was early shown to be complexed with an epithelial glycoprotein - the secretory component (SC). A common SC-dependent transport mechanism for pIgA and pentameric IgM was then proposed, implying that membrane SC acts as a receptor, now usually called the polymeric Ig receptor (pIgR). From the basolateral surface, pIg-pIgR complexes are taken up by endocytosis and then extruded into the lumen after apical cleavage of the receptor - bound SC having stabilizing and innate functions in the secretory antibodies. Mice deficient for pIgR show that this is the only receptor responsible for epithelial export of IgA and IgM. These knockout mice show a variety of defects in their mucosal defense and changes in their intestinal microbiota. In the gut, induction of B-cells occurs in gut-associated lymphoid tissue, particularly the Peyer's patches and isolated lymphoid follicles, but also in mesenteric lymph nodes. PC differentiation is accomplished in the lamina propria to which the activated memory/effector B-cells home. The airways also receive such cells from nasopharynx-associated lymphoid tissue but by different homing receptors. This compartmentalization is a challenge for mucosal vaccination, as are the mechanisms used by the mucosal immune system to discriminate between commensal symbionts (mutualism), pathobionts, and overt pathogens (elimination).

  3. Secretory immunity with special reference to the oral cavity

    PubMed Central

    Brandtzaeg, Per

    2013-01-01

    The two principal antibody classes present in saliva are secretory IgA (SIgA) and IgG; the former is produced as dimeric IgA by local plasma cells (PCs) in the stroma of salivary glands and is transported through secretory epithelia by the polymeric Ig receptor (pIgR), also named membrane secretory component (SC). Most IgG in saliva is derived from the blood circulation by passive leakage mainly via gingival crevicular epithelium, although some may be locally produced in the gingiva or salivary glands. Gut-associated lymphoid tissue (GALT) and nasopharynx-associated lymphoid tissue (NALT) do not contribute equally to the pool of memory/effector B cells differentiating to mucosal PCs throughout the body. Thus, enteric immunostimulation may not be the best way to activate the production of salivary IgA antibodies although the level of specific SIgA in saliva may still reflect an intestinal immune response after enteric immunization. It remains unknown whether the IgA response in submandibular/sublingual glands is better related to B-cell induction in GALT than the parotid response. Such disparity is suggested by the levels of IgA in submandibular secretions of AIDS patients, paralleling their highly upregulated intestinal IgA system, while the parotid IgA level is decreased. Parotid SIgA could more consistently be linked to immune induction in palatine tonsils/adenoids (human NALT) and cervical lymph nodes, as supported by the homing molecule profile observed after immune induction at these sites. Several other variables influence the levels of antibodies in salivary secretions. These include difficulties with reproducibility and standardization of immunoassays, the impact of flow rate, acute or chronic stress, protein loss during sample handling, and uncontrolled admixture of serum-derived IgG and monomeric IgA. Despite these problems, saliva is an easily accessible biological fluid with interesting scientific and clinical potentials. PMID:23487566

  4. Glycosylphosphatidylinositol-dependent secretory transport in Trypanosoma brucei.

    PubMed Central

    McDowell, M A; Ransom, D M; Bangs, J D

    1998-01-01

    We have investigated the role of glycosylphosphatidylinositol (GPI) anchors in forward secretory trafficking using African trypanosomes as a model system. Soluble GPI-minus forms of variant surface glycoprotein (VSG), in which the C-terminal GPI-addition peptide signal is deleted, are secreted from transformed procyclic trypanosomes with 5-fold reduced kinetics, relative to matched GPI-anchored constructs. Cell fractionation and immunofluorescence localization studies indicate that the GPI-minus VSG reporters accumulate in the endoplasmic reticulum (ER). This transport defect is specific, since overexpression of GPI-minus VSG has no effect on the rate of transport of a second soluble secretory reporter (BiPN) when co-expressed in the same cells. Two results suggest that delayed forward transport cannot be accounted for by failure to fold/assemble in the absence of a GPI anchor, thereby leading to prolonged association with ER quality-control machinery. First, no evidence was found for elevated association of GPI-minus VSG with the ER molecular chaperone, BiP. Secondly, newly synthesized GPI-minus VSG is dimerized efficiently, as judged by velocity-sedimentation analysis. GPI-dependent transport is not confined to the VSG reporters, because a similar dependence is found with another trypanosomal GPI-anchored protein, trans-sialidase. These findings suggest that GPI structures act in a positive manner to mediate efficient forward transport of some, and perhaps all, GPI-anchored proteins in the early secretory pathway of trypanosomes. Possible mechanisms for GPI-dependent transport are discussed with respect to current models of vesicular trafficking. PMID:9794811

  5. [Secretory immunoglobulin of the stomach in patients with chronic gastritis].

    PubMed

    Ostrovskiĭ, A B; Nikolaeva, O V; Isakova, V N

    1988-01-01

    The determination of the level of secretory IgA by a method of radial immunodiffusion after Mancini in the gastric juice of 48 patients with chronic gastritis in correlation with the status of the gastric mucosa, the level of acidification and the phase of exacerbation has shown diagnostic potentialities of the method. The highest IgA level was detected in patients with "rearrangement" gastritis and in patients with sharply suppressed gastric secretion. In marked atrophy of the gastric mucosa IgA secretion was significantly lowered. The period of remission was attended by a decrease in IgA secretion as compared with the phase of exacerbation.

  6. Orchestration of secretory protein folding by ER chaperones

    PubMed Central

    Gidalevitz, Tali; Stevens, Fred; Argon, Yair

    2013-01-01

    The endoplasmic reticulum is a major compartment of protein biogenesis in the cell, dedicated to production of secretory, membrane and organelle proteins. The secretome has distinct structural and post-translational characteristics, since folding in the ER occurs in an environment that is distinct in terms of its ionic composition, dynamics and requirements for quality contol. The folding machinery in the ER therefore includes chaperones and folding enzymes that introduce, monitor and react to disulfide bonds, glycans, and fluctuations of luminal calcium. We describe the major chaperone networks in the lumen and discuss how they have distinct modes of operation that enable cells to accomplish highly efficient production of the secretome. PMID:23507200

  7. Distribution of secretory component in hepatocytes and its mode of transfer into bile

    PubMed Central

    Mullock, Barbara M.; Hinton, Richard H.; Dobrota, Miloslav; Peppard, Jane; Orlans, Eva

    1980-01-01

    Immunoglobin A in bile and other external secretions is mostly bound to a glycoprotein known as secretory component. This glycoprotein is not synthesized by the same cells as immunoglobulin A and is not found in blood. We now report the mechanism by which secretory component reaches the bile and describe its function in immunoglobulin A transport across the hepatocyte. Fractionation of rat liver homogenates by zonal centrifugation was followed by measurement of the amounts of secretory component in the various fractions by rocket immunoelectrophoresis. Secretory component was found in two fractions. One of these was identified as containing Golgi vesicles from its isopycnic density and appearance in the electron microscope; the other contained principally fragments of the plasma membrane of the sinusoidal face of the hepatocyte, as shown by its particle size and content of marker enzymes. Only the latter fraction bound 125I-labelled immunoglobulin A added in vitro. At 5min after intravenous injection of [14C]fucose, the secretory component in the Golgi fraction was labelled, but not that in the plasma membrane. The secretory component in the sinusoidal plasma membrane did, however, become labelled before the first labelled secretory component appeared in bile, about 30min after injection. We suggest that fucose is added to the newly synthesized secretory component in the Golgi apparatus. The secretory component then passes, with the other newly secreted glycoproteins, to the sinusoidal plasma membrane. There it remains bound but exposed to the blood and able to bind any polymeric immunoglobulin A present in serum. The secretory component then moves across the hepatocyte to the bile-canalicular face in association with the endocytic-shuttle vesicles which carry immunoglobulin A. Hence there is a lag before newly synthesized secretory component appears in bile. ImagesPLATE 1Fig. 5. PMID:7470082

  8. The Senescence-Associated Secretory Phenotype Promotes Benign Prostatic Hyperplasia

    PubMed Central

    Vital, Paz; Castro, Patricia; Tsang, Susan; Ittmann, Michael

    2015-01-01

    Benign prostatic hyperplasia (BPH) is characterized by increased tissue mass in the transition zone of the prostate, which leads to obstruction of urine outflow and considerable morbidity in a majority of older men. Senescent cells accumulate in human tissues, including the prostate, with increasing age. Expression of proinflammatory cytokines is increased in these senescent cells, a manifestation of the senescence-associated secretory phenotype. Multiplex analysis revealed that multiple cytokines are increased in BPH, including GM-CSF, IL-1α, and IL-4, and that these are also increased in senescent prostatic epithelial cells in vitro. Tissue levels of these cytokines were correlated with a marker of senescence (cathepsin D), which was also strongly correlated with prostate weight. IHC analysis revealed the multifocal epithelial expression of cathepsin D and coexpression with IL-1α in BPH tissues. In tissue recombination studies in nude mice with immortalized prostatic epithelial cells expressing IL-1α and prostatic stromal cells, both epithelial and stromal cells exhibited increased growth. Expression of IL-1α in prostatic epithelial cells in a transgenic mouse model resulted in increased prostate size and bladder obstruction. In summary, both correlative and functional evidence support the hypothesis that the senescence-associated secretory phenotype can promote the development of BPH, which is the single most common age-related pathology in older men. PMID:24434012

  9. Assessing the biosynthetic capabilities of secretory glands in Citrus peel.

    PubMed

    Voo, Siau Sie; Grimes, Howard D; Lange, B Markus

    2012-05-01

    Epithelial cells (ECs) lining the secretory cavities of Citrus peel have been hypothesized to be responsible for the synthesis of essential oil, but direct evidence for such a role is currently sparse. We used laser-capture microdissection and pressure catapulting to isolate ECs and parenchyma cells (as controls not synthesizing oil) from the peel of young grapefruit (Citrus × paradisi 'Duncan'), isolated RNA, and evaluated transcript patterns based on oligonucleotide microarrays. A Gene Ontology analysis of these data sets indicated an enrichment of genes involved in the biosynthesis of volatile terpenoids and nonvolatile phenylpropanoids in ECs (when compared with parenchyma cells), thus indicating a significant metabolic specialization in this cell type. The gene expression patterns in ECs were consistent with the accumulation of the major essential oil constituents (monoterpenes, prenylated coumarins, and polymethoxylated flavonoids). Morphometric analyses demonstrated that secretory cavities are formed early during fruit development, whereas the expansion of cavities, and thus oil accumulation, correlates with later stages of fruit expansion. Our studies have laid the methodological and experimental groundwork for a vastly improved knowledge of the as yet poorly understood processes controlling essential oil biosynthesis in Citrus peel.

  10. Secretory proteins as potential semiochemical carriers in the horse.

    PubMed

    D'Innocenzo, Barbara; Salzano, Anna Maria; D'Ambrosio, Chiara; Gazzano, Angelo; Niccolini, Alberto; Sorce, Carlo; Dani, Francesca Romana; Scaloni, Andrea; Pelosi, Paolo

    2006-11-14

    Two soluble proteins were isolated as major secretory products of horse sweat and of the parotid gland and characterized for structural and functional properties. The first protein, lipocalin allergen EquC1, was characterized for its glycosylation sites and bound glycosidic moieties. Only one (Asn53) of the two putative glycosylation sites within the sequence was post-translationally modified; a different glycosylation pattern was determined with respect to data previously reported. When purified from horse sweat, this protein contained oleamide and other organic molecules as natural ligands. Ligand binding experiments indicated good protein selectivity toward volatile compounds having a straight chain structure of 9-11 carbon atoms, suggesting a role of this lipocalin in chemical communication. The second protein, here reported for the first time in the horse, belongs to the group of parotid secretory proteins, part of a large superfamily of binding proteins whose function in most cases is still unclear. This protein was sequenced and characterized for its post-translational modifications. Of the three cysteine residues present, two were involved in a disulfide bridge (Cys155-Cys198). A model, built up on the basis of similar proteins, indicated a general fold characterized by the presence of a long hydrophobic barrel. Binding experiments performed with a number of different organic compounds failed to identify ligands for this protein with a well-defined physiological role.

  11. Widespread occurrence of expressed fungal secretory peroxidases in forest soils.

    PubMed

    Kellner, Harald; Luis, Patricia; Pecyna, Marek J; Barbi, Florian; Kapturska, Danuta; Krüger, Dirk; Zak, Donald R; Marmeisse, Roland; Vandenbol, Micheline; Hofrichter, Martin

    2014-01-01

    Fungal secretory peroxidases mediate fundamental ecological functions in the conversion and degradation of plant biomass. Many of these enzymes have strong oxidizing activities towards aromatic compounds and are involved in the degradation of plant cell wall (lignin) and humus. They comprise three major groups: class II peroxidases (including lignin peroxidase, manganese peroxidase, versatile peroxidase and generic peroxidase), dye-decolorizing peroxidases, and heme-thiolate peroxidases (e.g. unspecific/aromatic peroxygenase, chloroperoxidase). Here, we have repeatedly observed a widespread expression of all major peroxidase groups in leaf and needle litter across a range of forest ecosystems (e.g. Fagus, Picea, Acer, Quercus, and Populus spp.), which are widespread in Europe and North America. Manganese peroxidases and unspecific peroxygenases were found expressed in all nine investigated forest sites, and dye-decolorizing peroxidases were observed in five of the nine sites, thereby indicating biological significance of these enzymes for fungal physiology and ecosystem processes. Transcripts of selected secretory peroxidase genes were also analyzed in pure cultures of several litter-decomposing species and other fungi. Using this information, we were able to match, in environmental litter samples, two manganese peroxidase sequences to Mycena galopus and Mycena epipterygia and one unspecific peroxygenase transcript to Mycena galopus, suggesting an important role of this litter- and coarse woody debris-dwelling genus in the disintegration and transformation of litter aromatics and organic matter formation.

  12. Salivary Secretory Disorders, Inducing Drugs, and Clinical Management

    PubMed Central

    Miranda-Rius, Jaume; Brunet-Llobet, Lluís; Lahor-Soler, Eduard; Farré, Magí

    2015-01-01

    Background: Salivary secretory disorders can be the result of a wide range of factors. Their prevalence and negative effects on the patient's quality of life oblige the clinician to confront the issue. Aim: To review the salivary secretory disorders, inducing drugs and their clinical management. Methods: In this article, a literature search of these dysfunctions was conducted with the assistance of a research librarian in the MEDLINE/PubMed Database. Results: Xerostomia, or dry mouth syndrome, can be caused by medication, systemic diseases such as Sjögren's Syndrome, glandular pathologies, and radiotherapy of the head and neck. Treatment of dry mouth is aimed at both minimizing its symptoms and preventing oral complications with the employment of sialogogues and topical acting substances. Sialorrhea and drooling, are mainly due to medication or neurological systemic disease. There are various therapeutic, pharmacologic, and surgical alternatives for its management. The pharmacology of most of the substances employed for the treatment of salivary disorders is well-known. Nevertheless, in some cases a significant improvement in salivary function has not been observed after their administration. Conclusion: At present, there are numerous frequently prescribed drugs whose unwanted effects include some kind of salivary disorder. In addition, the differing pathologic mechanisms, and the great variety of existing treatments hinder the clinical management of these patients. The authors have designed an algorithm to facilitate the decision making process when physicians, oral surgeons, or dentists face these salivary dysfunctions. PMID:26516310

  13. Pathogen-induced secretory diarrhea and its prevention.

    PubMed

    Anand, S; Mandal, S; Patil, P; Tomar, S K

    2016-11-01

    Secretory diarrhea is a historically known serious health implication around the world which primarily originates through pathogenic microorganisms rather than immunological or genetical disorders. This review highlights infective mechanisms of non-inflammatory secretory diarrhea causing pathogens, known therapeutics and their efficacy against them. These non-inflammatory diarrheal pathogens breach cell barriers, induce inflammation, disrupt fluid secretion across the epithelium by alteration in ion transport by faulting cystic fibrosis transmembrane conductance regulator (CFTR), calcium activated chloride channels and ion exchanger functions. Currently, a variety of prevention strategies have been used to treat these symptoms like use of antibacterial drugs, vaccines, fluid and nutritional therapy, probiotics and prebiotics as adjuncts. In progression of the need for a therapy having quick physiological effects, withdrawing the symptoms with a wide and safe therapeutic index, newer antisecretory agents like potent inhibitors, agonists and herbal remedies are some of the interventions which have come into light through greater understanding of the mechanisms and molecular targets involved in intestinal fluid secretion. Although these therapies have their own pros and cons inside the host, the quest for new antisecretory agents has been a successful elucidation to reduce burden of diarrheal disease.

  14. Cell-specific analysis of tracheobronchial secretory cells and secretions

    SciTech Connect

    Finkbeiner, W.E.

    1989-01-01

    In these studies, two methods (cell culture and monoclonal antibody production) that allowed cell-specific analysis of tracheobronchial secretion were used. Bovine tracheal submucosal gland cells were isolated, placed into culture and serially propagated. In culture, the cells maintained features of serous cells. The cells incorporated {sup 35}S into high molecular weight molecules. {beta}-adrenergic agonists stimulated release of radiolabeled molecules and elevations in intracellular cAMP levels, responses that could be blocked by the {beta}-adrenergic antagonist propranolol. Cyclic AMP appeared to be involved in the stimulus-secretion coupling events in serous cells since the phosphodiesterase inhibitor 1-methyl-3-isobutylxanthine potentiated the effects of isoproterenol on the secretory response and the elevation of intracellular cAMP levels. Furthermore, cAMP analogues elicited a secretory response in the absence of cAMP. The phosphorylation state of several cytosolic and particulate phosphoproteins was altered by cAMP-activated kinase activity. Monoclonal antibodies were produced against human airway secretions.

  15. Characterization of a Secretory Annexin in Echinococcus granulosus

    PubMed Central

    Song, Xingju; Hu, Dandan; Zhong, Xiuqin; Wang, Ning; Gu, Xiaobin; Wang, Tao; Peng, Xuerong; Yang, Guangyou

    2016-01-01

    Cystic echinococcosis, caused by Echinococcus granulosus, is a widespread parasitic zoonosis causing economic loss and public health problems. Annexins are important proteins usually present in the plasma membrane, but previous studies have shown that an annexin B33 protein of E. granulosus (Eg-ANX) could be detected in the excretory/secretory products and cyst fluid. In this study, we cloned and characterized Eg-ANX. In silico analysis showed that the amino acid sequence of Eg-ANX was conserved and lacked any signal peptides. The phospholipid-binding activity of recombinant Eg-ANX (rEg-ANX) was tested; liposomes could bind to rEg-ANX only in the presence of Ca2+. In addition, we performed western blotting and immunohistochemical analyses to further validate the secretory properties of Eg-ANX. The protein could be detected in the cyst fluid of E. granulosus and was also present in the intermediate host tissues, which suggested that Eg-ANX might play an important role in parasite–host interaction. PMID:26787154

  16. Detection of latent fingerprints using high-resolution 3D confocal microscopy in non-planar acquisition scenarios

    NASA Astrophysics Data System (ADS)

    Kirst, Stefan; Vielhauer, Claus

    2015-03-01

    In digitized forensics the support of investigators in any manner is one of the main goals. Using conservative lifting methods, the detection of traces is done manually. For non-destructive contactless methods, the necessity for detecting traces is obvious for further biometric analysis. High resolutional 3D confocal laser scanning microscopy (CLSM) grants the possibility for a detection by segmentation approach with improved detection results. Optimal scan results with CLSM are achieved on surfaces orthogonal to the sensor, which is not always possible due to environmental circumstances or the surface's shape. This introduces additional noise, outliers and a lack of contrast, making a detection of traces even harder. Prior work showed the possibility of determining angle-independent classification models for the detection of latent fingerprints (LFP). Enhancing this approach, we introduce a larger feature space containing a variety of statistical-, roughness-, color-, edge-directivity-, histogram-, Gabor-, gradient- and Tamura features based on raw data and gray-level co-occurrence matrices (GLCM) using high resolutional data. Our test set consists of eight different surfaces for the detection of LFP in four different acquisition angles with a total of 1920 single scans. For each surface and angles in steps of 10, we capture samples from five donors to introduce variance by a variety of sweat compositions and application influences such as pressure or differences in ridge thickness. By analyzing the present test set with our approach, we intend to determine angle- and substrate-dependent classification models to determine optimal surface specific acquisition setups and also classification models for a general detection purpose for both, angles and substrates. The results on overall models with classification rates up to 75.15% (kappa 0.50) already show a positive tendency regarding the usability of the proposed methods for LFP detection on varying surfaces in non-planar

  17. Excretion pattern of co-planar and non-planar tetra- and hexa-chlorobiphenyls in ovine milk and faeces

    SciTech Connect

    Vrecl, Milka . E-mail: milka.vrecl@vf.uni-lj.si; Ursic, Matjaz; Pogacnik, Azra; Zupancic-Kralj, Lucija; Jan, Janja

    2005-04-15

    This study employed the gas chromatography with electron capture detection to determine residual levels and excretion patterns of two pairs of structurally diverse polychlorinated biphenyl (PCB) congeners (IUPAC Nos. 54, 80, 155, and 169) administered to lactating sheep by intramuscular injection. PCB levels and excretion patterns in blood, milk, and faeces were time-dependent and differed from the composition of PCB congeners administered. Lactational transfer substantially exceeded the faecal transfer. Between days 3 and 7, the amount of PCB congeners 54 and 169 excreted in milk was around 50- and 800-fold higher than the amount of these two congeners excreted via faeces. During the same period, the relative contribution of co-planar PCB congeners (80 and 169) in PCB pattern decreased in blood and increased in milk and faeces compared with non-planar PCBs (54 and 155). On day 3, the ratio PCB 169 to 54 was 7-fold higher in milk than in faeces. PCB congeners with log K{sub ow} values under 6.5 reached peaks of their excretion in milk within the first three days after administration, while the super-lipophilic PCB 169 congener with log K{sub ow} value of over 7 has not reached the plateau until day 10, but afterwards, its level remained relatively high throughout the observation period. During the 57-day follow-up period, the excretion of PCB 80, 155, and 169 in milk was 4.5-, 14-, and 46-fold greater compared with PCB 54. Differences in levels and patterns were explained with some physico-chemical properties of individual PCB congeners, such as lipophilicity, planarity, metabolic stability, sorption/diffusion properties.

  18. Improvement of boundary conditions for non-planar boundaries represented by polygons with an initial particle arrangement technique

    NASA Astrophysics Data System (ADS)

    Zhang, Tiangang; Koshizuka, Seiichi; Murotani, Kohei; Shibata, Kazuya; Ishii, Eiji; Ishikawa, Masanori

    2016-02-01

    The boundary conditions represented by polygons in moving particle semi-implicit (MPS) method (Koshizuka and Oka, Nuclear Science and Engineering, 1996) have been widely used in the industry simulations since it can simply simulate complex geometry with high efficiency. However, the inaccurate particle number density near non-planar wall boundaries dramatically affects the accuracy of simulations. In this paper, we propose an initial boundary particle arrangement technique coupled with the wall weight function method (Zhang et al. Transaction of JSCES, 2015) to improve the particle number density near slopes and curved surfaces with boundary conditions represented by polygons in three dimensions. Two uniform grids are utilized in the proposed technique. The grid points in the first uniform grid are used to construct boundary particles, and the second uniform grid stores the same information as in the work by Zhang et al. The wall weight functions of the grid points in the second uniform grid are calculated by newly constructed boundary particles. The wall weight functions of the fluid particles are interpolated from the values stored on the grid points in the second uniform grid. Because boundary particles are located on the polygons, complex geometries can be accurately represented. The proposed method can dramatically improve the particle number density and maintain the high efficiency. The performance of the previously proposed wall weight function (Zhang et al.) with the boundary particle arrangement technique is verified in comparison with the wall weight function without boundary particle arrangement by investigating two example geometries. The simulations of a water tank with a wedge and a complex geometry show the general applicability of the boundary particle arrangement technique to complex geometries and demonstrate its improvement of the wall weight function near the slopes and curved surfaces.

  19. Twisted cyanines: a non-planar fluorogenic dye with superior photostability and its use in a protein-based fluoromodule.

    PubMed

    Shank, Nathaniel I; Pham, Ha H; Waggoner, Alan S; Armitage, Bruce A

    2013-01-09

    The cyanine dye thiazole orange (TO) is a well-known fluorogenic stain for DNA and RNA, but this property precludes its use as an intracellular fluorescent probe for non-nucleic acid biomolecules. Further, as is the case with many cyanines, the dye suffers from low photostability. Here, we report the synthesis of a bridge-substituted version of TO named α-CN-TO, where the central methine hydrogen of TO is replaced by an electron withdrawing cyano group, which was expected to decrease the susceptibility of the dye toward singlet oxygen-mediated degradation. An X-ray crystal structure shows that α-CN-TO is twisted drastically out of plane, in contrast to TO, which crystallizes in the planar conformation. α-CN-TO retains the fluorogenic behavior of the parent dye TO in viscous glycerol/water solvent, but direct irradiation and indirect bleaching studies showed that α-CN-TO is essentially inert to visible light and singlet oxygen. In addition, the twisted conformation of α-CN-TO mitigates nonspecific binding and fluorescence activation by DNA and a previously selected TO-binding protein and exhibits low background fluorescence in HeLa cell culture. α-CN-TO was then used to select a new protein that binds and activates fluorescence from the dye. The new α-CN-TO/protein fluoromodule exhibits superior photostability to an analogous TO/protein fluoromodule. These properties indicate that α-CN-TO will be a useful fluorogenic dye in combination with specific RNA and protein binding partners for both in vitro and cell-based applications. More broadly, structural features that promote nonplanar conformations can provide an effective method for reducing nonspecific binding of cationic dyes to nucleic acids and other biomolecules.

  20. Twisted Cyanines: A Non-Planar Fluorogenic Dye with Superior Photostability and its Use in a Protein-Based Fluoromodule

    PubMed Central

    Shank, Nathaniel I.; Pham, Ha; Waggoner, Alan S.; Armitage, Bruce A.

    2013-01-01

    The cyanine dye thiazole orange (TO) is a well-known fluorogenic stain for DNA and RNA, but this property precludes its use as an intracellular fluorescent probe for non-nucleic acid biomolecules. Further, as is the case with many cyanines, the dye suffers from low photostability. Here we report the synthesis of a bridge-substituted version of TO named α-CN-TO, where the central methine hydrogen of TO is replaced by an electron withdrawing cyano group, which was expected to decrease the susceptibility of the dye toward singlet oxygen-mediated degradation. An X-ray crystal structure shows that α-CN-TO is twisted drastically out of plane, in contrast to TO, which crystallizes in the planar conformation. α-CN-TO retains the fluorogenic behavior of the parent dye TO in viscous glycerol/water solvent, but direct irradiation and indirect bleaching studies showed that α-CN-TO is essentially inert to visible light and singlet oxygen. In addition, the twisted conformation of α-CN-TO mitigates non-specific binding and fluorescence activation by DNA and a previously selected TO-binding protein and exhibits low background fluorescence in HeLa cell culture. α-CN-TO was then used to select a new protein that binds and activates fluorescence from the dye. The new α-CN-TO/protein fluoromodule exhibits superior photostability to an analogous TO/protein fluoromodule. These properties indicate that α-CN-TO will be a useful fluorogenic dye in combination with specific RNA and protein binding partners for both in vitro and cell-based applications. More broadly, structural features that promote nonplanar conformations can provide an effective method for reducing nonspecific binding of cationic dyes to nucleic acids and other biomolecules. PMID:23252842

  1. Secretory Carcinoma of the Skin Harboring ETV6 Gene Fusions: A Cutaneous Analogue to Secretory Carcinomas of the Breast and Salivary Glands.

    PubMed

    Bishop, Justin A; Taube, Janis M; Su, Albert; Binder, Scott W; Kazakov, Dmitry V; Michal, Michal; Westra, William H

    2017-01-01

    Mammary analogue secretory carcinoma is a low-grade salivary gland carcinoma that exhibits analogous features to secretory carcinoma of the breast including the presence of a t(12;15) translocation resulting in the ETV6-NTRK3 gene fusion. Rare cases of purported secretory carcinoma of the skin adnexa have been reported, but their relationship to true secretory carcinoma of the breast and salivary glands is unclear, as they generally do not harbor ETV6 rearrangements. Cases of cutaneous neoplasms with histologic features identical to secretory carcinoma of the breast and salivary glands were identified from the consultation files of 3 academic medical institutions. Immunohistochemistry was performed for S100 protein, mammaglobin and STAT5a. Break-apart fluorescence in situ hybridization was used evaluate for disruption of the ETV6 gene. Six cases of cutaneous secretory carcinoma were identified. The tumors arose in 4 women and 2 men, ranging from 24 to 71 years in age (mean, 47 y). The carcinomas presented in the skin of the axilla (n=4), ventral neck (n=1), and cheek (n=1). The tumors arose in the superficial dermis in association with adnexal structures. None of the patients had a prior or concurrent breast or salivary gland tumor. They were histologically characterized by well-circumscribed but unencapsulated proliferations of bland, eosinophilic cells arranged in microcysts and follicles with intraluminal secretions. Ectopic breast or salivary gland tissue was not identified. The cases were diffusely positive for S100 protein (6 of 6), mammaglobin (6 of 6), and STAT5a (5 of 5). All 6 cases harbored rearrangements of ETV6. All tumors were treated by simple excision alone. No recurrences or metastases developed in the 2 cases with follow-up. Secretory carcinoma of the skin represents a phenotypic, immunohistochemical, and genetic counterpart to secretory carcinoma of the breast and salivary glands. This tumor entity is less anatomically restricted than previously

  2. Discovery and characterization of secretory IgD in rainbow trout: secretory IgD is produced through a novel splicing mechanism

    USGS Publications Warehouse

    Ramirez-Gomez, F.; Greene, W.; Rego, K.; Hansen, J.D.; Costa, G.; Kataria, P.; Bromage, E.S.

    2012-01-01

    The gene encoding IgH δ has been found in all species of teleosts studied to date. However, catfish (Ictalurus punctatus) is the only species of fish in which a secretory form of IgD has been characterized, and it occurs through the use of a dedicated δ-secretory exon, which is absent from all other species examined. Our studies have revealed that rainbow trout (Oncorhynchus mykiss) use a novel strategy for the generation of secreted IgD. The trout secretory δ transcript is produced via a run-on event in which the splice donor site at the end of the last constant domain exon (D7) is ignored and transcription continues until a stop codon is reached 33 nt downstream of the splice site, resulting in the production of an in-frame, 11-aa secretory tail at the end of the D7 domain. In silico analysis of several published IgD genes suggested that this unique splicing mechanism may also be used in other species of fish, reptiles, and amphibians. Alternative splicing of the secretory δ transcript resulted in two δ-H chains, which incorporated Cμ1 and variable domains. Secreted IgD was found in two heavily glycosylated isoforms, which are assembled as monomeric polypeptides associated with L chains. Secretory δ mRNA and IgD+ plasma cells were detected in all immune tissues at a lower frequency than secretory IgM. Our data demonstrate that secretory IgD is more prevalent and widespread across taxa than previously thought, and thus illustrate the potential that IgD may have a conserved role in immunity.

  3. Discovery and characterization of secretory IgD in rainbow trout: secretory IgD is produced through a novel splicing mechanism.

    PubMed

    Ramirez-Gomez, Francisco; Greene, Whitney; Rego, Katherine; Hansen, John D; Costa, Greg; Kataria, Priti; Bromage, Erin S

    2012-02-01

    The gene encoding IgH δ has been found in all species of teleosts studied to date. However, catfish (Ictalurus punctatus) is the only species of fish in which a secretory form of IgD has been characterized, and it occurs through the use of a dedicated δ-secretory exon, which is absent from all other species examined. Our studies have revealed that rainbow trout (Oncorhynchus mykiss) use a novel strategy for the generation of secreted IgD. The trout secretory δ transcript is produced via a run-on event in which the splice donor site at the end of the last constant domain exon (D7) is ignored and transcription continues until a stop codon is reached 33 nt downstream of the splice site, resulting in the production of an in-frame, 11-aa secretory tail at the end of the D7 domain. In silico analysis of several published IgD genes suggested that this unique splicing mechanism may also be used in other species of fish, reptiles, and amphibians. Alternative splicing of the secretory δ transcript resulted in two δ-H chains, which incorporated Cμ1 and variable domains. Secreted IgD was found in two heavily glycosylated isoforms, which are assembled as monomeric polypeptides associated with L chains. Secretory δ mRNA and IgD(+) plasma cells were detected in all immune tissues at a lower frequency than secretory IgM. Our data demonstrate that secretory IgD is more prevalent and widespread across taxa than previously thought, and thus illustrate the potential that IgD may have a conserved role in immunity.

  4. The secretory pathway of protists: spatial and functional organization and evolution.

    PubMed Central

    Becker, B; Melkonian, M

    1996-01-01

    All cells secrete a diversity of macromolecules to modify their environment or to protect themselves. Eukaryotic cells have evolved a complex secretory pathway consisting of several membrane-bound compartments which contain specific sets of proteins. Experimental work on the secretory pathway has focused mainly on mammalian cell lines or on yeasts. Now, some general principles of the secretory pathway have become clear, and most components of the secretory pathway are conserved between yeast cells and mammalian cells. However, the structure and function of the secretory system in protists have been less extensively studied. In this review, we summarize the current knowledge about the secretory pathway of five different groups of protists: Giardia lamblia, one of the earliest lines of eukaryotic evolution, kinetoplastids, the slime mold Dictyostelium discoideum, and two lineages within the "crown" of eukaryotic cell evolution, the alveolates (ciliates and Plasmodium species) and the green algae. Comparison of these systems with the mammalian and yeast system shows that most elements of the secretory pathway were presumably present in the earliest eukaryotic organisms. However, one element of the secretory pathway shows considerable variation: the presence of a Golgi stack and the number of cisternae within a stack. We suggest that the functional separation of the plasma membrane from the nucleus-endoplasmic reticulum system during evolution required a sorting compartment, which became the Golgi apparatus. Once a Golgi apparatus was established, it was adapted to the various needs of the different organisms. PMID:8987360

  5. Mammary analogue secretory carcinoma (MASC) of the salivary gland: A new tumor entity

    PubMed Central

    Damjanov, Ivan; Skenderi, Faruk; Vranic, Semir

    2016-01-01

    Mammary analogue secretory carcinoma (MASC) is a recently described low-grade malignant tumor of the salivary glands, biologically and morphologically equivalent to secretory breast carcinoma. We give a brief overview of this new entity, including morphological, immunohistochemical, molecular-genetic, clinical, epidemiologic features, differential diagnosis, and outcome results. PMID:27131022

  6. Mammary Analogue Secretory Carcinoma (MASC) of the salivary gland: A new tumor entity.

    PubMed

    Damjanov, Ivan; Skenderi, Faruk; Vranic, Semir

    2016-08-02

    Mammary analogue secretory carcinoma (MASC) is a recently described low-grade malignant tumor of the salivary glands, biologically and morphologically equivalent to secretory breast carcinoma. We give a brief overview of this new entity, including morphological, immunohistochemical, molecular-genetic, clinical, epidemiologic features, differential diagnosis, and outcome results.

  7. [Secretory immunoglobulin A in the amniotic fluid of healthy pregnant females].

    PubMed

    Briese, V; Straube, W; Brock, J; Stark, K H; Lorenz, U

    1983-01-01

    Amniotic fluid levels of secretory immunoglobulin A (S-AgA) were measured by simple radial immunodiffusion according to the method of Mancini using a monospecific antiserum against the human secretory component. 256 samples from healthy pregnant women were examined. Amniotic fluid S-IgA concentration increases significantly during normal pregnancy and shows a loose correlation to the phospholipid level.

  8. Intermediates in the constitutive and regulated secretory pathways released in vitro from semi-intact cells

    PubMed Central

    1992-01-01

    Regulated secretory cells have two pathways that transport secreted proteins from the Golgi complex to the cell surface. To identify carrier vesicles involved in regulated and constitutive secretion, PC12 pheochromocytoma cells were labeled with [35S]sulfate to identify markers for the two secretory pathways, then mechanically permeabilized and incubated in vitro. Small constitutive secretory vesicles, containing mostly sulfated proteoglycans, accumulated during an in vitro incubation with ATP. In the presence of GTP gamma S, the constitutive vesicles became significantly more dense, suggesting that a coated intermediate was stabilized. Larger immature regulated secretory granules, enriched in sulfated secretogranin II, also escaped from the permeabilized cells in vitro. During granule maturation, their density increased and the amount of cofractionating proteoglycans diminished. The data suggest that sorting continues during secretory granule maturation. PMID:1572894

  9. The evolution of plant secretory structures and emergence of terpenoid chemical diversity.

    PubMed

    Lange, Bernd Markus

    2015-01-01

    Secretory structures in terrestrial plants appear to have first emerged as intracellular oil bodies in liverworts. In vascular plants, internal secretory structures, such as resin ducts and laticifers, are usually found in conjunction with vascular bundles, whereas subepidermal secretory cavities and epidermal glandular trichomes generally have more complex tissue distribution patterns. The primary function of plant secretory structures is related to defense responses, both constitutive and induced, against herbivores and pathogens. The ability to sequester secondary (or specialized) metabolites and defense proteins in secretory structures was a critical adaptation that shaped plant-herbivore and plant-pathogen interactions. Although this review places particular emphasis on describing the evolution of pathways leading to terpenoids, it also assesses the emergence of other metabolite classes to outline the metabolic capabilities of different plant lineages.

  10. Development and Essential Oil Content of Secretory Glands of Sage (Salvia officinalis) 1

    PubMed Central

    Venkatachalam, K. V.; Kjonaas, Robert; Croteau, Rodney

    1984-01-01

    Scanning electron microscopy of sage (Salvia officinalis L.) leaves confirmed the presence of two basic types of glandular trichomes consisting of a capitate stalked form containing a multicellular stalk and surmounted by a unicellular secretory head, and a capitate sessile form containing a unicellular stalk and unicellular, or multicellular, secretory head. In the latter type, secretory activity and filling of the subcuticular cavity may begin at virtually any stage of the division cycle affording fully developed glands containing from one to twelve cells in the secretory head. Gas liquid chromatographic analysis of the oil content of the most numerous gland species (capitate stalked, capitate sessile with one and with eight secretory cells) indicated only minor quantitative differences in essential oil composition. Thus, each gland type is capable of producing the four major monoterpene families (p-menthanes, pinanes, bornanes and thujanes) characteristic of sage. Images Fig. 1 PMID:16663786

  11. Development and essential oil content of secretory glands of sage (Salvia officinalis)

    SciTech Connect

    Venkatachalam, K.V.; Kjonaas, R.; Croteau, R.

    1984-09-01

    Scanning electron microscopy of sage (Salvia officinalis L.) leave confirmed the presence of two basic types of glandular trichomes consisting of a capitate stalked form containing a multicellular stalk and surmounted by a unicellular secretory head, and a capitate sessile form containing a unicellular stalk and unicellular, or multicellular, secretory head. In the latter type, secretory activity and filling of the subcuticular cavity may begin at virtually any stage of the division cycle affording fully developed glands containing from one to twelve cells in the secretory head. Gas liquid chromatographic analysis of the oil content of the most numerous gland species (capitate stalked, capitate sessile with one and with eight secretory cells) indicated only minor quantitative differences in essential oil composition. Thus, each gland type is capable of producing the four major monoterpene families (p-menthanes, pinanes, bornanes and thujanes) characteristic of sage. 21 references, 2 figures.

  12. The secretory carrier membrane protein family: structure and membrane topology.

    PubMed

    Hubbard, C; Singleton, D; Rauch, M; Jayasinghe, S; Cafiso, D; Castle, D

    2000-09-01

    Secretory carrier membrane proteins (SCAMPs) are integral membrane proteins found in secretory and endocytic carriers implicated to function in membrane trafficking. Using expressed sequence tag database and library screens and DNA sequencing, we have characterized several new SCAMPs spanning the plant and animal kingdoms and have defined a broadly conserved protein family. No obvious fungal homologue has been identified, however. We have found that SCAMPs share several structural motifs. These include NPF repeats, a leucine heptad repeat enriched in charged residues, and a proline-rich SH3-like and/or WW domain-binding site in the N-terminal domain, which is followed by a membrane core containing four putative transmembrane spans and three amphiphilic segments that are the most highly conserved structural elements. All SCAMPs are 32-38 kDa except mammalian SCAMP4, which is approximately 25 kDa and lacks most of the N-terminal hydrophilic domain of other SCAMPs. SCAMP4 is authentic as determined by Northern and Western blotting, suggesting that this portion of the larger SCAMPs encodes the functional domain. Focusing on SCAMP1, we have characterized its structure further by limited proteolysis and Western blotting with the use of isolated secretory granules as a uniformly oriented source of antigen and by topology mapping through expression of alkaline phosphatase gene fusions in Escherichia coli. Results show that SCAMP1 is degraded sequentially from the N terminus and then the C terminus, yielding an approximately 20-kDa membrane core that contains four transmembrane spans. Using synthetic peptides corresponding to the three conserved amphiphilic segments of the membrane core, we have demonstrated their binding to phospholipid membranes and shown by circular dichroism spectroscopy that the central amphiphilic segment linking transmembrane spans 2 and 3 is alpha-helical. In the intact protein, these segments are likely to reside in the cytoplasm-facing membrane

  13. Fallopian tube secretory cell expansion: a sensitive biomarker for ovarian serous carcinogenesis.

    PubMed

    Wang, Yiying; Li, Li; Wang, Yue; Tang, Sarah Ngocvi; Zheng, Wenxin

    2015-01-01

    Recent advances suggest that precancerous lesions of pelvic serous carcinoma originate from tubal secretory cells. The purpose of our study was to determine if an increased number of secretory cells varies with age or location in the fallopian tube and to examine its association with serous neoplasia. Three groups (benign control, high-risk, and pelvic serous carcinoma) of age-matched patients were studied. The age data were stratified into 10-year intervals ranging from 20-29 to older than 80. The number of secretory and ciliated cells from both tubal fimbria and ampulla segments was counted by microscopy and immunohistochemical staining methods. The data were analyzed by standard contingency table and Poisson distribution methods after age justification. We found that the absolute number of tubal secretory cells increased significantly with age in all three groups. But a more dramatic increase of secretory cells was observed in high-risk and pelvic serous carcinoma patients. Secretory cell expansion is more prevalent than secretory cell outgrowth in both fimbria and ampulla tubal segments and is significantly associated with serous neoplasia (P < 0.001). Furthermore, age remained a significant risk factor for serous neoplasia after age adjustment. These findings suggest that secretory cell expansion could serve as a potential sensitive biomarker for early serous carcinogenesis within the fallopian tube. The study also supports a relationship between serous neoplasia and increased secretory to ciliated cell ratios, and the relationship between frequency of secretory cell expansion within the fallopian tube and increasing age and-more significantly-presence of high-risk factors or co-existing serous cancers.

  14. Fallopian tube secretory cell expansion: a sensitive biomarker for ovarian serous carcinogenesis.

    PubMed

    Wang, Yiying; Li, Li; Wang, Yue; Tang, Sarah Ngocvi; Zheng, Wenxin

    2016-01-01

    Recent advances suggest that precancerous lesions of pelvic serous carcinoma originate from tubal secretory cells. The purpose of our study was to determine if an increased number of secretory cells vary with age or location in the fallopian tube and to examine its association with serous neoplasia. Three groups (benign control, high-risk, and pelvic serous carcinoma) of age-matched patients were studied. The age data were stratified into 10-year intervals ranging from 20-29 to older than 80. The number of secretory and ciliated cells from both tubal fimbria and ampulla segments was counted by microscopy and immunohistochemical staining methods. The data were analyzed by standard contingency table and Poisson distribution methods after age justification. We found that the absolute number of tubal secretory cells increased significantly with age in all three groups. But a more dramatic increase of secretory cells was observed in high-risk and pelvic serous carcinoma patients. Secretory cell expansion is more prevalent than secretory cell outgrowth in both fimbria and ampulla tubal segments and is significantly associated with serous neoplasia (p < 0.001). Furthermore, age remained a significant risk factor for serous neoplasia after age adjustment. These findings suggest that secretory cell expansion could serve as a potential sensitive biomarker for early serous carcinogenesis within the fallopian tube. The study also supports a relationship between serous neoplasia and increased secretory to ciliated cell ratios, and the relationship between frequency of secretory cell expansion within the fallopian tube and increasing age and-more significantly-presence of high-risk factors or co-existing serous cancers.

  15. The secretory granule matrix: a fast-acting smart polymer.

    PubMed

    Nanavati, C; Fernandez, J M

    1993-02-12

    The secretory granule matrix is a miniature biopolymer that consists of a charged polymer network that traps peptides and transmitters when it condenses and releases them on exocytotic decondensation. Models of exocytotic fusion have treated this matrix as a short circuit and have neglected its electrical contributions. This matrix responded to negative voltages by swelling, which was accompanied by a large increase in conductance, and to positive voltages by condensing. Thus, the matrix resembled a diode. The swollen matrix exerted large pressures on the order of 12 bar. The responses took place within milliseconds of the application of the electric field. These findings suggest that matrix decondensation, and therefore product release, is controlled by potential gradients.

  16. Non-secretory multiple myeloma: from biology to clinical management

    PubMed Central

    Dupuis, Megan Murray; Tuchman, Sascha A

    2016-01-01

    Multiple myeloma (MM) is the second most common hematologic malignancy in the US. It is typically characterized by production of large amounts of defective immunoglobulin (Ig). Diagnosing MM and monitoring treatment response, including eventual relapse, are largely based on sequential measurements of Ig. However, a small subset of MM called non-secretory multiple myeloma (NSMM) produces no detectable Ig. This subset of true NSMM has become even smaller over time, as the advent of the serum free light chain assay has resulted in the majority of NSMM patients being recategorized as light-chain MM – that is, MM cells that produce only the light-chain component of Ig. True forms of NSMM, meaning MM that secretes no monoclonal proteins whatsoever, constitute a distinct entity that is reviewed; definition of NSMM using current detection methods, discuss the biology underpinning NSMM development, and share recommendations for how NSMM should be managed clinically with respect to detection, treatment, and monitoring. PMID:28008276

  17. Mammary analog secretory carcinoma of thyroid: A case report.

    PubMed

    Rupp, Aaron P; Bocklage, Thèrése J

    2017-01-01

    Mammary analog secretory carcinoma (MASC) is a recently described rare neoplasm that was first reported in the salivary gland with an associated ETV6-NTRK3 fusion. We present a case of MASC involving and presumably arising in the thyroid, which was originally diagnosed as papillary thyroid carcinoma on fine needle aspiration and surgical resection. The later correct diagnosis of MASC was confirmed by immunohistochemistry and molecular studies. The cytopathological features of MASC in the salivary gland are previously described; however, we present the first cytopathological description of MASC arising in the thyroid with the unique feature of prominent nuclear grooves. Differentiating MASC from overlapping features of cytopathologic mimics such as papillary thyroid carcinoma may carry crucial therapeutic implications. Diagn. Cytopathol. 2017;45:45-50. © 2016 Wiley Periodicals, Inc.

  18. Secretory expression of a heterologous nattokinase in Lactococcus lactis.

    PubMed

    Liang, Xiaobo; Zhang, Lixin; Zhong, Jin; Huan, Liandong

    2007-05-01

    Nattokinase has been reported as an oral health product for the prevention of atherosclerosis. We developed a novel strategy to express a nattokinase from Bacillus subtilis in a live delivery vehicle, Lactococcus lactis. Promoter P( nisZ) and signal peptide SP(Usp) were used for inducible and secretory expression of nattokinase in L. lactis. Western blotting analysis demonstrated that nattokinase was successfully expressed, and about 94% of the enzyme was secreted to the culture. The recombinant nattokinase showed potent fibrinolytic activity, equivalent to 41.7 urokinase units per milliliter culture. Expression and delivery of such a fibrinolytic enzyme in the food-grade vehicle L. lactis would facilitate the widespread application of nattokinase in the control and prevention of thrombosis diseases.

  19. Novel distribution of the secretory leucocyte proteinase inhibitor in kidney.

    PubMed Central

    Ohlsson, S; Ljungkrantz, I; Ohlsson, K; Segelmark, M; Wieslander, J

    2001-01-01

    The secretory leucocyte proteinase inhibitor (SLPI) is a low molecular weight, tissue-specific inhibitor of, for example, elastase and cathepsin G, which also have antimicrobial capacity. SLPI has been localised to the respiratory, gastrointestinal and genital tracts, but so far not to the kidney. The presence of SLPI in renal tubuli cells was demonstrated using immunohistochemistry and, by means of in situ hybridisation on human renal biopsies, we were able to demonstrate SLPI production. In various inflammatory conditions in the kidneys, the protease-antiprotease balance is disturbed. For this reason, as well as the possible role in the defence against ascending urinary tract infections, it is interesting to establish a source of SLPI in renal tubuli cells. PMID:11817677

  20. Characterization of mast cell secretory granules and their cell biology.

    PubMed

    Azouz, Nurit Pereg; Hammel, Ilan; Sagi-Eisenberg, Ronit

    2014-10-01

    Exocytosis and secretion of secretory granule (SG) contained inflammatory mediators is the primary mechanism by which mast cells exert their protective immune responses in host defense, as well as their pathological functions in allergic reactions and anaphylaxis. Despite their central role in mast cell function, the molecular mechanisms underlying the biogenesis and secretion of mast cell SGs remain largely unresolved. Early studies have established the lysosomal nature of the mast cell SGs and implicated SG homotypic fusion as an important step occurring during both their biogenesis and compound secretion. However, the molecular mechanisms that account for key features of this process largely remain to be defined. A novel high-resolution imaging based methodology allowed us to screen Rab GTPases for their phenotypic and functional impact and identify Rab networks that regulate mast cell secretion. This screen has identified Rab5 as a novel regulator of homotypic fusion of the mast cell SGs that thereby regulates their size and cargo composition.

  1. Silent reading and secretory otitis media in school children.

    PubMed

    Lous, J

    1993-01-01

    In an unselected cohort of 366 8-year-old children, the relationship between secretory otitis media and reading achievement was investigated. The children underwent 10 impedance audiometries and 5 pure tone audiometries during their first year at school. At the beginning of the second grade they all had a Silent Reading Word Test (OS-400). The background parameters were recorded by an interview with one of the parents. There was a significant but small correlation between type B tympanograms in the first grade and silent word reading. No association between silent reading score and otological history or pure tone screening was found. In a stepwise multiple regression model, 37% of the variance could be 'explained' by the included variables. The 'classroom factor' could 'explain' about 17% of the variance, followed by phonology at the start of school (6%), gender (5%), social group of the mother (4%), type B tympanogram (2%), absence from school (2%) and allergy (1%).

  2. Developmental genetics of secretory vesicle acidification during Caenorhabditis elegans spermatogenesis.

    PubMed

    Gleason, Elizabeth J; Hartley, Paul D; Henderson, Melissa; Hill-Harfe, Katherine L; Price, Paul W; Weimer, Robby M; Kroft, Tim L; Zhu, Guang-Dan; Cordovado, Suzanne; L'Hernault, Steven W

    2012-06-01

    Secretory vesicles are used during spermatogenesis to deliver proteins to the cell surface. In Caenorhabditis elegans, secretory membranous organelles (MO) fuse with the plasma membrane to transform spermatids into fertilization-competent spermatozoa. We show that, like the acrosomal vesicle of mammalian sperm, MOs undergo acidification during development. Treatment of spermatids with the V-ATPase inhibitor bafilomycin blocks both MO acidification and formation of functional spermatozoa. There are several spermatogenesis-defective mutants that cause defects in MO morphogenesis, including spe-5. We determined that spe-5, which is on chromosome I, encodes one of two V-ATPase B paralogous subunits. The spe-5 null mutant is viable but sterile because it forms arrested, multi-nucleate spermatocytes. Immunofluorescence with a SPE-5-specific monoclonal antibody shows that SPE-5 expression begins in spermatocytes and is found in all subsequent stages of spermatogenesis. Most SPE-5 is discarded into the residual body during spermatid budding, but a small amount remains in budded spermatids where it localizes to MOs as a discrete dot. The other V-ATPase B subunit is encoded by vha-12, which is located on the X chromosome. Usually, spe-5 mutants are self-sterile in a wild-type vha-12 background. However, an extrachromosomal transgene containing wild-type vha-12 driven by its own promoter allows spe-5 mutant hermaphrodites to produce progeny, indicating that VHA-12 can at least partially substitute for SPE-5. Others have shown that the X chromosome is transcriptionally silent in the male germline, so expression of the autosomally located spe-5 gene ensures that a V-ATPase B subunit is present during spermatogenesis.

  3. Thyroid hormone status regulates the expression of secretory phospholipases.

    PubMed

    Sharma, Pragya; Levesque, Tania; Boilard, Eric; Park, Edwards A

    2014-01-31

    Thyroid hormone (T3) stimulates various metabolic pathways and the hepatic actions of T3 are mediated primarily through the thyroid hormone receptor beta (TRβ). Hypothyroidism has been linked with low grade inflammation, elevated risk of hepatic steatosis and atherosclerosis. Secretory phospholipases (sPLA2) are associated with inflammation, hyperlipidemia and atherosclerosis. Due to potential linkage between thyroid hormone and sPLA2, we investigated the effect of thyroid hormone status on the regulation of secretory phospholipases in mice, rats and human liver. T3 suppressed the expression of the sPLA2 group IIa (PLA2g2a) gene in the liver of BALB/c mice and C57BL/6 transgenic mice expressing the human PLA2g2a. PLA2g2a was elevated with hypothyroidism and high fat diets which may contribute to the low grade inflammation associated with hypothyroidism and diet induced obesity. We also examined the effects of the TRβ agonist eprotirome on hepatic gene regulation. We observed that eprotirome inhibited the expression of selected sPLA2 genes and furthermore the cytokine mediated induction PLA2g2a was suppressed. In addition, eprotirome induced genes involved in fatty acid oxidation and cholesterol clearance while inhibiting lipogenic genes. Our results indicate that in vivo thyroid hormone status regulates the abundance of sPLA2 and the inhibition of PLA2g2a by T3 is conserved across species. By regulating sPLA2 genes, T3 may impact processes associated with atherosclerosis and inflammation and TRβ agonists may ameliorate inflammation and hyperlipidemia.

  4. Thyroid hormone status regulates the expression of secretory phospholipases

    PubMed Central

    Sharma, Pragya; Levesque, Tania; Boilard, Eric; Park, Edwards A.

    2014-01-01

    Thyroid hormone (T3) stimulates various metabolic pathways and the hepatic actions of T3 are mediated primarily through the thyroid hormone receptor beta (TRβ). Hypothyroidism has been linked with low grade inflammation, elevated risk of hepatic steatosis and atherosclerosis. Secretory phospholipases (sPLA2) are associated with inflammation, hyperlipidemia and atherosclerosis. Due to potential linkage between thyroid hormone and sPLA2, we investigated the effect of thyroid hormone status on the regulation of secretory phospholipases in mice, rats and human liver. T3 suppressed the expression of the sPLA2 group IIa (PLA2g2a) gene in the liver of BALB/c mice and C57BL/6 transgenic mice expressing the human PLA2g2a. PLA2g2a was elevated with hypothyroidism and high fat diets which may contribute to the low grade inflammation associated with hypothyroidism and diet induced obesity. We also examined the effects of the TRβ agonist eprotirome on hepatic gene regulation. We observed that eprotirome inhibited the expression of selected sPLA2 genes and furthermore the cytokine mediated induction PLA2g2a was suppressed. In addition, eprotirome induced genes involved in fatty acid oxidation and cholesterol clearance while inhibiting lipogenic genes. Our results indicate that in vivo thyroid hormone status regulates the abundance of sPLA2 and the inhibition of PLA2g2a by T3 is conserved across species. By regulating sPLA2 genes, T3 may impact processes associated with atherosclerosis and inflammation and TRβ agonists may ameliorate inflammation and hyperlipidemia. PMID:24440706

  5. Rac1 Regulates Endometrial Secretory Function to Control Placental Development.

    PubMed

    Davila, Juanmahel; Laws, Mary J; Kannan, Athilakshmi; Li, Quanxi; Taylor, Robert N; Bagchi, Milan K; Bagchi, Indrani C

    2015-08-01

    During placenta development, a succession of complex molecular and cellular interactions between the maternal endometrium and the developing embryo ensures reproductive success. The precise mechanisms regulating this maternal-fetal crosstalk remain unknown. Our study revealed that the expression of Rac1, a member of the Rho family of GTPases, is markedly elevated in mouse decidua on days 7 and 8 of gestation. To investigate its function in the uterus, we created mice bearing a conditional deletion of the Rac1 gene in uterine stromal cells. Ablation of Rac1 did not affect the formation of the decidua but led to fetal loss in mid gestation accompanied by extensive hemorrhage. To gain insights into the molecular pathways affected by the loss of Rac1, we performed gene expression profiling which revealed that Rac1 signaling regulates the expression of Rab27b, another GTPase that plays a key role in targeting vesicular trafficking. Consequently, the Rac1-null decidual cells failed to secrete vascular endothelial growth factor A, which is a critical regulator of decidual angiogenesis, and insulin-like growth factor binding protein 4, which regulates the bioavailability of insulin-like growth factors that promote proliferation and differentiation of trophoblast cell lineages in the ectoplacental cone. The lack of secretion of these key factors by Rac1-null decidua gave rise to impaired angiogenesis and dysregulated proliferation of trophoblast cells, which in turn results in overexpansion of the trophoblast giant cell lineage and disorganized placenta development. Further experiments revealed that RAC1, the human ortholog of Rac1, regulates the secretory activity of human endometrial stromal cells during decidualization, supporting the concept that this signaling G protein plays a central and conserved role in controlling endometrial secretory function. This study provides unique insights into the molecular mechanisms regulating endometrial secretions that mediate stromal

  6. Rac1 Regulates Endometrial Secretory Function to Control Placental Development

    PubMed Central

    Davila, Juanmahel; Laws, Mary J.; Kannan, Athilakshmi; Li, Quanxi; Taylor, Robert N.; Bagchi, Milan K.; Bagchi, Indrani C.

    2015-01-01

    During placenta development, a succession of complex molecular and cellular interactions between the maternal endometrium and the developing embryo ensures reproductive success. The precise mechanisms regulating this maternal-fetal crosstalk remain unknown. Our study revealed that the expression of Rac1, a member of the Rho family of GTPases, is markedly elevated in mouse decidua on days 7 and 8 of gestation. To investigate its function in the uterus, we created mice bearing a conditional deletion of the Rac1 gene in uterine stromal cells. Ablation of Rac1 did not affect the formation of the decidua but led to fetal loss in mid gestation accompanied by extensive hemorrhage. To gain insights into the molecular pathways affected by the loss of Rac1, we performed gene expression profiling which revealed that Rac1 signaling regulates the expression of Rab27b, another GTPase that plays a key role in targeting vesicular trafficking. Consequently, the Rac1-null decidual cells failed to secrete vascular endothelial growth factor A, which is a critical regulator of decidual angiogenesis, and insulin-like growth factor binding protein 4, which regulates the bioavailability of insulin-like growth factors that promote proliferation and differentiation of trophoblast cell lineages in the ectoplacental cone. The lack of secretion of these key factors by Rac1-null decidua gave rise to impaired angiogenesis and dysregulated proliferation of trophoblast cells, which in turn results in overexpansion of the trophoblast giant cell lineage and disorganized placenta development. Further experiments revealed that RAC1, the human ortholog of Rac1, regulates the secretory activity of human endometrial stromal cells during decidualization, supporting the concept that this signaling G protein plays a central and conserved role in controlling endometrial secretory function. This study provides unique insights into the molecular mechanisms regulating endometrial secretions that mediate stromal

  7. Calcium dynamics in bovine adrenal medulla chromaffin cell secretory granules.

    PubMed

    Santodomingo, Jaime; Vay, Laura; Camacho, Marcial; Hernández-Sanmiguel, Esther; Fonteriz, Rosalba I; Lobatón, Carmen D; Montero, Mayte; Moreno, Alfredo; Alvarez, Javier

    2008-10-01

    The secretory granules constitute one of the less well-known compartments in terms of Ca2+ dynamics. They contain large amounts of total Ca2+, but the free intragranular [Ca2+] ([Ca2+]SG), the mechanisms for Ca2+ uptake and release from the granules and their physiological significance regarding exocytosis are still matters of debate. We used in the present work an aequorin chimera targeted to the granules to investigate [Ca2+]SG homeostasis in bovine adrenal chromaffin cells. We found that most of the intracellular aequorin chimera is present in a compartment with 50-100 microM Ca2+. Ca2+ accumulation into this compartment takes place mainly through an ATP-dependent mechanism, namely, a thapsigargin-sensitive Ca2+-ATPase. In addition, fast Ca2+ release was observed in permeabilized cells after addition of inositol 1,4,5-trisphosphate (InsP3) or caffeine, suggesting the presence of InsP3 and ryanodine receptors in the vesicular membrane. Stimulation of intact cells with the InsP3-producing agonist histamine or with caffeine also induced Ca2+ release from the vesicles, whereas acetylcholine or high-[K+] depolarization induced biphasic changes in vesicular[Ca2+], suggesting heterogeneous responses of different vesicle populations, some of them releasing and some taking up Ca2+during stimulation. In conclusion, our data show that chromaffin cell secretory granules have the machinery required for rapid uptake and release of Ca2+, and this strongly supports the hypothesis that granular Ca2+ may contribute to its own secretion.

  8. Calcium dynamics in catecholamine-containing secretory vesicles.

    PubMed

    Moreno, Alfredo; Lobatón, Carmen D; Santodomingo, Jaime; Vay, Laura; Hernández-SanMiguel, Esther; Rizzuto, Rosario; Montero, Mayte; Alvarez, Javier

    2005-06-01

    We have used an aequorin chimera targeted to the membrane of the secretory granules to monitor the free [Ca(2+)] inside them in neurosecretory PC12 cells. More than 95% of the probe was located in a compartment with an homogeneous [Ca(2+)] around 40 microM. Cell stimulation with either ATP, caffeine or high-K(+) depolarization increased cytosolic [Ca(2+)] and decreased secretory granule [Ca(2+)] ([Ca(2+)](SG)). Inositol-(1,4,5)-trisphosphate, cyclic ADP ribose and nicotinic acid adenine dinucleotide phosphate were all ineffective to release Ca(2+) from the granules. Changes in cytosolic [Na(+)] (0-140 mM) or [Ca(2+)] (0-10 microM) did not modify either ([Ca(2+)](SG)). Instead, [Ca(2+)](SG) was highly sensitive to changes in the pH gradient between the cytosol and the granules. Both carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP) and nigericin, as well as cytosolic acidification, reversibly decreased [Ca(2+)](SG), while cytosolic alcalinization reversibly increased [Ca(2+)](SG). These results are consistent with the operation of a H(+)/Ca(2+) antiporter in the vesicular membrane. This antiporter could also mediate the effects of ATP, caffeine and high-K(+) on [Ca(2+)](SG), because all of them induced a transient cytosolic acidification. The FCCP-induced decrease in [Ca(2+)](SG) was reversible in 10-15 min even in the absence of cytosolic Ca(2+) or ATP, suggesting that most of the calcium content of the vesicles is bound to a slowly exchanging Ca(2+) buffer. This large store buffers [Ca(2+)](SG) changes in the long-term but allows highly dynamic free [Ca(2+)](SG) changes to occur in seconds or minutes.

  9. Sorting and storage during secretory granule biogenesis: looking backward and looking forward.

    PubMed Central

    Arvan, P; Castle, D

    1998-01-01

    Secretory granules are specialized intracellular organelles that serve as a storage pool for selected secretory products. The exocytosis of secretory granules is markedly amplified under physiologically stimulated conditions. While granules have been recognized as post-Golgi carriers for almost 40 years, the molecular mechanisms involved in their formation from the trans-Golgi network are only beginning to be defined. This review summarizes and evaluates current information about how secretory proteins are thought to be sorted for the regulated secretory pathway and how these activities are positioned with respect to other post-Golgi sorting events that must occur in parallel. In the first half of the review, the emerging role of immature secretory granules in protein sorting is highlighted. The second half of the review summarizes what is known about the composition of granule membranes. The numerous similarities and relatively limited differences identified between granule membranes and other vesicular carriers that convey products to and from the plasmalemma, serve as a basis for examining how granule membrane composition might be established and how its unique functions interface with general post-Golgi membrane traffic. Studies of granule formation in vitro offer additional new insights, but also important challenges for future efforts to understand how regulated secretory pathways are constructed and maintained. PMID:9620860

  10. Nonplanar Nanoscale Fin Field Effect Transistors on Textile, Paper, Wood, Stone, and Vinyl via Soft Material-Enabled Double-Transfer Printing.

    PubMed

    Rojas, Jhonathan P; Torres Sevilla, Galo A; Alfaraj, Nasir; Ghoneim, Mohamed T; Kutbee, Arwa T; Sridharan, Ashvitha; Hussain, Muhammad Mustafa

    2015-05-26

    The ability to incorporate rigid but high-performance nanoscale nonplanar complementary metal-oxide semiconductor (CMOS) electronics with curvilinear, irregular, or asymmetric shapes and surfaces is an arduous but timely challenge in enabling the production of wearable electronics with an in situ information-processing ability in the digital world. Therefore, we are demonstrating a soft-material enabled double-transfer-based process to integrate flexible, silicon-based, nanoscale, nonplanar, fin-shaped field effect transistors (FinFETs) and planar metal-oxide-semiconductor field effect transistors (MOSFETs) on various asymmetric surfaces to study their compatibility and enhanced applicability in various emerging fields. FinFET devices feature sub-20 nm dimensions and state-of-the-art, high-κ/metal gate stacks, showing no performance alteration after the transfer process. A further analysis of the transferred MOSFET devices, featuring 1 μm gate length, exhibits an ION value of nearly 70 μA/μm (VDS = 2 V, VGS = 2 V) and a low subthreshold swing of around 90 mV/dec, proving that a soft interfacial material can act both as a strong adhesion/interposing layer between devices and final substrate as well as a means to reduce strain, which ultimately helps maintain the device's performance with insignificant deterioration even at a high bending state.

  11. Non-Planar Structures of the High-Energy Rotational Conformers of 2-METHYLBUTA-1,3-DIENE (isoprene) and 2,3-DIMETHYLBUTA-1,3-DIENE

    NASA Astrophysics Data System (ADS)

    Panchenko, Yu. N.; Bock, Ch. W.; Larkin, J. D.; Abramenkov, A. V.; Kühnemann, F.

    2009-06-01

    Optimization of the geometrical parameters and determination of the force fields for rotamers of the title molecules were performed at the MP2(FC)/aug-cc-pVDZ//MP2(FC)/aug-cc-pVDZ computational level. The vibrational analyses of these conformers were carried out using scaled quantum-mechanical force field methodology. Recent experimental wavenumbers for these conformers and their deuteroisomers were incorporated into these analyses. The theoretical non-planar structures of the high-energy conformers of 2-methylbuta-1,3-diene (isoprene) and 2,3-dimethylbuta-1,3-diene were corroborated by good agreement between the experimental and theoretical wavenumbers of the molecules under investigation. The dihedral angles of the non-planar high-energy conformers for rotation around the =C-C= bond are as follows: 41.6^° for 2-methylbuta-1,3-diene (isoprene) and 47.0^° for 2,3-dimethylbuta-1,3-diene. Previous studies performed at the HF/6-31G level gave 41.0^° and 48.5^° for the first and second compounds, respectively. Yu. N. Panchenko, Ch. W. Bock, J. D. Larkin, A. V. Abramenkov, F. Kühnemann, Struct. Chem. 19, 421 (2008). Yu. N. Panchenko, Ch. W. Bock, J. D. Larkin, A. V. Abramenkov, Struct. Chem. 19, 793 (2008)

  12. Ca-dependent Nonsecretory Vesicle Fusion in a Secretory Cell

    PubMed Central

    Wang, Tzu-Ming; Hilgemann, Donald W.

    2008-01-01

    We have compared Ca-dependent exocytosis in excised giant membrane patches and in whole-cell patch clamp with emphasis on the rat secretory cell line, RBL. Stable patches of 2–4 pF are easily excised from RBL cells after partially disrupting actin cytoskeleton with latrunculin A. Membrane fusion is triggered by switching the patch to a cytoplasmic solution containing 100–200 μM free Ca. Capacitance and amperometric recording show that large secretory granules (SGs) containing serotonin are mostly lost from patches. Small vesicles that are retained (non-SGs) do not release serotonin or other substances detected by amperometry, although their fusion is reduced by tetanus toxin light chain. Non-SG fusion is unaffected by N-ethylmaleimide, phosphatidylinositol-4,5-bis-phosphate (PI(4,5)P2) ligands, such as neomycin, a PI-transfer protein that can remove PI from membranes, the PI(3)-kinase inhibitor LY294002 and PI(4,5)P2, PI(3)P, and PI(4)P antibodies. In patch recordings, but not whole-cell recordings, fusion can be strongly reduced by ATP removal and by the nonspecific PI-kinase inhibitors wortmannin and adenosine. In whole-cell recording, non-SG fusion is strongly reduced by osmotically induced cell swelling, and subsequent recovery after shrinkage is then inhibited by wortmannin. Thus, membrane stretch that occurs during patch formation may be a major cause of differences between excised patch and whole-cell fusion responses. Regarding Ca sensors for non-SG fusion, fusion remains robust in synaptotagmin (Syt) VII−/− mouse embryonic fibroblasts (MEFs), as well as in PLCδ1, PLC δ1/δ4, and PLCγ1−/− MEFs. Thus, Syt VII and several PLCs are not required. Furthermore, the Ca dependence of non-SG fusion reflects a lower Ca affinity (KD ∼71 μM) than expected for these C2 domain–containing proteins. In summary, we find that non-SG membrane fusion behaves and is regulated substantially differently from SG fusion, and we have identified an ATP

  13. Calsyntenins are secretory granule proteins in anterior pituitary gland and pancreatic islet alpha cells.

    PubMed

    Rindler, Michael J; Xu, Chong-Feng; Gumper, Iwona; Cen, Chuan; Sonderegger, Peter; Neubert, Thomas A

    2008-04-01

    Calsyntenins are members of the cadherin superfamily of cell adhesion molecules. They are present in postsynaptic membranes of excitatory neurons and in vesicles in transit to neuronal growth cones. In the current study, calsyntenin-1 (CST-1) and calsyntenin-3 (CST-3) were identified by mass spectrometric analysis (LC-MS/MS) of integral membrane proteins from highly enriched secretory granule preparations from bovine anterior pituitary gland. Immunofluorescence microscopy on thin frozen sections of rat pituitary revealed that CST-1 was present only in gonadotropes where it colocalized with follicle-stimulating hormone in secretory granules. In contrast, CST-3 was present not only in gonadotrope secretory granules but also in those of somatotropes and thyrotropes. Neither protein was detected in mammatropes. In addition, CST-1 was also localized to the glucagon-containing secretory granules of alpha cells in the pancreatic islets of Langerhans. Results indicate that calsyntenins function outside the nervous system and potentially are modulators of endocrine function.

  14. Astrocytes as secretory cells of the central nervous system: idiosyncrasies of vesicular secretion.

    PubMed

    Verkhratsky, Alexei; Matteoli, Michela; Parpura, Vladimir; Mothet, Jean-Pierre; Zorec, Robert

    2016-02-01

    Astrocytes are housekeepers of the central nervous system (CNS) and are important for CNS development, homeostasis and defence. They communicate with neurones and other glial cells through the release of signalling molecules. Astrocytes secrete a wide array of classic neurotransmitters, neuromodulators and hormones, as well as metabolic, trophic and plastic factors, all of which contribute to the gliocrine system. The release of neuroactive substances from astrocytes occurs through several distinct pathways that include diffusion through plasmalemmal channels, translocation by multiple transporters and regulated exocytosis. As in other eukaryotic cells, exocytotic secretion from astrocytes involves divergent secretory organelles (synaptic-like microvesicles, dense-core vesicles, lysosomes, exosomes and ectosomes), which differ in size, origin, cargo, membrane composition, dynamics and functions. In this review, we summarize the features and functions of secretory organelles in astrocytes. We focus on the biogenesis and trafficking of secretory organelles and on the regulation of the exocytotic secretory system in the context of healthy and diseased astrocytes.

  15. Early development of the secretory cavity of peltate glands in Humulus lupulus L. (Cannabaceae).

    PubMed

    Kim, E S; Mahlberg, P G

    2000-10-31

    Early development of the secretory cavity of chemically fixed peltate glands in Humulus lupulus L. showed secretions with different densities, light, gray and dark, in the cytoplasm of disc cells and in the periplasmic space adjacent to the developing secretory cavity. Secretions were detected in the disc cell wall and subsequently in the developing secretory cavity under the subcuticular wall of the sheath. Light and gray secretions in the cavity possessed a membrane-like surface feature. Secretions were in contact with the irregular inner surface of the cuticle. Secretions contributed to the thickening of the cuticle, whereas the membrane-like surface feature contributed to a network of Cannabis striae distributed throughout the cuticle. This study supports an early development and organization of the secretory cavity in H. lupulus, parallel to those in Cannabis, and may represent common features for lipophilic glands in angiosperms.

  16. Mapping organelle motion reveals a vesicular conveyor belt spatially replenishing secretory vesicles in stimulated chromaffin cells.

    PubMed

    Maucort, Guillaume; Kasula, Ravikiran; Papadopulos, Andreas; Nieminen, Timo A; Rubinsztein-Dunlop, Halina; Meunier, Frederic A

    2014-01-01

    How neurosecretory cells spatially adjust their secretory vesicle pools to replenish those that have fused and released their hormonal content is currently unknown. Here we designed a novel set of image analyses to map the probability of tracked organelles undergoing a specific type of movement (free, caged or directed). We then applied our analysis to time-lapse z-stack confocal imaging of secretory vesicles from bovine Chromaffin cells to map the global changes in vesicle motion and directionality occurring upon secretagogue stimulation. We report a defined region abutting the cortical actin network that actively transports secretory vesicles and is dissipated by actin and microtubule depolymerizing drugs. The directionality of this "conveyor belt" towards the cell surface is activated by stimulation. Actin and microtubule networks therefore cooperatively probe the microenvironment to transport secretory vesicles to the periphery, providing a mechanism whereby cells globally adjust their vesicle pools in response to secretagogue stimulation.

  17. Increased gastric secretory capacity in smokers without gastrointestinal lesions.

    PubMed Central

    Massarrat, S; Enschai, F; Pittner, P M

    1986-01-01

    The relationship between smoking and gastric secretory capacity was studied in 201 25-40 year old healthy subjects with normal laboratory data and no gastrointestinal lesions. Basal acid output (BAO) and peak acid output (PAO) were determined in all, and basal and stimulated pepsin outputs were measured in 85 participants. The accuracy of the patients' statements was checked by urinary nicotine assay. Basal acid output and PAO were significantly higher in male smokers (n = 55) than in male non-smokers (n = 49). In women PAO in smokers (n = 38) was higher than in non-smokers (n = 59). Female smokers (n = 38) had a higher pepsin output than female non-smokers (n = 23). Eight variables were considered in relation to BAO and PAO: age, height, weight, alcohol abuse, smoking habits, duration of smoking habit, number of cigarettes per day, and the product of years of smoking multiplied by daily number of cigarettes. The daily number of cigarettes X years of smoking was most closely correlated with BAO and PAO by stepwise multiple linear regression analysis. It was concluded that smoking is related to increased gastric acid capacity. PMID:3082724

  18. Cysteine rich secretory proteins in reproduction and venom.

    PubMed

    Gibbs, Gerard M; O'Bryan, Moira K

    2007-01-01

    The cysteine rich secretory proteins (Crisp) are predominantly found in the mammalian male reproductive tract and in the venom of reptiles. Crisps are two domain proteins with a structurally similar yet evolutionarily diverse N-terminal domain and a characteristic cysteine rich C-terminal domain which we refer to as the Crisp domain. Since their identification 30 years ago Crisp research in mammals has focused on the characterisation of their expression localization to infer function. While no doubt important observations, these have not substantially led to an understanding of the biochemical activity of the Crisps and their role in sperm function or fertilisation. Recently, we demonstrated that the Crisp-2 Crisp domain has a structure similar to ion channel toxins ShK and BgK and was itself able to regulate Ca2+ flux through ryanodine receptors. These data build upon the previous characterizations of reptile venom Crisps as regulators of several types of ion channels and permits for the first time a dissection of the biochemical activity of mammalian Crisps.

  19. Hepatitis C Virus Is Released via a Noncanonical Secretory Route.

    PubMed

    Bayer, Karen; Banning, Carina; Bruss, Volker; Wiltzer-Bach, Linda; Schindler, Michael

    2016-12-01

    We analyzed hepatitis C virus (HCV) morphogenesis using viral genomes encoding a mCherry-tagged E1 glycoprotein. HCV-E1-mCherry polyprotein expression, intracellular localization, and replication kinetics were comparable to those of untagged HCV, and E1-mCherry-tagged viral particles were assembled and released into cell culture supernatants. Expression and localization of structural E1 and nonstructural NS5A followed a temporospatial pattern with a succinct decrease in the number of replication complexes and the appearance of E1-mCherry punctae. Interaction of the structural proteins E1, Core, and E2 increased at E1-mCherry punctae in a time-dependent manner, indicating that E1-mCherry punctae represent assembled or assembling virions. E1-mCherry did not colocalize with Golgi markers. Furthermore, the bulk of viral glycoproteins within released particles revealed an EndoH-sensitive glycosylation pattern, indicating an absence of viral glycoprotein processing by the Golgi apparatus. In contrast, HCV-E1-mCherry trafficked with Rab9-positive compartments and inhibition of endosomes specifically suppressed HCV release. Our data suggest that assembled HCV particles are released via a noncanonical secretory route involving the endosomal compartment.

  20. Proteomic analysis of Toxocara canis excretory and secretory (TES) proteins.

    PubMed

    Sperotto, Rita Leal; Kremer, Frederico Schmitt; Aires Berne, Maria Elisabeth; Costa de Avila, Luciana F; da Silva Pinto, Luciano; Monteiro, Karina Mariante; Caumo, Karin Silva; Ferreira, Henrique Bunselmeyer; Berne, Natália; Borsuk, Sibele

    2017-01-01

    Toxocariasis is a neglected disease, and its main etiological agent is the nematode Toxocara canis. Serological diagnosis is performed by an enzyme-linked immunosorbent assay using T. canis excretory and secretory (TES) antigens produced by in vitro cultivation of larvae. Identification of TES proteins can be useful for the development of new diagnostic strategies since few TES components have been described so far. Herein, we report the results obtained by proteomic analysis of TES proteins using a liquid chromatography-tandem mass spectrometry (LC-MS/MS) approach. TES fractions were separated by one-dimensional SDS-PAGE and analyzed by LC-MS/MS. The MS/MS spectra were compared with a database of protein sequences deduced from the genome sequence of T. canis, and a total of 19 proteins were identified. Classification according to the signal peptide prediction using the SignalP server showed that seven of the identified proteins were extracellular, 10 had cytoplasmic or nuclear localization, while the subcellular localization of two proteins was unknown. Analysis of molecular functions by BLAST2GO showed that the majority of the gene ontology (GO) terms associated with the proteins present in the TES sample were associated with binding functions, including but not limited to protein binding (GO:0005515), inorganic ion binding (GO:0043167), and organic cyclic compound binding (GO:0097159). This study provides additional information about the exoproteome of T. canis, which can lead to the development of new strategies for diagnostics or vaccination.

  1. Excretory/secretory proteome of 14-day schistosomula, Schistosoma japonicum.

    PubMed

    Cao, Xiaodan; Fu, Zhiqiang; Zhang, Min; Han, Yanhui; Han, Qian; Lu, Ke; Li, Hao; Zhu, Chuangang; Hong, Yang; Lin, Jiaojiao

    2016-01-01

    Schistosomiasis remains a serious public health problem, with 200 million people infected and 779 million people at risk worldwide. The schistosomulum is the early stage of the complex lifecycle of Schistosoma japonicum in their vertebrate hosts, and is the main target of vaccine-induced protective immunity. Excretory/secretory (ES) proteins play a major role in host-parasite interactions and ES protein compositions of schistosomula of S. japonicum have not been characterized to date. In the present study, the proteome of ES proteins from 14 day schistosomula of S. japonicum was analyzed by liquid chromatography/tandem mass spectrometry and 713 unique proteins were finally identified. Gene ontology and pathway analysis revealed that identified proteins were mainly involved in carbohydrate metabolism, degradation, response to stimulus, oxidation-reduction, biological regulation and binding. Flow cytometry analysis demonstrated that thioredoxin peroxidase identified in this study had the effect on inhibiting MHCII and CD86 expression on LPS-activated macrophages. The present study provides insight into the growth and development of the schistosome in the final host and valuable information for screening vaccine candidates for schistosomiasis.

  2. Role of NBCe1 and AE2 in Secretory Ameloblasts

    PubMed Central

    Paine, Michael L.; Snead, Malcolm L.; Wang, HongJun; Abuladze, Natalia; Pushkin, Alexander; Liu, Weixin; Kao, Li Yo; Wall, Susan M.; Kim, Young-Hee; Kurtz, Ira

    2008-01-01

    The H+/base transport processes that control the pH of the microenvironment adjacent to ameloblasts are not currently well understood. Mice null for the AE2 anion exchanger have abnormal enamel. In addition, patients with mutations in the electrogenic sodium bicarbonate cotransporter NBCe1 and mice lacking NBCe1 have enamel abnormalities. These observations suggest that AE2 and NBCe1 play important roles in amelogenesis. The present study aimed to understand the roles of AE2 and NBC1 in ameloblasts. The data showed that NBCe1 is expressed at the basolateral membrane of secretory ameloblasts, whereas AE2 is expressed at the apical membrane. Transcripts for AE2a and NBCe1-B were detected in RNA isolated from cultured ameloblast-like LS8 cells. Our data are the first evidence that AE2 and NBCe1 are expressed in ameloblasts in vivo in a polarized fashion thereby providing a mechanism for ameloblast transcellular bicarbonate secretion in the process of enamel formation and maturation. PMID:18362326

  3. Secretory function of adrenal cortex in chronic alcoholis.

    PubMed

    Feher, I

    1999-01-01

    Three groups of male subjects (healthy subjects, chronic alcoholics with liver cirrhosis and patients with acute viral hepatitis) were included in a 24 hour pattern of excretion of the total and some fractions of 17-ketosteroids (KS), basal concentration of 11-hydroxycorticosteroids (11-OHCS) and dehydroepiandrosterone (DHEA) in plasma, as well as changes of concentration of the same steroids in plasma 15, 30 and 60 minutes after a single i.m. injection of insulin. In regard to healthy subjects and patients with acute viral hepatitis, chronic alcoholics with liver cirrhosis excrete decreased quantities of total and some fractions of 17-KS. In regard to healthy subjects, decreased excretion of the sum androsterone and etiocholanole was established as well as increased DHEA secretion in patients with acute viral hepatitis. In chronic alcoholics with liver cirrhosis basal concentrations of 11-OHCS in plasma and their increase after insulin administration are the same as in healthy subjects, but values of DHEA concentrations in plasma are decreased. It has been pointed to the possibility of damages of the secretory function of adrenal cortex in chronic alcoholics with liver cirrhosis. On the basis of above mentioned results, there is an assumption that adrenal gland primarily provides normal secretion of C21 steroid and thus, satisfying needs for these steroids, increases secretion of DHEA. Follow up of DHEA urinary secretion may provide insight into basal activity of adrenal cortex, whereas the functional state of the liver must be taken into account when interpreting the results.

  4. Detection of the senescence-associated secretory phenotype (SASP).

    PubMed

    Rodier, Francis

    2013-01-01

    Cellular senescence suppresses cancer by eliminating potentially oncogenic cells, participates in tissue repair, contributes to cancer therapy, and promotes organismal aging. Numerous activities of senescent cells depend on the aptitude of these cells to secrete myriads of bioactive molecules, a behavior termed the senescence-associated secretory phenotype (SASP). The SASP supports cell-autonomous functions like the senescence-associated growth arrest, and mediates paracrine interactions between senescent cells and their surrounding microenvironment. The biological functions and the regulation of the SASP are beginning to emerge, and current SASP assessment techniques include the analysis of SASP factors at the mRNA level, the direct measurement of factors inside or outside the cell (i.e., secreted), and the detection of SASP-provoked cellular responses. Here, we focus on a simple approach to collect SASP-conditioned media in order to directly measure secreted SASP factors using sandwich enzyme-linked immunosorbent assay. As an example, we discuss the assessment of the major SASP factor interleukin-6 in senescent human fibroblasts. Supplemental notes are provided to easily adapt this procedure to other SASP factors, change cell types, or scale the techniques for different volumes or high-throughput measurements. These techniques should facilitate the discovery of novel functions and regulators of the SASP.

  5. The complete general secretory pathway in gram-negative bacteria.

    PubMed Central

    Pugsley, A P

    1993-01-01

    The unifying feature of all proteins that are transported out of the cytoplasm of gram-negative bacteria by the general secretory pathway (GSP) is the presence of a long stretch of predominantly hydrophobic amino acids, the signal sequence. The interaction between signal sequence-bearing proteins and the cytoplasmic membrane may be a spontaneous event driven by the electrochemical energy potential across the cytoplasmic membrane, leading to membrane integration. The translocation of large, hydrophilic polypeptide segments to the periplasmic side of this membrane almost always requires at least six different proteins encoded by the sec genes and is dependent on both ATP hydrolysis and the electrochemical energy potential. Signal peptidases process precursors with a single, amino-terminal signal sequence, allowing them to be released into the periplasm, where they may remain or whence they may be inserted into the outer membrane. Selected proteins may also be transported across this membrane for assembly into cell surface appendages or for release into the extracellular medium. Many bacteria secrete a variety of structurally different proteins by a common pathway, referred to here as the main terminal branch of the GSP. This recently discovered branch pathway comprises at least 14 gene products. Other, simpler terminal branches of the GSP are also used by gram-negative bacteria to secrete a more limited range of extracellular proteins. PMID:8096622

  6. Gut Microbial Succession Follows Acute Secretory Diarrhea in Humans

    PubMed Central

    David, Lawrence A.; Weil, Ana; Ryan, Edward T.; Calderwood, Stephen B.; Harris, Jason B.; Chowdhury, Fahima; Begum, Yasmin; Qadri, Firdausi

    2015-01-01

    ABSTRACT Disability after childhood diarrhea is an important burden on global productivity. Recent studies suggest that gut bacterial communities influence how humans recover from infectious diarrhea, but we still lack extensive data and mechanistic hypotheses for how these bacterial communities respond to diarrheal disease and its treatment. Here, we report that after Vibrio cholerae infection, the human gut microbiota undergoes an orderly and reproducible succession that features transient reversals in relative levels of enteric Bacteroides and Prevotella. Elements of this succession may be a common feature in microbiota recovery from acute secretory diarrhea, as we observed similar successional dynamics after enterotoxigenic Escherichia coli (ETEC) infection. Our metagenomic analyses suggest that multiple mechanisms drive microbial succession after cholera, including bacterial dispersal properties, changing enteric oxygen and carbohydrate levels, and phage dynamics. Thus, gut microbiota recovery after cholera may be predictable at the level of community structure but is driven by a complex set of temporally varying ecological processes. Our findings suggest opportunities for diagnostics and therapies targeting the gut microbiota in humans recovering from infectious diarrhea. PMID:25991682

  7. Acidification in the epidermis and the role of secretory phospholipases

    PubMed Central

    Chan, Aegean

    2011-01-01

    The function of the epidermis is to form an effective barrier between the dry, external environment and the interior of the body. The barrier specifically resides in the extracellular lipid membranes of the stratum corneum (SC) and an acidic pH is necessary to maintain its competency against various insults. The purpose of this review is to explore the mechanisms which are postulated to contribute to the acidification of the stratum corneum, including both exogenous and endogenous sources. However, recent research as pointed to several endogenous mechanisms as the major source of acidification, including a sodium/proton pump (NHE1) and free fatty acid conversion from phospholipids by secretory phospholipase A2 (sPLA2). sPLA2 has been shown to play a central role in the formation of the SC “acid mantle” in the early maturation of the epidermis postnatally. Many aspects of this enzyme family are complex and still being elucidated in research and the most recent findings on the localization and functions of sPL A2-IB, -IIA, -IIC, -IID, -IIE, -IIF, -III, -V, -X and -XII in the epidermis are presented here. Given their role in inflammatory dermatoses, such as psoriasis and atopic dermatitis, understanding this complex enzyme family can lead to novel, life-changing therapies. PMID:21695017

  8. Synthesis of Prostaglandins and Eicosanoids by the Mast Cell Secretory Granule

    DTIC Science & Technology

    1988-01-01

    various lipid-derived mediators during exocytosis. MATERIALS AND METHODS The procedure for granule preparation is similar to that which has been described...Press, Inc. Printed in U.S.A. SYNTHESIS OF PROSTACLANDINS AND KICOSANOIDS BY THE M&ST CELL SECRETORY GRANULE Stephen P. Chock and Elsa A. Schmauder-Chock...SCISNTIFIC XeiOT Received September 30, 1988 SR88-32 The identification of a non-bilayer phospholipid storage in the secretory granule and the linking of

  9. Brevibacillus expression system: host-vector system for efficient production of secretory proteins.

    PubMed

    Mizukami, Makoto; Hanagata, Hiroshi; Miyauchi, Akira

    2010-04-01

    Brevibacillus expression system is an effective bacterial expression system for secretory proteins. The host bacterium, Brevibacillus choshinensis, a gram-positive bacterium, has strong capacity to secrete a large amount of proteins (approximately 30 g/L), which mostly consist of cell wall protein. A host-vector system that utilizes such high expression capacity has been constructed for the production of secretory proteins and tested for various heterologous proteins, including cytokines, enzymes, antigens, and adjuvants.

  10. Amniotic fluid secretory immunoglobulin A in normal pregnancy and in pregnancy complicated by rhesus isoimmunization.

    PubMed

    Briese, V; Straube, W; Brock, J; Stark, K H; Lorenz, U

    1982-01-01

    Amniotic fluid levels of secretory immunoglobulin A (S-IgA) were measured by simple radial immunodiffusion according to the method of Mancini using a monospecific antiserum against the human secretory component. We examined 256 samples from healthy pregnant women and 149 samples from mothers suffering from rhesus isoimmunization. During normal pregnancy amniotic fluid S-IgA increased significantly and showed a loose correlation with the phospholipid levels. This was not observed in pregnancies complicated by rhesus isoimmunization.

  11. Oil Secretory System in Vegetative Organs of Three Arnica Taxa: Essential Oil Synthesis, Distribution and Accumulation.

    PubMed

    Kromer, Krystyna; Kreitschitz, Agnieszka; Kleinteich, Thomas; Gorb, Stanislav N; Szumny, Antoni

    2016-05-01

    Arnica, a genus including the medicinal species A. montana, in its Arbo variety, and A. chamissonis, is among the plants richest in essential oils used as pharmaceutical materials. Despite its extensive use, the role of anatomy and histochemistry in the internal secretory system producing the essential oil is poorly understood. Anatomical sections allowed differentiation between two forms of secretory structures which differ according to their distribution in plants. The first axial type is connected to the vascular system of all vegetative organs and forms canals lined with epithelial cells. The second cortical type is represented by elongated intercellular spaces filled with oil formed only between the cortex cells of roots and rhizomes at maturity, with canals lacking an epithelial layer.Only in A. montana rhizomes do secretory structures form huge characteristic reservoirs. Computed tomography illustrates their spatial distribution and fusiform shape. The axial type of root secretory canals is formed at the interface between the endodermis and cortex parenchyma, while, in the stem, they are located in direct contact with veinal parenchyma. The peripheral phloem parenchyma cells are arranged in strands around sieve tube elements which possess a unique ability to accumulate large amounts of oil bodies. The cells of phloem parenchyma give rise to the aforementioned secretory structures while the lipid components (triacylglycerols) stored there support the biosynthesis of essential oils by later becoming a medium in which these oils are dissolved. The results indicate the integrity of axial secretory structures forming a continuous system in vegetative plant organs.

  12. Immunomodulatory potential of particular Trichinella spiralis muscle larvae excretory-secretory components.

    PubMed

    Cvetkovic, J; Sofronic-Milosavljevic, Lj; Ilic, N; Gnjatovic, M; Nagano, I; Gruden-Movsesijan, A

    2016-12-01

    Excretory-secretory antigens of Trichinella spiralis muscle larvae can induce the semi-matured status of rat dendritic cells. This may at least partly be the consequence of transient activation of extracellular signal-regulated kinases 1/2 (ERK1/2). Here we investigated the potential of several components of excretory-secretory antigens (native fraction containing 45, 49 and 53kDa proteins and recombinant Tsp53, representing one of the constituents of this fraction) to demonstrate previously observed effects of excretory-secretory antigens on dendritic cells in vitro, characterised by establishment of a particular phenotype (very low MHC II expression, moderate CD86 expression and significant ICAM-1 expression) and functional properties (low production of pro-inflammatory cytokine IL-12p70, and high production of IL-10 and TGF-β). Dendritic cells activated by these components were able to provoke proliferation of naïve T cells and their polarisation towards Th2 and anti-inflammatory responses. The investigated antigens had almost the same capacity to induce IL-4 and IL-10 production from T cells as excretory-secretory antigens, but failed to induce significant TGF-β synthesis. It could be concluded that the investigated excretory-secretory antigens components can largely reproduce the immunomodulatory effects of the complete excretory-secretory antigens and therefore may be considered as molecules important for creation of the anti-inflammatory milieu achieved by the parasite.

  13. Excretory/secretory products from in vitro-cultured Echinococcus granulosus protoscoleces.

    PubMed

    Virginio, Veridiana G; Monteiro, Karina M; Drumond, Fernanda; de Carvalho, Marcos O; Vargas, Daiani M; Zaha, Arnaldo; Ferreira, Henrique B

    2012-05-01

    Cystic hydatid disease (CHD) is caused by infection with Echinococcus granulosus metacestodes and affects humans and livestock. Proteins secreted or excreted by protoscoleces, pre-adult worms found in the metacestode, are thought to play fundamental roles in the host-parasite relationship. In this work, we performed an LC-MS/MS proteomic analysis of the excretory-secretory products obtained from the first 48 h of an in vitro culture of the protoscoleces. We identified 32 proteins, including 18 that were never detected previously in metacestode proteomic studies. Among the novel identified excretory-secretory products are antigenic proteins, such as EG19 and P-29 and a calpain protease. We also identified other important protoscolex excretory-secretory products, such as thioredoxin peroxidase and 14-3-3 proteins, which are potentially involved in evasion mechanisms adopted by parasites to establish infection. Several intracellular proteins were found in the excretory-secretory products, revealing a set of identified proteins not previously thought to be exposed at the host-parasite interface. Additionally, immunological analyses established the antigenic profiles of the newly identified excretory-secretory products and revealed, for the first time, the in vitro secretion of the B antigen by protoscoleces. Considering that the excretory-secretory products obtained in vitro might reflect the products released and exposed to the host in vivo, our results provide valuable information on parasite survival strategies in adverse host environments and on the molecular mechanisms underpinning CHD immunopathology.

  14. Epithelial Cell Culture from Human Adenoids: A Functional Study Model for Ciliated and Secretory Cells

    PubMed Central

    González, Claudia; Espinosa, Marisol; Sánchez, María Trinidad; Droguett, Karla; Ríos, Mariana; Fonseca, Ximena; Villalón, Manuel

    2013-01-01

    Background. Mucociliary transport (MCT) is a defense mechanism of the airway. To study the underlying mechanisms of MCT, we have both developed an experimental model of cultures, from human adenoid tissue of ciliated and secretory cells, and characterized the response to local chemical signals that control ciliary activity and the secretion of respiratory mucins in vitro. Materials and Methods. In ciliated cell cultures, ciliary beat frequency (CBF) and intracellular Ca2+ levels were measured in response to ATP, UTP, and adenosine. In secretory cultures, mucin synthesis and secretion were identified by using immunodetection. Mucin content was taken from conditioned medium and analyzed in the presence or absence of UTP. Results. Enriched ciliated cell monolayers and secretory cells were obtained. Ciliated cells showed a basal CBF of 10.7 Hz that increased significantly after exposure to ATP, UTP, or adenosine. Mature secretory cells showed active secretion of granules containing different glycoproteins, including MUC5AC. Conclusion. Culture of ciliated and secretory cells grown from adenoid epithelium is a reproducible and feasible experimental model, in which it is possible to observe ciliary and secretory activities, with a potential use as a model to understand mucociliary transport control mechanisms. PMID:23484122

  15. The organization of the secretory machinery in chromaffin cells as a major factor in modeling exocytosis

    PubMed Central

    Villanueva, José; Torregrosa-Hetland, Cristina J.; Gil, Amparo; González-Vélez, Virginia; Segura, Javier; Viniegra, Salvador; Gutiérrez, Luis M.

    2010-01-01

    The organization of cytoplasm in excitable cells was a largely ignored factor when mathematical models were developed to understand intracellular calcium and secretory behavior. Here we employed a combination of fluorescent evanescent and transmitted light microscopy to explore the F-actin cytoskeletal organization in the vicinity of secretory sites in cultured bovine chromaffin cells. This technique and confocal fluorescent microscopy show chromaffin granules associated with the borders of cortical cytoskeletal cages forming an intricate tridimensional network. Furthermore, the overexpression of SNAP-25 in these cells also reveals the association of secretory machinery clusters with the borders of these cytoskeletal cages. The importance of these F-actin cage borders is stressed when granules appear to interact and remain associated during exocytosis visualized in acridin orange loaded vesicles. These results will prompt us to propose a model of cytoskeletal cages, where the secretory machinery is associated with its borders. Both the calcium level and the secretory response are enhanced in this geometrical arrangement when compared with a random distribution of the secretory machinery that is not restricted to the borders of the cage. PMID:20885775

  16. The use of lectins as markers for differentiated secretory cells in planarians.

    PubMed

    Zayas, Ricardo M; Cebrià, Francesc; Guo, Tingxia; Feng, Junjie; Newmark, Phillip A

    2010-11-01

    Freshwater planarians have reemerged as excellent models to investigate mechanisms underlying regeneration. The introduction of molecular tools has facilitated the study of planarians, but cell- and tissue-specific markers are still needed to examine differentiation of most cell types. Here we report the utility of fluorescent lectin-conjugates to label tissues in the planarian Schmidtea mediterranea. We show that 16 lectin-conjugates stain planarian cells or tissues; 13 primarily label the secretory cells, their cytoplasmic projections, and terminal pores. Thus, we examined regeneration of the secretory system using lectin markers and functionally characterized two genes expressed in the secretory cells: marginal adhesive gland-1 (mag-1) and Smed-reticulocalbin1 (Smed-rcn1). RNAi knockdown of these genes caused a dramatic reduction of secretory cell lectin staining, suggesting a role for mag-1 and Smed-rcn1 in secretory cell differentiation. Our results provide new insights into planarian secretory system regeneration and add new markers for labeling several planarian tissues.

  17. HID-1 is required for homotypic fusion of immature secretory granules during maturation

    PubMed Central

    Du, Wen; Zhou, Maoge; Zhao, Wei; Cheng, Dongwan; Wang, Lifen; Lu, Jingze; Song, Eli; Feng, Wei; Xue, Yanhong; Xu, Pingyong; Xu, Tao

    2016-01-01

    Secretory granules, also known as dense core vesicles, are generated at the trans-Golgi network and undergo several maturation steps, including homotypic fusion of immature secretory granules (ISGs) and processing of prehormones to yield active peptides. The molecular mechanisms governing secretory granule maturation are largely unknown. Here, we investigate a highly conserved protein named HID-1 in a mouse model. A conditional knockout of HID-1 in pancreatic β cells leads to glucose intolerance and a remarkable increase in the serum proinsulin/insulin ratio caused by defective proinsulin processing. Large volume three-dimensional electron microscopy and immunofluorescence imaging reveal that ISGs are much more abundant in the absence of HID-1. We further demonstrate that HID-1 deficiency prevented secretory granule maturation by blocking homotypic fusion of immature secretory granules. Our data identify a novel player during the early maturation of immature secretory granules. DOI: http://dx.doi.org/10.7554/eLife.18134.001 PMID:27751232

  18. Non-planar Fault Model of the 2008 Yutian Normal Faulting Earthquake (M7.2), Xinjiang, China, and its implications

    NASA Astrophysics Data System (ADS)

    Furuya, M.; Yasuda, T.

    2010-12-01

    On 20 March 2008, a normal-faulting earthquake (M7.2/USGS) struck Yutian county, Xinjiang, China. The epicenter is close to the southern-edge of the Tarim basin, where three prominent fault systems, the Altyn-Tagh Fault, Karakax Fault, and Longmu-Gozha Co Fault, meet together at the northwest of the Tibetan plateau. The associated crustal deformation signals, if detected and examined in detail, will provide us with important constraints on how the continental crust in Tibetan plateau has been deforming, which has been contentious over the decades. Also, it could suggest clues on why normal faulting earthquakes prevail over the central to northern areas of Tibet. Here, based on our detected co-seismic crustal deformation signals by synthetic aperture radar (SAR) data, we report a non-planar kinematic fault source model that suggests the normal faulting rupture occurred in a dilatational step-over region of two oblique left-lateral strike slip faults. We used L-band ALOS/PALSAR data for the ascending track and C-band Envisat/ASAR data for the descending track. The observed data showed that the overall strike was from NE to SW, consistent with Shao and Ji (2008)'s preliminary source model. The largest slipped area in the InSAR data, however, indicates that the strike direction is almost NS, and that the rupture reached to the surface. The pixel-offset data also illustrate that the strike direction changed around the middle of the fault trace on the surface, suggesting that the fault slip occurred on a non-planar surface. In order to realistically represent the non-planar geometry, we employed an analytical formulation for the triangular dislocation element to invert the slip distribution (e.g., Mearten et al., 2005; Mead, 2007). In inverting the fault slip, both the non-negativity constraint on the slip direction and the smoothing constraint on the slip distribution were applied. Optimum fault source and its implications for regional tectonics and mechanism for the

  19. Atom Probe Tomography of Planar and Nonplanar InGaN Quantum Wells For Energy-Efficient Solid-State Lighting

    NASA Astrophysics Data System (ADS)

    Riley, James R.

    Atom probe tomography (APT) was used to characterize the composition and morphology of group-III-nitride nanowires, planar quantum wells (QWs), and nonplanar light-emitting diode (LED) heterostructures that all have applications for next-generation energy-efficient lighting. To demonstrate reliable APT analysis, the influence of extrinsic experimental factors and intrinsic materials characteristics on the field evaporation of group-III and nitrogen atoms was determined. A decrease in gallium ion detection due to uncorrelated evaporation events was shown to increase with increasing DC voltage, while a deficit in nitrogen ion detection was caused by uncorrelated evaporation of N2 ions following thermally-driven adatom diffusion. This nitrogen deficit was exacerbated on Ga-polar surfaces due to the variation in bonding coordination. Despite this complicated evaporation, the first evidence of reliable measurement of indium mole fraction regardless of surface polarity is provided. This result enabled comparative analysis of the interfacial abruptness and indium distribution in planar QWs grown along polar, nonpolar, and semipolar directions. In all cases the indium distribution did not exhibit statistically significant deviations from the atomic distribution expected for a random alloy, although nanometer-scale discontinuities in the QWs were observed in some cases. APT was used to map the morphology of QWs intentionally dosed with hydrogen to promote discontinuity formation and this information was utilized to improve state-of-the art high-resolution x-ray diffraction models which did not previously account for these types of morphological fluctuations. Finally, nonplanar LEDs were shown to exhibit broader light-emission spectra than conventional planar LEDs. To identify the origin of this spectral broadening, QWs grown on nanowire facets of different crystalline polarity were investigated with APT and correlated cathodoluminescence spectroscopy. The indium content in

  20. Resonantly pumped monolithic nonplanar Ho:YAG ring laser with high-power single-frequency laser output at 2122 nm.

    PubMed

    Wang, Lei; Gao, Chunqing; Gao, Mingwei; Li, Yan

    2013-04-22

    We demonstrated a stable single-frequency laser operating at 2122 nm from a monolithic nonplanar Ho:YAG ring oscillator (NPRO). The Ho:YAG NPRO was resonantly pumped by a 1907 nm Tm:YLF laser built up by ourselves. The maximum multimode output power from the Ho:YAG NPRO was 9.66 W and the slope efficiency was 71.7%. With accurate adjustment of the pump position to make the laser oscillate in single frequency condition, an output power of 8.0 W was obtained with a slope efficiency of 61.4% and an optical-optical efficiency of 50.0%. The power stability of the Ho:YAG NPRO laser was 0.29% at maximum single frequency output power. The beam quality M(2) factors were measured to be less than 1.1 in x- and y- directions.

  1. Nonplanar structure of C6H5SCF3 facilitates πσ∗-mediated photodissociation reaction on the S1 state

    NASA Astrophysics Data System (ADS)

    Kim, So-Yeon; Lee, Jeongmook; Kim, Sang Kyu; Choi, Young S.

    2016-08-01

    Vibrational structure of trifluoromethylthiobenzene (C6H5SCF3) on the S1 state has been investigated by resonance-enhanced two-photon ionization spectroscopy and nature of predissociation dynamics is inferred from homogeneously broadened spectral features. As C6H5SCF3 adopts a nonplanar structure in both the S0 and S1 states, the effective adiabatic barrier generated by avoided crossing of optically-bright bound S1 (ππ∗) and dark-repulsive S2 (πσ∗) surfaces along the reaction coordinate is significantly lowered, giving the S1 lifetime of ∼300 fs. This experiment demonstrates that the molecular structure spanned by the reactive flux near the curve-crossing region dictates reaction rate as well as nonadiabatic transition probability.

  2. White emission from non-planar InGaN/GaN MQW LEDs grown on GaN template with truncated hexagonal pyramids.

    PubMed

    Lee, Ming-Lun; Yeh, Yu-Hsiang; Tu, Shang-Ju; Chen, P C; Lai, Wei-Chih; Sheu, Jinn-Kong

    2015-04-06

    Non-planar InGaN/GaN multiple quantum well (MQW) structures are grown on a GaN template with truncated hexagonal pyramids (THPs) featuring c-plane and r-plane surfaces. The THP array is formed by the regrowth of the GaN layer on a selective-area Si-implanted GaN template. Transmission electron microscopy shows that the InGaN/GaN epitaxial layers regrown on the THPs exhibit different growth rates and indium compositions of the InGaN layer between the c-plane and r-plane surfaces. Consequently, InGaN/GaN MQW light-emitting diodes grown on the GaN THP array emit multiple wavelengths approaching near white light.

  3. Effects of non-extensive electrons and positive /negative dust particles on modulational instability of dust-ion-acoustic solitary waves in non-planar geometry

    NASA Astrophysics Data System (ADS)

    Eghbali, M.; Farokhi, B.; Eslamifar, M.

    2017-01-01

    The nonlinear propagation of cylindrical and spherical dust-ion-acoustic (DIA) envelope solitary waves in unmagnetized dusty plasma consisting of dust particles with opposite polarity and non-extensive distribution of electron is investigated. By using the reductive perturbation method, the modified nonlinear Schrödinger (NLS) equation in cylindrical and spherical geometry is obtained. The modulational instability (MI) of DIA waves governed by the NLS equation is also presented. The effects of different ranges of the non-extensive parameter q on the MI are studied. The growth rate of the MI is also given for different values of q. It is found that the basic features of the DIA waves are significantly modified by non-extensive electron distribution, polarity of the net dust-charge number density and non-planar geometry.

  4. Secretory pathway Ca2+/Mn2+-ATPase isoform 2 and lactation: specific localization of plasmalemmal and secretory pathway Ca2+ pump isoforms in the mammary gland

    SciTech Connect

    Faddy, Helen M.; Smart, Chanel E.; Xu, Ren; Lee, Genee Y.; Kenny, Paraic A.; Feng, Mingye; Rao, Rajini; Brown, Melissa A.; Bissell, Mina J.; Roberts-Thomson, Sarah J.; Monteith, Gregory R.

    2008-04-09

    The supply of calcium to the developing neonate via milk is an important physiological process. Until recently the mechanism for the enrichment of milk with calcium was thought to be almost entirely mediated via the secretory pathway. However, recent studies suggest that a specific isoform of the plasma membrane calcium ATPase, PMCA2, is the primary mechanism for calcium transport into milk, highlighting a major role for apical calcium transport. We compared the expression of the recently identified secretory calcium ATPase, SPCA2, and SPCA1, in the mouse mammary gland during different stages of development. SPCA2 levels increased over 35 fold during lactation, while SPCA1 increased only a modest two fold. The potential importance of SPCA2 in lactation was also highlighted by its localization to luminal secretory cells of the mammary gland during lactation, while SPCA1 was expressed throughout the cells of the mammary gland. We also observed major differences in the localization of PMCA2 and PMCA1 during lactation. Using the SCp2 mouse mammary epithelial cell 3D culture model, differences in the sub-cellular distribution of PMCA2 and PMCA1 were clear. These studies highlight the likely specific roles of PMCA2 and SPCA2 in lactation, and link the recently characterized SPCA2 calcium pump to the supply of calcium into milk and the regulation of Golgi resident enzymes important in lactation. They also indicate that calcium transport into milk is a complex interplay between apical and secretory pathways.

  5. Short-term cholinergic desensitization of rat pancreatic secretory response

    SciTech Connect

    Asselin, J.; Larose, L.; Morisset, J.

    1987-03-01

    Dispersed pancreatic acini were first exposed to carbamylcholine (10/sup -7/-10/sup -4/ M) for 60 min, washed, and reexposed to this same agonist (10/sup -8/-10/sup -3/ M) for 15 min. During this second incubation, the functional secretory capacity of these acini was evaluated by measuring amylase release. Acini preexposed to concentrations of carbamylcholine of 10/sup -6/ M or greater showed shifts to the right in the subsequent carbamylcholine dose-response curves of amylase release. A 3-h recovery period (without carbamylcholine) did not restore the altered carbamylcholine dose-response curve. Ca/sup 2 +/ concentrations of 10/sup -7/ M or 2.5 x 10/sup -3/ M instead of 0.5 x 10/sup -3/ M during the 60-min preincubation did not affect the desensitization process. With use of N-(/sup 3/H)methylscopolamine to evaluate muscarinic receptors, the only changes observed after desensitization were a significant decrease in the high-affinity and an equivalent increase in that of the low-affinity receptors. After cholinergic exposure amylase release stimulated by caerulein was only slightly modified, whereas amylase release in response to a phorbol ester 12-O-tetradecanoylphorbol-13-acetate and to the ionophore A23187 was not altered. These data indicate that short-term desensitization with a cholinergic agent is relatively specific to muscarinic agonists, causes changes in the muscarinic receptor high-and low-affinity concentration but does not alter intracellular steps after calcium mobilization or protein kinase C activation known to be involved in the secretion process.

  6. Postnatal development of bile secretory physiology in the dog

    SciTech Connect

    Tavoloni, N.; Jones, M.J.; Berk, P.D.

    1985-04-01

    To determine whether bile formation in the dog is an immature process at birth, several determinants of bile secretion were studied in anesthetized, bile duct-cannulated puppies of 0-42 days of age and adult dogs. Basal canalicular bile flow rate, estimated by /sup 14/C-erythritol biliary clearance, averaged 0.182 microliter/min/g liver in 0-3 day-old puppies and increased to 0.324 and 0.461 microliter/min/g in puppies 7-21 and 28-42 days of age, respectively. Calculated ductular bile water reabsorption (/sup 14/C-erythritol biliary clearance-bile flow) was virtually absent in 0-3 day-old puppies, and averaged 0.017 and 0.092 microliter/min/g in puppies of 7-21 and 28-42 days of age, respectively. In adult dogs, ductular bile water reabsorption was 0.132 microliter/min/g. These functional deficiencies of the newborn dog were associated with an increased biliary permeability to /sup 3/H-inulin which could not be accounted for solely by an increased solute diffusion due to the lower rate of canalicular bile flow. Administration of taurocholate up to 2000 nmol/min/kg produced in all animals a similar increase in canalicular bile flow and bile acid excretion, and was not associated with changes in ductular bile water reabsorption rate. These findings are interpreted to indicate that, in the dog, bile secretory function is immature at birth and develops during postnatal life.

  7. Systemic tolerance and secretory immunity after oral immunization

    PubMed Central

    1980-01-01

    Diminished systemic immune reaction after ingestion of antigen has been reported in several animal models. Conversely, it has been reported recently that oral immunization may lead to the production of secretory antibodies. To determine whether these events could occur concurrently, CBA/J mice were immunized intragastrically with varying doses of ovalbumin (OVA) and Streptococcus mutans. After 7 d, the animals were challenged systemically with antigen in complete adjuvant and 8 d later serum and saliva taken, and the draining lymph nodes assayed for a proliferative response. Intragastric doses of 1 mg OVA or 10(9) S. mutans led to significant suppression of the proliferative response, and intragastric doses of 10 mg OVA or 2.5 X 10(9) S. mutans led to the production of detectable salivary antibodies using hemagglutination. Serum antibodies were not detected after intragastric administration of OVA or S. mutans. Suppression of the proliferative response could be detected from 2-60 d after intragastric administration of OVA, and 2-21 d after S. mutans. Prior intragastric immunization with heterologous antigens did not suppress the response to OVA or S. mutans. Transfer of 40 X 10(6) mesenteric lymph node cells from mice given 20 mg OVA or 10(9) S. mutans led to suppression of the proliferative response in syngeneic recipients. Salivary antibodies wer removed by absorption with anti-IgA, but not anti-IgG or IgM, indicating that they were of the IgA class. It appears that intragastric administration of soluble or particulate antigens in mice may lead to the concurrent induction of salivary antibodies and systemic suppression. PMID:7452148

  8. [Clinicopathologic features of mammary analogue secretory carcinoma of salivary glands].

    PubMed

    Zhang, X P; Ni, H; Wang, X; Chen, H; Shi, S S; Yu, B; Zhou, X J; Rao, Q

    2017-01-08

    Objective: To investigate the clinicopathological features of mammary analogue secretory carcinoma (MASC) of salivary glands, and its diagnosis, differential diagnosis, immunohistochemistry and molecular pathology. Methods: Seventeen cases of MASC were enrolled, with 9 cases of salivary acinar cell carcinoma and 18 cases of adenoid cystic carcinoma as control groups from Nanjing General Hospital from 1997 to 2014 were included in this retrospective study, combined with immunohistochemistry and molecular detection of ETV6-NTRK3 gene fusion. All cases were histologically reviewed with immunohistochemical staining (EnVision) for S-100 protein, SOX10, GATA3, CD117 expression in each group. Fluorescence in situ hybridization (FISH) was used to detect the ETV6-NTRK3 gene fusion. Results: The age of MASC patients ranged from 27 to 74 years with mean age of 47 and ratio of male and female was 4∶3. All cases showed infiltrative growth and diverse cytology and histology, including lobular (8 cases), cystic papillary (3 cases), cribriform mixed with papillary and glandular structures (6 cases) at various proportions. Some tumors of MASC also exhibited solid growth areas with occasional microcystic honeycombed pattern composed of small cysts merged into larger cysts resembling thyroid follicles. S-100 protein and SOX10 were strongly positive in all MASC cases (17/17). In addition, there was insignificant positivity for GATA3 (3/17) and CD117 (4/17). ETV6 gene fusion detection was informative in 12 MASC cases by FISH with 10 positive cases and 2 negative cases. Conclusions: Combined immunohistochemical positivity of S-100 protein, CD117 and SOX10 are useful in the diagnosis and differential diagnosis of MASC. FISH detection of ETV6-NTRK3 fusion offers an additional molecular diagnostic marker for the diagnosis.

  9. Ultrastructural features of the early secretory pathway in Trichoderma reesei.

    PubMed

    Nykänen, Marko; Birch, Debra; Peterson, Robyn; Yu, Hong; Kautto, Liisa; Gryshyna, Anna; Te'o, Junior; Nevalainen, Helena

    2016-05-01

    We have systematically analysed the ultrastructure of the early secretory pathway in the Trichoderma reesei hyphae in the wild-type QM6a, cellulase-overexpressing Rut-C30 strain and a Rut-C30 transformant BV47 overexpressing a recombinant BiP1-VenusYFP fusion protein with an endoplasmic reticulum (ER) retention signal. The hyphae were studied after 24 h of growth using transmission electron microscopy, confocal microscopy and quantitative stereological techniques. All three strains exhibited different spatial organisation of the ER at 24 h in both a cellulase-inducing medium and a minimal medium containing glycerol as a carbon source (non-cellulase-inducing medium). The wild-type displayed a number of ER subdomains including parallel tubular/cisternal ER, ER whorls, ER-isolation membrane complexes with abundant autophagy vacuoles and dense bodies. Rut-C30 and its transformant BV47 overexpressing the BiP1-VenusYFP fusion protein also contained parallel tubular/cisternal ER, but no ER whorls; also, there were very few autophagy vacuoles and an increasing amount of punctate bodies where particularly the recombinant BiP1-VenusYFP fusion protein was localised. The early presence of distinct strain-specific features such as the dominance of ER whorls in the wild type and tub/cis ER in Rut-C30 suggests that these are inherent traits and not solely a result of cellular response mechanisms by the high secreting mutant to protein overload.

  10. Alterations in geometry, biomechanics, and mineral composition of juvenile rat femur induced by nonplanar PCB-155 and/or planar PCB-169.

    PubMed

    Brankovič, Jana; Jovanovski, Sašo; Jevnikar, Peter; Hofmeister, Alexander; Reininger-Gutmann, Birgit; Jan, Janja; Grošelj, Maja; Osredkar, Joško; Uršič, Matjaž; Fazarinc, Gregor; Pogačnik, Azra; Vrecl, Milka

    2017-04-01

    Exposure to widespread lipophilic and bioaccumulative polychlorinated biphenyls (PCBs) induces diverse biochemical and toxicological responses in various organs, including the bone. The aim of this study was to evaluate the changes in growth rate, geometry, serum, and bone biochemical parameters and biomechanics of juvenile rat femur induced by lactational exposure to nonplanar PCB-155 and planar PCB-169 individually and in combination. Fifteen lactating Wistar rats were divided into four groups (PCB-169, PCB-155, PCB-155+169, and control), and PCBs were administered intraperitoneally at different time points after delivery. Femurs from 22-day-old offspring were analyzed by microCT, three-point bending test and inductively coupled plasma-mass spectrometry (ICP-MS) to obtain data on bone geometry, biomechanics and mineral composition. The serum levels of calcium, phosphate and alkaline phosphatase were also determined. Lactational exposure to planar PCB-169 resulted in shorter and thinner femurs, reduced endosteal and periosteal perimeters, smaller total cross-sectional and medullary areas, and lowered serum bone marker levels and calcium levels in the bone, while femur mechanical properties were not significantly altered. The changes observed in the combination exposure (PCB-155+169) group were similar to those observed in the PCB-169 group but were less pronounced. In summary, our results demonstrate that alterations in lactationally exposed offspring were primarily induced by planar PCB-169. The milder outcome in the combined group suggested that the PCB-169-mediated toxic effects on the bone might be reduced by a nonplanar PCB-155 congener. © 2016 Wiley Periodicals, Inc. Environ Toxicol 32: 1135-1146, 2017.

  11. Multiphoton fluorescence lifetime imaging shows spatial segregation of secondary metabolites in Eucalyptus secretory cavities.

    PubMed

    Heskes, A M; Lincoln, C N; Goodger, J Q D; Woodrow, I E; Smith, T A

    2012-07-01

    Multiphoton fluorescence lifetime imaging provides an excellent tool for imaging deep within plant tissues while providing a means to distinguish between fluorophores with high spatial and temporal resolution. Ideal candidates for the application of multiphoton fluorescence lifetime imaging to plants are the embedded secretory cavities found in numerous species because they house complex mixtures of secondary metabolites within extracellular lumina. Previous investigations of this type of structure have been restricted by the use of sectioned material resulting in the loss of lumen contents and often disorganization of the delicate secretory cells; thus it is not known if there is spatial segregation of secondary metabolites within these structures. In this paper, we apply multiphoton fluorescence lifetime imaging to investigate the spatial arrangement of metabolites within intact secretory cavities isolated from Eucalyptus polybractea R.T. Baker leaves. The secretory cavities of this species are abundant (up to 10 000 per leaf), large (up to 6 nL) and importantly house volatile essential oil rich in the monoterpene 1,8-cineole, together with an immiscible, non-volatile component comprised largely of autofluorescent oleuropeic acid glucose esters. We have been able to optically section into the lumina of secretory cavities to a depth of ∼80 μm, revealing a unique spatial organization of cavity metabolites whereby the non-volatile component forms a layer between the secretory cells lining the lumen and the essential oil. This finding could be indicative of a functional role of the non-volatile component in providing a protective region of low diffusivity between the secretory cells and potentially autotoxic essential oil.

  12. Functional Characterization of Monomeric GTPase Rab1 in the Secretory Pathway of Leishmania*

    PubMed Central

    Bahl, Surbhi; Parashar, Smriti; Malhotra, Himanshu; Raje, Manoj; Mukhopadhyay, Amitabha

    2015-01-01

    Leishmania secretes a large number of its effectors to the extracellular milieu. However, regulation of the secretory pathway in Leishmania is not well characterized. Here, we report the cloning, expression, and characterization of the Rab1 homologue from Leishmania. We have found that LdRab1 localizes in Golgi in Leishmania. To understand the role of LdRab1 in the secretory pathway of Leishmania, we have generated transgenic parasites overexpressing GFP-LdRab1:WT, GFP-LdRab1:Q67L (a GTPase-deficient dominant positive mutant of Rab1), and GFP-LdRab1:S22N (a GDP-locked dominant negative mutant of Rab1). Surprisingly, our results have shown that overexpression of GFP-LdRab1:Q67L or GFP-LdRab1:S22N does not disrupt the trafficking and localization of hemoglobin receptor in Leishmania. To determine whether the Rab1-dependent secretory pathway is conserved in parasites, we have analyzed the role of LdRab1 in the secretion of secretory acid phosphatase and Ldgp63 in Leishmania. Our results have shown that overexpression of GFP-LdRab1:Q67L or GFP-LdRab1:S22N significantly inhibits the secretion of secretory acid phosphatase by Leishmania. We have also found that overexpression of GFP-LdRab1:Q67L or GFP-LdRab1:S22N retains RFP-Ldgp63 in Golgi and blocks the secretion of Ldgp63, whereas the trafficking of RFP-Ldgp63 in GFP-LdRab1:WT-expressing cells is unaltered in comparison with control cells. Taken together, our results have shown that the Rab1-regulated secretory pathway is well conserved, and hemoglobin receptor trafficking follows an Rab1-independent secretory pathway in Leishmania. PMID:26499792

  13. Human Toxoplasma gondii-specific secretory immunoglobulin A reduces T. gondii infection of enterocytes in vitro.

    PubMed Central

    Mack, D G; McLeod, R

    1992-01-01

    Whey from 17 women (four acutely infected with Toxoplasma gondii, eight chronically infected, and five uninfected) was studied. T. gondii-specific secretory IgA antibodies were demonstrated by ELISA in whey from acutely infected and one of eight chronically infected women. Such antibodies to tachyzoite proteins of < or = 14, 22, 26-28, 30, 46, 60, 70-80, and > 100 kD (eliminated by protease but not periodate or neuraminidase treatment) were demonstrated in whey from acutely infected subjects when Western blots were probed with their whey and antibodies to human secretory IgA or IgA or secretory piece. Secretory IgA from four of eight chronically infected women recognized the 46- and 69-kD epitopes. Other whey samples were negative. Incubation of T. gondii tachyzoites with whey or purified secretory IgA from acutely infected (but not seronegative) women caused 50-75% reduction in infection of enterocytes in vitro. Whey reactive with the 46-kD epitope from three of six chronically infected women caused less (> or = 40%) inhibition. Whey and purified secretory IgA from two of three acutely infected women agglutinated tachyzoites. Whey did not result in complement-dependent lysis of T. gondii. These results indicate that it may be possible to produce human secretory IgA to T. gondii capable of reducing initial infection of enterocytes, as such IgA is present during natural infection. They also demonstrate candidate epitopes for such protection. Images PMID:1469104

  14. Estradiol increases cAMP in the oviductal secretory cells through a nongenomic mechanism.

    PubMed

    Oróstica, María L; Lopez, John; Rojas, Israel; Rocco, Jocelyn; Díaz, Patricia; Reuquén, Patricia; Cardenas, Hugo; Parada-Bustamante, Alexis; Orihuela, Pedro A

    2014-09-01

    In the rat oviduct, estradiol (E2) accelerates egg transport by a nongenomic action that requires previous conversion of E2 to methoxyestrogens via catechol-O-methyltranferase (COMT) and activation of estrogen receptor (ER) with subsequent production of cAMP and inositol triphosphate (IP3). However, the role of the different oviductal cellular phenotypes on this E2 nongenomic pathway remains undetermined. The aim of this study was to investigate the effect of E2 on the levels of cAMP and IP3 in primary cultures of secretory and smooth muscle cells from rat oviducts and determine the mechanism by which E2 increases cAMP in the secretory cells. In the secretory cells, E2 increased cAMP but not IP3, while in the smooth muscle cells E2 decreased cAMP and increased IP3. Suppression of protein synthesis by actinomycin D did not prevent the E2-induced cAMP increase, but this was blocked by the ER antagonist ICI 182 780 and the inhibitors of COMT OR 486, G protein-α inhibitory (Gαi) protein pertussis toxin and adenylyl cyclase (AC) SQ 22536. Expression of the mRNA for the enzymes that metabolizes estrogens, Comt, Cyp1a1, and Cyp1b1 was found in the secretory cells, but this was not affected by E2. Finally, confocal immunofluorescence analysis showed that E2 induced colocalization between ESR1 (ERα) and Gαi in extranuclear regions of the secretory cells. We conclude that E2 differentially regulates cAMP and IP3 in the secretory and smooth muscle cells of the rat oviduct. In the secretory cells, E2 increases cAMP via a nongenomic action that requires activation of COMT and ER, coupling between ESR1 and Gαi, and stimulation of AC.

  15. A hydrophobic patch in a charged alpha-helix is sufficient to target proteins to dense core secretory granules.

    PubMed

    Dikeakos, Jimmy D; Lacombe, Marie-Josée; Mercure, Chantal; Mireuta, Matei; Reudelhuber, Timothy L

    2007-01-12

    Many endocrine and neuroendocrine cells contain specialized secretory organelles called dense core secretory granules. These organelles are the repository of proteins and peptides that are secreted in a regulated manner when the cell receives a physiological stimulus. The targeting of proteins to these secretory granules is crucial for the generation of certain peptide hormones, including insulin and ACTH. Although previous work has demonstrated that proteins destined to a variety of cellular locations, including secretory granules, contain targeting sequences, no single consensus sequence for secretory granule-sorting signals has emerged. We have shown previously that alpha-helical domains in the C-terminal tail of the prohormone convertase PC1/3 play an important role in the ability of this region of the protein to direct secretory granule targeting (Jutras, I. Seidah, N. G., and Reudelhuber, T. L. (2000) J. Biol. Chem. 275, 40337-40343). In this study, we show that a variety of alpha-helical domains are capable of directing a heterologous secretory protein to granules. By testing a series of synthetic alpha-helices, we also demonstrate that the presence of charged (either positive or negative) amino acids spatially segregated from a hydrophobic patch in the alpha-helices of secretory proteins likely plays a critical role in the ability of these structures to direct secretory granule sorting.

  16. EFTEM cytochemistry and sexual dimorphism of secretory granules in male and female hamster submandibular glands.

    PubMed

    Moriguchi, Keiichi; Utsumi, Michiya; Ohno, Norikazu

    2011-05-01

    After glutaraldehyde fixation followed by osmium tetroxide postfixing, the secretory granules of acinar cells in male hamster submandibular glands (SGs) exhibit a characteristic bipartite substructure, with an electron-lucid rim and a more electron-dense central core. In female hamsters, the reverse is seen, with the larger portion of the granules forming an electron-lucid core and an outer electron-dense crescent rim. In the present study of endogenous peroxidase (PO) activity of male and female hamster SGs, secretory granules in the acinar cells were studied by DAB cytochemical technique. Individual granules showed bipartite substructure with the PO activity in a positive center core and unreacted lucid rim in both the male and the female acinar cells. Through isolation of granular fractions, the male and the female granules exhibited the same bipartite structure. We also examined the relation between the PO activity and counterstained areas in male and female hamster SGs, and the secretory granules of acinar cells by using EFTEM. In the male SG, the secretory granules exhibited the characteristic bipartite substructure to carry out parallel-EELS, nitrogen reflecting the presence of DAB moieties and uranium from counterstaing the presence the central core but not in the rim. On the other hand, the female bipartite secretory granules of the SG, exhibit the nitrogen reflecting the presence in the central core and uranium in the rim.

  17. Taenia taeniaeformis: inhibition of rat testosterone production by excretory-secretory product of the cultured metacestode.

    PubMed

    Rikihisa, Y; Lin, Y C; Fukaya, T

    1985-06-01

    In 3- to 5-month-old male Sprague-Dawley rats infected with the hepatic metacestode, Taenia taeniaeformis, the serum testosterone level was significantly lower than in comparable uninfected controls. By transmission electron microscopy, testicular Leydig cells of infected rats had less smooth endoplasmic reticulum than control Leydig cells. Cultured metacestodes isolated from the hepatic cysts secreted or excreted substances into the incubation medium. The effect of the excretory-secretory product on testosterone concentration in the sera and testes of 15-day-old rats was examined. Subcutaneous injection of 50-200 micrograms of excretory-secretory product/0.1 ml saline/rat for 2 days significantly reduced human chorionic gonadotropin-stimulated serum and testicular testosterone concentrations. Furthermore, the effect of the excretory-secretory product on isolated rat Leydig cell testosterone production was examined. Rat Leydig cells produced testosterone in vitro and, in the presence of 50 IU human chorionic gonadotropin/ml incubation medium, they responded with approximately 100% increase in testosterone production. Addition of 2-10 micrograms excretory-secretory product protein/ml of culture medium significantly reduced the testosterone production by rat Leydig cells in vitro. These results indicate that excretory-secretory product of cultured T. taeniaeformis metacestodes has a direct inhibitory effect on Leydig cell testosterone production under stimulation with human chorionic gonadotropin.

  18. Cofilin recruits F-actin to SPCA1 and promotes Ca2+-mediated secretory cargo sorting.

    PubMed

    Kienzle, Christine; Basnet, Nirakar; Crevenna, Alvaro H; Beck, Gisela; Habermann, Bianca; Mizuno, Naoko; von Blume, Julia

    2014-09-01

    The actin filament severing protein cofilin-1 (CFL-1) is required for actin and P-type ATPase secretory pathway calcium ATPase (SPCA)-dependent sorting of secretory proteins at the trans-Golgi network (TGN). How these proteins interact and activate the pump to facilitate cargo sorting, however, is not known. We used purified proteins to assess interaction of the cytoplasmic domains of SPCA1 with actin and CFL-1. A 132-amino acid portion of the SPCA1 phosphorylation domain (P-domain) interacted with actin in a CFL-1-dependent manner. This domain, coupled to nickel nitrilotriacetic acid (Ni-NTA) agarose beads, specifically recruited F-actin in the presence of CFL-1 and, when expressed in HeLa cells, inhibited Ca(2+) entry into the TGN and secretory cargo sorting. Mutagenesis of four amino acids in SPCA1 that represent the CFL-1 binding site also affected Ca(2+) import into the TGN and secretory cargo sorting. Altogether, our findings reveal the mechanism of CFL-1-dependent recruitment of actin to SPCA1 and the significance of this interaction for Ca(2+) influx and secretory cargo sorting.

  19. Ionizing radiation effects on the secretory-stage ameloblasts and enamel organic extracellular matrix.

    PubMed

    de Moraes Ramos-Perez, Flávia Maria; do Espírito Santo, Alexandre Ribeiro; da Cruz Perez, Danyel Elias; Novaes, Pedro Duarte; Bóscolo, Frab Norberto; Line, Sérgio Roberto Peres; de Almeida, Solange Maria

    2014-08-01

    This study assessed the effects of high doses of ionizing radiation on eruption rate, odontogenic region morphology, secretory-stage ameloblasts, and enamel organic extracellular matrix (EOECM) of rat maxillary incisors. For the study, 30 male rats were divided into three experimental groups: control (non-irradiated), irradiated by 15 Gy, and irradiated by 25 Gy. Irradiated groups received a single dose of 15 or 25 Gy of X-rays in the head and neck region. The maxillary incisor eruption rate was measured. Sections of 5-µm thickness of the maxillary incisor odontogenic regions were evaluated using bright field light microscopy. Ultrathin sections of secretory ameloblasts and their EOECM were analyzed by transmission electron microscopy (TEM). Irradiated groups showed significantly diminished eruption rate values at the 4th and at the 6th day after irradiation. Reduced optical retardation values were observed in the irradiated groups. The odontogenic region of maxillary incisors from irradiated rats exhibited altered and poorly organized preameloblasts. TEM showed degeneration areas in the secretory-stage EOECM and several autophagosomes in the secretory ameloblasts from irradiated animals. In conclusion, high radiation doses delay eruption and induce disturbances in secretory ameloblasts and EOECM of rat maxillary incisors. These findings may be associated with structural defects of mature enamel.

  20. Secretory granules of mast cells accumulate mature and immature MHC class II molecules.

    PubMed

    Vincent-Schneider, H; Théry, C; Mazzeo, D; Tenza, D; Raposo, G; Bonnerot, C

    2001-01-01

    Bone marrow-derived mast cells as well as dendritic cells, macrophages and B lymphocytes express major histocompatibility complex (MHC) class II molecules. In mast cells, the majority of MHC class II molecules reside in intracellular cell type-specific compartments, secretory granules. To understand the molecular basis for the localisation of MHC class II molecules in secretory granules, MHC class II molecules were expressed, together with the invariant chain, in the mast cell line, RBL-2H3. Using electron and confocal microscopy, we observed that in RBL-2H3 cells, mature and immature class II molecules accumulate in secretory granules. Two particular features of class II transport accounted for this intracellular localization: first, a large fraction of newly synthesized MHC class II molecules remained associated with invariant chain fragments. This defect, resulting in a slower rate of MHC class II maturation, was ascribed to a low cathepsin S activity. Second, although a small fraction of class II dimers matured (i.e. became free of invariant chain), allowing their association with antigenic peptides, they were retained in secretory granules. As a consequence of this intracellular localization, cell surface expression of class II molecules was strongly increased by cell activation stimuli which induced the release of the contents of secretory granules. Our results suggest that antigen presentation, and thereby antigen specific T cell stimulation, are regulated in mast cells by stimuli which induce mast cell activation.

  1. Lumenal protein within secretory granules affects fusion pore expansion.

    PubMed

    Weiss, Annita Ngatchou; Anantharam, Arun; Bittner, Mary A; Axelrod, Daniel; Holz, Ronald W

    2014-07-01

    It is often assumed that upon fusion of the secretory granule membrane with the plasma membrane, lumenal contents are rapidly discharged and dispersed into the extracellular medium. Although this is the case for low-molecular-weight neurotransmitters and some proteins, there are numerous examples of the dispersal of a protein being delayed for many seconds after fusion. We have investigated the role of fusion-pore expansion in determining the contrasting discharge rates of fluorescent-tagged neuropeptide-Y (NPY) (within 200 ms) and tissue plasminogen activator (tPA) (over many seconds) in adrenal chromaffin cells. The endogenous proteins are expressed in separate chromaffin cell subpopulations. Fusion pore expansion was measured by two independent methods, orientation of a fluorescent probe within the plasma membrane using polarized total internal reflection fluorescence microscopy and amperometry of released catecholamine. Together, they probe the continuum of the fusion-pore duration, from milliseconds to many seconds after fusion. Polarized total internal reflection fluorescence microscopy revealed that 71% of the fusion events of tPA-cer-containing granules maintained curvature for >10 s, with approximately half of the structures likely connected to the plasma membrane by a short narrow neck. Such events were not commonly observed upon fusion of NPY-cer-containing granules. Amperometry revealed that the expression of tPA-green fluorescent protein (GFP) prolonged the duration of the prespike foot ∼2.5-fold compared to NPY-GFP-expressing cells and nontransfected cells, indicating that expansion of the initial fusion pore in tPA granules was delayed. The t1/2 of the main catecholamine spike was also increased, consistent with a prolonged delay of fusion-pore expansion. tPA added extracellularly bound to the lumenal surface of fused granules. We propose that tPA within the granule lumen controls its own discharge. Its intrinsic biochemistry determines not only

  2. Secretory immunoglobulin A and immunoglobulin G in horse saliva.

    PubMed

    Palm, Anna-Karin E; Wattle, Ove; Lundström, Torbjörn; Wattrang, Eva

    2016-11-01

    This study aimed to increase the knowledge on salivary antibodies in the horse since these constitute an important part of the immune defence of the oral cavity. For that purpose assays to detect horse immunoglobulin A (IgA) including secretory IgA (SIgA) were set up and the molecular weights of different components of the horse IgA system were estimated. Moreover, samples from 51 clinically healthy horses were tested for total SIgA and IgG amounts in saliva and relative IgG3/5 (IgG(T)) and IgG4/7 (IgGb) content were tested in serum and saliva. Results showed a mean concentration of 74μg SIgA/ml horse saliva and that there was a large inter-individual variation in salivary SIgA concentration. For total IgG the mean concentration was approx. 5 times lower than that of SIgA, i.e. 20μg IgG/ml saliva and the inter-individual variation was lower than that observed for SIgA. The saliva-serum ratio for IgG isotypes IgG3/5 and IgG4/7 was also assessed in the sampled horses and this analysis showed that the saliva-serum ratio of IgG4/7 was in general approximately 4 times higher than that of IgG3/5. The large inter-individual variation in salivary SIgA levels observed for the normal healthy horses in the present study emphasises the need for a large number of observations when studying this parameter especially in a clinical setting. Moreover, our results also indicated that some of the salivary IgG does not originate from serum but may be produced locally. Thus, these results provide novel insight, and a base for further research, into salivary antibody responses of horses.

  3. Preservation of Facial Nerve With Adjuvant Radiotherapy for Recurrent Mammary Analogue Secretory Carcinoma of Parotid Gland.

    PubMed

    Jin, Shufang; Ma, Hailong; He, Yue

    2016-06-01

    Mammary analogue secretory carcinoma of salivary glands harbors the recurrent ETV6-NTRK3 gene fusion because of the translocation t (12; 15) (p13; q25) and resembles breast secretory carcinoma. This tumor composed of papillary, cystic, solid, and cribriform patterns. Immunohistochemically, the tumors are positive for mammaglobin, CK7, CK8, STAT5a, vimentin, and S100. In this report, the authors presented a patient of recurrent parotid gland mammary analogue secretory carcinoma in a 22-year-old woman. The patient received extended parotidectomy with partial adhesive masseter surgery. The facial nerve was preserved during the surgery and adjuvant radiotherapy was performed postoperation. The patient did not suffer local recurrence and facial paralysis in the 18 months follow-up period.

  4. [Secretory type and the X-sex chromatin characteristics of persons with peptic ulcer].

    PubMed

    Tsoneva, M; Brailski, Kh; Ilieva, P; Popov, P

    1976-01-01

    Blood groups according to ABO and Rh systems were tested in 137 ulcer patients as well as the secretory state. The titre of blood group ABH antigens was determined in the same patients, secreted in saliva and gastric juice. The results are compared with those obtained from the examination of a control group of 30 clinically healthy subjects. X-sex chromatin in the cells of biopsy mucosa was examined in 40 ulcer patients, taken from the ulcer area after gastroscopy. As a control, X-sex chromatin was studied in smears from exfoliated oral mucosa in the same subjects. The data confirm the presence of genetic predisposition to ulcer disease development in subjects from blood group O (alpha, beta) and non-secretory type. Low titres of blood group antigens were found in the secretory patients. Differences in X-sex chromating characteristics were observed between bucal mucosa and the mucosa from the ulcer area.

  5. Segregation of sphingolipids and sterols during formation of secretory vesicles at the trans-Golgi network

    PubMed Central

    Klemm, Robin W.; Ejsing, Christer S.; Surma, Michal A.; Kaiser, Hermann-Josef; Gerl, Mathias J.; Sampaio, Julio L.; de Robillard, Quentin; Ferguson, Charles; Proszynski, Tomasz J.; Shevchenko, Andrej

    2009-01-01

    The trans-Golgi network (TGN) is the major sorting station in the secretory pathway of all eukaryotic cells. How the TGN sorts proteins and lipids to generate the enrichment of sphingolipids and sterols at the plasma membrane is poorly understood. To address this fundamental question in membrane trafficking, we devised an immunoisolation procedure for specific recovery of post-Golgi secretory vesicles transporting a transmembrane raft protein from the TGN to the cell surface in the yeast Saccharomyces cerevisiae. Using a novel quantitative shotgun lipidomics approach, we could demonstrate that TGN sorting selectively enriched ergosterol and sphingolipid species in the immunoisolated secretory vesicles. This finding, for the first time, indicates that the TGN exhibits the capacity to sort membrane lipids. Furthermore, the observation that the immunoisolated vesicles exhibited a higher membrane order than the late Golgi membrane, as measured by C-Laurdan spectrophotometry, strongly suggests that lipid rafts play a role in the TGN-sorting machinery. PMID:19433450

  6. Kinetics of early TCR signaling regulate the pathway of lytic granule delivery to the secretory domain

    PubMed Central

    Beal, Allison M.; Anikeeva, Nadia; Varma, Rajat; Cameron, Thomas O.; Vasiliver-Shamis, Gaia; Norris, Philip J.; Dustin, Michael L.; Sykulev, Yuri

    2009-01-01

    SUMMARY Cytolytic granule mediated killing of virus-infected cells is an essential function of cytotoxic T lymphocytes. Analysis of lytic granule delivery shows that the granules can take long or short paths to the secretory domain where they are released. Both paths utilize the same intracellular molecular events, which have different spatial and temporal arrangements in each path and are regulated by the kinetics of downstream Ca2+ mediated signaling. Rapid and robust signaling causes swift granule concentration near the MTOC and subsequent delivery by the polarized MTOC directly to the secretory domain - the shortest and fastest path. Indolent signaling leads to late recruitment of granules that move along microtubules to the periphery of the synapse and then move tangentially to fuse at the outer edge of the secretory domain - a longer path. The short pathway is associated with faster granule release and more efficient killing than the long pathway. PMID:19833088

  7. Regulated and constitutive protein targeting can be distinguished by secretory polarity in thyroid epithelial cells

    PubMed Central

    1991-01-01

    We have studied concurrent apical/basolateral and regulated/constitutive secretory targeting in filter-grown thyroid epithelial monolayers in vitro, by following the exocytotic routes of two newly synthesized endogenous secretory proteins, thyroglobulin (Tg) and p500. Tg is a regulated secretory protein as indicated by its acute secretory response to secretagogues. Without stimulation, pulse-labeled Tg exhibits primarily two kinetically distinct routes: less than or equal to 80% is released in an apical secretory phase which is largely complete by 6-10 h, with most of the remaining Tg retained in intracellular storage from which delayed apical discharge is seen. The rapid export observed for most Tg is unlikely to be because of default secretion, since its apical polarity is preserved even during the period (less than or equal to 10 h) when p500 is released basolaterally by a constitutive pathway unresponsive to secretagogues. p500 also exhibits a second, kinetically distinct secretory route: at chase times greater than 10 h, a residual fraction (less than or equal to 8%) of p500 is secreted with an apical preponderance similar to that of Tg. It appears that this fraction of p500 has failed to be excluded from the regulated pathway, which has a predetermined apical polarity. From these data we hypothesize that a targeting hierarchy may exist in thyroid epithelial cells such that initial sorting to the regulated pathway may be a way of insuring apical surface delivery from one of two possible exocytotic routes originating in the immature storage compartment. PMID:1991788

  8. Mammary analogue secretory carcinoma of the salivary glands: a diagnostic dilemma.

    PubMed

    Hindocha, N; Wilson, M H; Pring, M; Hughes, C W; Thomas, S J

    2016-08-12

    Mammary analogue secretory carcinoma (MASC) is a recently identified salivary gland neoplasm that can mimic other salivary gland tumours such as acinic cell carcinoma and cystadenocarcinoma. It is distinguished from these by differences in immunohistochemical profile and the identification of an ETV6-NTRK3 translocation (12;15)(p13;q25), which is also found in secretory carcinomas of the breast. Previous publications have suggested that MASC tumours have similar biological behaviour to acinic cell carcinoma. We report two cases of MASC that affected the upper lip, and showed an infiltrative and locally aggressive growth pattern that required several operations to ensure clearance of microscopic tumour cells.

  9. Cytologic findings of mammary analogue secretory carcinoma arising in the thyroid.

    PubMed

    Rodríguez-Urrego, Paula A; Dogan, Snjezana; Lin, Oscar

    2017-03-02

    Mammary analogue secretory carcinoma (MASC) of the salivary gland, first described by Skálová et al in 2010, is a tumor that morphologically and genetically resembles breast secretory carcinoma harboring ETV6-NTRK3 fusion gene. To date, only seven cases of primary thyroid MASC have been described. The overall findings are similar to those seen in the salivary gland counterpart including the ETV6-NTRK3 fusion gene. This is the second report describing the cytologic features of MASC at this primary location, which also showed a classical type papillary carcinoma component. Diagn. Cytopathol. 2017. © 2017 Wiley Periodicals, Inc.

  10. Large lipid-rich mammary analogue secretory carcinoma of parotid gland: An unusual case.

    PubMed

    Joshi, Prashant; Mridha, Asit Ranjan; Singh, Shuchita; Kinra, Prateek; Ray, Ruma; Thakar, Alok

    2015-01-01

    Mammary analogue secretory carcinoma (MASC) of the salivary gland is a malignant tumor which bears morphologic, immunohistochemical and molecular features similar to those of mammary secretory carcinoma. The tumor is considered as a low-grade malignancy perhaps slightly more aggressive than acinic cell carcinoma. High-grade transformation with recurrences, regional nodal involvement, metastases, and cancer-related death has been reported in a few cases. We report an unusual case of large MASC of the parotid gland in a young patient without regional lymph node involvement. To the best of our knowledge till date such a large MASC of the salivary gland has not been reported in the English literature.

  11. Secretory activity of mast cell during stress: effect of prolyl-glycyl-proline and Semax.

    PubMed

    Umarova, B A; Kopylova, G N; Smirnova, E A; Guseva, A A; Zhuikova, S E

    2003-10-01

    Stress increased secretory activity of mast cells in the mesentery and subcutaneous fat of rats. Intraperitoneal injection of Semax and prolyl-glycyl-proline in doses of 0.05 and 1 mg/kg, respectively, 1 h before stress abolished this effect. The test preparations did not modulate secretory activity of mast cells in unstressed animals. Semax and prolyl-glycyl-proline in vitro prevented activation of mast cells with synacten and acetylcholine. The stabilizing effect of peptides on mast cells probably determines their antiulcer activity.

  12. A study of the nonlinear aerodynamics of bodies in nonplanar motion. Ph.D. Thesis - Stanford Univ., Calif.; [numerical analysis of aerodynamic force and moment systems during large amplitude, arbitrary motions

    NASA Technical Reports Server (NTRS)

    Schiff, L. B.

    1974-01-01

    Concepts from the theory of functionals are used to develop nonlinear formulations of the aerodynamic force and moment systems acting on bodies in large-amplitude, arbitrary motions. The analysis, which proceeds formally once the functional dependence of the aerodynamic reactions upon the motion variables is established, ensures the inclusion, within the resulting formulation, of pertinent aerodynamic terms that normally are excluded in the classical treatment. Applied to the large-amplitude, slowly varying, nonplanar motion of a body, the formulation suggests that the aerodynamic moment can be compounded of the moments acting on the body in four basic motions: steady angle of attack, pitch oscillations, either roll or yaw oscillations, and coning motion. Coning, where the nose of the body describes a circle around the velocity vector, characterizes the nonplanar nature of the general motion.

  13. ETV6 rearrangement in a case of mammary analogue secretory carcinoma of the skin.

    PubMed

    Chang, Michael D; Arthur, Allison K; García, Joaquín J; Sukov, William R; Shon, Wonwoo

    2016-11-01

    Mammary analog secretory carcinoma of salivary glands is a relatively recently recognized entity that harbors the ETV6-NTRK3 fusion transcript. To date, only rare cases of mammary analog secretory carcinoma of the skin have been reported. A 57-year-old man presented with a 6.0 cm cystic mass in the axilla, involving the dermis and superficial subcutis. Microscopically, the tumor exhibited nodular aggregation of tubular and microcystic structures embedded in the dense fibrotic and hyalinized stroma. Characteristic 'colloid-like' eosinophilic secretory material was present within intraluminal spaces. Tumor cells were largely characterized by vesicular nuclei with inconspicuous nucleoli and pink vacuolated cytoplasm. With respect to immunohistochemistry, tumor cells were intensely positive for AE1/AE3, Cam 5.2, and CK7, whereas Ber-EP4 and CEA were completely negative. A dual color break-apart fluorescence in situ hybridization probe identified rearrangement of the ETV6 gene locus on chromosome 12. The patient is alive with no evidence of recurrent disease or metastasis 3 years after the initial surgery. In conclusion, we report a rare example of mammary analog secretory carcinoma of the skin with ETV6 rearrangement. Awareness of this unique cutaneous tumor and subsequent reporting of additional cases is necessary for better characterization of its completely clinicopathologic spectrum.

  14. Secretory cargo sorting by Ca2+-dependent Cab45 oligomerization at the trans-Golgi network

    PubMed Central

    Blank, Birgit; Maiser, Andreas; Emin, Derya; Prescher, Jens; Beck, Gisela; Kienzle, Christine; Bartnik, Kira; Habermann, Bianca; Pakdel, Mehrshad; Leonhardt, Heinrich; Lamb, Don C.

    2016-01-01

    Sorting and export of transmembrane cargoes and lysosomal hydrolases at the trans-Golgi network (TGN) are well understood. However, elucidation of the mechanism by which secretory cargoes are segregated for their release into the extracellular space remains a challenge. We have previously demonstrated that, in a reaction that requires Ca2+, the soluble TGN-resident protein Cab45 is necessary for the sorting of secretory cargoes at the TGN. Here, we report that Cab45 reversibly assembles into oligomers in the presence of Ca2+. These Cab45 oligomers specifically bind secretory proteins, such as COMP and LyzC, in a Ca2+-dependent manner in vitro. In intact cells, mutation of the Ca2+-binding sites in Cab45 impairs oligomerization, as well as COMP and LyzC sorting. Superresolution microscopy revealed that Cab45 colocalizes with secretory proteins and the TGN Ca2+ pump (SPCA1) in specific TGN microdomains. These findings reveal that Ca2+-dependent changes in Cab45 mediate sorting of specific cargo molecules at the TGN. PMID:27138253

  15. Intestinal Mucus Gel and Secretory Antibody are Barriers to Campylobacter jejuni Adherence to INT 407 Cells

    DTIC Science & Technology

    1987-06-01

    An in vitro mucus assay was developed to study the role of mucus gel and secretory immunoglobulin A (sIgA) in preventing attachment of Campylobacter ... jejuni to INT 407 cells. An overlay of rabbit small intestinal mucus was found to impede the attachment of C. jejuni to a monolayer of INT 407 cells

  16. Phosphorylation of αSNAP is Required for Secretory Organelle Biogenesis in Toxoplasma gondii.

    PubMed

    Stewart, Rebecca J; Ferguson, David J P; Whitehead, Lachlan; Bradin, Clare H; Wu, Hong J; Tonkin, Christopher J

    2016-02-01

    Upon infection, apicomplexan parasites quickly invade host cells and begin a replicative cycle rapidly increasing in number over a short period of time, leading to tissue lysis and disease. The secretory pathway of these highly polarized protozoan parasites tightly controls, in time and space, the biogenesis of specialized structures and organelles required for invasion and intracellular survival. In other systems, regulation of protein trafficking can occur by phosphorylation of vesicle fusion machinery. Previously, we have shown that Toxoplasma gondii αSNAP - a protein that controls the disassembly of cis-SNARE complexes--is phosphorylated. Here, we show that this post-translational modification is required for the correct function of αSNAP in controlling secretory traffic. We demonstrate that during intracellular development conditional expression of a non-phosphorylatable form of αSNAP results in Golgi fragmentation and vesiculation of all downstream secretory organelles. In addition, we show that the vestigial plastid (termed apicoplast), although reported not to be reliant on Golgi trafficking for biogenesis, is also affected upon overexpression of αSNAP and is much more sensitive to the levels of this protein than targeting to other organelles. This work highlights the importance of αSNAP and its phosphorylation in Toxoplasma organelle biogenesis and exposes a hereto fore-unexplored mechanism of regulation of vesicle fusion during secretory pathway trafficking in apicomplexan parasites.

  17. Regulated phosphorylation of secretory granule membrane proteins of the rat parotid gland

    SciTech Connect

    Marino, C.R.; Castle, J.D.; Gorelick, F.S. )

    1990-07-01

    An antiserum raised against purified rat parotid secretory granule membrane proteins has been used to identify organelle-specific protein phosphorylation events following stimulation of intact cells from the rat parotid gland. After lobules were prelabeled with ({sup 32}P)orthophosphate and exposed to secretagogues, phosphoproteins were immunoprecipitated with the granule membrane protein antiserum, separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and visualized by autoradiography. Parallel studies of stimulated amylase release were performed. Isoproterenol treatment of parotid lobules resulted in an increase in the phosphate content of immunoprecipitable 60- and 72-kDa proteins that correlated with amylase release in a time-dependent manner. Forskolin addition mimicked these effects, but only the isoproterenol effects were reversed by propranolol treatment. To confirm the specificity of the antiserum to the secretory granule membrane fraction, subcellular isolation techniques were employed following in situ phosphorylation. The 60- and 72-kDa phosphoproteins were immunoprecipitated from both a particulate fraction and a purified secretory granule fraction. Furthermore, the extraction properties of both species suggest that they are integral membrane proteins. These findings support the possibility that stimulus-regulated secretion may involve phosphorylation of integral membrane proteins of the exocrine secretory granule.

  18. HMGB2 holds the key to the senescence-associated secretory phenotype.

    PubMed

    Guerrero, Ana; Gil, Jesús

    2016-11-07

    The senescence-associated secretory phenotype (SASP) is a hallmark of senescence with an important physiological impact, but how it is established is unclear. In this issue, Aird et al. (2016. J. Cell Biol. https://doi.org/10.1083/jcb.201608026) describe how chromatin-bound HMGB2 fine tunes SASP expression by avoiding heterochromatin spreading.

  19. The secretory patterns of relaxin and human chorionic gonadotropin in human pregnancy.

    PubMed

    Seki, K; Uesato, T; Tabei, T; Kato, K

    1985-10-01

    Relaxin and human chorionic gonadotropin (hCG) were simultaneously determined in the same serum samples obtained from pregnant women. Although the secretory pattern of relaxin, in general, appeared to parallel that of hCG during human pregnancy, several discrepancies were discerned in the secretory patterns of the two hormones. The mean hCG concentration significantly differed between weeks 4-7 and 8-11 of pregnancy, but the mean relaxin concentration did not. The mean relaxin concentration began to decrease at weeks 16-19 whereas that of hCG did so at weeks 12-15. The mean relaxin concentration at weeks 4-7 was significantly higher than that at weeks 24-27, though there was no significant difference between the mean hCG concentrations in the two periods. These differences in the secretory pattern of relaxin from that of hCG indicate that relaxin secretion in pregnancy is not determined only by the circulating level of hCG. The responsiveness of the corpus luteum of pregnancy to hCG stimulation of relaxin secretion may vary as a function of the age of the corpus luteum, and this may partially account for the differences between the secretory pattern of relaxin and that of hCG observed in the present study.

  20. Homotypic Fusion of Immature Secretory Granules during Maturation in a Cell-free Assay

    PubMed Central

    Urbé, Sylvie; Page, Lesley J.; Tooze, Sharon A.

    1998-01-01

    The biogenesis of secretory granules embodies several morphological and biochemical changes. In particular, in neuroendocrine cells maturation of secretory granules is characterized by an increase in size which has been proposed to reflect homotypic fusion of immature secretory granules (ISGs). Here we describe an assay that provides the first biochemical evidence for such a fusion event and allows us to analyze its regulation. The assay reconstitutes homotypic fusion between one population of ISGs containing a [35S]sulfate-labeled substrate, secretogranin II (SgII), and a second population containing the prohormone convertase PC2. Both substrate and enzyme are targeted exclusively to ISGs. Fusion is measured by quantification of a cleavage product of SgII produced by PC2. With this assay we show that fusion only occurs between ISGs and not between ISGs and MSGs, is temperature dependent, and requires ATP and GTP and cytosolic proteins. NSF (N-ethylmaleimide–sensitive fusion protein) is amongst the cytosolic proteins required, whereas we could not detect a requirement for p97. The ability to reconstitute ISG fusion in a cell-free assay is an important advance towards the identification of molecules involved in the maturation of secretory granules and will increase our understanding of this process. PMID:9864358

  1. Analysis of Protein Localization and Secretory Pathway Function Using the Yeast "Saccharomyces Cerevisiae"

    ERIC Educational Resources Information Center

    Vallen, Elizabeth

    2002-01-01

    The isolation and characterization of mutants has been crucial in understanding a number of processes in the field of cell biology. In this exercise, students examine the effects of mutations in the secretory pathway on protein localization. Yeast strains deficient for synthesis of histidinol dehydrogenase are transformed with a plasmid encoding a…

  2. 21 CFR 866.5380 - Free secretory component immuno-logical test system.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... system. 866.5380 Section 866.5380 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Immunological Test Systems § 866.5380 Free secretory component immuno-logical test system. (a) Identification. A...

  3. 21 CFR 866.5380 - Free secretory component immuno-logical test system.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... system. 866.5380 Section 866.5380 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Immunological Test Systems § 866.5380 Free secretory component immuno-logical test system. (a) Identification. A...

  4. In vitro fusion between Saccharomyces cerevisiae secretory vesicles and cytoplasmic-side-out plasma membrane vesicles.

    PubMed Central

    Arrastua, Lorena; San Sebastian, Eider; Quincoces, Ana F; Antony, Claude; Ugalde, Unai

    2003-01-01

    The final step in the secretory pathway, which is the fusion event between secretory vesicles and the plasma membrane, was reconstructed using highly purified secretory vesicles and cytoplasmic-side-out plasma membrane vesicles from the yeast Saccharomyces cerevisiae. Both organelle preparations were obtained from a sec 6-4 temperature-sensitive mutant. Fusion was monitored by means of a fluorescence assay based on the dequenching of the lipophilic fluorescent probe octadecylrhodamine B-chloride (R18). The probe was incorporated into the membrane of secretory vesicles, and it diluted in unlabelled cytoplasmic-side-out plasma membrane vesicles as the fusion process took place. The obtained experimental dequenching curves were found by mathematical analysis to consist of two independent but simultaneous processes. Whereas one of them reflected the fusion process between both vesicle populations as confirmed by its dependence on the assay conditions, the other represented a non-specific transfer of the probe. The fusion process may now be examined in detail using the preparation, validation and analytical methods developed in this study. PMID:12435271

  5. Secretory COPII Protein SEC31B Is Required for Pollen Wall Development1[OPEN

    PubMed Central

    Zhao, Bingchun; Shi, Haidan; Wang, Wanlei; Liu, Xiaoyu; Gao, Hui; Wang, Xiaoxiao; Zhang, Yinghui; Yang, Meidi; Li, Rui

    2016-01-01

    The pollen wall protects pollen grains from abiotic and biotic stresses. During pollen wall development, tapetal cells play a vital role by secreting proteins, signals, and pollen wall material to ensure microspore development. But the regulatory mechanism underlying the secretory pathway of the tapetum is largely unknown. Here, we characterize the essential role of the Arabidopsis (Arabidopsis thaliana) COPII protein SECRETORY31B (SEC31B) in pollen wall development and the secretory activity of tapetal cells. The sporophyte-controlled atsec31b mutant exhibits severe pollen and seed abortion. Transmission electron microscopy observation indicates that pollen exine formation in the atsec31b mutant is disrupted significantly. AtSEC31B is a functional COPII protein revealed by endoplasmic reticulum (ER) exit site localization, interaction with AtSEC13A, and retarded ER-Golgi protein trafficking in the atsec31b mutant. A genetic tapetum-specific rescue assay indicates that AtSEC31B functions primarily in the tapetum. Moreover, deletion of AtSEC31B interrupted the formation of the ER-derived tapetosome and altered the location of the ATP-BINDING CASSETTE TRANSPORTER9 protein in the tapetum. Therefore, this work demonstrates that AtSEC31B plays a vital role in pollen wall development by regulating the secretory pathway of the tapetal cells. PMID:27634427

  6. Interactions between Melanin Enzymes and Their Atypical Recruitment to the Secretory Pathway by Palmitoylation

    PubMed Central

    Upadhyay, Srijana; Xu, Xinping

    2016-01-01

    ABSTRACT Melanins are biopolymers that confer coloration and protection to the host organism against biotic or abiotic insults. The level of protection offered by melanin depends on its biosynthesis and its subcellular localization. Previously, we discovered that Aspergillus fumigatus compartmentalizes melanization in endosomes by recruiting all melanin enzymes to the secretory pathway. Surprisingly, although two laccases involved in the late steps of melanization are conventional secretory proteins, the four enzymes involved in the early steps of melanization lack a signal peptide or a transmembrane domain and are thus considered “atypical” secretory proteins. In this work, we found interactions among melanin enzymes and all melanin enzymes formed protein complexes. Surprisingly, the formation of protein complexes by melanin enzymes was not critical for their trafficking to the endosomal system. By palmitoylation profiling and biochemical analyses, we discovered that all four early melanin enzymes were strongly palmitoylated during conidiation. However, only the polyketide synthase (PKS) Alb1 was strongly palmitoylated during both vegetative hyphal growth and conidiation when constitutively expressed alone. This posttranslational lipid modification correlates the endosomal localization of all early melanin enzymes. Intriguingly, bioinformatic analyses predict that palmitoylation is a common mechanism for potential membrane association of polyketide synthases (PKSs) and nonribosomal peptide synthetases (NRPSs) in A. fumigatus. Our findings indicate that protein-protein interactions facilitate melanization by metabolic channeling, while posttranslational lipid modifications help recruit the atypical enzymes to the secretory pathway, which is critical for compartmentalization of secondary metabolism. PMID:27879337

  7. PICK1 Deficiency Impairs Secretory Vesicle Biogenesis and Leads to Growth Retardation and Decreased Glucose Tolerance

    PubMed Central

    Jansen, Anna M.; Jin, Chunyu; Rickhag, Mattias; Lund, Viktor K.; Jensen, Morten; Bhatia, Vikram; Sørensen, Gunnar; Madsen, Andreas N.; Xue, Zhichao; Møller, Siri K.; Woldbye, David; Qvortrup, Klaus; Huganir, Richard; Stamou, Dimitrios; Kjærulff, Ole; Gether, Ulrik

    2013-01-01

    Secretory vesicles in endocrine cells store hormones such as growth hormone (GH) and insulin before their release into the bloodstream. The molecular mechanisms governing budding of immature secretory vesicles from the trans-Golgi network (TGN) and their subsequent maturation remain unclear. Here, we identify the lipid binding BAR (Bin/amphiphysin/Rvs) domain protein PICK1 (protein interacting with C kinase 1) as a key component early in the biogenesis of secretory vesicles in GH-producing cells. Both PICK1-deficient Drosophila and mice displayed somatic growth retardation. Growth retardation was rescued in flies by reintroducing PICK1 in neurosecretory cells producing somatotropic peptides. PICK1-deficient mice were characterized by decreased body weight and length, increased fat accumulation, impaired GH secretion, and decreased storage of GH in the pituitary. Decreased GH storage was supported by electron microscopy showing prominent reduction in secretory vesicle number. Evidence was also obtained for impaired insulin secretion associated with decreased glucose tolerance. PICK1 localized in cells to immature secretory vesicles, and the PICK1 BAR domain was shown by live imaging to associate with vesicles budding from the TGN and to possess membrane-sculpting properties in vitro. In mouse pituitary, PICK1 co-localized with the BAR domain protein ICA69, and PICK1 deficiency abolished ICA69 protein expression. In the Drosophila brain, PICK1 and ICA69 co-immunoprecipitated and showed mutually dependent expression. Finally, both in a Drosophila model of type 2 diabetes and in high-fat-diet-induced obese mice, we observed up-regulation of PICK1 mRNA expression. Our findings suggest that PICK1, together with ICA69, is critical during budding of immature secretory vesicles from the TGN and thus for vesicular storage of GH and possibly other hormones. The data link two BAR domain proteins to membrane remodeling processes in the secretory pathway of peptidergic endocrine

  8. "Abnormal" bromination reaction selectivity of 5-diarylamino-2-methylbenzo[b]thiophene caused by a "non-planar" conjugated model: Synthesis and theoretical calculation

    NASA Astrophysics Data System (ADS)

    Wu, Bian-Peng; Pang, Mei-Li; Tan, Ting-Feng; Meng, Ji-Ben

    2013-01-01

    5-Diarylamino-2-methylbenzo[b]thiophene was a new kind of triphenylamine-based charge-transporting material. For further modification of the compounds, bromination selectivity was studied through experiments and molecular simulation using Gaussian 09 program under B3LYP/6-311G (d, p) aided by Gaussian View 05 and Multiwfn Program. The results showed that bromination of 5-diarylamino-2-methyl-benzo[b]thiophene would occurred at an "abnormal" positions (4- and/or 4'-position) rather than at the "normal" position (3-position), which was different from those benzo[b]thiophene derivatives with simple 5-substitutes reported in literatures. The "abnormal" selectivity resulted from a special electron structure in which there was an "interfinger-like" frontier orbital or a special "non-planar" conjugated model. Electrons would be donated to the o- and p-positions of linked aromatic rings by this manner of electron delocalization, so that TPA unit rather than thiophene ring became the main factor in the selectivity, and reaction active energy at 4-position was lowest. The results were confirmed by the synthesis of three 4-Br and/or 4'-Br derivatives, compound 4, 5 and 6. Single crystal X-ray diffraction of compound 6 gave conclusive evidence on the abnormal selectivity.

  9. Tissue distribution of co-planar and non-planar tetra- and hexa-chlorobiphenyl isomers in guinea pigs after oral ingestion

    SciTech Connect

    Jan, J.; Logar, B.; Jan, J.

    1996-03-01

    Food ingestion is the most important route for the uptake of lipophilic organochlorine contaminants. Uptake and transfer of the contaminants from the digestive tract to target organs can be used for risk evaluation. The bioconcentration and migration of polychlorobiphenyls (PCBs) is highly structure - dependent. Bioconcentration is correlated with lipophilicity on the basis of the n-octanol/water partition coefficient in its logarithmic form - logKow. However, some factors e.g. diffusion through cell membranes, accumulation in specific organs and tissues, uptake and deputation kinetics and metabolism can also influence the bioconcentration. Individual PCB compounds of commercial PCB preparation are taken up by organisms to markedly different extents. Until now little is known about the distribution of non-planar and co-planar PCBs in different tissues. Co-planar PCBs have dioxin - like toxicity. This study examines differences in the bioconcentration of two pairs of tetra and hexa chlorobiphenyls from the digestive tract and their distribution in different tissues of guinea pigs.

  10. Relationship between High-frequency Radiation and Asperity Ruptures, Revealed by Hybrid Back-projection with a Non-planar Fault Model

    PubMed Central

    Okuwaki, Ryo; Yagi, Yuji; Hirano, Shiro

    2014-01-01

    High-frequency seismic waves are generated by abrupt changes of rupture velocity and slip-rate during an earthquake. Therefore, analysis of high-frequency waves is crucial to understanding the dynamic rupture process. Here, we developed a hybrid back-projection method that considers variations in focal mechanisms by introducing a non-planar fault model that reflects the subducting slab geometry. We applied it to teleseismic P-waveforms of the Mw 8.8 2010 Chile earthquake to estimate the spatiotemporal distribution of high-frequency (0.5–2.0 Hz) radiation. By comparing the result with the coseismic slip distribution obtained by waveform inversion, we found that strong high-frequency radiation can precede and may trigger a large asperity rupture. Moreover, in between the large slip events, high-frequency radiation of intermediate strength was concentrated along the rupture front. This distribution suggests that by bridging the two large slips, this intermediate-strength high-frequency radiation might play a key role in the interaction of the large slip events. PMID:25406638

  11. Impact of the non-planar morphology of pre-patterned substrates on the structural and electronic properties of embedded site-controlled InAs quantum dots

    SciTech Connect

    Hakkarainen, T. V.; Tommila, J.; Schramm, A.; Guina, M.; Luna, E.

    2013-11-07

    We present an experimental and theoretical analysis of the influence of a surface nanopattern on the properties of embedded InAs/GaAs quantum dots (QD). In particular, we analyze QDs grown on nanoimprint lithography (NIL) patterned grooves and investigate the influence of the non-planar surface morphology on the size, shape, strain distribution, and electronic structure of the embedded QDs. We show that the height reduction of InAs QDs during GaAs capping is significantly less pronounced for the QDs grown on the pattern than for the self-assembled QDs. Furthermore, the pattern has a strong impact on the strain and composition profile within the QD. The experimentally observed strain distribution was successfully reproduced with a three-dimensional model assuming an inverse-cone type composition gradient. Moreover, we show that the specific morphology of the QDs grown in the grooves gives rise to an increase of the vertically polarized photoluminescence emission which was explained by employing 8-band k.p calculations. Our findings emphasize that the surface curvature of the pattern not only determines the nucleation sites of the QDs but also has a strong impact on their morphological properties including shape, size, composition profile, and strain distribution. These properties are strongly cross-correlated and determine the electronic and optical characteristics of the QDs.

  12. Tracheobronchial epithelium of the sheep: IV. Lectin histochemical characterization of secretory epithelial cells.

    PubMed

    Mariassy, A T; Plopper, C G; St George, J A; Wilson, D W

    1988-09-01

    Conventional histochemical characterization of the mucus secretory apparatus is often difficult to reconcile with the biochemical analysis of respiratory secretions. This study was designed to examine the secretory glycoconjugates in airways using lectins with biochemically defined affinities for main sugar residues of mucus. We used five biotinylated lectins--DBA (Dolichos biflorus) and SBA (Glycine max) for N-acetyl galactosamine (galNAc), BSA I (Bandeiraea simplicifolia) and PNA (Arachis hypogea) for galactose (gal), and UEA I (Ulex europeus)--for detection of fucose (fuc) in HgCl2-fixed, paraffin-embedded, serially sectioned trachea, lobar and segmental bronchi and bronchioles of nine sheep. Lectins selectively localized the carbohydrate residues in luminal secretions, on epithelial cell surfaces, and in secretory cells. In proximal airways, the major carbohydrate residues in luminal secretions, cell surfaces, goblet cells, and glands were fuc and gal-NAc. PNA reacted mainly with apical granules of less than 10% of goblet cells, and gal residues were only detected in some of the mucous cells and on basolateral cell surfaces. Distal airways contained sparse secretion in the lumen, mucous cells contained weakly reactive fuc and gal-NAc, and the epithelial surfaces of Clara cells contained gal. Sugars abundant in the airway secretions were also the major component of cells in glands. We conclude that there is a correlation between specific sugar residues in secretory cells, glycocalyx, and luminal secretions in proximal and distal airways. This suggests that lectins may be used to obtain information about airway secretory cell composition from respiratory secretions.

  13. Mammary analogue secretory carcinoma of parotid: Is preoperative cytological diagnosis possible?

    PubMed

    Oza, Nikita; Sanghvi, Kintan; Shet, Tanuja; Patil, Asawari; Menon, Santosh; Ramadwar, Mukta; Kane, Shubhada

    2016-06-01

    Mammary analogue secretory carcinoma is a recently recognized tumor of salivary gland with characteristic t(12;15)(q13;q25) that results in ETV6-NTRK3 fusion product. Distinguishing mammary analogue secretory carcinoma from other salivary gland tumors is important. Present study highlights cytologic findings in three cases of mammary analogue secretory carcinoma of parotid which facilitate preoperative diagnosis with the aid of ancillary diagnostic techniques. Fine needle aspiration cytology of parotid was performed on three cases after clinical examination. Immunocytochemistry for mammoglobin and S100 were performed. Parotidectomy was done in all cases. The corresponding hematoxylin and eosin stained slides and blocks of all cases were studied. Molecular analysis was done in one of the cases. Cases 1 and 3 revealed uniform atypical epithelial cells arranged in branching papillary pattern with few cells in microcystic pattern. Case 2 showed atypical cells arranged mainly in loose clusters and few singly dissociated. Individual cells revealed round nuclei, vesicular chromatin, prominent nucleoli and abundant finely vacuolated cytoplasm with metachromasia prominent in May-Grunwald-Giemsa smear (case 3). Characteristic hob-nail cells covering papillae were observed in cases 1 and 3. Immunocytochemistry showed strong positivity for mammoglobin and S100 thereby confirming the diagnosis of mammary analogue secretory carcinoma preoperatively. The diagnosis was in concordance with surgical specimen. Also, characteristic ETV6-NTRK3 translocation was confirmed in case 1. Increased awareness and high index of suspicion is necessary for the upfront diagnosis, more so for the papillary variant of mammary analogue secretory carcinoma. Immunocytochemistry aids in confirming this preoperative diagnosis, based on which treatment can be planned. Diagn. Cytopathol. 2016;44:519-525. © 2016 Wiley Periodicals, Inc.

  14. Secretory-granule dynamics visualized in vivo with a phogrin-green fluorescent protein chimaera.

    PubMed Central

    Pouli, A E; Emmanouilidou, E; Zhao, C; Wasmeier, C; Hutton, J C; Rutter, G A

    1998-01-01

    To image the behaviour in real time of single secretory granules in neuroendocrine cells we have expressed cDNA encoding a fusion construct between the dense-core secretory-granule-membrane glycoprotein, phogrin (phosphatase on the granule of insulinoma cells), and enhanced green fluorescent protein (EGFP). Expressed in INS-1 beta-cells and pheochromocytoma PC12 cells, the chimaera was localized efficiently (up to 95%) to dense-core secretory granules (diameter 200-1000 nm), identified by co-immunolocalization with anti-(pro-)insulin antibodies in INS-1 cells and dopamine beta-hydroxylase in PC12 cells. Using laser-scanning confocal microscopy and digital image analysis, we have used this chimaera to monitor the effects of secretagogues on the dynamics of secretory granules in single living cells. In unstimulated INS-1 beta-cells, granule movement was confined to oscillatory movement (dithering) with period of oscillation 5-10 s and mean displacement <1 microm. Both elevated glucose concentrations (30 mM), and depolarization of the plasma membrane with K+, provoked large (5-10 microm) saltatory excursions of granules across the cell, which were never observed in cells maintained at low glucose concentration. By contrast, long excursions of granules occurred in PC12 cells without stimulation, and occurred predominantly from the cell body towards the cell periphery and neurite extensions. Purinergic-receptor activation with ATP provoked granule movement towards the membrane of PC12 cells, resulting in the transfer of fluorescence to the plasma membrane consistent with fusion of the granule and diffusion of the chimaera in the plasma membrane. These results illustrate the potential use of phogrin-EGFP chimeras in the study of secretory-granule dynamics, the regulation of granule-cytoskeletal interactions and the trafficking of a granule-specific transmembrane protein during the cycle of exocytosis and endocytosis. PMID:9639579

  15. Combination of keratins and alpha-smooth muscle actin distinguishes secretory coils from ducts of eccrine sweat glands.

    PubMed

    Li, Haihong; Zhang, Xiang; Zeng, Shaopeng; Li, Xuexue; Zhang, Bingna; Chen, Lu; Lin, Changmin; Zhang, Mingjun; Tang, Shijie; Fu, Xiaobing

    2015-04-01

    Eccrine sweat glands are comprised of secretory coils and ducts, which are distinct in morphology and function. To better understand the roles of the two parts in development, homeostasis, wound repair and regeneration of eccrine sweat glands, we must distinguish between them. In this study, the localization of keratins and alpha-SMA in human eccrine sweat glands was examined by immunofluorescence staining. Based on the differential localization of keratins and alpha-SMA in different cell types, four pairs of antibodies (K5/K7, K5/alpha-SMA, K14/K7 and K14/alpha-SMA) were used to differentiate secretory coils from ducts by double-immunofluorescence staining. Immunofluorescence staining showed that myoepithelial cells of secretory coils expressed K5, K14 and alpha-SMA, whereas secretory cells of secretory coils expressed K7, K8, K15, K18 and K19. Ductal cells expressed K5, K8, K14 and K19. Double-staining showed that the secretory coils were K5(+)/K7(+), K5(+)/alpha-SMA(+), K14(+)/K7(+) and K14(+)/alpha-SMA(+), whereas ducts were K5(+)/K7(-), K5(+)/alpha-SMA(-), K14(+)/K7(-) and K14(+)/alpha-SMA(-). In conclusion, by combining use of keratins and alpha-SMA antibodies, secretory coils can be easily differentiated from ducts in morphology.

  16. A secretory protease inhibitor requires androgens for its expression in male sex accessory tissues but is expressed constitutively in pancreas.

    PubMed Central

    Mills, J S; Needham, M; Parker, M G

    1987-01-01

    A full length cDNA clone encoding a mouse prostatic secretory glycoprotein (p12) whose synthesis is dependent upon testicular androgens has been cloned and characterized. The predicted amino acid sequence of p12 shares extensive homology with several members of the Kazal family of secretory protease inhibitors, in particular the pancreatic secretory trypsin inhibitors. In agreement with sequence data, prostatic secretory p12, purified from mouse ventral prostate secretion, exhibits anti-trypsin activity. Steady-state levels of protease inhibitor mRNA in ventral prostate are reduced from approximately 0.06% in normal mice to undetectable after androgen withdrawal but are inducible within 4 h by re-administration of testosterone. Androgen-dependent expression of the secretory protease inhibitor mRNA was also observed in coagulating gland and seminal vesicle. In seminal vesicle, a tissue of different embryonic origin to the prostate, the kinetics of secretory protease inhibitor mRNA loss after castration are not as rapid as in the ventral prostate and coagulating gland. Low-level androgen independent expression was also observed in the pancreas. There appears to be a single gene for this secretory protease inhibitor and yet expression is markedly stimulated by testosterone in the sex accessory tissues and unaffected by this hormone in the pancreas. Images Fig. 3. Fig. 4. Fig. 5. Fig. 6. Fig. 7. PMID:3428272

  17. Secretory component delays the conversion of secretory IgA into antigen-binding competent F(ab')2: a possible implication for mucosal defense.

    PubMed

    Crottet, P; Corthésy, B

    1998-11-15

    Secretory component (SC) represents the soluble ectodomain of the polymeric Ig receptor, a membrane protein that transports mucosal Abs across epithelial cells. In the protease-rich environment of the intestine, SC is thought to stabilize the associated IgA by unestablished molecular mechanisms. To address this question, we reconstituted SC-IgA complexes in vitro by incubating dimeric IgA (IgAd) with either recombinant human SC (rSC) or SC isolated from human colostral milk (SCm). Both complexes exhibited an identical degree of covalency when exposed to redox agents, peptidyl disulfide isomerase, and temperature changes. In cross-competition experiments, 50% inhibition of binding to IgAd was achieved at approximately 10 nM SC competitor. Western blot analysis of IgAd digested with intestinal washes indicated that the alpha-chain in IgAd was primarily split into a 40-kDa species, a phenomenon delayed in rSC- or SCm-IgAd complexes. In the same assay, either of the SCs was resistant to degradation only if complexed with IgAd. In contrast, the kappa light chain was not digested at all, suggesting that the F(ab')2 region was left intact. Accordingly, IgAd and SC-IgAd digestion products retained functionality as indicated by Ag reactivity in ELISA. Size exclusion chromatography under native conditions of digested IgAd and rSC-IgAd demonstrates that SC exerts its protective role in secretory IgA by delaying cleavage in the hinge/Fc region of the alpha-chain, not by holding together degraded fragments. The function of integral secretory IgA and F(ab')2 is discussed in terms of mucosal immune defenses.

  18. Cab45 is required for Ca2+-dependent secretory cargo sorting at the trans-Golgi network

    PubMed Central

    Alleaume, Anne-Marie; Kienzle, Christine; Carreras-Sureda, Amado; Valverde, Miguel

    2012-01-01

    Ca2+ import into the lumen of the trans-Golgi network (TGN) by the secretory pathway calcium ATPase1 (SPCA1) is required for the sorting of secretory cargo. How is Ca2+ retained in the lumen of the Golgi, and what is its role in cargo sorting? We show here that a soluble, lumenal Golgi resident protein, Cab45, is required for SPCA1-dependent Ca2+ import into the TGN; it binds secretory cargo in a Ca2+-dependent reaction and is required for its sorting at the TGN. PMID:23266954

  19. MASEP gamma knife radiosurgery for secretory pituitary adenomas: experience in 347 consecutive cases

    PubMed Central

    Wan, Heng; Chihiro, Ohye; Yuan, Shubin

    2009-01-01

    Background Secretory pituitary adenomas are very common brain tumors. Historically, the treatment armamentarium for secretory pituitary adenomas included neurosurgery, medical management, and fractionated radiotherapy. In recent years, MASEP gamma knife radiosurgery (MASEP GKRS) has emerged as an important treatment modality in the management of secretory pituitary adenomas. The goal of this research is to define accurately the efficacy, safety, complications, and role of MASEP GKRS for treatment of secretory pituitary adenomas. Methods Between 1997 and 2007 a total of 347 patients with secretory pituitary adenomas treated with MASEP GKRS and with at least 60 months of follow-up data were identified. In 47 of these patients some form of prior treatment such as transsphenoidal resection, or craniotomy and resection had been conducted. The others were deemed ineligible for microsurgery because of body health or private choice, and MASEP GKRS served as the primary treatment modality. Endocrinological, ophthalmological, and neuroradiological responses were evaluated. Results MASEP GKRS was tolerated well in these patients under the follow-up period ranged from 60 to 90 months; acute radioreaction was rare and 17 patients had transient headaches with no clinical significance. Late radioreaction was noted in 1 patient and consisted of consistent headache. Of the 68 patients with adrenocorticotropic hormone-secreting(ACTH) adenomas, 89.7% showed tumor volume decrease or remain unchanged and 27.9% experienced normalization of hormone level. Of the 176 patients with prolactinomas, 23.3% had normalization of hormone level and 90.3% showed tumor volume decrease or remain unchanged. Of the 103 patients with growth hormone-secreting(GH) adenomas, 95.1% experienced tumor volume decrease or remain unchanged and 36.9% showed normalization of hormone level. Conclusion MASEP GKRS is safe and effective in treating secretory pituitary adenomas. None of the patients in our study

  20. Airway Secretory microRNAome Changes during Rhinovirus Infection in Early Childhood

    PubMed Central

    Gutierrez, Maria J.; Gomez, Jose L.; Perez, Geovanny F.; Pancham, Krishna; Val, Stephanie; Pillai, Dinesh K.; Giri, Mamta; Ferrante, Sarah; Freishtat, Robert; Rose, Mary C.; Preciado, Diego; Nino, Gustavo

    2016-01-01

    Background Innate immune responses are fine-tuned by small noncoding RNA molecules termed microRNAs (miRs) that modify gene expression in response to the environment. During acute infections, miRs can be secreted in extracellular vesicles (EV) to facilitate cell-to-cell genetic communication. The purpose of this study was to characterize the baseline population of miRs secreted in EVs in the airways of young children (airway secretory microRNAome) and examine the changes during rhinovirus (RV) infection, the most common cause of asthma exacerbations and the most important early risk factor for the development of asthma beyond childhood. Methods Nasal airway secretions were obtained from children (≤3 yrs. old) during PCR-confirmed RV infections (n = 10) and age-matched controls (n = 10). Nasal EVs were isolated with polymer-based precipitation and global miR profiles generated using NanoString microarrays. We validated our in vivo airway secretory miR data in an in vitro airway epithelium model using apical secretions from primary human bronchial epithelial cells (HBEC) differentiated at air-liquid interface (ALI). Bioinformatics tools were used to determine the unified (nasal and bronchial) signature airway secretory miRNAome and changes during RV infection in children. Results Multiscale analysis identified four signature miRs comprising the baseline airway secretory miRNAome: hsa-miR-630, hsa-miR-302d-3p, hsa- miR-320e, hsa-miR-612. We identified hsa-miR-155 as the main change in the baseline miRNAome during RV infection in young children. We investigated the potential biological relevance of the airway secretion of hsa-mir-155 using in silico models derived from gene datasets of experimental in vivo human RV infection. These analyses confirmed that hsa-miR-155 targetome is an overrepresented pathway in the upper airways of individuals infected with RV. Conclusions Comparative analysis of the airway secretory microRNAome in children indicates that RV infection

  1. Fast Domain Partitioning Method for dynamic boundary integral equations applicable to non-planar faults dipping in 3-D elastic half-space

    NASA Astrophysics Data System (ADS)

    Ando, Ryosuke

    2016-11-01

    The elastodynamic boundary integral equation method (BIEM) in real space and in the temporal domain is an accurate semi-analytical tool to investigate the earthquake rupture dynamics on non-planar faults. However, its heavy computational demand for a historic integral generally increases with a time complexity of O(MN3)for the number of time steps N and elements M due to volume integration in the causality cone. In this study, we introduce an efficient BIEM, termed the `Fast Domain Partitioning Method' (FDPM), which enables us to reduce the computation time to the order of the surface integral, O(MN2), without degrading the accuracy. The memory requirement is also reduced to O(M2) from O(M2N). FDPM uses the physical nature of Green's function for stress to partition the causality cone into the domains of the P and S wave fronts, the domain in-between the P and S wave fronts, and the domain of the static equilibrium, where the latter two domains exhibit simpler dependences on time and/or space. The scalability of this method is demonstrated on the large-scale parallel computing environments of distributed memory systems. It is also shown that FDPM enables an efficient use of memory storage, which makes it possible to reduce computation times to a previously unprecedented level. We thus present FDPM as a powerful tool to break through the current fundamental difficulties in running dynamic simulations of coseismic ruptures and earthquake cycles under realistic conditions of fault geometries.

  2. Growth-induced non-planar magnetic anisotropy in FeCoZr-CaF{sub 2} nanogranular films: Structural and magnetic characterization

    SciTech Connect

    Kasiuk, J. V.; Fedotova, J. A.; Przewoznik, J. Zukrowski, J.; Sikora, M.; Kapusta, Cz.; Grce, A.; Milosavljević, M.

    2014-07-28

    The relation between nanoscale structure, local atomic order and magnetic properties of (FeCoZr){sub x}(CaF{sub 2}){sub 100−x} (29 ≤ x ≤ 73 at. %) granular films is studied as a function of metal/insulator fraction ratio. The films of a thickness of 1–6 μm were deposited on Al-foils and glass-ceramic substrates, by ion sputtering of targets of different metal/insulator contents. Structural characterization with X-ray and electron diffraction as well as transmission electron microscopy revealed that the films are composed of isolated nanocrystalline bcc α-FeCo(Zr) alloy and insulating fcc CaF{sub 2} matrix. They grow in a columnar structure, where elongated metallic nanograins are arranged on top of each other within the columns almost normal to the substrate surface. Mössbauer spectroscopy and magnetometry results indicate that their easy magnetization axes are oriented at an angle of 65°–74° to the surface in films with x between 46 and 74, above the electrical percolation threshold, which is attributed to the growth-induced shape anisotropy. Interatomic distances characteristic for metallic state of α-FeCo(Zr) nanograins were revealed by X-ray Absorption Spectroscopy. The results show a lack of surface oxidation of the alloy nanograins, so the growth-induced orientation of nanograins in the films cannot be attributed to this effect. The study is among the first to report a growth-induced non-planar magnetic anisotropy in metal/insulator granular films above the percolation threshold and to reveal the origin of it.

  3. [Advantages of the use of Andro-Gin device for treatment of secretory infertility].

    PubMed

    Aleksandrov, V P; Koren'kov, D G; Nikolaeva, E V

    2006-01-01

    The trial included 144 patients aged from 24 to 46 years with secretory infertility in the presence of chronic prostatitis of 2 to 13 year duration. The study group consisted of 96 patients who received conventional treatment in combination with physical factors provided by the unit APK-ELM-01 Andro-Gin. The control group consisted of 48 patients on conventional treatment alone. The results were processed mathematically and statistically. The patients were examined before, 2 weeks, 1 and 6 months after the treatment. Positive response was greater in the study group: better parameters of prostatic secretion, spermogram. Normalization of all the parameters occurred in 95 and 80% patients of the study and control groups, respectively. Thus, Andro-Gin is effective for treatment of chronic prostatitis with secretory infertility.

  4. Cysteine-rich secretory proteins are not exclusively expressed in the male reproductive tract.

    PubMed

    Reddy, Thulasimala; Gibbs, Gerard M; Merriner, D Jo; Kerr, Jeffrey B; O'Bryan, Moira K

    2008-11-01

    The Cysteine-RIch Secretory Proteins (CRISPs) are abundantly produced in the male reproductive tract of mammals and within the venom of reptiles and have been shown to regulate ion channel activity. CRISPs, along with the Antigen-5 proteins and the Pathogenesis related-1 (Pr-1) proteins, form the CAP superfamily of proteins. Analyses of EST expression databases are increasingly suggesting that mammalian CRISPs are expressed more widely than in the reproductive tract. We, therefore, conducted a reverse transcription PCR expression profile and immunohistochemical analyses of 16 mouse tissues to define the sites of production of each of the four murine CRISPs. These data showed that each of the CRISPs have distinct and sometimes overlapping expression profiles, typically associated with the male and female reproductive tract, the secretory epithelia of exocrine glands, and immune tissues including the spleen and thymus. These investigations raise the potential for a role for CRISPs in general mammalian physiology.

  5. Cytopathological features of mammary analogue secretory carcinoma--review of literature.

    PubMed

    Takeda, Maiko; Kasai, Takahiko; Morita, Kohei; Takeuchi, Mao; Nishikawa, Takeshi; Yamashita, Akinori; Mikami, Shinji; Hosoi, Hiroshi; Ohbayashi, Chiho

    2015-02-01

    Mammary analogue secretory carcinoma (MASC) is a recently described salivary gland tumor that morphologically resembles mammary secretory carcinoma and carries the identical ETV6-NTRK3 fusion gene. We report a surgical resected case of MASC in the parotid gland of a 41-year-old man. The cytological smears of a preoperative fine-needle aspiration showed many sheets and crowded clusters of monotonous epithelioid cells with mild atypia, suggestive of monomorphic tumor. Histologically, the tumor was composed of cuboidal cells with follicular, tubular, and solid structures, reminiscent of acinic cell carcinoma of follicular variant, which had been previously classified. This case had ETV6-NTRK3 fusion gene transcript confirmed by fluorescence in situ hybridization and reverse transcription polymerase chain reaction. In the cytological and histopathological diagnosis of monomorphic tumor of salivary gland, MASC needs to be taken into consideration as a differential diagnosis. Further immunohistochemical and gene analyses are needed in diagnosis of MASC.

  6. [Detection of secretory immunoglobulin A in cervical secretion and in cervicovaginal irrigation fluids].

    PubMed

    Briese, V; Hofmann, R; Meissner, J; Straube, W

    1983-01-01

    Secretory immunoglobulin A (S-IgA) is an essential part of local immune system of female genital tract. Accordingly the estimation of S-IgA in female genital secretions is important for judgement of immunologic problems in female genital tract. The estimation of S-IgA was carried out by means of an antiserum against human secretory component (anti SC) and S-IgA standard. It was used the single radial immunodiffusion according to Mancini and co-workers. Our results refer to changes in the quantity of S-IgA locally produced in the mucous membranes of female genital tract correlated to the menstrual cycle. The essential results are a decrease of S-IgA in midcycle in cervical mucus and vaginal fluids as well as of fertile and sterile patients.

  7. Electron microprobe analysis of human labial gland secretory granules in cystic fibrosis

    SciTech Connect

    Izutsu, K.; Johnson, D.; Schubert, M.; Wang, E.; Ramsey, B.; Tamarin, A.; Truelove, E.; Ensign, W.; Young, M.

    1985-06-01

    X-ray microanalysis of freeze-dried labial gland cryosections revealed that Na concentration was doubled and the Ca/S concentration ratio was decreased in secretory granules of labial glands from patients with cystic fibrosis (CF) when compared with glands from normal subjects. Other results suggested that the decrease in the Ca/S concentration ratio resulted from an increase in S concentration. These findings imply that mucous granules in labial saliva showed a CF-related increase in Na and S content, and such changes would be expected to affect the rheology of the mucus after exocytosis. In contrast with a previous study in human parotid glands, no evidence was found for CF-related changes in cytoplasmic or nuclear Na, K, and Ca concentrations. Significant elemental differences were found between secretory granules and nuclei and cytoplasm of control cells.

  8. Pancreatic endoproteases and pancreatic secretory trypsin inhibitor immunoreactivity in human Paneth cells.

    PubMed Central

    Bohe, M; Borgström, A; Lindström, C; Ohlsson, K

    1986-01-01

    Normal and metaplastic gastrointestinal mucosa obtained at surgical resection were studied by light microscopy, using the unlabelled antibody enzyme method for immunohistochemical staining of lysozyme, pancreatic endoproteases, and pancreatic secretory trypsin inhibitor (PSTI). Paneth cells in the mucosa of normal small intestine, gastric mucosa with intestinal metaplasia, and colonic metaplastic mucosa were found to contain anionic trypsin, cationic trypsin, lysozyme, and PSTI immunoreactivity, but not chymotrypsin and elastase immunoreactivity. Normal gastric and colonic mucosa and some goblet cells in the small intestine showed positive PSTI immunoreactivity but no endoprotease immunoreactivity. The presence of immunoreactive trypsin and immunoreactive PSTI in the Paneth cells, which are of secretory type, probably indicates an important extrapancreatic source of these proteins rather than a storage of endocytosed material. Images PMID:3525612

  9. Serous cutaneous glands in anurans: Fourier transform analysis of the repeating secretory granule substructure

    NASA Astrophysics Data System (ADS)

    Nosi, D.; Delfino, G.; Quercioli, F.

    2013-03-01

    A combined transmission electron microscopy (TEM) and Fourier transform analysis has been performed on the secretory granules storing active peptides/proteins in serous cutaneous glands of n = 12 anuran species. Previous TEM investigation showed that the granules are provided with remarkable repeating substructures based on discrete subunits, arranged into a consistent framework. Furthermore, TEM findings revealed that this recurrent arrangement is acquired during a prolonged post-Golgian (or maturational) processing that affects the secretory product. Maturation leads to a variety of patterns depending on the degree of subunit clustering. This variety of recurrent patterns has been plotted into a range of frequency spectra. Through this quantitative approach, we found that the varying granule substructure can be reduced to a single mechanism of peptide/protein aggregation.

  10. Degradation of immunoglobulins, protease inhibitors, and interleukin-1 by a secretory proteinase of Acanthamoeba castellanii

    PubMed Central

    Na, Byoung-Kuk; Cho, Jong-Hwa; Song, Chul-Yong; Kim, Tong-Soo

    2002-01-01

    The effect of a secretory proteinase from the pathogenic amoebae Acanthamoeba castellanii on host's defense-oriented or regulatory proteins such as immunoglobulins, interleukin-1, and protease inhibitors was investigated. The enzyme was found to degrade secretory immunoglobulin A (sIgA), IgG, and IgM. It also degraded interleukin-1α (IL-1α) and IL-1β. Its activity was not inhibited by endogenous protease inhibitors, such as α2-macroglobulin, α1-trypsin inhibitor, and α2-antiplasmin. Furthermore, the enzyme rapidly degraded those endogenous protease inhibitors as well. The degradation of host's defense-oriented or regulatory proteins by the Acanthamoeba proteinase suggested that the enzyme might be an important virulence factor in the pathogenesis of Acanthamoeba infection. PMID:12073735

  11. Secretory breast carcinoma in a 41-year-old man with long-term follow-up: a special report.

    PubMed

    Li, Guanqiao; Zhong, Xiaojie; Yao, Jia; Chen, Jimin; Wang, Mei; Liu, Haiying; Yang, Shiping

    2015-01-01

    Secretory breast carcinoma (SBC) is a rare tumor that is particularly rare in male adults. To our knowledge, only 28 previous male cases of secretory breast carcinoma have been reported. The present a case of secretory breast carcinoma has the longest symptom duration of (40 years) in a male individual until now. Typically, the clinical features and treatment of male SBC are similar with female SBC. The ETV6-NTRK3 fusion gene is a specific genetic alteration in SBC. When compared to other types of male breast cancer, patients with male secretory breast cancer are much younger, and have a lower rate of estrogen/progesterone hormone receptor positivity. Modified radical mastectomy has been favored as a therapeutic approach in all female SBC, male SBC and other types of male breast cancer. [corrected].

  12. A Critical Role for Toxoplasma gondii Vacuolar Protein Sorting VPS9 in Secretory Organelle Biogenesis and Host Infection

    PubMed Central

    Sakura, Takaya; Sindikubwabo, Fabien; Oesterlin, Lena K.; Bousquet, Hugo; Slomianny, Christian; Hakimi, Mohamed-Ali; Langsley, Gordon; Tomavo, Stanislas

    2016-01-01

    Accurate sorting of proteins to the three types of parasite-specific secretory organelles namely rhoptry, microneme and dense granule in Toxoplasma gondii is crucial for successful host cell invasion by this obligate intracellular parasite. Despite its tiny body architecture and limited trafficking machinery, T. gondii relies heavily on transport of vesicles containing proteins, lipids and important virulence-like factors that are delivered to these secretory organelles. However, our understanding on how trafficking of vesicles operates in the parasite is still limited. Here, we show that the T. gondii vacuolar protein sorting 9 (TgVps9), has guanine nucleotide exchange factor (GEF) activity towards Rab5a and is crucial for sorting of proteins destined to secretory organelles. Our results illuminate features of TgVps9 protein as a key trafficking facilitator that regulates protein maturation, secretory organelle formation and secretion, thereby ensuring a primary role in host infection by T. gondii. PMID:27966671

  13. Multiplex Sequential Immunoprecipitation of Insulin Secretory Granule Proteins from Radiolabeled Pancreatic Islets.

    PubMed

    Guest, Paul C

    2017-01-01

    Pulse radiolabeling of cells with radioactive amino acids is a common method for tracking the biosynthesis of proteins. Specific proteins can then be immunoprecipitated and analyzed by electrophoresis and imaging techniques. This chapter presents a protocol for the biosynthetic labeling of pancreatic islets with (35)S-methionine, followed by multiplex sequential immunoprecipitation of insulin and three other secretory granule accessory proteins. This provided a means of distinguishing those pancreatic islet proteins with different biosynthetic rates in response to the media glucose concentrations.

  14. An Alternative Terminal Step of the General Secretory Pathway in Staphylococcus aureus

    PubMed Central

    Craney, Arryn; Dix, Melissa M.; Adhikary, Ramkrishna; Cravatt, Benjamin F.

    2015-01-01

    ABSTRACT Type I signal peptidase (SPase) is essential for viability in wild-type bacteria because the terminal step of the bacterial general secretory pathway requires its proteolytic activity to release proteins from their membrane-bound N-terminal leader sequences after translocation across the cytoplasmic membrane. Here, we identify the Staphylococcus aureus operon ayrRABC (SA0337 to SA0340) and show that once released from repression by AyrR, the protein products AyrABC together confer resistance to the SPase inhibitor arylomycin M131 by providing an alternate and novel method of releasing translocated proteins. Thus, the derepression of ayrRABC allows cells to bypass the essentiality of SPase. We demonstrate that AyrABC functionally complements SPase by mediating the processing of the normally secreted proteins, albeit in some cases with reduced efficiency and either without cleavage or via cleavage at a site N-terminal to the canonical SPase cleavage site. Thus, ayrRABC encodes a secretion stress-inducible alternate terminal step of the general secretory pathway. Importance  Addressing proteins for proper localization within or outside a cell in both eukaryotes and prokaryotes is often accomplished with intrinsic signals which mediate membrane translocation and which ultimately must be removed. The canonical enzyme responsible for the removal of translocation signals is bacterial type I signal peptidase (SPase), which functions at the terminal step of the general secretory pathway and is thus essential in wild-type bacteria. Here, we identify a four-gene operon in S. aureus that encodes an alternate terminal step of the general secretory pathway and thus makes SPase nonessential. The results have important implications for protein secretion in bacteria and potentially for protein trafficking in prokaryotes and eukaryotes in general. PMID:26286693

  15. α-Synuclein binds the KATP channel at insulin-secretory granules and inhibits insulin secretion

    PubMed Central

    Geng, Xuehui; Lou, Haiyan; Wang, Jian; Li, Lehong; Swanson, Alexandra L.; Sun, Ming; Beers-Stolz, Donna; Watkins, Simon; Perez, Ruth G.

    2011-01-01

    α-Synuclein has been studied in numerous cell types often associated with secretory processes. In pancreatic β-cells, α-synuclein might therefore play a similar role by interacting with organelles involved in insulin secretion. We tested for α-synuclein localizing to insulin-secretory granules and characterized its role in glucose-stimulated insulin secretion. Immunohistochemistry and fluorescent sulfonylureas were used to test for α-synuclein localization to insulin granules in β-cells, immunoprecipitation with Western blot analysis for interaction between α-synuclein and KATP channels, and ELISA assays for the effect of altering α-synuclein expression up or down on insulin secretion in INS1 cells or mouse islets, respectively. Differences in cellular phenotype between α-synuclein knockout and wild-type β-cells were found by using confocal microscopy to image the fluorescent insulin biosensor Ins-C-emGFP and by using transmission electron microscopy. The results show that anti-α-synuclein antibodies labeled secretory organelles within β-cells. Anti-α-synuclein antibodies colocalized with KATP channel, anti-insulin, and anti-C-peptide antibodies. α-Synuclein coimmunoprecipitated in complexes with KATP channels. Expression of α-synuclein downregulated insulin secretion at 2.8 mM glucose with little effect following 16.7 mM glucose stimulation. α-Synuclein knockout islets upregulated insulin secretion at 2.8 and 8.4 mM but not 16.7 mM glucose, consistent with the depleted insulin granule density at the β-cell surface membranes observed in these islets. These findings demonstrate that α-synuclein interacts with KATP channels and insulin-secretory granules and functionally acts as a brake on secretion that glucose stimulation can override. α-Synuclein might play similar roles in diabetes as it does in other degenerative diseases, including Alzheimer's and Parkinson's diseases. PMID:20858756

  16. Structure and function of snake venom cysteine-rich secretory proteins.

    PubMed

    Yamazaki, Yasuo; Morita, Takashi

    2004-09-01

    Cysteine-rich secretory proteins (CRISPs) are primarily found in the epididymis of mammals and are expressed in diverse organisms. However, the functions of most CRISPs remain unknown. Recent studies reveal that CRISPs are widely distributed in snake venoms and that they inhibit smooth muscle contraction and cyclic nucleotide-gated ion channels. In this review, we discuss recent findings on several snake venom-derived CRISPs.

  17. Brucella Modulates Secretory Trafficking via Multiple Type IV Secretion Effector Proteins

    PubMed Central

    Myeni, Sebenzile; Child, Robert; Ng, Tony W.; Kupko, John J.; Wehrly, Tara D.; Porcella, Stephen F.; Knodler, Leigh A.; Celli, Jean

    2013-01-01

    The intracellular pathogenic bacterium Brucella generates a replicative vacuole (rBCV) derived from the endoplasmic reticulum via subversion of the host cell secretory pathway. rBCV biogenesis requires the expression of the Type IV secretion system (T4SS) VirB, which is thought to translocate effector proteins that modulate membrane trafficking along the endocytic and secretory pathways. To date, only a few T4SS substrates have been identified, whose molecular functions remain unknown. Here, we used an in silico screen to identify putative T4SS effector candidate proteins using criteria such as limited homology in other bacterial genera, the presence of features similar to known VirB T4SS effectors, GC content and presence of eukaryotic-like motifs. Using β-lactamase and CyaA adenylate cyclase reporter assays, we identified eleven proteins translocated into host cells by Brucella, five in a VirB T4SS-dependent manner, namely BAB1_0678 (BspA), BAB1_0712 (BspB), BAB1_0847 (BspC), BAB1_1671 (BspE) and BAB1_1948 (BspF). A subset of the translocated proteins targeted secretory pathway compartments when ectopically expressed in HeLa cells, and the VirB effectors BspA, BspB and BspF inhibited protein secretion. Brucella infection also impaired host protein secretion in a process requiring BspA, BspB and BspF. Single or combined deletions of bspA, bspB and bspF affected Brucella ability to replicate in macrophages and persist in the liver of infected mice. Taken together, these findings demonstrate that Brucella modulates secretory trafficking via multiple T4SS effector proteins that likely act coordinately to promote Brucella pathogenesis. PMID:23950720

  18. Proteomic analysis of plasma membrane and secretory vesicles from human neutrophils

    PubMed Central

    Jethwaney, Deepa; Islam, Md Rafiqul; Leidal, Kevin G; de Bernabe, Daniel Beltran-Valero; Campbell, Kevin P; Nauseef, William M; Gibson, Bradford W

    2007-01-01

    Background Polymorphonuclear neutrophils (PMN) constitute an essential cellular component of innate host defense against microbial invasion and exhibit a wide array of responses both to particulate and soluble stimuli. As the cells recruited earliest during acute inflammation, PMN respond rapidly and release a variety of potent cytotoxic agents within minutes of exposure to microbes or their products. PMN rely on the redistribution of functionally important proteins, from intracellular compartments to the plasma membrane and phagosome, as the means by which to respond quickly. To determine the range of membrane proteins available for rapid recruitment during PMN activation, we analyzed the proteins in subcellular fractions enriched for plasma membrane and secretory vesicles recovered from the light membrane fraction of resting PMN after Percoll gradient centrifugation and free-flow electrophoresis purification using mass spectrometry-based proteomics methods. Results To identify the proteins light membrane fractions enriched for plasma membrane vesicles and secretory vesicles, we employed a proteomic approach, first using MALDI-TOF (peptide mass fingerprinting) and then by HPLC-MS/MS using a 3D ion trap mass spectrometer to analyze the two vesicle populations from resting PMN. We identified several proteins that are functionally important but had not previously been recovered in PMN secretory vesicles. Two such proteins, 5-lipoxygenase-activating protein (FLAP) and dysferlin were further validated by immunoblot analysis. Conclusion Our data demonstrate the broad array of proteins present in secretory vesicles that provides the PMN with the capacity for remarkable and rapid reorganization of its plasma membrane after exposure to proinflammatory agents or stimuli. PMID:17692124

  19. Development of a new platform for secretory production of recombinant proteins in Corynebacterium glutamicum.

    PubMed

    Yim, Sung Sun; Choi, Jae Woong; Lee, Roo Jin; Lee, Yong Jae; Lee, Se Hwa; Kim, So Young; Jeong, Ki Jun

    2016-01-01

    Corynebacterium glutamicum, which has been for long an industrial producer of various L-amino acids, nucleic acids, and vitamins, is now also regarded as a potential host for the secretory production of recombinant proteins. To harness its potential as an industrial platform for recombinant protein production, the development of an efficient secretion system is necessary. Particularly, regarding protein production in large-scale bioreactors, it would be appropriate to develop a secretory expression system that is specialized for high cell density cultivation conditions. Here we isolated a new signal peptide that mediates the efficient secretion of recombinant proteins under high cell density cultivation conditions. The secretome of C. glutamicum ATCC 13032 under high cell density cultivation conditions was initially investigated, and one major protein was identified as a hypothetical protein encoded by cg1514. Novel secretory production systems were then developed using the Cg1514 signal peptide and its own promoter. Efficient protein secretion was demonstrated using three protein models: endoxylanase, α-amylase, and camelid antibody fragment (VHH). For large-scale production, fed-batch cultivations were also conducted and high yields were successfully achieved--as high as 1.07 g/L (endoxylanase), 782.6 mg/L (α-amylase), and 1.57 g/L (VHH)--in the extracellular medium. From the culture media, all model proteins could be simply purified by one-step column chromatography with high purities and recovery yields. To the best of our knowledge, this is the first report of the development of an efficient secretory expression system by secretome analysis under high cell density cultivation conditions in C. glutamicum.

  20. Searching for mammary analogue [corrected] secretory carcinoma of salivary gland among its mimics.

    PubMed

    Pinto, Andre; Nosé, Vania; Rojas, Claudia; Fan, Yao-Shan; Gomez-Fernandez, Carmen

    2014-01-01

    Mammary analog secretory carcinoma of salivary gland is a recently described entity with unique morphologic, clinical, and genetic characteristics, including the characteristic t(12;15)(p13;q25) with ETV6-NTRK3 translocation found in secretory carcinomas of the breast. Before their initial description, these salivary gland tumors were generally diagnosed as acinic cell carcinoma or adenocarcinoma. For the purpose of this study, all cases of salivary gland acinic cell carcinoma, cribriform cystadenocarcinoma, and adenocarcinoma, not otherwise specified (NOS), diagnosed over a 10-year period were retrieved from our surgical pathology files. There were a total of 11 cases diagnosed as acinic cell carcinoma, 10 cases of adenocarcinoma, NOS, and 6 cases of cribriform cystadenocarcinoma. All slides were reviewed by two pathologists (AP, CGF) and tumors that show morphologic features of mammary analog secretory carcinoma according to the recent literature were selected. This process narrowed down the initial number to six cases originally diagnosed as acinic cell carcinoma, three cases originally diagnosed as adenocarcinoma, NOS, and one case originally diagnosed as cribriform cystadenocarcinoma. The 10 cases were subjected to immunohistochemistry for S-100, mammaglobin, and ANO1, as well as fluorescence in situ hybridization analysis for t(12;15)(p13;q25) with ETV6-NTRK3 fusion rearrangement. The ETV6-NTRK3 gene rearrangement was detected in three tumors. These three tumors, initially diagnosed as acinic cell carcinomas, stained positive for S-100 and mammaglobin, and negative for ANO1 by immunohistochemistry. Two of the three patients were male (2/3). In summary, mammary analog secretory carcinoma is a newly described diagnostic entity that should be in the differential diagnosis of salivary gland tumors that morphologically mimic other neoplasms, mainly acinic cell carcinomas. They differ from conventional acinic cell tumors immunohistochemically and molecularly

  1. Signaling from the Secretory Granule to the Nucleus: Uhmk1 and PAM

    PubMed Central

    Francone, Victor P.; Ifrim, Marius F.; Rajagopal, Chitra; Leddy, Christopher J.; Wang, Yanping; Carson, John H.; Mains, Richard E.; Eipper, Betty A.

    2010-01-01

    Neurons and endocrine cells package peptides in secretory granules (large dense-core vesicles) for storage and stimulated release. Studies of peptidylglycine α-amidating monooxygenase (PAM), an essential secretory granule membrane enzyme, revealed a pathway that can relay information from secretory granules to the nucleus, resulting in alterations in gene expression. The cytosolic domain (CD) of PAM, a type 1 membrane enzyme essential for the production of amidated peptides, is basally phosphorylated by U2AF homology motif kinase 1 (Uhmk1) and other Ser/Thr kinases. Proopiomelanocortin processing in AtT-20 corticotrope tumor cells was increased when Uhmk1 expression was reduced. Uhmk1 was concentrated in the nucleus, but cycled rapidly between nucleus and cytosol. Endoproteolytic cleavage of PAM releases a soluble CD fragment that localizes to the nucleus. Localization of PAM-CD to the nucleus was decreased when PAM-CD with phosphomimetic mutations was examined and when active Uhmk1 was simultaneously overexpressed. Membrane-tethering Uhmk1 did not eliminate its ability to exclude PAM-CD from the nucleus, suggesting that cytosolic Uhmk1 could cause this response. Microarray analysis demonstrated the ability of PAM to increase expression of a small subset of genes, including aquaporin 1 (Aqp1) in AtT-20 cells. Aqp1 mRNA levels were higher in wild-type mice than in mice heterozygous for PAM, indicating that a similar relationship occurs in vivo. Expression of PAM-CD also increased Aqp1 levels whereas expression of Uhmk1 diminished Aqp1 expression. The outlines of a pathway that ties secretory granule metabolism to the transcriptome are thus apparent. PMID:20573687

  2. Discovery of lansoprazole and its unique pharmacological properties independent from anti-secretory activity.

    PubMed

    Satoh, Hiroshi

    2013-01-01

    The proton pump inhibitors (PPIs) lansoprazole (LPZ) and omeprazole (OPZ) have been widely used for more than 20 years in the treatment of acid-related diseases such as gastro-duodenal ulcers and reflux esophagitis. Both LPZ and OPZ are derivatives of 2-[(2- pyridylmethyl)sulfinyl]-1H-benzimidazole, but LPZ has a trifluoroethoxy group in the molecule which seems to provide unique pharmacological properties in addition to its anti-secretory effect. For example, the anti-secretory effect of LPZ in rats was roughly 2 times greater than that of OPZ but the anti-ulcer effects were more than 10 times stronger than those of OPZ in rat models of reflux esophagitis, indomethacin-induced gastric antral ulcers and mepirizole-induced duodenal ulcers. It has also been reported that LPZ has acid-independent protective effects on the gastrointestinal mucosa, anti-inflammatory effects, and anti-bacterial effects on Helicobacter pylori. In contrast, recent advances in endoscopy have revealed that non-steroidal anti-inflammatory drugs (NSAIDs) often cause ulcers not only in the stomach and duodenum, but also in the small intestine in humans. Anti-secretory drugs such as PPIs and histamine H(2)-receptor antagonists (H(2)-RAs) are commonly used for the treatment of upper gastrointestinal mucosal lesions induced by NSAIDs. However, the effects of these drugs on NSAID-induced small intestinal lesions are still not fully understood. In this article, both a brief history of the discovery of LPZ and the unique pharmacological properties of LPZ independent from its anti-secretory action are reviewed, and the effects of PPIs and H(2)-RAs on NSAID-induced small intestinal lesions are discussed.

  3. Ca2+ dynamics in the secretory vesicles of neurosecretory PC12 and INS1 cells.

    PubMed

    SantoDomingo, Jaime; Fonteriz, Rosalba I; Lobatón, Carmen D; Montero, Mayte; Moreno, Alfredo; Alvarez, Javier

    2010-11-01

    We have investigated the dynamics of the free [Ca(2+)] inside the secretory granules of neurosecretory PC12 and INS1 cells using a low-Ca(2+)-affinity aequorin chimera fused to synaptobrevin-2. The steady-state secretory granule [Ca(2+)] ([Ca(2+)](SG)] was around 20-40 μM in both cell types, about half the values previously found in chromaffin cells. Inhibition of SERCA-type Ca(2+) pumps with thapsigargin largely blocked Ca(2+) uptake by the granules in Ca(2+)-depleted permeabilized cells, and the same effect was obtained when the perfusion medium lacked ATP. Consistently, the SERCA-type Ca(2+) pump inhibitor benzohydroquinone induced a rapid release of Ca(2+) from the granules both in intact and permeabilized cells, suggesting that the continuous activity of SERCA-type Ca(2+) pumps is essential to maintain the steady-state [Ca(2+)](SG). Both inositol 1,4,5-trisphosphate (InsP(3)) and caffeine produced a rapid Ca(2+) release from the granules, suggesting the presence of InsP(3) and ryanodine receptors in the granules. The response to high-K(+) depolarization was different in both cell types, a decrease in [Ca(2+)](SG) in PC12 cells and an increase in [Ca(2+)](SG) in INS1 cells. The difference may rely on the heterogeneous response of different vesicle populations in each cell type. Finally, increasing the glucose concentration triggered a decrease in [Ca(2+)](SG) in INS1 cells. In conclusion, our data show that the secretory granules of PC12 and INS1 cells take up Ca(2+) through SERCA-type Ca(2+) pumps and can release it through InsP(3) and ryanodine receptors, supporting the hypothesis that secretory granule Ca(2+) may be released during cell stimulation and contribute to secretion.

  4. Secretory Carcinoma in a 79- Year-Old Woman: An Exceptionally Rare Type of Breast Carcinoma

    PubMed Central

    Posso, Veronica; Redrobán, Ligia

    2016-01-01

    Secretory breast carcinoma is an exceptionally rare mammary gland neoplasia described mainly in adult females and children of both sexes, and very rarely in the elderly. It has particular histopathological and immunohistochemical features and a favorable prognosis. We report the case of a 79-year-old Hispanic woman with a palpable breast mass. Currently, the patient is disease free after a followup period of 6 years without local recurrence or axillary lymph-nodes nor distant metastases. PMID:28058101

  5. Development of secretory cells and crystal cells in Eichhornia crassipes ramet shoot apex.

    PubMed

    Xu, Guo Xin; Tan, Chao; Wei, Xiao Jing; Gao, Xiao Yan; Zheng, Hui Qiong

    2011-04-01

    The distribution and development of secretory cells and crystal cells in young shoot apexes of water hyacinth were investigated through morphological and cytological analysis. The density of secretory cells and crystal cells were high in parenchyma tissues around the vascular bundles of shoot apexes. Three developmental stages of the secretory cells can be distinguished under transmission electron microscopy. Firstly, a large number of electron-dense vesicles formed in the cytoplasm, then fused with the tonoplast and released into the vacuole in the form of electron-dense droplets. As these droplets fused together, a large mass of dark material completely filled the vacuole. To this end, a secretion storage vacuole (SSV) formed. Secondly, an active secretion stage accompanied with degradation of the large electron-dense masses through an ill-defined autophagic process at periphery and in the limited internal regions of the SSV. Finally, after most storage substances were withdrawn, the materials remaining in the spent SSV consisted of an electron-dense network structure. The distribution and development of crystal cells in shoot apical tissue of water hyacinth were also studied by light and electron microscopy. Crystals initially formed at one site in the vacuole, where tube-like membrane structures formed crystal chambers. The chamber enlarged as the crystal grew in bidirectional manner and formed needle-shaped raphides. Most of these crystals finally occurred as raphide bundles, and the others appeared as block-like rhombohedral crystals in the vacuole. These results suggest that the formation of both secretory cells and crystal cells are involved in the metamorphosis of vacuoles and a role for vacuoles in water hyacinth rapid growth and tolerance.

  6. Selective Targeting of Proteins within Secretory Pathway for Endoplasmic Reticulum-associated Degradation

    PubMed Central

    Vecchi, Lara; Petris, Gianluca; Bestagno, Marco; Burrone, Oscar R.

    2012-01-01

    The endoplasmic reticulum-associated degradation (ERAD) is a cellular quality control mechanism to dispose of misfolded proteins of the secretory pathway via proteasomal degradation. SEL1L is an ER-resident protein that participates in identification of misfolded molecules as ERAD substrates, therefore inducing their ER-to-cytosol retrotranslocation and degradation. We have developed a novel class of fusion proteins, termed degradins, composed of a fragment of SEL1L fused to a target-specific binding moiety located on the luminal side of the ER. The target-binding moiety can be a ligand of the target or derived from specific mAbs. Here, we describe the ability of degradins with two different recognition moieties to promote degradation of a model target. Degradins recognize the target protein within the ER both in secretory and membrane-bound forms, inducing their degradation following retrotranslocation to the cytosol. Thus, degradins represent an effective technique to knock-out proteins within the secretory pathway with high specificity. PMID:22523070

  7. Novel N-Benzoyl-2-Hydroxybenzamide Disrupts Unique Parasite Secretory Pathway

    PubMed Central

    Fomovska, Alina; Huang, Qingqing; El Bissati, Kamal; Mui, Ernest J.; Witola, William H.; Cheng, Gang; Zhou, Ying; Sommerville, Caroline; Roberts, Craig W.; Bettis, Sam; Prigge, Sean T.; Afanador, Gustavo A.; Hickman, Mark R.; Lee, Patty J.; Leed, Susan E.; Auschwitz, Jennifer M.; Pieroni, Marco; Stec, Jozef; Muench, Stephen P.; Rice, David W.; Kozikowski, Alan P.

    2012-01-01

    Toxoplasma gondii is a protozoan parasite that can damage the human brain and eyes. There are no curative medicines. Herein, we describe our discovery of N-benzoyl-2-hydroxybenzamides as a class of compounds effective in the low nanomolar range against T. gondii in vitro and in vivo. Our lead compound, QQ-437, displays robust activity against the parasite and could be useful as a new scaffold for development of novel and improved inhibitors of T. gondii. Our genome-wide investigations reveal a specific mechanism of resistance to N-benzoyl-2-hydroxybenzamides mediated by adaptin-3β, a large protein from the secretory protein complex. N-Benzoyl-2-hydroxybenzamide-resistant clones have alterations of their secretory pathway, which traffics proteins to micronemes, rhoptries, dense granules, and acidocalcisomes/plant-like vacuole (PLVs). N-Benzoyl-2-hydroxybenzamide treatment also alters micronemes, rhoptries, the contents of dense granules, and, most markedly, acidocalcisomes/PLVs. Furthermore, QQ-437 is active against chloroquine-resistant Plasmodium falciparum. Our studies reveal a novel class of compounds that disrupts a unique secretory pathway of T. gondii, with the potential to be used as scaffolds in the search for improved compounds to treat the devastating diseases caused by apicomplexan parasites. PMID:22354304

  8. A genome-wide RNA interference screen identifies two novel components of the metazoan secretory pathway

    PubMed Central

    Wendler, Franz; Gillingham, Alison K; Sinka, Rita; Rosa-Ferreira, Cláudia; Gordon, David E; Franch-Marro, Xavier; Peden, Andrew A; Vincent, Jean-Paul; Munro, Sean

    2010-01-01

    Genetic screens in the yeast Saccharomyces cerevisiae have identified many proteins involved in the secretory pathway, most of which have orthologues in higher eukaryotes. To investigate whether there are additional proteins that are required for secretion in metazoans but are absent from yeast, we used genome-wide RNA interference (RNAi) to look for genes required for secretion of recombinant luciferase from Drosophila S2 cells. This identified two novel components of the secretory pathway that are conserved from humans to plants. Gryzun is distantly related to, but distinct from, the Trs130 subunit of the TRAPP complex but is absent from S. cerevisiae. RNAi of human Gryzun (C4orf41) blocks Golgi exit. Kish is a small membrane protein with a previously uncharacterised orthologue in yeast. The screen also identified Drosophila orthologues of almost 60% of the yeast genes essential for secretion. Given this coverage, the small number of novel components suggests that contrary to previous indications the number of essential core components of the secretory pathway is not much greater in metazoans than in yeasts. PMID:19942856

  9. The structure and dynamics of secretory component and its interactions with polymeric immunoglobulins

    PubMed Central

    Stadtmueller, Beth M; Huey-Tubman, Kathryn E; López, Carlos J; Yang, Zhongyu; Hubbell, Wayne L; Bjorkman, Pamela J

    2016-01-01

    As a first-line vertebrate immune defense, the polymeric immunoglobulin receptor (pIgR) transports polymeric IgA and IgM across epithelia to mucosal secretions, where the cleaved ectodomain (secretory component; SC) becomes a component of secretory antibodies, or when unliganded, binds and excludes bacteria. Here we report the 2.6Å crystal structure of unliganded human SC (hSC) and comparisons with a 1.7Å structure of teleost fish SC (tSC), an early pIgR ancestor. The hSC structure comprises five immunoglobulin-like domains (D1-D5) arranged as a triangle, with an interface between ligand-binding domains D1 and D5. Electron paramagnetic resonance measurements confirmed the D1-D5 interface in solution and revealed that it breaks upon ligand binding. Together with binding studies of mutant and chimeric SCs, which revealed domain contributions to secretory antibody formation, these results provide detailed models for SC structure, address pIgR evolution, and demonstrate that SC uses multiple conformations to protect mammals from pathogens. DOI: http://dx.doi.org/10.7554/eLife.10640.001 PMID:26943617

  10. Circadian pancreatic enzyme pattern and relationship between secretory and motor activity in fasting humans.

    PubMed

    Keller, Jutta; Layer, Peter

    2002-08-01

    It is unknown whether nonparallel pancreatic enzyme output occurs under basal conditions in humans. We aimed to determine whether the circadian or wake-sleep cycle influences the relationship among pancreatic enzymes or between pancreatic secretory and jejunal motor activity. Using orojejunal multilumen intubation, we measured enzyme outputs and proximal jejunal motility index during consecutive daytime and nighttime periods in each of seven fasting, healthy volunteers. Enzyme outputs were correlated tightly during daytime phases of wakefulness and nighttime phases of sleep (r > 0.72, P < 0.001). During nocturnal phases of wakefulness, output of proteases (r = 0.84, P < 0.001), but not of amylase and trypsin (r = 0.12), remained associated. Nocturnally, particularly during sleep, pancreatic secretory activity was directly correlated with jejunal motility index (r > 0.50, P < 0.001). In conclusion, parallel secretion of pancreatic enzymes dominates throughout the circadian cycle. Nonparallel secretion during nocturnal phases of wakefulness may be due to merely circadian effects or to the coupling of the wake-sleep and the circadian cycle. The association between fluctuations of secretory and motor activity appears to be particularly tight during the night.

  11. Augmentation of arginase Ⅱ expression in the human endometrial epithelium in the secretory phase.

    PubMed

    Tajima, Makiko; Harada, Tatsuya; Ishikawa, Tomonori; Iwahara, Yuki; Kubota, Toshiro

    2012-12-03

    L-arginine is the common substrate for arginase and nitric oxide synthase (NOS). Arginase converts L-arginine to urea and L-ornithine. L-Ornithine is the principal precursor for the production of polyamines and L-proline, which are required for cell proliferation and collagen synthesis. Endothelial NOS is expressed in the human endometrial glandular epithelium, but the expression and physiological roles of arginase in the human endometrium are not clear. The objective of this study was to investigate the expression and distribution patterns of arginases Ⅰ (A-Ⅰ) and Ⅱ (A-Ⅱ) in the human endometrium by using immunohistochemistry, reverse transcription-polymerase chain reaction (RTPCR), and western blotting. A-Ⅰ and A-Ⅱ were detected by immunohistochemistry in human endometrial epithelial cells during the proliferative and secretory phases of the menstrual cycle. RT-PCR showed that A-Ⅰ and A-Ⅱ mRNA were expressed in human endometrial tissue. Western blotting analysis results showed the expression of A-Ⅱ protein. Immunohistochemistry and western blotting results showed that expression levels of A-Ⅱ were significantly higher in the secretory phase than in the proliferative phase. Increased A-Ⅱ levels in the secretory phase may be responsible for endometrial growth by increasing polyamines and proline products.

  12. Avian minor salivary glands: an ultrastructural study of the secretory granules in mucous and seromucous cells.

    PubMed

    Olmedo, L A; Samar, M E; Avila, R E; de Crosa, M G; Dettin, L

    2000-01-01

    Ultrastructural descriptions in birds are scarce thus, in this study we have characterized the secretory granules of mucous and seromucous cells from the palatine and lingual salivary glands of birds with different diets. The samples were taken from the tongue and palatine mucosa of chicken (Gallus gallus), quail (Coturnix coturnix), chimango (Milvago chimango) and white heron (Egretta thula). The samples were processed for observation by transmission electron microscopy (TEM) employing 4% Karnovsky solution for fixation. The most noteworthy finding was the heterogeneous ultrastructural appearance of the secretory granules. Differences in substructure were found between the four species, between the palatine and lingual glands in the same species and even within the same acinus and the same cell. At variance with other authors, these differences cannot be attributed to the type of fixative solution used taking into account that all the samples were processed in the same way. Previous histochemical studies have shown the presence of sulfated and non sulfated glycoconjugates in these glands which can be associated to the maturation of the granules. These granules are probably representative of peculiar storage of the secretory products that would give rise to a heterogeneous and complex ultrastructural pattern of granules in the mucosa and seromucosa cells of these avian species.

  13. A pediatric case of mammary analogue secretory carcinoma within the parotid.

    PubMed

    Quattlebaum, S Craig; Roby, Brianne; Dishop, Megan K; Said, M Sherif; Chan, Kenny

    2015-01-01

    Mammary analogue secretory carcinoma (MASC) is a recently described entity in the differential diagnosis of salivary gland tumors. It is notable for a characteristic t(12;15)(p13;q25) translocation that results in a unique fusion protein, ETV6-NTRK3. While several studies have retrospectively identified this translocation in cases previously diagnosed as a different salivary malignancy, there have been relatively few cases where this translocation was identified on initial pathology results, and fewer still in a pediatric population. We present a case of a 15 year old female with a slowly enlarging, painless, left facial mass. MRI demonstrated a cystic mass extending into the deep lobe of the parotid, and she underwent parotidectomy. The tumor cells stained positive for S100 and CK19. ETV6 translocation was present, confirming the diagnosis. Mammary analogue secretory carcinoma is a recently described tumor of the salivary glands, which often masquerades as more common primary salivary gland tumors and cysts. More research is needed to characterize the typical behavior of this neoplasm and the optimal treatment regimen. With identification of its characteristic translocation, mammary analogue secretory carcinoma can be easily differentiated from its more prevalent counterparts, and should therefore remain within the differential of the pathologist and head and neck surgeon.

  14. Novel N-benzoyl-2-hydroxybenzamide disrupts unique parasite secretory pathway.

    PubMed

    Fomovska, Alina; Huang, Qingqing; El Bissati, Kamal; Mui, Ernest J; Witola, William H; Cheng, Gang; Zhou, Ying; Sommerville, Caroline; Roberts, Craig W; Bettis, Sam; Prigge, Sean T; Afanador, Gustavo A; Hickman, Mark R; Lee, Patty J; Leed, Susan E; Auschwitz, Jennifer M; Pieroni, Marco; Stec, Jozef; Muench, Stephen P; Rice, David W; Kozikowski, Alan P; McLeod, Rima

    2012-05-01

    Toxoplasma gondii is a protozoan parasite that can damage the human brain and eyes. There are no curative medicines. Herein, we describe our discovery of N-benzoyl-2-hydroxybenzamides as a class of compounds effective in the low nanomolar range against T. gondii in vitro and in vivo. Our lead compound, QQ-437, displays robust activity against the parasite and could be useful as a new scaffold for development of novel and improved inhibitors of T. gondii. Our genome-wide investigations reveal a specific mechanism of resistance to N-benzoyl-2-hydroxybenzamides mediated by adaptin-3β, a large protein from the secretory protein complex. N-Benzoyl-2-hydroxybenzamide-resistant clones have alterations of their secretory pathway, which traffics proteins to micronemes, rhoptries, dense granules, and acidocalcisomes/plant-like vacuole (PLVs). N-Benzoyl-2-hydroxybenzamide treatment also alters micronemes, rhoptries, the contents of dense granules, and, most markedly, acidocalcisomes/PLVs. Furthermore, QQ-437 is active against chloroquine-resistant Plasmodium falciparum. Our studies reveal a novel class of compounds that disrupts a unique secretory pathway of T. gondii, with the potential to be used as scaffolds in the search for improved compounds to treat the devastating diseases caused by apicomplexan parasites.

  15. Differentiation of osmotic and secretory diarrhoea by stool carbohydrate and osmolar gap measurements

    PubMed Central

    Castro-Rodriguez, J. A.; Salazar-Lindo, E.; Leon-Barua, R.

    1997-01-01

    

 Clinical features and laboratory tests that determine carbohydrate in faeces were evaluated to determine which was best able to distinguish between osmotic and secretory diarrhoea in infants and children. For this purpose 80 boys aged 3 to 24 months, with acute watery diarrhoea, were studied prospectively. The faecal osmolar gap (FOG) was calculated as: serum osmolarity − [2 × (faecal sodium + potassium concentration)]. Fifty eight patients were classified as having predominantly osmotic diarrhoea (FOG >100 mosmol/l), and 22 as having predominantly secretory diarrhoea (FOG ⩽100 mosmol/l). The two groups were comparable in their clinical features on admission, in the results of blood and urine tests, and in the evolution of their diarrhoeal illness. Evidence of steatorrhoea (by positive Sudan III test) and of acid faecal pH on admission were significantly more frequent in patients with osmotic diarrhoea. Mean (SD) faecal osmolarity was not significantly different between the two groups (319 (80) mosmol/l in secretory diarrhoea v 361 (123) mosmol/l in osmotic diarrhoea). Tests for reducing substances in faeces such as Benedict's test—with and without hydrolysis—and glucose strip, all showed a positive and significant association with osmotic diarrhoea (p <0.05, <0.025, <0.05, respectively). The presence of excess reducing substances (Benedict's test with hydrolysis >++) on admission was the most sensitive and specific test with the best predictive value for differentiating between the two types of watery diarrhoea.

 PMID:9370895

  16. The plant secretory pathway seen through the lens of the cell wall.

    PubMed

    van de Meene, A M L; Doblin, M S; Bacic, Antony

    2017-01-01

    Secretion in plant cells is often studied by looking at well-characterised, evolutionarily conserved membrane proteins associated with particular endomembrane compartments. Studies using live cell microscopy and fluorescent proteins have illuminated the highly dynamic nature of trafficking, and electron microscopy studies have resolved the ultrastructure of many compartments. Biochemical and molecular analyses have further informed about the function of particular proteins and endomembrane compartments. In plants, there are over 40 cell types, each with highly specialised functions, and hence potential variations in cell biological processes and cell wall structure. As the primary function of secretion in plant cells is for the biosynthesis of cell wall polysaccharides and apoplastic transport complexes, it follows that utilising our knowledge of cell wall glycosyltransferases (GTs) and their polysaccharide products will inform us about secretion. Indeed, this knowledge has led to novel insights into the secretory pathway, including previously unseen post-TGN secretory compartments. Conversely, our knowledge of trafficking routes of secretion will inform us about polarised and localised deposition of cell walls and their constituent polysaccharides/glycoproteins. In this review, we look at what is known about cell wall biosynthesis and the secretory pathway and how the different approaches can be used in a complementary manner to study secretion and provide novel insights into these processes.

  17. The structure and dynamics of secretory component and its interactions with polymeric immunoglobulins.

    PubMed

    Stadtmueller, Beth M; Huey-Tubman, Kathryn E; López, Carlos J; Yang, Zhongyu; Hubbell, Wayne L; Bjorkman, Pamela J

    2016-03-04

    As a first-line vertebrate immune defense, the polymeric immunoglobulin receptor (pIgR) transports polymeric IgA and IgM across epithelia to mucosal secretions, where the cleaved ectodomain (secretory component; SC) becomes a component of secretory antibodies, or when unliganded, binds and excludes bacteria. Here we report the 2.6Å crystal structure of unliganded human SC (hSC) and comparisons with a 1.7Å structure of teleost fish SC (tSC), an early pIgR ancestor. The hSC structure comprises five immunoglobulin-like domains (D1-D5) arranged as a triangle, with an interface between ligand-binding domains D1 and D5. Electron paramagnetic resonance measurements confirmed the D1-D5 interface in solution and revealed that it breaks upon ligand binding. Together with binding studies of mutant and chimeric SCs, which revealed domain contributions to secretory antibody formation, these results provide detailed models for SC structure, address pIgR evolution, and demonstrate that SC uses multiple conformations to protect mammals from pathogens.

  18. The Actomyosin Ring Recruits Early Secretory Compartments to the Division Site in Fission Yeast

    PubMed Central

    Vjestica, Aleksandar; Tang, Xin-Zi

    2008-01-01

    The ultimate goal of cytokinesis is to establish a membrane barrier between daughter cells. The fission yeast Schizosaccharomyces pombe utilizes an actomyosin-based division ring that is thought to provide physical force for the plasma membrane invagination. Ring constriction occurs concomitantly with the assembly of a division septum that is eventually cleaved. Membrane trafficking events such as targeting of secretory vesicles to the division site require a functional actomyosin ring suggesting that it serves as a spatial landmark. However, the extent of polarization of the secretion apparatus to the division site is presently unknown. We performed a survey of dynamics of several fluorophore-tagged proteins that served as markers for various compartments of the secretory pathway. These included markers for the endoplasmic reticulum, the COPII sites, and the early and late Golgi. The secretion machinery exhibited a marked polarization to the division site. Specifically, we observed an enrichment of the transitional endoplasmic reticulum (tER) accompanied by Golgi cisternae biogenesis. These processes required actomyosin ring assembly and the function of the EFC-domain protein Cdc15p. Cdc15p overexpression was sufficient to induce tER polarization in interphase. Thus, fission yeast polarizes its entire secretory machinery to the cell division site by utilizing molecular cues provided by the actomyosin ring. PMID:18184749

  19. Effect of oral acetylcysteine on tobacco smoke-induced secretory cell hyperplasia.

    PubMed

    Jeffery, P K; Rogers, D F; Ayers, M M

    1985-01-01

    The present investigation explores whether N-acetylcysteine (NAC) inhibits the secretory cell hyperplasia known to occur experimentally in specific pathogen-free (SPF) bronchitic rats. The animals were divided into 4 groups: no tobacco smoke (TS), no drug, no TS but NAC (1040 mg/kg body weight), TS but no drug, and TS plus NAC. NAC-treated animals showed no ill effects, TS exposed animals showed an initial fall in weight gain which never fully recovered (P less than 0.01): NAC did not protect. TS caused a significant increase (62-421%) in secretory cell number at all airway levels distal to the upper trachea (P less than 0.01) and NAC significantly inhibited it (P less than 0.01-0.05) in all, mostly in secretory cells containing acidic glycoprotein. TS exposure also induced a significant rise in epithelial cell concentration and of ciliated, mucous and especially basal cell number (P less than 0.001). NAC inhibited the mucous cell increase (P less than 0.001) and had 3 effects on the peak of dividing cells: it was (a) delayed until 3 days (b) greatly reduced in size and (c) prolonged at a lower level until its return to control values at 10 days of TS exposure.

  20. Cysteine-Rich Atrial Secretory Protein from the Snail Achatina achatina: Purification and Structural Characterization

    PubMed Central

    Shabelnikov, Sergey; Kiselev, Artem

    2015-01-01

    Despite extensive studies of cardiac bioactive peptides and their functions in molluscs, soluble proteins expressed in the heart and secreted into the circulation have not yet been reported. In this study, we describe an 18.1-kDa, cysteine-rich atrial secretory protein (CRASP) isolated from the terrestrial snail Achatina achatina that has no detectable sequence similarity to any known protein or nucleotide sequence. CRASP is an acidic, 158-residue, N-glycosylated protein composed of eight alpha-helical segments stabilized with five disulphide bonds. A combination of fold recognition algorithms and ab initio folding predicted that CRASP adopts an all-alpha, right-handed superhelical fold. CRASP is most strongly expressed in the atrium in secretory atrial granular cells, and substantial amounts of CRASP are released from the heart upon nerve stimulation. CRASP is detected in the haemolymph of intact animals at nanomolar concentrations. CRASP is the first secretory protein expressed in molluscan atrium to be reported. We propose that CRASP is an example of a taxonomically restricted gene that might be responsible for adaptations specific for terrestrial pulmonates. PMID:26444993

  1. Saturated fatty acids alter the late secretory pathway by modulating membrane properties.

    PubMed

    Payet, Laurie-Anne; Pineau, Ludovic; Snyder, Ellen C R; Colas, Jenny; Moussa, Ahmed; Vannier, Brigitte; Bigay, Joelle; Clarhaut, Jonathan; Becq, Frédéric; Berjeaud, Jean-Marc; Vandebrouck, Clarisse; Ferreira, Thierry

    2013-12-01

    Saturated fatty acids (SFA) have been reported to alter organelle integrity and function in many cell types, including muscle and pancreatic β-cells, adipocytes, hepatocytes and cardiomyocytes. SFA accumulation results in increased amounts of ceramides/sphingolipids and saturated phospholipids (PL). In this study, using a yeast-based model that recapitulates most of the trademarks of SFA-induced lipotoxicity in mammalian cells, we demonstrate that these lipid species act at different levels of the secretory pathway. Ceramides mostly appear to modulate the induction of the unfolded protein response and the transcription of nutrient transporters destined to the cell surface. On the other hand, saturated PL, by altering membrane properties, directly impact vesicular budding at later steps in the secretory pathway, i.e. at the trans-Golgi Network level. They appear to do so by increasing lipid order within intracellular membranes which, in turn, alters the recruitment of loose lipid packing-sensing proteins, required for optimal budding, to nascent vesicles. We propose that this latter general mechanism could account for the well-documented deleterious impacts of fatty acids on the last steps of the secretory pathway in several cell types.

  2. Vacuolar H+-ATPase activity is required for endocytic and secretory trafficking in Arabidopsis.

    PubMed

    Dettmer, Jan; Hong-Hermesdorf, Anne; Stierhof, York-Dieter; Schumacher, Karin

    2006-03-01

    In eukaryotic cells, compartments of the highly dynamic endomembrane system are acidified to varying degrees by the activity of vacuolar H(+)-ATPases (V-ATPases). In the Arabidopsis thaliana genome, most V-ATPase subunits are encoded by small gene families, thus offering potential for a multitude of enzyme complexes with different kinetic properties and localizations. We have determined the subcellular localization of the three Arabidopsis isoforms of the membrane-integral V-ATPase subunit VHA-a. Colocalization experiments as well as immunogold labeling showed that VHA-a1 is preferentially found in the trans-Golgi network (TGN), the main sorting compartment of the secretory pathway. Uptake experiments with the endocytic tracer FM4-64 revealed rapid colocalization with VHA-a1, indicating that the TGN may act as an early endosomal compartment. Concanamycin A, a specific V-ATPase inhibitor, blocks the endocytic transport of FM4-64 to the tonoplast, causes the accumulation of FM4-64 together with newly synthesized plasma membrane proteins, and interferes with the formation of brefeldin A compartments. Furthermore, nascent cell plates are rapidly stained by FM4-64, indicating that endocytosed material is redirected into the secretory flow after reaching the TGN. Together, our results suggest the convergence of the early endocytic and secretory trafficking pathways in the TGN.

  3. Engineered tobacco etch virus (TEV) protease active in the secretory pathway of mammalian cells.

    PubMed

    Cesaratto, Francesca; López-Requena, Alejandro; Burrone, Oscar R; Petris, Gianluca

    2015-10-20

    Tobacco etch virus protease (TEVp) is a unique endopeptidase with stringent substrate specificity. TEVp has been widely used as a purified protein for in vitro applications, but also as a biological tool directly expressing it in living cells. To adapt the protease to diverse applications, several TEVp mutants with different stability and enzymatic properties have been reported. Herein we describe the development of a novel engineered TEVp mutant designed to be active in the secretory pathway. While wild type TEVp targeted to the secretory pathway of mammalian cells is synthetized as an N-glycosylated and catalytically inactive enzyme, a TEVp mutant with selected mutations at two verified N-glycosylation sites and at an exposed cysteine was highly efficient. This mutant was very active in the endoplasmic reticulum (ER) of living cells and can be used as a biotechnological tool to cleave proteins within the secretory pathway. As an immediate practical application we report the expression of a complete functional monoclonal antibody expressed from a single polypeptide, which was cleaved by our TEVp mutant into the two antibody chains and secreted as an assembled and functional molecule. In addition, we show active TEVp mutants lacking auto-cleavage activity.

  4. Intracellular and transcellular transport of secretory and membrane proteins in the rat hepatocyte

    SciTech Connect

    Sztul, E.S.

    1984-01-01

    The intra- and transcellular transport of hepatic secretory and membrane proteins was studied in rats in vivo using (/sup 3/H)fucose and (/sup 35/S)cyteine as metabolic precursors. Incorporated radioactivity in plasma, bile, and liver subcellular fractions was measured and the labeled proteins of the Golgi complex, bile and plasma were separated by SDS-PAGE and identified by fluorography. /sup 3/H-radioactivity in Golgi fractions peaked at 10 min post injection (p.i.) and then declined concomitantly with the appearance of labeled glycoproteins in plasma. Maximal secretion of secretory fucoproteins from the Golgi complex occurred between 10 and 20 min p.i. In contrast, the clearance of labeled proteins from Golgi membrane subfractions occurred past 30 min p.i., indicating that membrane proteins leave the Golgi complex at least 10 min later than the bulk of content proteins. A major 80K form of Secretory Component (SC) was identified in the bile by precipitation with an anti IgA antibody. A comparative study of kinetics of transport of /sup 35/S-labeled SC and /sup 35/S-labeled albumin showed that albumin peaked in bile at approx.45 min p.i., whereas the SC peak occurred at 80 min p.i., suggesting that the transit time differs for plasma and membrane proteins which are delivered to the bile canaliculus (BC).

  5. Natural polyreactive secretory immunoglobulin A autoantibodies as a possible barrier to infection in humans.

    PubMed Central

    Quan, C P; Berneman, A; Pires, R; Avrameas, S; Bouvet, J P

    1997-01-01

    Secretory immunoglobulin A (S-IgA) was investigated in human secretions for the presence of natural antibodies (Abs) acting as the first "immune barrier" to infection before induction or boosting of specific responses. These molecules could be the secretory counterpart of the natural Abs in serum that were previously shown by our laboratory to be polyreactive to autoantigens. Significant levels of S-IgA Abs to human actin, myosin, tubulin, and spectrin were detected in 10 saliva and 8 colostrum samples from normal subjects. Computer-assisted analysis of immunoblots of extracts from human muscles showed these Abs to react with a large number of autoantigens. Their polyreactivity was confirmed by cross-inhibition and by immunoblotting studies of affinity-purified natural Abs, assayed against a large variety of surface or secreted antigens from Streptococcus pyogenes. The thiocyanate elution method showed that functional affinities of some natural Abs can be of the same order of magnitude as those of tetanus vaccine antitoxins. Moreover, nonimmune binding of these natural Abs to the gut protein Fv (Fv-fragment binding protein) can enhance their effector functions. This demonstrates that human secretions contain polyreactive auto-Abs which can also react with pathogens. These secretory Abs of "skeleton key" specificities are possibly produced by a primordial B-1-cell-associated immune system and can be involved in a plurispecific mucosal protection against pathogens, irrespective of the conventional immune response. PMID:9316998

  6. The key target of neuroprotection after the onset of ischemic stroke: secretory pathway Ca2+-ATPase 1

    PubMed Central

    Li, Li-hua; Tian, Xiang-rong; Hu, Zhi-ping

    2015-01-01

    The regulatory mechanisms of cytoplasmic Ca2+ after myocardial infarction-induced Ca2+ overload involve secretory pathway Ca2+-ATPase 1 and the Golgi apparatus and are well understood. However, the effect of Golgi apparatus on Ca2+ overload after cerebral ischemia and reperfusion remains unclear. Four-vessel occlusion rats were used as animal models of cerebral ischemia. The expression of secretory pathway Ca2+-ATPase 1 in the cortex and hippocampus was detected by immunoblotting, and Ca2+ concentrations in the cytoplasm and Golgi vesicles were determined. Results showed an overload of cytoplasmic Ca2+ during ischemia and reperfusion that reached a peak after reperfusion. Levels of Golgi Ca2+ showed an opposite effect. The expression of Golgi-specific secretory pathway Ca2+-ATPase 1 in the cortex and hippocampus decreased before ischemia and reperfusion, and increased after reperfusion for 6 hours. This variation was similar to the alteration of calcium in separated Golgi vesicles. These results indicate that the Golgi apparatus participates in the formation and alleviation of calcium overload, and that secretory pathway Ca2+-ATPase 1 tightly responds to ischemia and reperfusion in nerve cells. Thus, we concluded that secretory pathway Ca2+-ATPase 1 plays an essential role in cytosolic calcium regulation and its expression can be used as a marker of Golgi stress, responding to cerebral ischemia and reperfusion. The secretory pathway Ca2+-ATPase 1 can be an important neuroprotective target of ischemic stroke. PMID:26487855

  7. Non-planar interconnects in double-sided flexible Cu-PET substrates using a laser-assisted maskless microdeposition process: 3D finite element modeling and experimental analysis

    NASA Astrophysics Data System (ADS)

    Jabari, Elahe; Tong, Steven; Azhari, Amir; Toyserkani, Ehsan

    2014-03-01

    Non-planar (3D) interconnects have an important role in the electronic packaging industry these days. These unconventional interconnects allow manufacturers to save materials and space while connecting circuit components on flexible and non-planar substrates. Among a variety of flexible boards, double-sided flexible substrates have attracted the electronic industry to effectively and compactly develop miniaturized flexible devices such as sensors-on-chips. This study reports our developmental procedure for the creation of non-planar silver interconnects on the edge of double-sided copper substrates separated by a layer of polyethylene terephthalate (PET) using laser-assisted maskless microdeposition (LAMM). The article consists of the characterization of the LAMM process to effectively deposit Ag nanoparticles for production of conductive interconnects. Several parameters, including the deposition and laser processing parameters, are optimized to achieve interconnects free of pores, cracks and delamination. For investigating the topography and microstructure of interconnects, various analytical tools, such as SEM, XRD, Profilometery, and EDS were used. Furthermore, a 3D finite element numerical model was developed to predict the laser processing of silver nanoparticles on the substrate. The model includes a coupled thermal and structural governing physics to derive the temperature history throughout the simulation as well as strain/displacement within the substrate, which is identified the major source of cark formation in Ag tracks. The SEM micrographs of the laser processed nanoparticles suggest that a minimum of 1.24 W laser power was needed for an effective nanoparticles sintering to obtain conductive 3D interconnects with minimum amount of cracks whereas a 1.7 W laser power caused PET to decompose.

  8. A parametric study of planform and aeroelastic effects on aerodynamic center, alpha- and q- stability derivatives. Appendix C: Method for computing the aerodynamic influence coefficient matrix of nonplanar wing-body-tail configurations

    NASA Technical Reports Server (NTRS)

    Roskam, J.

    1972-01-01

    Expressions are derived for computing the aerodynamic influence coefficient matrix for nonplanar wing-body-tail configurations. An aerodynamic influence coefficient is defined as the load in lbs. induced on a panel as a result of a unit angle of attack on another panel. Fuselage, wing and tail thickness are assumed to be small with the result that the thickness effect on the flow-field is negligible. The method for determining the aerodynamic influence coefficient matrix is based on the lifting solution to the small perturbation, steady potential flow equation.

  9. Ranking Gene Ontology terms for predicting non-classical secretory proteins in eukaryotes and prokaryotes.

    PubMed

    Huang, Wen-Lin

    2012-11-07

    Protein secretion is an important biological process for both eukaryotes and prokaryotes. Several sequence-based methods mainly rely on utilizing various types of complementary features to design accurate classifiers for predicting non-classical secretory proteins. Gene Ontology (GO) terms are increasing informative in predicting protein functions. However, the number of used GO terms is often very large. For example, there are 60,020 GO terms used in the prediction method Euk-mPLoc 2.0 for subcellular localization. This study proposes a novel approach to identify a small set of m top-ranked GO terms served as the only type of input features to design a support vector machine (SVM) based method Sec-GO to predict non-classical secretory proteins in both eukaryotes and prokaryotes. To evaluate the Sec-GO method, two existing methods and their used datasets are adopted for performance comparisons. The Sec-GO method using m=436 GO terms yields an independent test accuracy of 96.7% on mammalian proteins, much better than the existing method SPRED (82.2%) which uses frequencies of tri-peptides and short peptides, secondary structure, and physicochemical properties as input features of a random forest classifier. Furthermore, when applying to Gram-positive bacterial proteins, the Sec-GO with m=158 GO terms has a test accuracy of 94.5%, superior to NClassG+ (90.0%) which uses SVM with several feature types, comprising amino acid composition, di-peptides, physicochemical properties and the position specific weighting matrix. Analysis of the distribution of secretory proteins in a GO database indicates the percentage of the non-classical secretory proteins annotated by GO is larger than that of classical secretory proteins in both eukaryotes and prokaryotes. Of the m top-ranked GO features, the top-four GO terms are all annotated by such subcellular locations as GO:0005576 (Extracellular region). Additionally, the method Sec-GO is easily implemented and its web tool of

  10. Episodic slow slip process in a non-planar fault model constrained by non-volcanic tremor locations along Cascadia subduction zone

    NASA Astrophysics Data System (ADS)

    Li, D.; Liu, Y.

    2015-12-01

    Slow slip events (SSEs) are observed in the circum-Pacific subduction zones and exhibit a wide diversity of source parameters (including equivalent moment, duration and recurrence interval). Gao et al [2012] compiled source parameters of SSEs around the world and revealed their empirical scaling relations distinct from those of regular earthquakes. However, the nature of this diversity is unclear. Previous 3-D numerical simulations in a simplified plate model have reproduced the along-strike segmentation of episodic SSEs in Cascadia margin and their source scaling relations in the framework of rate- and state- friction law [Liu, 2014]. But the planar fault model is inefficient to investigate the effect of the fault geometry on the source characteristics of SSEs in a specific subduction zone. In this study, we adopt a non-planar Cascadia subduction fault geometry constrained by relocated seismicity [McCrory et al. 2012] in the rate-state friction model. We have reproduced episodic SSEs beneath Vancouver Island and Washington arising every ~1.5 year with a maximum cumulative slip of ~2.5 cm. We find three phases characteristic in cumulative moment rate and slip rate in each episode. These three phases, defined as nucleation, fast-spreading and healing, lasting for ~160, ~60 and ~140 days, respectively. Both the nucleation and healing phases are beneath GPS detection threshold but the nucleation phase is spatially correlated with the small tremor activities arising deeper than episodic tremor and slip (ETS). To further investigate the diversity of SSEs in Cascadia, we introduce the recent 6-year tremor locations in Cascadia (http://pnsn.org/tremor) to constrain the near-lithostatic pore pressure distribution at the SSE depths in an 800km-long Cascadia fault model. Additionally, we set a step change of effective normal stress in SSE region from 1.5 MPa in Vancouver Island and Washington to 2.0 MPa in Oregon. The results show that the modeled SSEs exhibit distinct slip

  11. Antigenic homogeneity of male Müllerian gland (MG) secretory proteins of a caecilian amphibian with secretory proteins of the mammalian prostate gland and seminal vesicles: evidence for role of the caecilian MG as a male accessory reproductive gland.

    PubMed

    Radha, Arumugam; Sree, Sreesha; Faisal, Kunnathodi; Kumar, G Pradeep; Oommen, Oommen V; Akbarsha, Mohammad A

    2014-10-01

    Whereas in all other vertebrates the Müllerian ducts of genetic males are aborted during development, under the influence of Müllerian-inhibiting substance, in the caecilian amphibians they are retained as a pair of functional glands. It has long been speculated that the Müllerian gland might be the male accessory reproductive gland but there has been no direct evidence to this effect. The present study was undertaken to determine whether the caecilian Müllerian gland secretory proteins would bear antigenic similarity to secretory proteins of the prostate gland and/or the seminal vesicles of a mammal. The secretory proteins of the Müllerian gland of Ichthyophis tricolor were evaluated for cross-reactivity with antisera raised against rat ventral prostate and seminal vesicle secretory proteins, adopting SDS-PAGE, two-dimensional electrophoresis and immunoblot techniques. Indeed there was a cross-reaction of five Müllerian gland secretory protein fractions with prostatic protein antiserum and of three with seminal vesicle protein antiserum. A potential homology exists because in mammals the middle group of the prostate primordia is derived from a diverticulum of the Müllerian duct. Thus this study, by providing evidence for expression of prostatic and seminal vesicle proteins in the Müllerian gland, substantiates the point that in caecilians the Müllerian glands are the male accessory reproductive glands.

  12. Large-scale identification of endogenous secretory peptides using electron transfer dissociation mass spectrometry.

    PubMed

    Sasaki, Kazuki; Osaki, Tsukasa; Minamino, Naoto

    2013-03-01

    Mass spectrometry-based unbiased analysis of the full complement of secretory peptides is expected to facilitate the identification of unknown biologically active peptides. However, tandem MS sequencing of endogenous peptides in their native form has proven difficult because they show size heterogeneity and contain multiple internal basic residues, the characteristics not found in peptide fragments produced by in vitro digestion. Endogenous peptides remain largely unexplored by electron transfer dissociation (ETD), despite its widespread use in bottom-up proteomics. We used ETD, in comparison to collision induced dissociation (CID), to identify endogenous peptides derived from secretory granules of a human endocrine cell line. For mass accuracy, both MS and tandem MS were analyzed on an Orbitrap. CID and ETD, performed in different LC-MS runs, resulted in the identification of 795 and 569 unique peptides (ranging from 1000 to 15000 Da), respectively, with an overlap of 397. Peptides larger than 3000 Da accounted for 54% in CID and 46% in ETD identifications. Although numerically outperformed by CID, ETD provided more extensive fragmentation, leading to the identification of peptides that are not reached by CID. This advantage was demonstrated in identifying a new antimicrobial peptide from neurosecretory protein VGF (non-acronymic), VGF[554-577]-NH2, or in differentiating nearly isobaric peptides (mass difference less than 2 ppm) that arise from alternatively spliced exons of the gastrin-releasing peptide gene. CID and ETD complemented each other to add to our knowledge of the proteolytic processing sites of proteins implicated in the regulated secretory pathway. An advantage of the use of both fragmentation methods was also noted in localization of phosphorylation sites. These findings point to the utility of ETD mass spectrometry in the global study of endogenous peptides, or peptidomics.

  13. The sorting of proglucagon to secretory granules is mediated by carboxypeptidase E and intrinsic sorting signals.

    PubMed

    McGirr, Rebecca; Guizzetti, Leonardo; Dhanvantari, Savita

    2013-05-01

    Proglucagon is expressed in pancreatic alpha cells, intestinal L cells and brainstem neurons. Tissue-specific processing of proglucagon yields the peptide hormones glucagon in the alpha cell and glucagon-like peptide (GLP)-1 and GLP-2 in L cells. Both glucagon and GLP-1 are secreted in response to nutritional status and are critical for regulating glycaemia. The sorting of proglucagon to the dense-core secretory granules of the regulated secretory pathway is essential for the appropriate secretion of glucagon and GLP-1. We examined the roles of carboxypeptidase E (CPE), a prohormone sorting receptor, the processing enzymes PC1/3 and PC2 and putative intrinsic sorting signals in proglucagon sorting. In Neuro 2a cells that lacked CPE, PC1/3 and PC2, proglucagon co-localised with the Golgi marker p115 as determined by quantitative immunofluorescence microscopy. Expression of CPE, but not of PC1/3 or PC2, enhanced proglucagon sorting to granules. siRNA-mediated knockdown of CPE disrupted regulated secretion of glucagon from pancreatic-derived alphaTC1-6 cells, but not of GLP-1 from intestinal cell-derived GLUTag cells. Mutation of the PC cleavage site K70R71, the dibasic R17R18 site within glucagon or the alpha-helix of glucagon, all significantly affected the sub-cellular localisation of proglucagon. Protein modelling revealed that alpha helices corresponding to glucagon, GLP-1 and GLP-2, are arranged within a disordered structure, suggesting some flexibility in the sorting mechanism. We conclude that there are multiple mechanisms for sorting proglucagon to the regulated secretory pathway, including a role for CPE in pancreatic alpha cells, initial cleavage at K70R71 and multiple sorting signals.

  14. Axons provide the secretory machinery for trafficking of voltage-gated sodium channels in peripheral nerve

    PubMed Central

    González, Carolina; Cánovas, José; Fresno, Javiera; Couve, Eduardo; Court, Felipe A.; Couve, Andrés

    2016-01-01

    The regulation of the axonal proteome is key to generate and maintain neural function. Fast and slow axoplasmic waves have been known for decades, but alternative mechanisms to control the abundance of axonal proteins based on local synthesis have also been identified. The presence of the endoplasmic reticulum has been documented in peripheral axons, but it is still unknown whether this localized organelle participates in the delivery of axonal membrane proteins. Voltage-gated sodium channels are responsible for action potentials and are mostly concentrated in the axon initial segment and nodes of Ranvier. Despite their fundamental role, little is known about the intracellular trafficking mechanisms that govern their availability in mature axons. Here we describe the secretory machinery in axons and its contribution to plasma membrane delivery of sodium channels. The distribution of axonal secretory components was evaluated in axons of the sciatic nerve and in spinal nerve axons after in vivo electroporation. Intracellular protein trafficking was pharmacologically blocked in vivo and in vitro. Axonal voltage-gated sodium channel mRNA and local trafficking were examined by RT-PCR and a retention-release methodology. We demonstrate that mature axons contain components of the endoplasmic reticulum and other biosynthetic organelles. Axonal organelles and sodium channel localization are sensitive to local blockade of the endoplasmic reticulum to Golgi transport. More importantly, secretory organelles are capable of delivering sodium channels to the plasma membrane in isolated axons, demonstrating an intrinsic capacity of the axonal biosynthetic route in regulating the axonal proteome in mammalian axons. PMID:26839409

  15. Effects of ozone on the cholinergic secretory responsiveness of ferret tracheal glands

    SciTech Connect

    McBride, R.K.; Oberdoerster, G.; Marin, M.G. )

    1991-06-01

    Oxidant air pollutants exacerbate several pulmonary diseases. Inhalation of ozone has been shown to induce airway smooth muscle hyperresponsiveness. Oxidant injury could also affect airway secretory mechanisms. The authors postulated that oxidant exposure would alter the glycoconjugate secretory function of airway submucosal glands. To test this hypothesis they examined the effects of in vivo ozone exposure on the in vitro secretory responsiveness of ferret tracheal glands. Ferrets were exposed to 1 ppm ozone, 24 hr/day for 3 or 7 days. Following exposure, glandular explants, denuded of surface epithelial cells, were prepared and incubated in medium containing 3H-glucosamine for 18 hr. Basal secretion of labeled glycoconjugates was significantly increased 31% following 3 days of ozone exposure (P less than or equal to 0.05) and remained elevated 11% after 7 days of exposure compared to the air-exposed group. After 3 or 7 days of exposure to ozone, tracheal gland responsiveness to carbachol was increased as indicated by significantly lower EC50 values (log molar concentration) of -6.43 {plus minus} 0.04 (n = 6) and -6.50 {plus minus} 0.11 (n = 5), respectively; compared to -6.20 {plus minus} 0.08 (n = 6) for the air-exposed group. There was no difference in carbachol EC50 values for air and 7-day ozone-exposed animals treated with dexamethasone. Dexamethasone did not attenuate the ozone-induced increase in basal secretion. Tracheal gland responsiveness to {alpha}- or {beta}-adrenergic agonists was not changed by oxidant exposure. These experiments suggest that oxidant injury not only increases basal secretion of respiratory glycoconjugates but also increases tracheal gland sensitivity to a cholinergic agonist.

  16. Modulation of Sertoli cell secretory function by rat round spermatid protein(s).

    PubMed

    Onoda, M; Djakiew, D

    1990-10-01

    The influence of rat round spermatid protein(s) (RSP) on protein synthesis and secretory function of Sertoli cells was used in the bicameral chamber system. Round spermatids (RS) were purified from 90-day-old rats by centrifugal elutriation. RS were incubated in a supplement-enriched culture medium that lacked exogenous proteins. The RS-conditioned media were dialysed and lyophilized to obtain RSP. Most de novo protein synthesized under basal conditions by Sertoli cells (18-day-old) was secreted into the apical chamber (apical/basal ratio: 3.42). Follicle-stimulating hormone (FSH, 100 ng/ml) stimulated total protein secretion from Sertoli cells by a factor of 1.54. The RSP (100 micrograms/ml) stimulated total protein secretion from Sertoli cells by a factor of 2.33. The enhancement of total Sertoli cell protein secretion by FSH and RSP additively increased by a factor of 2.82. The combined effect of FSH and RSP on total protein secretion from Sertoli cells was dose dependent and saturated at approximately 200 micrograms/ml of RSP. Polarity of total protein secretion from Sertoli cells (apical/basal ratio: 3.42) was stimulated by RSP predominantly in the apical direction (apical/basal ratio: 8.48). The modulation of radiolabeled Sertoli cell secretory proteins (ceruloplasmin, CP; sulfated glycoprotein-2, SGP-2; testins and transferrin, Tf) by cold (non-labeled) RSP was investigated by immunoprecipitation followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The secretion of CP, SGP-2 and Tf was stimulated in a dose-dependent manner by the addition of RSP up to a saturating concentration of between 200 and 300 micrograms/ml, whereas the secretion of Sertoli cell testins did not reach saturation at 300 micrograms/ml RSP. These results indicate that FSH and RSP independently modulate Sertoli cell protein secretion, and that Sertoli cell secretory proteins may differentially respond to RSP stimulation.

  17. Secretory glands and microvascular systems imaged in aqueous solution by atmospheric scanning electron microscopy (ASEM).

    PubMed

    Yamazawa, Toshiko; Nakamura, Naotoshi; Sato, Mari; Sato, Chikara

    2016-12-01

    Exocrine glands, e.g., salivary and pancreatic glands, play an important role in digestive enzyme secretion, while endocrine glands, e.g., pancreatic islets, secrete hormones that regulate blood glucose levels. The dysfunction of these secretory organs immediately leads to various diseases, such as diabetes or Sjögren's syndrome, by poorly understood mechanisms. Gland-related diseases have been studied by optical microscopy (OM), and at higher resolution by transmission electron microscopy (TEM) of Epon embedded samples, which necessitates hydrophobic sample pretreatment. Here, we report the direct observation of tissue in aqueous solution by atmospheric scanning electron microscopy (ASEM). Salivary glands, lacrimal glands, and pancreas were fixed, sectioned into slabs, stained with phosphotungstic acid (PTA), and inspected in radical scavenger d-glucose solution from below by an inverted scanning electron microscopy (SEM), guided by optical microscopy from above to target the tissue substructures. A 2- to 3-µm specimen thickness was visualized by the SEM. In secretory cells, cytoplasmic vesicles and other organelles were clearly imaged at high resolution, and the former could be classified according to the degree of PTA staining. In islets of Langerhans, the microvascular system used as an outlet by the secretory cells was also clearly observed. Microvascular system is also critically involved in the onset of diabetic complications and was clearly visible in subcutaneous tissue imaged by ASEM. The results suggest the use of in-solution ASEM for histology and to study vesicle secretion systems. Further, the high-throughput of ASEM makes it a potential tool for the diagnosis of exocrine and endocrine-related diseases.

  18. Large-scale Identification of Endogenous Secretory Peptides Using Electron Transfer Dissociation Mass Spectrometry*

    PubMed Central

    Sasaki, Kazuki; Osaki, Tsukasa; Minamino, Naoto

    2013-01-01

    Mass spectrometry-based unbiased analysis of the full complement of secretory peptides is expected to facilitate the identification of unknown biologically active peptides. However, tandem MS sequencing of endogenous peptides in their native form has proven difficult because they show size heterogeneity and contain multiple internal basic residues, the characteristics not found in peptide fragments produced by in vitro digestion. Endogenous peptides remain largely unexplored by electron transfer dissociation (ETD), despite its widespread use in bottom-up proteomics. We used ETD, in comparison to collision induced dissociation (CID), to identify endogenous peptides derived from secretory granules of a human endocrine cell line. For mass accuracy, both MS and tandem MS were analyzed on an Orbitrap. CID and ETD, performed in different LC-MS runs, resulted in the identification of 795 and 569 unique peptides (ranging from 1000 to 15000 Da), respectively, with an overlap of 397. Peptides larger than 3000 Da accounted for 54% in CID and 46% in ETD identifications. Although numerically outperformed by CID, ETD provided more extensive fragmentation, leading to the identification of peptides that are not reached by CID. This advantage was demonstrated in identifying a new antimicrobial peptide from neurosecretory protein VGF (non-acronymic), VGF[554–577]-NH2, or in differentiating nearly isobaric peptides (mass difference less than 2 ppm) that arise from alternatively spliced exons of the gastrin-releasing peptide gene. CID and ETD complemented each other to add to our knowledge of the proteolytic processing sites of proteins implicated in the regulated secretory pathway. An advantage of the use of both fragmentation methods was also noted in localization of phosphorylation sites. These findings point to the utility of ETD mass spectrometry in the global study of endogenous peptides, or peptidomics. PMID:23250050

  19. Regulation by L channels of Ca(2+)-evoked secretory responses in ouabain-treated chromaffin cells.

    PubMed

    De Pascual, Ricardo; Colmena, Inés; Ruiz-Pascual, Lucía; Baraibar, Andrés Mateo; Egea, Javier; Gandía, Luis; García, Antonio G

    2016-10-01

    It is known that the sustained depolarisation of adrenal medullary bovine chromaffin cells (BCCs) with high K(+) concentrations produces an initial sharp catecholamine release that subsequently fades off in spite depolarisation persists. Here, we have recreated a sustained depolarisation condition of BCCs by treating them with the Na(+)/K(+) ATPase blocker ouabain; in doing so, we searched experimental conditions that permitted the development of a sustained long-term catecholamine release response that could be relevant during prolonged stress. BCCs were perifused with nominal 0Ca(2+) solution, and secretion responses were elicited by intermittent application of short 2Ca(2+) pulses (Krebs-HEPES containing 2 mM Ca(2+)). These pulses elicited a biphasic secretory pattern with an initial 30-min period with secretory responses of increasing amplitude and a second 30-min period with steady-state, non-inactivating responses. The initial phase was not due to gradual depolarisation neither to gradual increases of the cytosolic calcium transients ([Ca(2+)]c) elicited by 2Ca(2+) pulses in BBCs exposed to ouabain; both parameters increased soon after ouabain addition. Νifedipine blocked these responses, and FPL64176 potentiated them, suggesting that they were triggered by Ca(2+) entry through non-inactivating L-type calcium channels. This was corroborated by nifedipine-evoked blockade of the L-type Ca(2+) channel current and the [Ca(2+)]c transients elicited by 2Ca(2+) pulses. Furthermore, the plasmalemmal Na(+)/Ca(2+) exchanger (NCX) blocker SEA0400 caused a mild inhibition followed by a large rebound increase of the steady-state secretory responses. We conclude that these two phases of secretion are mostly contributed by Ca(2+) entry through L calcium channels, with a minor contribution of Ca(2+) entry through the reverse mode of the NCX.

  20. [Effect of plant hormones on the components of secretory pathway in human normal and tumor cells].

    PubMed

    Vil'danova, M S; Savitskaia, M A; Onishchenko, G E; Smirnova, E A

    2014-01-01

    Plant hormones play a key role in plant growth and differentiation. Many hormones are known as potential antitumor agents, yet others appear to affect the secretory activity and are produced by mammalian cells as pro-inflammatory cytokines. The goal of this research was to study the effect of abscisic and gibberellic acids on the secretory system of human cultured epidermoid carcinoma cells A431 and keratinocytes HaCat. Immunocytochemical and morphometric analysis demonstrated that subtoxic concentration of plant hormones induced the broadening of the ER network and increased the size of Golgi complex. Electron microscopy studies confirmed the hypertrophic changes of the Golgi apparatus, specifically, the swelling of cisternae in the trans-compartment of dictyosomes after exposure to abscisic acid, and swelling of cis- and trans-compartment of dictyosomes after exposure to abscisic acid, and swelling of cis- and trans-compartments of dictyosomes after exposure to gibberellic acid. Using of Click-iT technique allowed to detect the elevation of the total protein synthesis only in A431 cells exposed to abscisic acid. Cumulative data suggests that, under these conditions, the hypertrophy of Golgi apparatus may reflect the enhanced secretory activity of cells. In other experiments, the hypertrophy of Golgi is not related to increased protein synthesis and therefore may suggest the stress-related changes of ER and Golgi apparatus. Our results demonstrate that morphologically similar reaction of cellular organelles, such as hypertrophy of Golgi apparatus, is the result of different functional activities, and that molecular mechanisms underlying the changes induced in cells need further investigations.

  1. Leptin secretory dynamics and associated disordered eating psychopathology across the weight spectrum

    PubMed Central

    Baskaran, Charumathi; Eddy, Kamryn T.; Miller, Karen K.; Meenaghan, Erinne; Misra, Madhusmita; Lawson, Elizabeth A.

    2016-01-01

    Leptin secretory dynamics across the weight spectrum and their relationship with disordered eating psychopathology have not been studied. Our objective was to compare leptin secretory dynamics in 13 anorexia nervosa (AN), 12 overweight/obese (OB) and 12 normal-weight women using deconvolution analysis. Methods In this cross-sectional study conducted at a tertiary referral center, serum leptin levels were obtained every 20 minutes from 2000-0800h. Dual energy X-ray absorptiometry was used to measure %body fat. Disordered eating psychopathology was assessed by the Eating Disorders Examination-Questionnaire (EDE-Q) and Eating Disorders Inventory-2 (EDI-2). Results The groups differed for basal leptin secretion (BASAL) (p=0.02). Mean leptin pulse amplitude, pulse mass, total pulsatile secretion (TPS) and area under the curve (AUC) were significantly different between groups before and after adjustment for BASAL (p<0.0001 for all). Leptin AUC correlated strongly with TPS (r=0.97, p<0.0001) and less with BASAL (r=0.35, p=0.03). On multivariate analysis, only TPS was a significant predictor of leptin AUC (p<0.0001). TPS was inversely associated with most EDE-Q and EDI-2 parameters and the associations remained significant for EDE-Q eating concern (p=0.01), and EDI-2 asceticism, ineffectiveness and social insecurity (p<0.05) after adjusting for BASAL. These relationships were not significant when controlled for %body fat. Conclusion Secretory dynamics of leptin differ across weight spectrum, with mean pulse amplitude, mean pulse mass and TPS being low in AN and high in OB. Pulsatile, rather than basal secretion, is the major contributor to leptin AUC. Decreased pulsatile leptin is associated with disordered eating psychopathology, possibly reflecting low %body fat in AN. PMID:26903591

  2. Secretion of sulfated and nonsulfated forms of parathyroid chromogranin A (secretory protein-I)

    SciTech Connect

    Gorr, S.U.; Cohn, D.V. )

    1990-02-25

    Chromogranin A (secretory protein-I) is an acidic, sulfated glycoprotein found in secretory granules of most endocrine cells but not in exocrine or epithelial cells. Parathyroid chromogranin A is sulfated on tyrosine residues, whereas adrenal chromogranin A appears to be sulfated mainly on oligosaccharide residues. Chromogranin B, on the other hand, is tyrosine-sulfated in the bovine adrenal whereas this protein is absent from the parathyroid. The role of this tissue- or species-specific sulfation of chromogranin is not known. Tyrosine sulfation is a common post-translational modification of proteins in the exocytotic pathway and has been suggested to play a role in the sorting or intracellular transport of secretory proteins. To test this, porcine parathyroid tissue slices were metabolically labeled with 35SO4 and (3H)Lys, and the tissue and incubation medium analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, immunoblotting, and immunoprecipitation with chromogranin A-specific antiserum or by radioimmunoassay for parathormone. Secretion of total and 3H-labeled chromogranin A was about 3- and 7-fold higher, respectively, at 0.5 mM than at 3.0 mM Ca2+, and secretion of 35SO4-labeled chromogranin A was 67-fold higher. This indicates that either sulfated chromogranin A is directed primarily to the Ca2+-regulated pathway or that sulfation occurs following sorting to this pathway. Sodium chlorate (1-10 mM) inhibited sulfation in a dose-dependent manner by up to 95% but it had no effect on the onset or rate of chromogranin A secretion. These data indicate that regulated secretion of parathyroid chromogranin A does not require sulfation of tyrosine residues.

  3. Secretory clusterin inhibits osteoclastogenesis by attenuating M-CSF-dependent osteoclast precursor cell proliferation

    SciTech Connect

    Choi, Bongkun; Kang, Soon-Suk; Kang, Sang-Wook; Min, Bon-Hong; Lee, Eun-Jin; Song, Da-Hyun; Kim, Sang-Min; Song, Youngsup; Yoon, Seung-Yong; Chang, Eun-Ju

    2014-07-18

    Highlights: • We describe the expression and secretion of clusterin in osteoclasts. • Endogenous clusterin deficiency does not affect osteoclast formation. • Exogenous treatment with secretory clusterin decreases osteoclast differentiation. • Secretory clusterin attenuates osteoclast precursor cell proliferation by inhibiting M-CSF-mediated ERK activation. - Abstract: Secretory clusterin (sCLU)/apolipoprotein J is a multifunctional glycoprotein that is ubiquitously expressed in various tissues. Reduced sCLU in the joints of patients with bone erosive disease is associated with disease activity; however, its exact role has yet to be elucidated. Here, we report that CLU is expressed and secreted during osteoclastogenesis in mouse bone marrow-derived macrophages (BMMs) that are treated with receptor activator of nuclear factor kappa-B ligand (RANKL) and macrophage colony-stimulating factor (M-CSF). CLU-deficient BMMs obtained from CLU{sup −/−} mice exhibited no significant alterations in OC differentiation in comparison with BMMs obtained from wild-type mice. In contrast, exogenous sCLU treatment significantly inhibited OC formation in both BMMs and OC precursor cultures. The inhibitory effect of sCLU was more prominent in BMMs than OC precursor cultures. Interestingly, treating BMMs with sCLU decreased the proliferative effects elicited by M-CSF and suppressed M-CSF-induced ERK activation of OC precursor cells without causing apoptotic cell death. This study provides the first evidence that sCLU reduces OC formation by inhibiting the actions of M-CSF, thereby suggesting its protective role in bone erosion.

  4. Damascene fabrication of nonplanar microcoils

    DOEpatents

    Adams, David P.; Vasile, Michael J.

    2003-06-17

    A process for fabricating coils using a Damascene process uses a curved substrate having a surface extending along and about an axis made of a first material. A groove is formed in the curved surface along and around said axis, and the groove is filled with a second material that is different from the first material to form a coil of second material in said first material. Excess second material is then removed from the surface of the first material, leaving the coil of second material in the groove.

  5. Photolithography simulation on nonplanar substrates

    NASA Astrophysics Data System (ADS)

    Yeung, Michael S.

    1990-06-01

    A iod1 of oJ. )tica. I hthograpliy suitable for certain types of onedimensional 1)eriOdic tOpogra)hy including birds beaks afl(L reflowed BISC4 structures is described. it is based on a formalism of diffraction grating theory which uses a. coordinate t. raiisforna. tiou to iap all the nonpla. uar surfaces oiit. o parallel planes a. iid it ca. ii be used for the rigorous simulation of photoresist. latent images of oiiedimensional mask patterns with periodicity fuller j)a. ra. lleI or I)(rI)e11(Iicu1a. r to that of the topogra)hy. Effects of t. opogra)11y scattering 1)111k illiaging and )11otob1eachu1Ig are fully ta. keii into a. ccounl. for both types of mask patterns. Simulation results illustrate the combined effects of topography scattering and I) ulk iiiaging in iioiipla. na. r pliotoli tliogra. phiy using high iiuinerical aperture optics.

  6. Two Dimensional Gel Electrophoresis of Insulin Secretory Granule Proteins from Biosynthetically-Labeled Pancreatic Islets.

    PubMed

    Guest, Paul C

    2017-01-01

    Pulse-chase radiolabeling of cells with radioactive amino acids is a common method for tracking the biosynthesis of proteins. Radiolabeled newly synthesized proteins can be analyzed by a number of techniques such as two dimensional gel electrophoresis (2DE). This chapter presents a protocol for the biosynthetic labeling of pancreatic islets with (35)S-methionine in the presence of basal and stimulatory concentrations of glucose, followed by subcellular fractionation to produce a secretory granule fraction and analysis of the granule protein contents by 2DE. This provides a means of determining whether or not the biosynthetic rates of the entire granule constituents are coordinately regulated.

  7. 2D Gel Electrophoresis of Insulin Secretory Granule Proteins from Biosynthetically Labelled Pancreatic Islets.

    PubMed

    Guest, Paul C

    2017-01-01

    Pulse radiolabelling of cells with radioactive amino acids such is a common method for investigating the biosynthetic rates of proteins. In this way, the abundance of newly synthesized proteins can be determined by several proteomic techniques including 2D gel electrophoresis (2DE). This chapter describes a protocol for labelling pancreatic islets with (35)S-methionine in the presence of low and high concentrations of glucose, followed by subcellular fractionation enrichment of secretory granule proteins and analysis of the granule protein contents by 2DE. This demonstrated that the biosynthetic rates of most of the granule proteins are co-ordinately regulated in the presence of stimulatory glucose concentrations.

  8. Are there salvage routes within the general secretory pathway in yeast?

    PubMed

    Gozalbo, D; Martínez, J P; Sentandreu, R

    1992-04-01

    It is generally accepted that both extracellular protein secretion and plasma membrane expansion in yeast occur basically as in higher eukaryotic cells. In addition to the constitutive (default) secretory pathway, some specialized mammalian cells possess a regulated route which at present has not been detected in yeast. However, there is a body of experimental results suggesting that under certain circumstances export of integral plasma membrane and exocellular proteins may take place through alternative (salvage) pathways. The existence of these latter routes would enable the yeast cell to adapt more efficiently to distinct or adverse conditions requiring the secretion of discrete amounts of specific sets of proteins.

  9. Sending proteins to dense core secretory granules: still a lot to sort out.

    PubMed

    Dikeakos, Jimmy D; Reudelhuber, Timothy L

    2007-04-23

    The intracellular sorting of peptide hormone precursors to the dense core secretory granules (DCSGs) is essential for their bioactivation. Despite the fundamental importance of this cellular process, the nature of the sorting signals for entry of proteins into DCSGs remains a source of vigorous debate. This review highlights recent discoveries that are consistent with a model in which several protein domains, acting in a cell-specific fashion and at different steps in the sorting process, act in concert to regulate the entry of proteins into DCSGs.

  10. Pancreatic vasoactive intestinal polypeptide-oma as a cause of secretory diarrhoea.

    PubMed

    Masel, S L; Brennan, B A; Turner, J H; Cullingford, G L; Cullen, D J

    2000-04-01

    A 42-year-old woman presented with a 4-year history of worsening diarrhoea that was watery, profuse and confirmed to be secretory in nature. She had tested positive for phenolphthalein on urinary laxative screening but continued to deny laxative usage. Her vasoactive intestinal polypeptide (VIP) level was subsequently found to be markedly elevated. Despite a normal abdominal ultrasound, a computed tomography scan revealed a 5-cm pancreatic tail mass. Octreotide scanning was used to exclude metastatic disease and she went on to have surgical removal of a localized pancreatic vasoactive intestinal polypeptide-oma which resulted in the complete resolution of her diarrhoea.

  11. Minor salivary glands as a major source of secretory immunoglobin A in the human oral cavity.

    PubMed

    Crawford, J M; Taubman, M A; Smith, D J

    1975-12-19

    Secretory immunoglobulin A is the predominant immunoglobulin in labial minor salivary gland secretions. Its mean concentration is four times higher in these secretions than in parotid gland secretion. The minor salivary glands can produce 30 to 35 percent of the immunoglobulin A that enters the oral cavity. This, together with the potential accessibility of these glands to antigenic stimulation, suggest that they may be an important source of the immune factors that are involved in the regulation of the microorganisms in the oral environment.

  12. In vitro production of Toxocara canis excretory-secretory (TES) antigen.

    PubMed

    Thomas, Divyamol; Jeyathilakan, N; Abdul Basith, S; Senthilkumar, T M A

    2016-09-01

    Toxocara canis is a widespread gastrointestinal nematode parasite of dogs and cause Toxocara larva migrans, an important zoonotic disease in humans on ingestion of infective eggs. Toxocarosis is one of the few human parasitic diseases whose serodiagnosis uses a standardized antigen, T. canis excretory secretory antigen (TES). The present study describes collection of T. canis adult worm, collection and embryonation of T. canis eggs, hatching and separation of T. canis larvae, in vitro maintenance of T. canis second stage larvae for production of TES, concentration of culture fluid TES and yield of TES in correlation with various methods cited in literature.

  13. Transport and quality control of MHC class I molecules in the early secretory pathway.

    PubMed

    Springer, Sebastian

    2015-06-01

    Folding and peptide binding of major histocompatibility complex (MHC) class I molecules have been thoroughly researched, but the mechanistic connection between these biochemical events and the progress of class I through the early secretory pathway is much less well understood. This review focuses on the question how the partially assembled forms of class I (which lack high-affinity peptide and/or the light chain beta-2 microglobulin) are retained inside the cell. Such investigations offer researchers exciting chances to understand the connections between class I structure, conformational dynamics, peptide binding kinetics and thermodynamics, intracellular transport, and antigen presentation.

  14. Ataxia and secretory diarrhea: two unusual paraneoplastic syndromes occurring concurrently in the same patient with ganglioneuroblastoma.

    PubMed

    Gesundheit, Benjamin; Smith, Charles R; Gerstle, J Ted; Weitzman, Sheila S; Chan, Helen S L

    2004-09-01

    The presence of rare paraneoplastic syndromes, the opsoclonus-myoclonus-ataxia syndrome (OMA), presumably caused by antineuronal antibody production, and diarrhea, caused by vasoactive intestinal peptide (VIP) secreted by neuroblastoma, may strongly signal the presence of neuroblastoma. The authors describe a child who presented with both syndromes concurrently; this has never been described previously in the same patient. However, diagnosis of neuroblastoma was delayed by a workup focused on the prolonged diarrhea rather than the ataxia. The diarrhea resolved after tumor resection, whereas OMA required further therapy. Increased awareness of VIP-secretory diarrhea, especially in an ataxic child, might contribute to an earlier diagnosis of neuroblastoma.

  15. Clinical and biological role of secretory phospholipase A2 in acute respiratory distress syndrome infants

    PubMed Central

    2013-01-01

    Introduction Secretory phospholipase A2 is supposed to play a role in acute lung injury but no data are available for pediatric acute respiratory distress syndrome (ARDS). It is not clear which enzyme subtypes are secreted and what the relationships are between enzyme activity, biophysical and biochemical parameters, and clinical outcomes. We aimed to measure the enzyme and identify its subtypes and to study its biochemical and biophysical effect. The secondary aim was to correlate enzyme activity with clinical outcome. Methods Bronchoalveolar lavage was performed in 24 infants with ARDS and 14 controls with no lung disease. Samples were assayed for secretory phospholipase A2 and molecules related to its activity and expression. Western blotting and captive bubble surfactometry were also performed. Clinical data were real time downloaded. Results Tumor necrosis factor-α (814 (506-2,499) vs. 287 (111-1,315) pg/mL; P = 0.04), enzyme activity (430 (253-600) vs. 149 (61-387) IU/mL; P = 0.01), free fatty acids (4.3 (2.8-8.6) vs. 2 (0.8-4.6) mM; P = 0.026), and minimum surface tension (25.6 ± 6.1 vs. 18 ± 1.8 mN/m; P = 0.006) were higher in ARDS than in controls. Phospholipids are lower in ARDS than in controls (76.5 (54-100) vs. 1,094 (536-2,907) μg/mL; P = 0.0001). Three enzyme subtypes were identified (-IIA, -V, -X), although in lower quantities in controls; another subtype (-IB) was mainly detected in ARDS. Significant correlations exist between enzyme activity, free fatty acids (ρ = 0.823; P < 0.001), and surface tension (ρ = 0.55; P < 0.028). Correlations also exist with intensive care stay (ρ = 0.54; P = 0.001), PRISM-III24 (ρ = 0.79; P< 0.001), duration of ventilation (ρ = 0.53; P = 0.002), and oxygen therapy (ρ = 0.54; P = 0.001). Conclusions Secretory phospholipase A2 activity is raised in pediatric ARDS and constituted of four subtypes. Enzyme correlates with some inflammatory mediators, surface tension, and major clinical outcomes. Secretory

  16. Secretory leukocyte protease inhibitor (SLPI) might contaminate murine monoclonal antibodies after purification on protein G.

    PubMed

    Schenk, Jörg A; Fettke, Joerg; Lenz, Christine; Albers, Katharina; Mallwitz, Frank; Gajovic-Eichelmann, Nenad; Ehrentreich-Förster, Eva; Kusch, Emely; Sellrie, Frank

    2012-03-31

    The large scale production of a monoclonal anti-progesterone antibody in serum free medium followed by affinity chromatography on protein G lead to a contamination of the antibody sample with a protein of about 14 kDa. This protein was identified by mass spectrometry as secretory leukocyte protease inhibitor (SLPI). This SLPI contamination lead to a failure of the fiber-optic based competitive fluorescence assay to detect progesterone in milk. Purification of the monoclonal antibody using protein A columns circumvented this problem.

  17. Abnormal ion content, hydration and granule expansion of the secretory granules from cystic fibrosis airway glandular cells

    SciTech Connect

    Baconnais, S.; Delavoie, F. |; Zahm, J.M.; Milliot, M.; Castillon, N.; Terryn, C.; Banchet, V.; Michel, J.; Danos, O.; Merten, M.; Chinet, T.; Zierold, K.; Bonnet, N.; Puchelle, E. , E-Mail: edith.puchelle@univ-reims.fr; Balossier, G.

    2005-10-01

    The absence or decreased expression of cystic fibrosis transmembrane conductance regulator (CFTR) induces increased Na{sup +} absorption and hyperabsorption of the airway surface liquid (ASL) resulting in a dehydrated and hyperviscous ASL. Although the implication of abnormal airway submucosal gland function has been suggested, the ion and water content in the Cystic Fibrosis (CF) glandular secretory granules, before exocytosis, is unknown. We analyzed, in non-CF and CF human airway glandular cell lines (MM-39 and KM4, respectively), the ion content in the secretory granules by electron probe X-ray microanalysis and the water content by quantitative dark field imaging on freeze-dried cryosections. We demonstrated that the ion content (Na{sup +}, Mg{sup 2+}, P, S and Cl{sup -}) is significantly higher and the water content significantly lower in secretory granules from the CF cell line compared to the non-CF cell line. Using videomicroscopy, we observed that the secretory granule expansion was deficient in CF glandular cells. Transfection of CF cells with CFTR cDNA or inhibition of non-CF cells with CFTR{sub inh}-172, respectively restored or decreased the water content and granule expansion, in parallel with changes in ion content. We hypothesize that the decreased water and increased ion content in glandular secretory granules may contribute to the dehydration and increased viscosity of the ASL in CF.

  18. Accumulation of Major Histocompatibility Complex Class II Molecules in Mast Cell Secretory Granules and Their Release upon Degranulation

    PubMed Central

    Raposo, Graça; Tenza, Danielle; Mecheri, Salahedine; Peronet, Roger; Bonnerot, Christian; Desaymard, Catherine

    1997-01-01

    To investigate the relationship between major histocompatibility complex (MHC) class II compartments, secretory granules, and secretory lysosomes, we analyzed the localization and fate of MHC class II molecules in mast cells. In bone marrow-derived mast cells, the bulk of MHC class II molecules is contained in two distinct compartments, with features of both lysosomal compartments and secretory granules defined by their protein content and their accessibility to endocytic tracers. Type I granules display internal membrane vesicles and are accessed by exogenous molecules after a time lag of 20 min; type II granules are reached by the endocytic tracer later and possess a serotonin-rich electron-dense core surrounded by a multivesicular domain. In these type I and type II granules, MHC class II molecules, mannose-6-phosphate receptors and lysosomal membrane proteins (lamp1 and lamp2) localize to small intralumenal vesicles. These 60–80-nm vesicles are released along with inflammatory mediators during mast cell degranulation triggered by IgE-antigen complexes. These observations emphasize the intimate connection between the endocytic and secretory pathways in cells of the hematopoietic lineage which allows regulated secretion of the contents of secretory lysosomes, including membrane proteins associated with small vesicles. PMID:9398681

  19. Accumulation of major histocompatibility complex class II molecules in mast cell secretory granules and their release upon degranulation.

    PubMed

    Raposo, G; Tenza, D; Mecheri, S; Peronet, R; Bonnerot, C; Desaymard, C

    1997-12-01

    To investigate the relationship between major histocompatibility complex (MHC) class II compartments, secretory granules, and secretory lysosomes, we analyzed the localization and fate of MHC class II molecules in mast cells. In bone marrow-derived mast cells, the bulk of MHC class II molecules is contained in two distinct compartments, with features of both lysosomal compartments and secretory granules defined by their protein content and their accessibility to endocytic tracers. Type I granules display internal membrane vesicles and are accessed by exogenous molecules after a time lag of 20 min; type II granules are reached by the endocytic tracer later and possess a serotonin-rich electron-dense core surrounded by a multivesicular domain. In these type I and type II granules, MHC class II molecules, mannose-6-phosphate receptors and lysosomal membrane proteins (lamp1 and lamp2) localize to small intralumenal vesicles. These 60-80-nm vesicles are released along with inflammatory mediators during mast cell degranulation triggered by IgE-antigen complexes. These observations emphasize the intimate connection between the endocytic and secretory pathways in cells of the hematopoietic lineage which allows regulated secretion of the contents of secretory lysosomes, including membrane proteins associated with small vesicles.

  20. Kinesin-related Smy1 enhances the Rab-dependent association of myosin-V with secretory cargo

    PubMed Central

    Lwin, Kyaw Myo; Li, Donghao; Bretscher, Anthony

    2016-01-01

    The mechanisms by which molecular motors associate with specific cargo is a central problem in cell organization. The kinesin-like protein Smy1 of budding yeast was originally identified by the ability of elevated levels to suppress a conditional myosin-V mutation (myo2-66), but its function with Myo2 remained mysterious. Subsequently, Myo2 was found to provide an essential role in delivery of secretory vesicles for polarized growth and in the transport of mitochondria for segregation. By isolating and characterizing myo2 smy1 conditional mutants, we uncover the molecular function of Smy1 as a factor that enhances the association of Myo2 with its receptor, the Rab Sec4, on secretory vesicles. The tail of Smy1—which binds Myo2—its central dimerization domain, and its kinesin-like head domain are all necessary for this function. Consistent with this model, overexpression of full-length Smy1 enhances the number of Sec4 receptors and Myo2 motors per transporting secretory vesicle. Rab proteins Sec4 and Ypt11, receptors for essential transport of secretory vesicles and mitochondria, respectively, bind the same region on Myo2, yet Smy1 functions selectively in the transport of secretory vesicles. Thus a kinesin-related protein can function intimately with a myosin-V and its receptor in the transport of a specific cargo. PMID:27307583

  1. Overproduction of a Model Sec- and Tat-Dependent Secretory Protein Elicits Different Cellular Responses in Streptomyces lividans.

    PubMed

    Gullón, Sonia; Marín, Silvia; Mellado, Rafael P

    2015-01-01

    Streptomyces lividans is considered an efficient host for the secretory production of homologous and heterologous proteins. To identify possible bottlenecks in the protein production process, a comparative transcriptomic approach was adopted to study cellular responses during the overproduction of a Sec-dependent model protein (alpha-amylase) and a Tat-dependent model protein (agarase) in Streptomyces lividans. The overproduction of the model secretory proteins via the Sec or the Tat route in S. lividans does elicit a different major cell response in the bacterium. The stringent response is a bacterial response to nutrients' depletion, which naturally occurs at late times of the bacterial cell growth. While the induction of the stringent response at the exponential phase of growth may limit overall productivity in the case of the Tat route, the induction of that response does not take place in the case of the Sec route, which comparatively is an advantage in secretory protein production processes. Hence, this study identifies a potential major drawback in the secretory protein production process depending on the secretory route, and provides clues to improving S. lividans as a protein production host.

  2. Overproduction of a Model Sec- and Tat-Dependent Secretory Protein Elicits Different Cellular Responses in Streptomyces lividans

    PubMed Central

    Gullón, Sonia; Marín, Silvia; Mellado, Rafael P.

    2015-01-01

    Streptomyces lividans is considered an efficient host for the secretory production of homologous and heterologous proteins. To identify possible bottlenecks in the protein production process, a comparative transcriptomic approach was adopted to study cellular responses during the overproduction of a Sec-dependent model protein (alpha-amylase) and a Tat-dependent model protein (agarase) in Streptomyces lividans. The overproduction of the model secretory proteins via the Sec or the Tat route in S. lividans does elicit a different major cell response in the bacterium. The stringent response is a bacterial response to nutrients’ depletion, which naturally occurs at late times of the bacterial cell growth. While the induction of the stringent response at the exponential phase of growth may limit overall productivity in the case of the Tat route, the induction of that response does not take place in the case of the Sec route, which comparatively is an advantage in secretory protein production processes. Hence, this study identifies a potential major drawback in the secretory protein production process depending on the secretory route, and provides clues to improving S. lividans as a protein production host. PMID:26200356

  3. A novel imaging method for quantitative Golgi localization reveals differential intra-Golgi trafficking of secretory cargoes

    PubMed Central

    Tie, Hieng Chiong; Mahajan, Divyanshu; Chen, Bing; Cheng, Li; VanDongen, Antonius M. J.; Lu, Lei

    2016-01-01

    Cellular functions of the Golgi are determined by the unique distribution of its resident proteins. Currently, electron microscopy is required for the localization of a Golgi protein at the sub-Golgi level. We developed a quantitative sub-Golgi localization method based on centers of fluorescence masses of nocodazole-induced Golgi ministacks under conventional optical microscopy. Our method is rapid, convenient, and quantitative, and it yields a practical localization resolution of ∼30 nm. The method was validated by the previous electron microscopy data. We quantitatively studied the intra-Golgi trafficking of synchronized secretory membrane cargoes and directly demonstrated the cisternal progression of cargoes from the cis- to the trans-Golgi. Our data suggest that the constitutive efflux of secretory cargoes could be restricted at the Golgi stack, and the entry of the trans-Golgi network in secretory pathway could be signal dependent. PMID:26764092

  4. Morphological and morphometric study of atrial specific granules and other secretory components in dogs experimentally infected with Trypanosoma cruzi.

    PubMed Central

    Caliari, M. V.; Lana, M.; Leite, V. H.; Tafuri, W. L.

    1995-01-01

    Changes in blood volume can induce morphometric and morphological alterations in the secretory complex of the myoendocrine cells due to the stretching of atrial walls. These alterations were studied by electron microscopy, using dogs infected intraperitonially with Trypanosoma cruzi and necropsied during the acute phase of the infection when congestive heart failure was present. Several changes were observed in the myoendocrine cells of the heart: hypertrophy and hyperplasia of rough endoplasmic reticulum and Golgi complex, increase in telenuclear secretory complex, increase in fusion of type B atrial specific granules (ASG), decrease of the total number of ASG, enlargement of the maximum diameter of type A ASG and a relative increase in the number of type B ASG. These alterations suggest a larger secretory activity of the atrial myoendocrine cells with a larger secretion of atrial natriuretic peptide (ANP). Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 PMID:7547444

  5. Morphology and histology of secretory setae in terrestrial larvae of biting midges of the genus Forcipomyia (Diptera: Ceratopogonidae).

    PubMed

    Urbanek, Aleksandra; Richert, Malwina; Giłka, Wojciech; Szadziewski, Ryszard

    2011-11-01

    Apneustic larvae of the genus Forcipomyia possess unique secretory setae located on the dorsal surface along the body in two rows, one pair on each thoracic and abdominal segment and two pairs on the head. Morphological and histological studies of secretory setae in fourth instar larvae of Forcipomyia nigra (Winnertz) and Forcipomyia nigrans Remm indicate they are modified mechanoreceptors (sensilla trichodea) in which the trichogen cell is a glandular cell producing a hygroscopic secretion. The cytoplasm of the glandular trichogen cell fills the lumen of a secretory seta, which shows one or more pores on the apex. The cytoplasm contains numerous microtubules responsible for transportation of proteinaceous vesicles, and an extremely large polyploid nucleus typical of gland cells. The main role of the hygroscopic secretion is to moist the body and thus facilitate cuticular respiration.

  6. The novel secretory protein CGREF1 inhibits the activation of AP-1 transcriptional activity and cell proliferation.

    PubMed

    Deng, Weiwei; Wang, Lan; Xiong, Ying; Li, Jing; Wang, Ying; Shi, Taiping; Ma, Dalong

    2015-08-01

    The transcription factor AP-1 plays an important role in inflammation and cell survival. Using a dual-luciferase reporter assay system and a library of 940 candidate human secretory protein cDNA clones, we identified that CGREF1 can inhibit the transcriptional activity of AP-1. We demonstrated that CGREF1 is secreted via the classical secretory pathway through the ER-to-Golgi apparatus. Functional investigations revealed that overexpression of CGREF1 can significantly inhibit the phosphorylation of ERK and p38 MAPK, and suppress the proliferation of HEK293T and HCT116 cells. Conversely, specific siRNAs against CGREF1 can increase the transcriptional activity of AP-1. These results clearly indicated that CGREF1 is a novel secretory protein, and plays an important role in regulation of AP-1 transcriptional activity and cell proliferation.

  7. Amyloid formation of growth hormone in presence of zinc: Relevance to its storage in secretory granules

    PubMed Central

    Jacob, Reeba S.; Das, Subhadeep; Ghosh, Saikat; Anoop, Arunagiri; Jha, Narendra Nath; Khan, Tuhin; Singru, Praful; Kumar, Ashutosh; Maji, Samir K.

    2016-01-01

    Amyloids are cross-β-sheet fibrillar aggregates, associated with various human diseases and native functions such as protein/peptide hormone storage inside secretory granules of neuroendocrine cells. In the current study, using amyloid detecting agents, we show that growth hormone (GH) could be stored as amyloid in the pituitary of rat. Moreover, to demonstrate the formation of GH amyloid in vitro, we studied various conditions (solvents, glycosaminoglycans, salts and metal ions) and found that in presence of zinc metal ions (Zn(II)), GH formed short curvy fibrils. The amyloidogenic nature of these fibrils was examined by Thioflavin T binding, Congo Red binding, transmission electron microscopy and X-ray diffraction. Our biophysical studies also suggest that Zn(II) initiates the early oligomerization of GH that eventually facilitates the fibrillation process. Furthermore, using immunofluorescence study of pituitary tissue, we show that GH in pituitary significantly co-localizes with Zn(II), suggesting the probable role of zinc in GH aggregation within secretory granules. We also found that GH amyloid formed in vitro is capable of releasing monomers. The study will help to understand the possible mechanism of GH storage, its regulation and monomer release from the somatotrophs of anterior pituitary. PMID:27004850

  8. A parafusin-related Toxoplasma protein in Ca2+-regulated secretory organelles.

    PubMed

    Matthiesen, S H; Shenoy, S M; Kim, K; Singer, R H; Satir, B H

    2001-12-01

    We cloned a gene, PRPI, of Toxoplasma gondii encoding a 637-amino-acids protein having a calculated mass of 70 kDa. The sequence showed high homology to parafusin, a protein that in Paramecium tetraurelia participates in Ca2+-regulated exocytosis and is a paralog of phosphoglucomutase. We show that Toxoplasma gondii homogenate and an expressed recombinant PRP1 fusion protein cross-react with a specific peptide-derived antibody to parafusin in Western blots. Antibodies to the recombinant PRP1 showed cross-reaction with parafusin and recognized PRP1, as bands at M, 63 x 10(3) and 68 x 10(3), respectively. PRP1 is labeled when Toxoplasma gondii cells are incubated with inorganic 32P and appears as the major band on autoradiograms of SDS-PAGE gels. The localization of PRP1 was examined in secretory organelles of Toxoplasma gondii by deconvolution light microscopy followed by three dimensional reconstruction using pairwise combinations of specific antibodies. PRP1 localized to the apical third of the cell. It co-localized with micronemes, the only secretory organelle the secretion of which is Ca2+ dependent. Quantification of the co-localized stain suggests that only mature micronemes ready for exocytosis have PRP1. These findings suggest that PRP1, parafusin and other members of the phosphoglucomutase superfamily have a conserved role in Ca2+-regulated exocytic processes.

  9. BACE2 is stored in secretory granules of mouse and rat pancreatic beta cells.

    PubMed

    Finzi, Giovanna; Franzi, Francesca; Placidi, Claudia; Acquati, Francesco; Palumbo, Elisa; Russo, Antonella; Taramelli, Roberto; Sessa, Fausto; La Rosa, Stefano

    2008-01-01

    BACE2 is a protease homologous to BACE1 protein, an enzyme involved in the amyloid formation of Alzheimer disease (AD). However, despite the high homology between these two proteins, the biological role of BACE2 is still controversial, even though a few studies have suggested a pathogenetic role in sporadic inclusion-body myositis and hereditary inclusion-body myopathy, which are characterized by vacuolization of muscular fibers with intracellular deposits of proteins similar to those found in the brain of AD patients. Although BACE2 has also been identified in the pancreas, its function remains unknown and its specific localization in different pancreatic cell types has not been definitively ascertained. For these reasons, the authors have investigated the cellular and subcellular localization of BACE2 in normal rodent pancreases. BACE2 immunoreactivity was found in secretory granules of beta cells, co-stored with insulin and IAPP, while it was lacking in the other endocrine and exocrine cell types. The presence of BACE2 in secretory granules of beta cells suggests that it may play a role in diabetes-associated amyloidogenesis.

  10. Flow cytometry-assisted purification and proteomic analysis of the corticotropes dense-core secretory granules.

    PubMed

    Gauthier, Daniel J; Sobota, Jacqueline A; Ferraro, Francesco; Mains, Richard E; Lazure, Claude

    2008-09-01

    The field of organellar proteomics has emerged as an attempt to minimize the complexity of the proteomics data obtained from whole cell and tissue extracts while maximizing the resolution on the protein composition of a single subcellular compartment. Standard methods involve lengthy density-based gradient and/or immunoaffinity purification steps followed by extraction, 1-DE or 2-DE, gel staining, in-gel tryptic digestion, and protein identification by MS. In this paper, we present an alternate approach to purify subcellular organelles containing a fluorescent reporter molecule. The gel-free procedure involves fluorescence-assisted sorting of the secretory granules followed by gentle extraction in a buffer compatible with tryptic digestion and MS. Once the subcellular organelle labeled, this procedure can be done in a single day, requires no major modification to any instrumentation and can be readily adapted to the study of other organelles. When applied to corticotrope secretory granules, it led to a much enriched granular fraction from which numerous proteins could be identified through MS.

  11. Platelet secretory behaviour: as diverse as the granules … or not?

    PubMed

    Heijnen, H; van der Sluijs, P

    2015-12-01

    Platelets play a central role in the arrest of bleeding after damage to a blood vessel and in the development of thrombosis. Platelets rapidly respond after interaction with sub-endothelial components and release cargo from their storage granules. The three principal granule types of platelets are α-granules, dense granules and lysosomes. Timed release of granule contents and regulated expression of critical receptors are essential for maintenance of the platelet thrombus, yet also have important functions beyond hemostasis (i.e. inflammatory reactions and immune responses). α-granules store adhesive molecules such as von Willebrand factor and fibrinogen, growth factors and inflammatory and angiogenic mediators, which play crucial roles in inflammatory responses and tumor genesis. The α-granules comprise a group of subcellular compartments with a unique composition and ultrastructure. Recent studies have suggested that differential secretory kinetics of α-granule subtypes is responsible for a thematic release of adhesive and inflammatory mediators. In addition, new results indicate that activation-dependent synthesis and release of cytokines also contribute to the inflammatory role of platelets. We will discuss the various methods that platelets use to regulate secretory processes and how these relate to potential differential secretion patterns, thereby promoting adhesiveness and/or inflammatory functions. We will focus on the heterogenic granule population, open canalicular system (OCS) plasticity, the role of contractile and mechanobiological forces, and the fusogenic machinery.

  12. Identification and Characterization of an Escorter for Two Secretory Adhesins in Toxoplasma gondii

    PubMed Central

    Reiss, Matthias; Viebig, Nicola; Brecht, Susan; Fourmaux, Marie-Noelle; Soete, Martine; Di Cristina, Manlio; Dubremetz, Jean François; Soldati, Dominique

    2001-01-01

    The intracellular protozoan parasite Toxoplasma gondii shares with other members of the Apicomplexa a common set of apical structures involved in host cell invasion. Micronemes are apical secretory organelles releasing their contents upon contact with host cells. We have identified a transmembrane micronemal protein MIC6, which functions as an escorter for the accurate targeting of two soluble proteins MIC1 and MIC4 to the micronemes. Disruption of MIC1, MIC4, and MIC6 genes allowed us to precisely dissect their contribution in sorting processes. We have mapped domains on these proteins that determine complex formation and targeting to the organelle. MIC6 carries a sorting signal(s) in its cytoplasmic tail whereas its association with MIC1 involves a lumenal EGF-like domain. MIC4 binds directly to MIC1 and behaves as a passive cargo molecule. In contrast, MIC1 is linked to a quality control system and is absolutely required for the complex to leave the early compartments of the secretory pathway. MIC1 and MIC4 bind to host cells, and the existence of such a complex provides a plausible mechanism explaining how soluble adhesins act. We hypothesize that during invasion, MIC6 along with adhesins establishes a bridge between the host cell and the parasite. PMID:11157983

  13. Identification of the Moving Junction Complex of Toxoplasma gondii: A Collaboration between Distinct Secretory Organelles

    PubMed Central

    2005-01-01

    Apicomplexan parasites, including Toxoplasma gondii and Plasmodium sp., are obligate intracellular protozoa. They enter into a host cell by attaching to and then creating an invagination in the host cell plasma membrane. Contact between parasite and host plasma membranes occurs in the form of a ring-shaped moving junction that begins at the anterior end of the parasite and then migrates posteriorly. The resulting invagination of host plasma membrane creates a parasitophorous vacuole that completely envelops the now intracellular parasite. At the start of this process, apical membrane antigen 1 (AMA1) is released onto the parasite surface from specialized secretory organelles called micronemes. The T. gondii version of this protein, TgAMA1, has been shown to be essential for invasion but its exact role has not previously been determined. We identify here a trio of proteins that associate with TgAMA1, at least one of which associates with TgAMA1 at the moving junction. Surprisingly, these new proteins derive not from micronemes, but from the anterior secretory organelles known as rhoptries and specifically, for at least two, from the neck portion of these club-shaped structures. Homologues for these AMA1-associated proteins are found throughout the Apicomplexa strongly suggesting that this moving junction apparatus is a conserved feature of this important class of parasites. Differences between the contributing proteins in different species may, in part, be the result of selective pressure from the different niches occupied by these parasites. PMID:16244709

  14. Characterization of excretory-secretory antigens of adult Toxocara canis by western blotting.

    PubMed

    Sudhakar, N R; Samanta, S; Sahu, Shivani; Raina, O K; Gupta, S C; Goswami, T K; Lokesh, K M; Kumar, Ashok

    2014-06-01

    Toxocara canis is one of the most common helminth worm of dogs which continues to stimulate both public health concern alongside the higher scientific interest. It may cause visceral and ocular damage in humans especially in children. The identification of specific antigens of T. canis is important so as to develop better diagnostic techniques. Excretory-secretory (ES) antigens were prepared by culturing the adult T. canis worms in RPMI 1640 medium without serum supplementation followed by ammonium sulphate precipitation. These antigens were separated using sodium dodecyl sulphate-electrophoresis (SDS-PAGE). Recovered proteins ranged from 30 to 384 kDa. The specific reactivity of the T. canis excretory-secretory (TC-ES) proteins was checked by western blotting. The immuno-reactivity of the naturally infected dog sera with the TC-ES antigens showed five bands at 43, 57,105, 139 and 175 kDa. The immuno-reactivity of the hyper immune serum raised in rabbits against TC-ES antigens was observed with ten polypeptides of 21, 25, 30, 37, 45, 50, 57, 69, 77 and 105 kDa. Common antigens band were observed at 57 and 105 KDa. These antigens merit further evaluation as candidate for use in diagnosis of toxocariasis in humans and adult dogs.

  15. Sphingomyelin is sorted at the trans Golgi network into a distinct class of secretory vesicle

    PubMed Central

    Deng, Yongqiang; Rivera-Molina, Felix E.; Toomre, Derek K.; Burd, Christopher G.

    2016-01-01

    One of the principal functions of the trans Golgi network (TGN) is the sorting of proteins into distinct vesicular transport carriers that mediate secretion and interorganelle trafficking. Are lipids also sorted into distinct TGN-derived carriers? The Golgi is the principal site of the synthesis of sphingomyelin (SM), an abundant sphingolipid that is transported. To address the specificity of SM transport to the plasma membrane, we engineered a natural SM-binding pore-forming toxin, equinatoxin II (Eqt), into a nontoxic reporter termed Eqt-SM and used it to monitor intracellular trafficking of SM. Using quantitative live cell imaging, we found that Eqt-SM is enriched in a subset of TGN-derived secretory vesicles that are also enriched in a glycophosphatidylinositol-anchored protein. In contrast, an integral membrane secretory protein (CD8α) is not enriched in these carriers. Our results demonstrate the sorting of native SM at the TGN and its transport to the plasma membrane by specific carriers. PMID:27247384

  16. Engineering Yarrowia lipolytica to express secretory invertase with strong FBA1IN promoter.

    PubMed

    Hong, Seung-Pyo; Seip, John; Walters-Pollak, Dana; Rupert, Ross; Jackson, Raymond; Xue, Zhixiong; Zhu, Quinn

    2012-02-01

    Oleaginous yeast Yarrowia lipolytica is an important host for the production of lipid-derived compounds or heterologous proteins. Selection of strong promoters and effective expression systems is critical for heterologous protein secretion. To search for a strong promoter in Y. lipolytica, activities of FBA1, TDH1 and GPM1 promoters were compared to that of TEF1 promoter by constructing GUS reporter fusions. The FBA1 promoter activity was 2.2 and 5.5 times stronger than the TDH1 and GPM1 promoters, respectively. The FBA1IN promoter (FBA1 sequence of -826 to +169) containing an intron (+64 to +165) showed five-fold higher expression than the FBA1 promoter (-831 to -1). The transcriptional enhancement by the 5'-region within the FBA1 gene was confirmed by GPM1::FBA1 chimeric promoter construction. Using the strong FBA1IN promoter, four different S. cerevisiae SUC2 expression cassettes were tested for the SUC+ phenotype in Y. lipolytica. Functional invertase secretion was facilitated by the Xpr2 prepro-region with an additional 13 amino acids of mature Xpr2, or by the native Suc2 signal sequence. However, these two secretory signals in tandem, or the mature Suc2 with no secretory signal, did not direct secretion of functional invertase. Unlike previously reported Y. lipolytica SUC+ strains, our engineered stains secreted most of invertase into the medium.

  17. A micromethod for the assay of cellular secretory physiology: Application to rabbit parietal cells

    SciTech Connect

    Adrian, T.E.; Goldenring, J.R.; Oddsdottir, M.; Zdon, M.J.; Zucker, K.A.; Lewis, J.J.; Modlin, I.M. )

    1989-11-01

    A micromethod for investigating secretory physiology in isolated cells was evaluated. The method utilized a specially designed polycarbonate incubation chamber to provide constant oxygenation to cells incubating in a 96-well microtiter plate. Cells were rapidly separated from media by vacuum filtration. Isolated parietal cells were utilized to demonstrate the versatility of the method for assay of intracellular accumulation of ({sup 14}C)-aminopyrine, secretion of intrinsic factor into the medium, and assay of intracellular cAMP. Histamine stimulated the uptake of ({sup 14}C)aminopyrine and intrinsic factor secretion in a sustained and linear fashion. At the end of the 2-h period uptake of aminopyrine and secretion of intrinsic factor were increased 17- and 5-fold, respectively. This response to histamine was accompanied by a rapid and sustained 3-fold rise in intracellular cyclic AMP. In contrast, carbamylcholine caused a transient increase in ({sup 14}C)aminopyrine accumulation and intrinsic factor secretion which was most pronounced during the first 10 min and had almost ceased by 30 min. Carbamylcholine had no effect on intracellular cAMP levels. This new method, which can handle 400 replicates using parietal cells from the fundic mucosa of a single rabbit, is suitable for studying the time course of intracellular events which accompany general secretory processes.

  18. Expression of transcripts for cysteine-rich secretory proteins (CRISPs) in the murine lacrimal gland.

    PubMed

    Haendler, B; Toda, I; Sullivan, D A; Schleuning, W D

    1999-03-01

    Cysteine-rich secretory proteins (CRISPs) represent a family of evolutionarily conserved proteins which may play a role in the innate immune system and are transcriptionally regulated by androgens in several tissues. Transcripts for all three members of the CRISP family have now been identified in the murine lacrimal gland. RT-PCR using primers able to discriminate between the related CRISP forms allowed the amplification of fragments with the expected length. DNA sequencing revealed a complete identity with the hitherto characterized epididymal CRISP-1, testicular CRISP-2, and salivary gland CRISP-3. An analysis of several mouse strains indicated that all expressed the three CRISP forms, but in differing amounts. RT-PCR analysis of RNA isolated from acinar cells of lacrimal glands revealed that they expressed CRISP-1 and CRISP-2. Semiquantitative and quantitative analyses furthermore showed higher CRISP-1 and CRISP-3 mRNA levels in the lacrimal glands of male BALB/c and NOD mice when compared to females. Testosterone treatment of C3H/HeJ female mice was followed by an upregulation of the steady-state CRISP-1 but not CRISP-2 transcript levels. A comparable stimulation was observed for the mRNAs coding for parotid secretory protein (PSP), a factor previously shown to exhibit sexual dimorphism in the murine lacrimal gland. The expression of CRISP transcripts in the lacrimal gland is consistent with a function in the innate immune system.

  19. Exocytosis of Alphaherpesvirus Virions, Light Particles, and Glycoproteins Uses Constitutive Secretory Mechanisms

    PubMed Central

    Hogue, Ian B.; Scherer, Julian

    2016-01-01

    ABSTRACT Many molecular and cell biological details of the alphaherpesvirus assembly and egress pathway remain unclear. Recently we developed a live-cell fluorescence microscopy assay of pseudorabies virus (PRV) exocytosis, based on total internal reflection fluorescence (TIRF) microscopy and a virus-encoded pH-sensitive fluorescent probe. Here, we use this assay to distinguish three classes of viral exocytosis in a nonpolarized cell type: (i) trafficking of viral glycoproteins to the plasma membrane, (ii) exocytosis of viral light particles, and (iii) exocytosis of virions. We find that viral glycoproteins traffic to the cell surface in association with constitutive secretory Rab GTPases and exhibit free diffusion into the plasma membrane after exocytosis. Similarly, both virions and light particles use these same constitutive secretory mechanisms for egress from infected cells. Furthermore, we show that viral light particles are distinct from cellular exosomes. Together, these observations shed light on viral glycoprotein trafficking steps that precede virus particle assembly and reinforce the idea that virions and light particles share a biogenesis and trafficking pathway. PMID:27273828

  20. Matrix-dependent local retention of secretory vesicle cargo in cortical neurons.

    PubMed

    de Wit, Joris; Toonen, Ruud F; Verhage, Matthijs

    2009-01-07

    Neurons secrete many diffusible signals from synaptic and other secretory vesicles. We characterized secretion of guidance cues, neuropeptides, neurotrophins, and proteases from single secretory vesicles using pHluorin-tagged cargo in cortical neurons. Stimulation triggered transient and persistent fusion events. Transient events represented full release followed by cargo diffusion or incomplete release followed by vesicle retrieval, as previously observed in neuroendocrine cells. Unexpectedly, we also observed that certain cargo, such as Semaphorin 3A (Sema3A), was delivered at the cell surface as stable deposits. Stable deposits and transient events were observed for single cargo and both were SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) and calcium dependent. The ratio between stable and transient events did not depend on cargo size, subcellular localization (synaptic vs extrasynaptic secretion), or the presence of the extracellular matrix. Instead, the ratio is cargo specific and depends on an interaction with the vesicle matrix through a basic domain in the cargo protein. Inhibition of this interaction through deletion of the basic domain in Sema3A abolished stable deposits and rendered all events transient. Strikingly, cargo favoring transient release was stably deposited after corelease with cargo favoring stable deposit. These data argue against cargo diffusion after exocytosis as a general principle. Instead, the vesicle matrix retains secreted signals, probably for focal signaling at the cell surface.

  1. Primary Cutaneous Mammary Analog Secretory Carcinoma With ETV6-NTRK3 Translocation.

    PubMed

    Amin, Sapna M; Beattie, Adam; Ling, Xia; Jennings, Lawrence J; Guitart, Joan

    2016-11-01

    Mammary analog secretory carcinoma (MASC) is a recently described tumor of the salivary glands named for its morphological and molecular similarity to secretory carcinoma of the breast. Many primary carcinomas arising from the adnexal glands also share similar morphology to those arising from the breast. Brandt et al first described primary cutaneous MASC in 2009 and since then only 2 other cases have been reported. Herein, we describe a long-standing mass on the arm of an otherwise healthy 40-year-old female. Histologic examination revealed a circumscribed but unencapsulated, nodular tumor composed of bland epithelial cells arranged in solid and microcystic growth patterns. The cells showed vacuolated cytoplasm and round to oval nuclei with vesicular chromatin. Intraluminal homogenous eosinophilic secretions were present. Mitotic figures were not identified. The tumor cells stained positive for CK8/18, CK7, and S100 but were negative for other markers performed, including estrogen receptor, progesterone receptor, HER2/neu, paired box 8 (PAX8), and thyroid transcription factor 1 (TTF1). As the patient clinically had no other masses or known carcinomas, a diagnosis of primary cutaneous MASC was rendered. The ETV6-NTRK3 fusion transcript was subsequently detected by reverse transcriptase polymerase chain reaction amplification, further supporting the diagnosis. We present this case to review the histologic features of MASC and highlight the importance of recognizing this lesion not only as a possible cutaneous metastasis but also as a primary cutaneous tumor.

  2. Non Secretory Multiple Myeloma With Extensive Extramedullary Plasmacytoma: A Diagnostic Dilemma

    PubMed Central

    Low, Soo Fin; Mohd Tap, Nor Hanani; Kew, Thean Yean; Ngiu, Chai Soon; Sridharan, Radhika

    2015-01-01

    Multiple myeloma (MM) is characterized by progressive proliferation of malignant plasma cells, usually initiating in the bone marrow. MM can affect any organ; a total of 7 - 18% of patients with MM demonstrate extramedullary involvement at diagnosis. Non-secretory multiple myeloma (NSMM) is a rare variant that accounts for 1 - 5% of all cases of multiple myeloma. The disease is characterized by the absence of monoclonal gammopathy in serum and urine electrophoresis. Our case report highlights the diagnostic challenge of a case of NSMM with extensive extramedullary involvement in a young female patient who initially presented with right shoulder pain and bilateral breasts lumps. Skeletal survey showed multiple lytic bony lesions. The initial diagnosis was primary breast carcinoma with osseous metastases. No monoclonal gammopathy was found in the serum or urine electrophoresis. Bone marrow and breast biopsies revealed marked plasmacytosis. The diagnosis was delayed for a month in view of the lack of clinical suspicion of multiple myeloma in a young patient and scant biochemical expression of non-secretory type of multiple myeloma. PMID:26528383

  3. Enhanced and Secretory Expression of Human Granulocyte Colony Stimulating Factor by Bacillus subtilis SCK6

    PubMed Central

    Bashir, Shaista; Sadaf, Saima; Ahmad, Sajjad; Akhtar, Muhammad Waheed

    2015-01-01

    This study describes a simplified approach for enhanced expression and secretion of a pharmaceutically important human cytokine, that is, granulocyte colony stimulating factor (GCSF), in the culture supernatant of Bacillus subtilis SCK6 cells. Codon optimized GCSF and pNWPH vector containing SpymwC signal sequence were amplified by prolonged overlap extension PCR to generate multimeric plasmid DNA, which was used directly to transform B. subtilis SCK6 supercompetent cells. Expression of GCSF was monitored in the culture supernatant for 120 hours. The highest expression, which corresponded to 17% of the total secretory protein, was observed at 72 hours of growth. Following ammonium sulphate precipitation, GCSF was purified to near homogeneity by fast protein liquid chromatography on a QFF anion exchange column. Circular dichroism spectroscopic analysis showed that the secondary structure contents of the purified GCSF are similar to the commercially available GCSF. Biological activity, as revealed by the regeneration of neutrophils in mice treated with ifosfamine, was also similar to the commercial preparation of GCSF. This, to our knowledge, is the first study that reports secretory expression of human GCSF in B. subtilis SCK6 with final recovery of up to 96 mg/L of the culture supernatant, without involvement of any chemical inducer. PMID:26881203

  4. Localization of Oleuropeyl Glucose Esters and a Flavanone to Secretory Cavities of Myrtaceae

    PubMed Central

    Heskes, Allison M.; Goodger, Jason Q. D.; Tsegay, Sammi; Quach, Tim; Williams, Spencer J.; Woodrow, Ian E.

    2012-01-01

    We report the widespread occurrence of structurally diverse oleuropeyl glucose esters, including the new diester eucaglobulin B, localized specifically to the essential oil secretory cavities of myrtaceous species. Clear taxonomic patterns in the composition of cavity extracts within the genus Eucalyptus are shown with species from subgenus Symphyomyrtus dominated by oleuropeyl glucose esters and species from subgenus Eucalyptus dominated instead by the flavanone, pinocembrin. We also examined the intra-species occurrence of oleuropeyl glucose esters by quantifying the abundant constituents cuniloside B and froggattiside A in trees from two populations of Eucalyptus polybractea R.T. Baker. All trees contained both compounds, which were positively correlated with total essential oil concentration. This apparent ubiquity of oleuropeyl glucose esters at both intra- and inter-specific levels in Eucalyptus is indicative of important physiological or ecological functions. The significance of their prevalence and the sequestration of these esters and also pinocembrin to the extracellular domain of secretory cavities is discussed in light of their potential biological activities and our findings that they are spatially segregated to the exterior of cavity lumina. The localization of oleuropeyl glucose esters to a specific and isolatable tissue type has the potential to aid in future elucidation of function and biosynthesis. PMID:22911712

  5. Bacterial mimetics of endocrine secretory granules as immobilized in vivo depots for functional protein drugs

    PubMed Central

    Céspedes, María Virtudes; Fernández, Yolanda; Unzueta, Ugutz; Mendoza, Rosa; Seras-Franzoso, Joaquin; Sánchez-Chardi, Alejando; Álamo, Patricia; Toledo-Rubio, Verónica; Ferrer-Miralles, Neus; Vázquez, Esther; Schwartz, Simó; Abasolo, Ibane; Corchero, José Luis; Mangues, Ramon; Villaverde, Antonio

    2016-01-01

    In the human endocrine system many protein hormones including urotensin, glucagon, obestatin, bombesin and secretin, among others, are supplied from amyloidal secretory granules. These granules form part of the so called functional amyloids, which within the whole aggregome appear to be more abundant than formerly believed. Bacterial inclusion bodies (IBs) are non-toxic, nanostructured functional amyloids whose biological fabrication can be tailored to render materials with defined biophysical properties. Since under physiological conditions they steadily release their building block protein in a soluble and functional form, IBs are considered as mimetics of endocrine secretory granules. We have explored here if the in vivo implantation of functional IBs in a given tissue would represent a stable local source of functional protein. Upon intratumoral injection of bacterial IBs formed by a potent protein ligand of CXCR4 we have observed high stability and prevalence of the material in absence of toxicity, accompanied by apoptosis of CXCR4+ cells and tumor ablation. Then, the local immobilization of bacterial amyloids formed by therapeutic proteins in tumors or other tissues might represent a promising strategy for a sustained local delivery of protein drugs by mimicking the functional amyloidal architecture of the mammals’ endocrine system. PMID:27775083

  6. Mammary analogue secretory carcinoma: the first submandibular case reported including findings on fine needle aspiration cytology.

    PubMed

    Petersson, Fredrik; Lian, Derrick; Chau, Yuk Ping; Yan, Benedict

    2012-03-01

    We present the first case (male, 35 years old) of a mammary analogue secretory carcinoma occurring in a submandibular gland and document findings on fine needle aspiration cytology. On histology, the tumor displayed characteristic features: circumscribed nodules composed of bland, pink to light red neoplastic cells with low proliferative/mitotic activity arranged in tubular, vaguely cribriform, and microcystic structures containing Periodic acid Schiff-positive, diastase-resistant secretory material. Immunohistochemistry showed strong and diffuse positivity for cytokeratin 7, S100 protein, and vimentin, as well as moderate to strong immunoreactivity for c-kit in the majority of tumor cells. A rearrangement of the ETV6 gene on fluorescence in situ hybridization was documented. The patient underwent an ipsilateral selective (levels I-IV) neck dissection which showed metastasis in 3 out of 36 lymph nodes (levels 1-3). Adjuvant radiotherapy was administered. No local recurrence or metastatic disease has been detected during a follow up period of 28 months.

  7. Immunoprotection in sheep against Haemonchus contortus using its thiol-purified excretory/secretory proteins.

    PubMed

    Arunkumar, Selvarayar

    2012-01-01

    Excretory/Secretory antigen was prepared by culturing live adult worms of Haemonchus contortus in RPMI 1640 medium at a concentration of 50 worms per mL in a culture-flask at 37 ˚C for 24 hr and the culture supernatant was used as antigen. The E/S antigen was purified by thiol-sepharose affinity chromatography. On western blot analysis, it was demonstrated that thiol-purified antigen showed a single reactive band at 66 kDa. In immunization trial, sheep were administered intramuscularly with 500 µg of thiol-purified excretory/secretory antigen along with montanide as adjuvant on day 0, 30 and 60. On ELISA, it was observed that the mean absorbance values were significantly (p ≤ 0.01) higher up to 20 weeks post immunization in Group-I (purified antigen) compared to Group- II (unimmunized control). Further, the mean EPG values was lower in Group I (200.00 ± 40.82 to 400.00 ± 91.29) than Group II (2200.00 ± 108.01 to 5100.00 ± 169.56) and the percentage reduction in mean fecal egg counts was 88.50%. Similarly, the mean abomasal worm counts was lower in Group I (808.33 ± 78.29) than Group II (3280.00 ± 147.19) and the percentage reduction in mean abomasal worm count was 75.40%.

  8. Haemonchus contortus excretory and secretory proteins (HcESPs) suppress functions of goat PBMCs in vitro.

    PubMed

    Gadahi, Javaid Ali; Yongqian, Bu; Ehsan, Muhammad; Zhang, Zhen Chao; Wang, Shuai; Yan, Ruo Feng; Song, Xiao Kai; Xu, Li Xin; Li, Xiang Rui

    2016-06-14

    Excretory and secretory products (ESPs) of nematode contain various proteins which are capable of inducing the instigation or depression of the host immune response and are involved in the pathogenesis of the worms. In the present study, Haemonchus contortus excretory and secretory products (HcESPs) were collected from the adult worms. Binding of HcESPs to goat peripheral blood mononuclear cells (PBMCs) was confirmed by immune-fluorescence assay. Effects of the HcESPs on cytokine production, cell proliferation, cell migration and nitric oxide (NO) production of PBMCs were checked by co-incubation of HcESPs with goat PBMCs. The results indicated that the production of IL-4 and IFN-γ were significantly decreased by HcESPs in dose dependent manner. On the contrary, the production of IL-10 and IL-17 were increased. Cell migration was significantly enhanced by HcESPs, whereas, HcESPs treatment significantly suppressed the cell proliferation and NO production. These results indicated that the HcESPs played important suppressive regulatory roles on PBMCs and provided highlights to the understanding of the host-parasite interactions.

  9. Effects of photoperiod on secretory patterns of growth hormone in adult male goats.

    PubMed

    Jin, Jin; Sawai, Ken; Hashizume, Tsutomu

    2013-12-01

    The aim of the present study was to clarify the effect of photoperiod on secretory patterns of growth hormone (GH) in male goats. Adult male goats were kept at 20°C with an 8-h or 16-h light photoperiod, and secretory patterns of GH secretion were compared. In addition, plasma profiles of prolactin (PRL), insulin-like growth factor-I (IGF-I) and testosterone (T) were also examined to characterize GH secretion. GH was secreted in a pulsatile manner. There was no significant difference in pulse frequency between the 8-h and 16-h photoperiods. However, GH pulse amplitude tended to be greater in the group with the 16-h photoperiod (P = 0.1), and mean GH concentrations were significantly greater in the 16-h photoperiod (P < 0.05). The GH-releasing response to GH releasing hormone was greater in the 16-h than 8-h photoperiod (P < 0.05). Plasma PRL and IGF-I levels were higher in the 16-h than 8-h photoperiod (P < 0.05). In contrast, plasma T levels were lower in the 16-h photoperiod (P < 0.05). These results show that a long light photoperiod enhances the secretion of GH as well as PRL and IGF-I, but reduces plasma T concentrations in male goats.

  10. Neutral sphingomyelinase (SMPD3) deficiency disrupts the Golgi secretory pathway and causes growth inhibition

    PubMed Central

    Stoffel, Wilhelm; Hammels, Ina; Jenke, Bitta; Binczek, Erika; Schmidt-Soltau, Inga; Brodesser, Susanne; Schauss, Astrid; Etich, Julia; Heilig, Juliane; Zaucke, Frank

    2016-01-01

    Systemic loss of neutral sphingomyelinase (SMPD3) in mice leads to a novel form of systemic, juvenile hypoplasia (dwarfism). SMPD3 deficiency in mainly two growth regulating cell types contributes to the phenotype, in chondrocytes of skeletal growth zones to skeletal malformation and chondrodysplasia, and in hypothalamic neurosecretory neurons to systemic hypothalamus–pituitary–somatotropic hypoplasia. The unbiased smpd3−/− mouse mutant and derived smpd3−/− primary chondrocytes were instrumental in defining the enigmatic role underlying the systemic and cell autonomous role of SMPD3 in the Golgi compartment. Here we describe the unprecedented role of SMPD3. SMPD3 deficiency disrupts homeostasis of sphingomyelin (SM), ceramide (Cer) and diacylglycerol (DAG) in the Golgi SMPD3-SMS1 (SM-synthase1) cycle. Cer and DAG, two fusogenic intermediates, modify the membrane lipid bilayer for the initiation of vesicle formation and transport. Dysproteostasis, unfolded protein response, endoplasmic reticulum stress and apoptosis perturb the Golgi secretory pathway in the smpd3−/− mouse. Secretion of extracellular matrix proteins is arrested in chondrocytes and causes skeletal malformation and chondrodysplasia. Similarly, retarded secretion of proteo-hormones in hypothalamic neurosecretory neurons leads to hypothalamus induced combined pituitary hormone deficiency. SMPD3 in the regulation of the protein vesicular secretory pathway may become a diagnostic target in the etiology of unknown forms of juvenile growth and developmental inhibition. PMID:27882938

  11. Extracellular superoxide dismutase is present in secretory vesicles of human neutrophils and released upon stimulation.

    PubMed

    Iversen, Marie B; Gottfredsen, Randi H; Larsen, Ulrike G; Enghild, Jan J; Praetorius, Jeppe; Borregaard, Niels; Petersen, Steen V

    2016-08-01

    Extracellular superoxide dismutase (EC-SOD) is an antioxidant enzyme present in the extracellular matrix (ECM), where it provides protection against oxidative degradation of matrix constituents including type I collagen and hyaluronan. The enzyme is known to associate with macrophages and polymorphonuclear leukocytes (neutrophils) and increasing evidence supports a role for EC-SOD in the development of an inflammatory response. Here we show that human EC-SOD is present at the cell surface of isolated neutrophils as well as stored within secretory vesicles. Interestingly, we find that EC-SOD mRNA is absent throughout neutrophil maturation indicating that the protein is synthesized by other cells and subsequently endocytosed by the neutrophil. When secretory vesicles were mobilized by neutrophil stimulation using formyl-methionyl-leucyl-phenylalanine (fMLF) or phorbol 12-myristate 13-acetate (PMA), the protein was released into the extracellular space and found to associate with DNA released from stimulated cells. The functional consequences were evaluated by the use of neutrophils isolated from wild-type and EC-SOD KO mice, and showed that EC-SOD release significantly reduce the level of superoxide in the extracellular space, but does not affect the capacity to generate neutrophil extracellular traps (NETs). Consequently, our data signifies that EC-SOD released from activated neutrophils affects the redox conditions of the extracellular space and may offer protection against highly reactive oxygen species such as hydroxyl radicals otherwise generated as a result of respiratory burst activity of activated neutrophils.

  12. Secretory COPII coat component Sec23a is essential for craniofacial chondrocyte maturation.

    PubMed

    Lang, Michael R; Lapierre, Lynne A; Frotscher, Michael; Goldenring, James R; Knapik, Ela W

    2006-10-01

    An increasing number of human disorders have been linked to mutations in genes of the secretory pathway. The chemically induced zebrafish crusher variant results in malformed craniofacial skeleton, kinked pectoral fins and a short body length. By positional cloning, we identified a nonsense mutation converting leucine to a stop codon (L402X) in the sec23a gene, an integral component of the COPII complex, which is critical for anterograde protein trafficking between endoplasmic reticulum and Golgi apparatus. Zebrafish crusher mutants develop normally until the onset of craniofacial chondrogenesis. crusher chondrocytes accumulate proteins in a distended endoplasmic reticulum, resulting in severe reduction of cartilage extracellular matrix (ECM) deposits, including type II collagen. We demonstrate that the paralogous gene sec23b is also an essential component of the ECM secretory pathway in chondrocytes. In contrast, knockdown of the COPI complex does not hinder craniofacial morphogenesis. As SEC23A lesions cause the cranio-lenticulo-sutural dysplasia syndrome, crusher provides the first vertebrate model system that links the biology of endoplasmic reticulum to Golgi trafficking with a clinically relevant dysmorphology.

  13. [Secretory immunoglobulin A (S-IgA) in the amniotic fluid supports the fetal maturity profile].

    PubMed

    Briese, V; Brock, J; Stark, K H

    1987-01-01

    Amniotic fluid levels of secretory immunoglobulin A(S-IgA) were measured by single radial immunodiffusion according to the method of Mancini using a monospecific antiserum against the human secretory component. 114 amniotic fluid samples were examined. The S-IgA values showed a correlation (r = 0.49) with the phospholipid levels. Additionally, it was observed no correlation between Serum and amniotic fluid S-IgA concentrations estimated as parallel investigations in 20 patients. S-IgA values greater than or equal to 82 mg/l were used to diagnose a sufficient fetal lung maturity. False negative values of S-IgA are possible. In 19 cases of newborns without signs of respiratory distress syndrome amniotic fluid S-IgA estimations were performed up to 32 hours before the delivery. False negative phospholipid levels were estimated in 2 cases, false negative S-IgA levels in 4 cases. The measurement of S-IgA in amniotic fluid will serve as an additional parameter for the estimation of fetal lung maturity.

  14. Differential Maturation of the Two Regulated Secretory Pathways in Human iPSC-Derived Neurons.

    PubMed

    Emperador Melero, Javier; Nadadhur, Aishwarya G; Schut, Desiree; Weering, Jan V; Heine, Vivi M; Toonen, Ruud F; Verhage, Matthijs

    2017-03-14

    Neurons communicate by regulated secretion of chemical signals from synaptic vesicles (SVs) and dense-core vesicles (DCVs). Here, we investigated the maturation of these two secretory pathways in micro-networks of human iPSC-derived neurons. These micro-networks abundantly expressed endogenous SV and DCV markers, including neuropeptides. DCV transport was microtubule dependent, preferentially anterograde in axons, and 2-fold faster in axons than in dendrites. SV and DCV secretion were strictly Ca(2+) and SNARE dependent. DCV secretion capacity matured until day in vitro (DIV) 36, with intense stimulation releasing 6% of the total DCV pool, and then plateaued. This efficiency is comparable with mature mouse neurons. In contrast, SV secretion capacity continued to increase until DIV50, with substantial further increase in secretion efficiency and decrease in silent synapses. These data show that the two secretory pathways can be studied in human neurons and that they mature differentially, with DCV secretion reaching maximum efficiency when that of SVs is still low.

  15. Secretory mechanism of fibroin, a silk protein, in the posterior silk gland cells of Bombyx mori.

    PubMed

    Sasaki, S; Nakagaki, I

    1980-01-01

    There are two microtubule-microfilament systems in the posterior silk gland cells of Bombyx mori. One is a radial microtubule system; the other is a circular microtubule-microfilament system. These two systems are presumably concerned with the intracellular transport of secretory granules of fibroin and the secretion of fibroin into the lumen, respectively. Conventional and scanning electron microscopic observations of the two microtubule-microfilament systems in the posterior silk gland cells are reported. Scanning electron micrographs showed that a number of parallel linear cytoplasmic processes ran circularly on the luminal surface of the posterior silk gland cells. These processes were assumed to correspond to the circular microtubule-microfilament systems. The effects of cytochalasin (B or D), a secretion stimulating agent of fibroin, on the intracellular recording of membrane potential from the posterior silk gland cells are also reported. Exposure to cytochalasin resulted in depolarization of the membrane potential of the gland cells. Possible functional roles of the two microtubule-microfilament systems in the secretory mechanism of fibroin are discussed with reference to the effects of antimitotic reagents and cytochalasin on these two systems.

  16. Divalent cations increase lipid order in erythrocytes and susceptibility to secretory phospholipase A2.

    PubMed

    Vest, Rebekah S; Gonzales, Laurie J; Permann, Seth A; Spencer, Emily; Hansen, Lee D; Judd, Allan M; Bell, John D

    2004-04-01

    Elevated concentrations of intracellular calcium in erythrocytes increase membrane order and susceptibility to secretory phospholipase A2. We hypothesize that calcium aids the formation of domains of ordered lipids within erythrocyte membranes by interacting directly with the inner leaflet of the cell membrane. The interface of these domains with regions of more fluid lipids may create an environment with weakened neighbor-neighbor interactions that would facilitate phospholipid migration into the active site of bound secretory phospholipase A2. This hypothesis was investigated by determining the effects of seven other divalent ions on erythrocyte membrane properties. Changes in membrane order were assessed with steady-state fluorescence spectroscopy and two-photon microscopy with an environment-sensitive probe, laurdan. Each ion increased apparent membrane order in model membranes and in erythrocytes when introduced with an ionophore, suggesting that direct binding to the inner face of the membrane accounts for the effects of calcium on membrane fluidity. Furthermore, the degree to which ions affected membrane properties correlated with the ionic radius and electronegativity of the ions. Lastly, erythrocytes became more susceptible to enzyme hydrolysis in the presence of elevated intracellular levels of nickel and manganese, but not magnesium. These differences appeared related to the ability of the ions to induce a transition in erythrocyte shape.

  17. Immunodominant antigens in Naegleria fowleri excretory--secretory proteins were potential pathogenic factors.

    PubMed

    Kim, Jong-Hyun; Yang, Ae-Hee; Sohn, Hae-Jin; Kim, Daesik; Song, Kyoung-Ju; Shin, Ho-Joon

    2009-11-01

    Naegleria fowleri, a ubiquitous pathogenic free-living amoeba, is the most virulent species and causes primary amoebic meningoencephalitis in laboratory animals and humans. The parasite secretes various inducing molecules as biological responses, which are thought to be involved in pathophysiological and immunological events during infection. To investigate what molecules of N. fowleri excretory-secretory proteins (ESPs) are related with amoebic pathogenicity, N. fowleri ESPs fractionated by two-dimensional electrophoresis were reacted with N. fowleri infection or immune sera. To identify immunodominant ESPs, six major protein spots were selected and analyzed by N-terminal sequencing. Finally, six proteins, 58, 40, 24, 21, 18, and 16 kDa of molecular weight, were partially cloned and matched with reference proteins as follow: 58 kDa of exendin-3 precursor, 40 kDa of secretory lipase, 24 kDa of cathepsin B-like proteases and cysteine protease, 21 kDa of cathepsin B, 18 kDa of peroxiredoxin, and 16 kDa of thrombin receptor, respectively. These results suggest that N. fowleri ESPs contained important proteins, which may play an important role in the pathogenicity of N. fowleri.

  18. Secretory vesicle priming by CAPS is independent of its SNARE-binding MUN domain.

    PubMed

    Nguyen Truong, Cuc Quynh; Nestvogel, Dennis; Ratai, Olga; Schirra, Claudia; Stevens, David R; Brose, Nils; Rhee, JeongSeop; Rettig, Jens

    2014-11-06

    Priming of secretory vesicles is a prerequisite for their Ca(2+)-dependent fusion with the plasma membrane. The key vesicle priming proteins, Munc13s and CAPSs, are thought to mediate vesicle priming by regulating the conformation of the t-SNARE syntaxin, thereby facilitating SNARE complex assembly. Munc13s execute their priming function through their MUN domain. Given that the MUN domain of Ca(2+)-dependent activator protein for secretion (CAPS) also binds syntaxin, it was assumed that CAPSs prime vesicles through the same mechanism as Munc13s. We studied naturally occurring splice variants of CAPS2 in CAPS1/CAPS2-deficient cells and found that CAPS2 primes vesicles independently of its MUN domain. Instead, the pleckstrin homology domain of CAPS2 seemingly is essential for its priming function. Our findings indicate a priming mode for secretory vesicles. This process apparently requires membrane phospholipids, does not involve the binding or direct conformational regulation of syntaxin by MUN domains of CAPSs, and is therefore not redundant with Munc13 action.

  19. Mutants defective in secretory/vacuolar pathways in the EUROFAN collection of yeast disruptants.

    PubMed

    Avaro, Sandrine; Belgareh-Touzé, Naïma; Sibella-Argüelles, Carla; Volland, Christiane; Haguenauer-Tsapis, Rosine

    2002-03-15

    We have screened the EUROFAN (European Functional Analysis Network) deletion strain collection for yeast mutants defective in secretory/vacuolar pathways and/or associated biochemical modifications. We used systematic Western immunoblotting to analyse the electrophoretic pattern of several markers of the secretory/vacuolar pathways, the soluble alpha-factor, the periplasmic glycoprotein invertase, the plasma membrane GPI-anchored protein Gas1p, and two vacuolar proteins, the soluble carboxypeptidase Y and the membrane-bound alkaline phosphatase, which are targeted to the vacuole by different pathways. We also used colony immunoblotting to monitor the secretion of carboxypeptidase Y into the medium, to identify disruptants impaired in vacuolar targeting. We identified 25 mutants among the 631 deletion strains. Nine of these mutants were disrupted in genes identified in recent years on the basis of their involvement in trafficking (VPS53, VAC7, VAM6, APM3, SYS1), or glycosylation (ALG12, ALG9, OST4, ROT2). Three of these genes were identified on the basis of trafficking defects by ourselves and others within the EUROFAN project (TLG2, RCY1, MON2). The deletion of ERV29, which encodes a COPII vesicle protein, impaired carboxypeptidase Y trafficking from the endoplasmic reticulum to the Golgi apparatus. We also identified eight unknown ORFs, the deletion of which reduced Golgi glycosylation or impaired the Golgi to vacuole trafficking of carboxypeptidase Y. YJR044c, which we identified as a new VPS gene, encodes a protein with numerous homologues of unknown function in sequence databases.

  20. Identification of Secretory Proteins in Mycobacterium tuberculosis Using Pseudo Amino Acid Composition

    PubMed Central

    Yang, Huan; Tang, Hua; Chen, Xin-Xin; Zhang, Chang-Jian; Zhu, Pan-Pan; Ding, Hui

    2016-01-01

    Tuberculosis is killing millions of lives every year and on the blacklist of the most appalling public health problems. Recent findings suggest that secretory protein of Mycobacterium tuberculosis may serve the purpose of developing specific vaccines and drugs due to their antigenicity. Responding to global infectious disease, we focused on the identification of secretory proteins in Mycobacterium tuberculosis. A novel method called MycoSec was designed by incorporating g-gap dipeptide compositions into pseudo amino acid composition. Analysis of variance-based technique was applied in the process of feature selection and a total of 374 optimal features were obtained and used for constructing the final predicting model. In the jackknife test, MycoSec yielded a good performance with the area under the receiver operating characteristic curve of 0.93, demonstrating that the proposed system is powerful and robust. For user's convenience, the web server MycoSec was established and an obliging manual on how to use it was provided for getting around any trouble unnecessary. PMID:27597968

  1. An overview of computer algorithms for deconvolution-based assessment of in vivo neuroendocrine secretory events.

    PubMed

    Veldhuis, J D; Johnson, M L

    1990-06-01

    The availability of increasingly efficient computational systems has made feasible the otherwise burdensome analysis of complex neurobiological data, such as in vivo neuroendocrine glandular secretory activity. Neuroendocrine data sets are typically sparse, noisy and generated by combined processes (such as secretion and metabolic clearance) operating simultaneously over both short and long time spans. The concept of a convolution integral to describe the impact of two or more processes acting jointly has offered an informative mathematical construct with which to dissect (deconvolve) specific quantitative features of in vivo neuroendocrine phenomena. Appropriate computer-based deconvolution algorithms are capable of solving families of 100-300 simultaneous integral equations for a large number of secretion and/or clearance parameters of interest. For example, one application of computer technology allows investigators to deconvolve the number, amplitude and duration of statistically significant underlying secretory episodes of algebraically specifiable waveform and simultaneously estimate subject- and condition-specific neurohormone metabolic clearance rates using all observed data and their experimental variances considered simultaneously. Here, we will provide a definition of selected deconvolution techniques, review their conceptual basis, illustrate their applicability to biological data and discuss new perspectives in the arena of computer-based deconvolution methodologies for evaluating complex biological events.

  2. Pasteurella multocida toxin: Targeting mast cell secretory granules during kiss-and-run secretion.

    PubMed

    Danielsen, Elisabeth M; Christiansen, Nina; Danielsen, E Michael

    2016-02-01

    Pasteurella multocida toxin (PMT), a virulence factor of the pathogenic Gram-negative bacterium P. multocida, is a 146 kDa protein belonging to the A-B class of toxins. Once inside a target cell, the A domain deamidates the α-subunit of heterotrimeric G-proteins, thereby activating downstream signaling cascades. However, little is known about how PMT selects and enters its cellular targets. We therefore studied PMT binding and uptake in porcine cultured intestinal mucosal explants to identify susceptible cells in the epithelium and underlying lamina propria. In comparison with Vibrio cholera B-subunit, a well-known enterotoxin taken up by receptor-mediated endocytosis, PMT binding to the epithelial brush border was scarce, and no uptake into enterocytes was detected by 2h, implying that none of the glycolipids in the brush border are a functional receptor for PMT. However, in the lamina propria, PMT distinctly accumulated in the secretory granules of mast cells. This also occurred at 4 °C, ruling out endocytosis, but suggestive of uptake via pores that connect the granules to the cell surface. Mast cell granules are known to secrete their contents by a "kiss-and-run" mechanism, and we propose that PMT may exploit this secretory mechanism to gain entry into this particular cell type.

  3. Assessing the Biosynthetic Capabilities of Secretory Glands in Citrus Peel1[W][OA

    PubMed Central

    Voo, Siau Sie; Grimes, Howard D.; Lange, B. Markus

    2012-01-01

    Epithelial cells (ECs) lining the secretory cavities of Citrus peel have been hypothesized to be responsible for the synthesis of essential oil, but direct evidence for such a role is currently sparse. We used laser-capture microdissection and pressure catapulting to isolate ECs and parenchyma cells (as controls not synthesizing oil) from the peel of young grapefruit (Citrus × paradisi ‘Duncan’), isolated RNA, and evaluated transcript patterns based on oligonucleotide microarrays. A Gene Ontology analysis of these data sets indicated an enrichment of genes involved in the biosynthesis of volatile terpenoids and nonvolatile phenylpropanoids in ECs (when compared with parenchyma cells), thus indicating a significant metabolic specialization in this cell type. The gene expression patterns in ECs were consistent with the accumulation of the major essential oil constituents (monoterpenes, prenylated coumarins, and polymethoxylated flavonoids). Morphometric analyses demonstrated that secretory cavities are formed early during fruit development, whereas the expansion of cavities, and thus oil accumulation, correlates with later stages of fruit expansion. Our studies have laid the methodological and experimental groundwork for a vastly improved knowledge of the as yet poorly understood processes controlling essential oil biosynthesis in Citrus peel. PMID:22452856

  4. PLURIVESICULAR SECRETORY PROCESSES AND NERVE ENDINGS IN THE PINEAL GLAND OF THE RAT

    PubMed Central

    De Robertis, Eduardo; de Iraldi, Amanda Pellegrino

    1961-01-01

    The pineal body of white normal rats, 1.5 to 3 months old, was studied under the electron microscope. A single type of parenchymal cell—the pinealocyte—is recognized as the main component of the tissue, and some of the structural characteristics of the nucleus and cytoplasm are described. The main morphological characteristic of the pinealocytes is represented by club-shaped perivascular expansions connected to the cell by thin pedicles. They are found lying in a large, clear space surrounding the blood capillaries. The name plurivesicular secretory processes is proposed, to emphasize the main structural feature and the probable function of these cellular expansions. A tubulofibrillar component is mainly found in the pedicle, and within the expansion there are numerous small mitochondria and densily packed vesicles of about 425 A. Two types of vesicles, one with a homogeneous content and another with a very dense osmium deposit, are described. Between the two types there are intermediary forms. In these processes, mitochondria show profound changes which may lead to complete vacuolization. The significance of this plurivesicular secretory component is discussed in the light of recent work on the biogenic amines of the pineal body and preliminary experiments showing the release of the vesicles containing dense granules after treatment with reserpine. These vesicles are interpreted as the site of storage of some of the biogenic amines. Bundles of unmyelinated nerve fibers and endings on large blood vessels which also contain a plurivesicular content are described and tentatively interpreted as adrenergic nerve terminals. PMID:13720811

  5. Multiple effects of the phenylhydrazone derivative FCCP on the secretory pathway in rat plasma cells.

    PubMed

    Antoine, J C; Jouanne, C

    1986-10-01

    We studied the sensitivity of the last steps of the secretory process of antibody-producing cells to carbonylcyanide-p-trifluoromethoxyphenylhydrazone (FCCP) and sodium azide (NaN3), agents which lower the cellular ATP content by inhibiting oxidative phosphorylation and mitochondrial electron transport, respectively. Popliteal lymph node cells or purified plasma cells from rats immunized against horseradish peroxidase were incubated with the drugs. The rate of secretion of anti-HRP antibodies was measured by an enzyme-linked immunoadsorbent assay or after biosynthetic labeling with L-[3H]fucose. FCCP as well as NaN3 were shown to rapidly inhibit (in less than 5 min) the secretion of immunoglobulins (Ig) and to partially block the release of fucosylated Ig. This indicates that the drugs inhibit the transport of Ig from the Golgi apparatus (GA) (where fucose is added to Ig) to the plasma membrane. However, the degree of inhibition reached 40 to 50% with NaN3 and 70 to 80% with FCCP, whereas both drugs similarly depleted ATP stores by 45 to 55%. These results are consistent with multiple effects of FCCP on the secretion pathway of Ig. As a tentative explanation, we suggest that FCCP, because of its protonophore properties, not only reduces cellular ATP levels but may also neutralize the Golgi or post-Golgi acidic compartments recently shown to be involved in the transport of plasma membrane and secretory proteins.

  6. The Fas pathway is involved in pancreatic β cell secretory function

    PubMed Central

    Schumann, Desiree M.; Maedler, Kathrin; Franklin, Isobel; Konrad, Daniel; Størling, Joachim; Böni-Schnetzler, Marianne; Gjinovci, Asllan; Kurrer, Michael O.; Gauthier, Benoit R.; Bosco, Domenico; Andres, Axel; Berney, Thierry; Greter, Melanie; Becher, Burkhard; Chervonsky, Alexander V.; Halban, Philippe A.; Mandrup-Poulsen, Thomas; Wollheim, Claes B.; Donath, Marc Y.

    2007-01-01

    Pancreatic β cell mass and function increase in conditions of enhanced insulin demand such as obesity. Failure to adapt leads to diabetes. The molecular mechanisms controlling this adaptive process are unclear. Fas is a death receptor involved in β cell apoptosis or proliferation, depending on the activity of the caspase-8 inhibitor FLIP. Here we show that the Fas pathway also regulates β cell secretory function. We observed impaired glucose tolerance in Fas-deficient mice due to a delayed and decreased insulin secretory pattern. Expression of PDX-1, a β cell-specific transcription factor regulating insulin gene expression and mitochondrial metabolism, was decreased in Fas-deficient β cells. As a consequence, insulin and ATP production were severely reduced and only partly compensated for by increased β cell mass. Up-regulation of FLIP enhanced NF-κB activity via NF-κB-inducing kinase and RelB. This led to increased PDX-1 and insulin production independent of changes in cell turnover. The results support a previously undescribed role for the Fas pathway in regulating insulin production and release. PMID:17299038

  7. Neuronal porosome - The secretory portal at the nerve terminal: Its structure-function, composition, and reconstitution

    NASA Astrophysics Data System (ADS)

    Jena, Bhanu P.

    2014-09-01

    Cup-shaped secretory portals at the cell plasma membrane called porosomes mediate secretion from cells. Membrane bound secretory vesicles transiently dock and fuse at the cytosolic compartment of the porosome base to expel intravesicular contents to the outside during cell secretion. In the past decade, the structure, isolation, composition, and functional reconstitution of the neuronal porosome complex has been accomplished providing a molecular understanding of its structure-function. Neuronal porosomes are 15 nm cup-shaped lipoprotein structures composed of nearly 40 proteins; compared to the 120 nm nuclear pore complex comprised of over 500 protein molecules composed of 30 different proteins. Being a membrane-associated supramolecular complex has precluded determination of the atomic structure of the porosome. However recent studies using small-angle X-ray solution scattering (SAXS), provide at sub-nanometer resolution, the native 3D structure of the neuronal porosome complex associated with docked synaptic vesicle at the nerve terminal. Additionally, results from the SAXS study and earlier studies using atomic force microscopy, provide the possible molecular mechanism involved in porosome-mediated neurotransmitter release at the nerve terminal.

  8. Bacterial mimetics of endocrine secretory granules as immobilized in vivo depots for functional protein drugs.

    PubMed

    Céspedes, María Virtudes; Fernández, Yolanda; Unzueta, Ugutz; Mendoza, Rosa; Seras-Franzoso, Joaquin; Sánchez-Chardi, Alejando; Álamo, Patricia; Toledo-Rubio, Verónica; Ferrer-Miralles, Neus; Vázquez, Esther; Schwartz, Simó; Abasolo, Ibane; Corchero, José Luis; Mangues, Ramon; Villaverde, Antonio

    2016-10-24

    In the human endocrine system many protein hormones including urotensin, glucagon, obestatin, bombesin and secretin, among others, are supplied from amyloidal secretory granules. These granules form part of the so called functional amyloids, which within the whole aggregome appear to be more abundant than formerly believed. Bacterial inclusion bodies (IBs) are non-toxic, nanostructured functional amyloids whose biological fabrication can be tailored to render materials with defined biophysical properties. Since under physiological conditions they steadily release their building block protein in a soluble and functional form, IBs are considered as mimetics of endocrine secretory granules. We have explored here if the in vivo implantation of functional IBs in a given tissue would represent a stable local source of functional protein. Upon intratumoral injection of bacterial IBs formed by a potent protein ligand of CXCR4 we have observed high stability and prevalence of the material in absence of toxicity, accompanied by apoptosis of CXCR4(+) cells and tumor ablation. Then, the local immobilization of bacterial amyloids formed by therapeutic proteins in tumors or other tissues might represent a promising strategy for a sustained local delivery of protein drugs by mimicking the functional amyloidal architecture of the mammals' endocrine system.

  9. Feedback inhibition of thymic secretory activity in mice treated by the thymic extract TP-1 (thymostimulin).

    PubMed Central

    Shoham, J; Ben-David, E; Sandbank, U

    1982-01-01

    The ultrastructural changes occurring in the medullary epithelium of the thymus of young mice, as a result of repeated injections of thymic extract, TP-1 (thymostimulin) was investigated. After daily injection of TP-1 for 3 weeks, no changes in thymus architecture could be observed by light microscopy. However, by electron microscopy, specific changes were noticed in the epithelial cells. The secretory granules became dilated and engorged; diameter of granules in normal control thymus was approximately 200-250 nm, but reached 1000 nm in treated mice. Degenerative changes appeared in some of these granules, including myelin bodies, distorted configuration and fat droplets. Signs of involution of whole cells and presence of cellular debri within macrophages were observed. Acid phosphatase staining disclosed many lysosomes containing ingested granules. No such findings were observed in control untreated mice, or in mice treated by a heart extract similarly prepared to TP-1. All these findings can be taken as ultrastructural evidence for feedback inhibition of thymic secretory activity, in analogy to the changes occurring other feedback inhibited, peptide hormone secreting glands. The data indicate that (i) the thymus respond to feedback inhibitory stimuli, as other endocrine glands do; (ii)TP-1, the thymic extract under study, contains a physiologically significant thymic hormone, which, when introduced in high doses can exert specific feedback inhibition. This can be taken as an additional, new criterion for the definition of thymic hormones. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 PMID:7056566

  10. A pH-regulated quality control cycle for surveillance of secretory protein assembly.

    PubMed

    Vavassori, Stefano; Cortini, Margherita; Masui, Shoji; Sannino, Sara; Anelli, Tiziana; Caserta, Imma R; Fagioli, Claudio; Mossuto, Maria F; Fornili, Arianna; van Anken, Eelco; Degano, Massimo; Inaba, Kenji; Sitia, Roberto

    2013-06-27

    To warrant the quality of the secretory proteome, stringent control systems operate at the endoplasmic reticulum (ER)-Golgi interface, preventing the release of nonnative products. Incompletely assembled oligomeric proteins that are deemed correctly folded must rely on additional quality control mechanisms dedicated to proper assembly. Here we unveil how ERp44 cycles between cisGolgi and ER in a pH-regulated manner, patrolling assembly of disulfide-linked oligomers such as IgM and adiponectin. At neutral, ER-equivalent pH, the ERp44 carboxy-terminal tail occludes the substrate-binding site. At the lower pH of the cisGolgi, conformational rearrangements of this peptide, likely involving protonation of ERp44's active cysteine, simultaneously unmask the substrate binding site and -RDEL motif, allowing capture of orphan secretory protein subunits and ER retrieval via KDEL receptors. The ERp44 assembly control cycle couples secretion fidelity and efficiency downstream of the calnexin/calreticulin and BiP-dependent quality control cycles.

  11. A pH-Regulated Quality Control Cycle for Surveillance of Secretory Protein Assembly

    PubMed Central

    Vavassori, Stefano; Cortini, Margherita; Masui, Shoji; Sannino, Sara; Anelli, Tiziana; Caserta, Imma R.; Fagioli, Claudio; Mossuto, Maria F.; Fornili, Arianna; van Anken, Eelco; Degano, Massimo; Inaba, Kenji; Sitia, Roberto

    2013-01-01

    Summary To warrant the quality of the secretory proteome, stringent control systems operate at the endoplasmic reticulum (ER)-Golgi interface, preventing the release of nonnative products. Incompletely assembled oligomeric proteins that are deemed correctly folded must rely on additional quality control mechanisms dedicated to proper assembly. Here we unveil how ERp44 cycles between cisGolgi and ER in a pH-regulated manner, patrolling assembly of disulfide-linked oligomers such as IgM and adiponectin. At neutral, ER-equivalent pH, the ERp44 carboxy-terminal tail occludes the substrate-binding site. At the lower pH of the cisGolgi, conformational rearrangements of this peptide, likely involving protonation of ERp44’s active cysteine, simultaneously unmask the substrate binding site and −RDEL motif, allowing capture of orphan secretory protein subunits and ER retrieval via KDEL receptors. The ERp44 assembly control cycle couples secretion fidelity and efficiency downstream of the calnexin/calreticulin and BiP-dependent quality control cycles. PMID:23685074

  12. Toxoplasma gondii Vps11, a subunit of HOPS and CORVET tethering complexes, is essential for the biogenesis of secretory organelles.

    PubMed

    Morlon-Guyot, Juliette; Pastore, Sandra; Berry, Laurence; Lebrun, Maryse; Daher, Wassim

    2015-08-01

    Apicomplexan parasites harbour unique secretory organelles (dense granules, rhoptries and micronemes) that play essential functions in host infection. Toxoplasma gondii parasites seem to possess an atypical endosome-like compartment, which contains an assortment of proteins that appear to be involved in vesicular sorting and trafficking towards secretory organelles. Recent studies highlighted the essential roles of many regulators such as Rab5A, Rab5C, sortilin-like receptor and syntaxin-6 in secretory organelle biogenesis. However, little is known about the protein complexes that recruit Rab-GTPases and SNAREs for membrane tethering in Apicomplexa. In mammals and yeast, transport, tethering and fusion of vesicles from early endosomes to lysosomes and the vacuole, respectively, are mediated by CORVET and HOPS complexes, both built on the same Vps-C core that includes Vps11 protein. Here, we show that a T. gondii Vps11 orthologue is essential for the biogenesis or proper subcellular localization of secretory organelle proteins. TgVps11 is a dynamic protein that associates with Golgi endosomal-related compartments, the vacuole and immature apical secretory organelles. Conditional knock-down of TgVps11 disrupts biogenesis of dense granules, rhoptries and micronemes. As a consequence, parasite motility, invasion, egress and intracellular growth are affected. This phenotype was confirmed with additional knock-down mutants of the HOPS complex. In conclusion, we show that apicomplexan parasites use canonical regulators of the endolysosome system to accomplish essential parasite-specific functions in the biogenesis of their unique secretory organelles.

  13. Beta-agonists and secretory cell number and intracellular glycoproteins in airway epithelium. The effect of isoproterenol and salbutamol.

    PubMed Central

    Jones, R.; Reid, L.

    1979-01-01

    This study describes the effect of systemic administration of the beta-adrenergic agonists isoproterenol and salbutamol on the secretory cell populations in seven regions of rat airway epithelium (three extrapulmonary and four intrapulmonary) and on the size of salivary glands and heart. Isoproterenol (a nonselective beta-adrenergic agonist) significantly increases secretory cell number in all airway regions except the midtrachea; salbutamol (a selective beta 2 agonist) increases secretory cell number only in proximal and peripheral regions. The absolute number of secretory cells is greatest in the most peripheral region after isoproterenol administration and in the most proximal region after salbutamol, although both drugs produce the greatest relative increase at the periphery. In proximal and, particularly, peripheral regions, the increase by isoproterenol (less than 3- and 14-fold, respectively) is greater than by salbutamol (less than 2- and less than 3-fold, respectively). In all airway regions, both drugs modify intracellular glycoprotein in the secretory cell population; within a given region, modification is much the same. In the most proximal region, the population of cells synthesizing only granules of neutral glycoprotein significantly increases while in other regions increase is in cells synthesizing only granules of acid. A significant shift in glycoprotein synthesis occurs whether or not the secretory cell population is increased, which suggests that existing as well as newly appearing cells modify their product. Isoproterenol significantly increases the size of the parotid and submaxillary glands; salbutamol increases the size of the parotid only. Isoproterenol significantly increases the weight of both ventricles of the heart; salbutamol has no such effect. PMID:36762

  14. Dense-core granules in neuroendocrine cells and neurons release their secretory constituents by piecemeal degranulation (review).

    PubMed

    Crivellato, Enrico; Nico, Beatrice; Bertelli, Eugenio; Nussdorfer, Gastone G; Ribatti, Domenico

    2006-12-01

    The term piecemeal degranulation (PMD) refers to a slow releasing process mediated by vesicular transport of stored secretory granule contents. This form of cell secretion was first proposed for basophils, mast cells and eosinophils, but evidence has begun to accumulate that PMD also occurs in dense-core granules of neuroendocrine cells and neurons. This review summarizes the electron-microscopic evidence that has been gathered in support of this view and also discusses the possible physiological significance of PMD in this class of secretory organelles in comparison with 'full fusion' and 'kiss-and-run' exocytosis.

  15. Survival, activity and release of antigenic excretory secretory products and microfilariae of Setaria digitata maintained in artificial media.

    PubMed

    Sundar, S T B; D'Souza, Placid E

    2015-03-01

    The survival, activity and release of excretory secretory products from Setaria digitata, the filarial worm of cattle was studied. Adult female worms were maintained in vitro in DMEM and Tyrode solution. Worms incubated in DMEM were alive and very active for 2 days. The activity was moderate for another 2 days and after the fourth day increased mortality was observed. Antigenic excretory secretory products were also released. Worms incubated in Tyrode solution were very active without any mortality up to 4 h of incubation. Copious amount of ova and microfilaria were shed by the incubated worms in a time-dependent manner.

  16. Unmasking MASC: bringing to light the unique morphologic, immunohistochemical and genetic features of the newly recognized mammary analogue secretory carcinoma of salivary glands.

    PubMed

    Bishop, Justin A

    2013-03-01

    Mammary analogue secretory carcinoma (MASC) is a recently described salivary gland neoplasm that is characterized by its striking morphologic and molecular similarities to secretory carcinoma of the breast. This review highlights the characteristic clinical, histologic, immunophenotypic, and molecular features of MASC, and draws attention to the differential diagnosis of this increasingly recognized tumor.

  17. The Ca2+/H+ antiporter TMEM165 expression, localization in the developing, lactating and involuting mammary gland parallels the secretory pathway Ca2+ATPase (SPCA1)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Plasma membrane Ca2+-ATPase 2 (PMCA2) knockout mice showed that ~ 60 % of calcium in milk is transported across the mammary cells apical membrane by PMCA2. The remaining milk calcium is thought to arrive via the secretory pathway through the actions of secretory pathway Ca2+-ATPase’s 1 and/or 2 (SP...

  18. [Clinical importance of tympanometry in the diagnosis of chronic secretory otitis].

    PubMed

    Spremo, S; Markić, Z; Kurbalija

    1998-01-01

    Secretory otitis media is defined as a fluid in the middle ear without signs or symptoms of infection. As the aetiology and pathogenesis of the disease are unknown, and as it affects children aged from 3 to 12 years, treatment procedures proposed for management of secretory otitis media, are not uniform. Some authors [1, 4, 6] consider that functional or mechanical obstructions of the Eustachian tube could provoke secretory otitis. The purpose of the treatment is to remove exudate from the middle ear and appropriately ventilate it for a longer period. That could instantly normalize the hearing and exclude the appearance of late complications of secretory otitis. Although the disease could heal spontaneously, the treatment should be performed immediately for preventing sequelae of secretory otitis. The aim of the study was to evaluate possible aetiologic factors of secretory otitis in our population, and to evaluate results of lympanometry in children with exudate in the middle ear. There were 65 children, aged from 3 to 12 years (Table 1), who complained of deafness and were examined at the ORL Department in Banja Luka. The clinical examination revealed the integrity and color of tympanic membrane, scars, adhesions and atrophic areas. Audiometry and tympanometry had been performed in addition. Patients who proved to have exudate in the middle ear received nasal decongestants and mucolitics during three months, and were evaluated every three weeks by audiometry and tympanometry. Pathologic findings in the nose and epipharynx were the most common findings: enlarged adenoids in 38 (58%) patients, hypetrophic rhinitis in 15 (23%) and allergic rhinitis in 5 (8%) patients. Frequent relapses of middle ear infection in the first three years of life were found in 26 (40%) patients and early first attacks in the first year of life in 15 (23%) patients (Table 2). Premature onset (15%) and allergy (21%) had also been frequently found. Results of tympanometry and audiometry are

  19. Characterisation of a secretory serine protease inhibitor (SjB6) from Schistosoma japonicum

    PubMed Central

    2014-01-01

    Background Proteins belonging to the serine protease inhibitor (serpin) superfamily play essential physiological roles in many organisms. In pathogens, serpins are thought to have evolved specifically to limit host immune responses by interfering with the host immune-stimulatory signals. Serpins are less well characterised in parasitic helminths, although some are thought to be involved in mechanisms associated with host immune modulation. In this study, we cloned and partially characterised a secretory serpin from Schistosoma japonicum termed SjB6, these findings provide the basis for possible functional roles. Methods SjB6 gene was identified through database mining of our previously published microarray data, cloned and detailed sequence and structural analysis and comparative modelling carried out using various bioinformatics and proteomics tools. Gene transcriptional profiling was determined by real-time PCR and the expression of native protein determined by immunoblotting. An immunological profile of the recombinant protein produced in insect cells was determined by ELISA. Results SjB6 contains an open reading frame of 1160 base pairs that encodes a protein of 387 amino acid residues. Detailed sequence analysis, comparative modelling and structural-based alignment revealed that SjB6 contains the essential structural motifs and consensus secondary structures typical of inhibitory serpins. The presence of an N-terminal signal sequence indicated that SjB6 is a secretory protein. Real-time data indicated that SjB6 is expressed exclusively in the intra-mammalian stage of the parasite life cycle with its highest expression levels in the egg stage (p < 0.0001). The native protein is approximately 60 kDa in size and recombinant SjB6 (rSjB6) was recognised strongly by sera from rats experimentally infected with S. japonicum. Conclusions The significantly high expression of SjB6 in schistosome eggs, when compared to other life cycle stages, suggests a possible

  20. Bcl-2 promoter sequence G-quadruplex interactions with three planar and non-planar cationic porphyrins: TMPyP4, TMPyP3, and TMPyP2.

    PubMed

    Le, Vu H; Nagesh, Narayana; Lewis, Edwin A

    2013-01-01

    The interactions of three related cationic porphyrins, TMPyP4, TMPyP3 and TMPyP2, with a WT 39-mer Bcl-2 promoter sequence G-quadruplex were studied using Circular Dichroism, ESI mass spectrometry, Isothermal Titration Calorimetry, and Fluorescence spectroscopy. The planar cationic porphyrin TMPyP4 (5, 10, 15, 20-meso-tetra (N-methyl-4-pyridyl) porphine) is shown to bind to a WT Bcl-2 G-quadruplex via two different binding modes, an end binding mode and a weaker mode attributed to intercalation. The related non-planar ligands, TMPyP3 and TMPyP2, are shown to bind to the Bcl-2 G-quadruplex by a single mode. ESI mass spectrometry experiments confirmed that the saturation stoichiometry is 4:1 for the TMPyP4 complex and 2:1 for the TMPyP2 and TMPyP3 complexes. ITC experiments determined that the equilibrium constant for formation of the (TMPyP4)1/DNA complex (K1 = 3.7 × 10(6)) is approximately two orders of magnitude greater than the equilibrium constant for the formation of the (TMPyP2)1/DNA complex, (K1 = 7.0 × 10(4)). Porphyrin fluorescence is consistent with intercalation in the case of the (TMPyP4)3/DNA and (TMPyP4)4/DNA complexes. The non-planar shape of the TMPyP2 and TMPyP3 molecules results in both a reduced affinity for the end binding interaction and the elimination of the intercalation binding mode.

  1. [Restoration of secretory activity of digestive glands in conditions of acute hyperkinesis at persons with different levels of motor activity].

    PubMed

    Griaznykh, A V

    2012-01-01

    The dynamics' features of restoration reactions of the secretory function of gastric glands have been studied at patients with differences in the level and specificity of daily physical activity. The dependence between the level and characteristics of daily physical activity and reactivity of the secretory apparatus of the gastric glands in the recovery period after the muscle load has been established. The high reactivity of the secretory activity of digestive glands is typical for individuals with high level of daily physical activity. The differences of the functional stability of the secretion's mechanisms of various components of gastric juice under the influence of physical exercise and in recovery have been revealed. The greatest stability of the secretory mechanisms of digestive glands has been discovered at athletes practicing with the development of endurance--at skiers. Heterochronous reducing reactions of gastric and pancreatic secretion after physical stress have been defined. The reduction of secretion's mechanisms of gastric juice ingredients and the electrolyte and acid composition of digestive juices isn't simultaneous: the first order is for ferment's secretion. The inverse relationship between the content of digestive enzymes pepsinogen-1 and -2 in blood's serum and the concentration of proteolytic enzymes in the gastric content has been found at persons with various degrees of adaptation to the muscular tension.

  2. Mammary analogue secretory carcinoma of salivary gland origin: an update and expanded morphologic and immunohistochemical spectrum of recently described entity.

    PubMed

    Skalova, Alena

    2013-07-01

    Mammary analogue secretory carcinoma of salivary gland origin (MASC) is a recently described tumor with ETV6 translocation. Akin to secretory breast cancer, MASC expresses S-100 protein, mammaglobin, vimentin, and harbors a t(12;15) (p13;q25) translocation which leads to ETV6-NTRK3 fusion product. Histologically, MASC displays a lobulated growth pattern and is often composed of microcystic, tubular, and solid structures with abundant eosinophilic homogeneous or bubbly secretions. Colloid-like secretory material stains positive for periodic acid-Schiff (PAS) with and without diastase and for Alcian blue. The cells of MASC are devoid of PAS-positive secretory zymogen granules. These features help to exclude the most important differential diagnostic considerations, namely acinic cell carcinoma, low-grade cribriform cystadenocarcinoma, cystadenocarcinoma (not otherwise specified), and low-grade mucoepidermoid carcinoma. To date the presence of the ETV6-NTRK3 fusion gene has not been demonstrated in any other salivary gland tumor than MASC. It is likely that MASC is more common than currently recognized and with further studies, the clinical need for molecular studies of the ETV6-NTRK3 fusion may diminish. However, molecular testing is recommended at this time to arrive at the diagnosis of MASC.

  3. Determination of the electrophoretic pattern of somatic and excretory-secretory proteins of Ligula intestinalis parasite in spirlin (Alburnoides bipunctatus).

    PubMed

    Youssefi, M R; Hosseinifard, S M; Halimi, M; Kordafshari, S

    2012-12-01

    Ligula intestinalis parasite is a cestodes that causes remarkable damages to fish. It is also of prime importance in economic and hygienic aspects. SDS-PAGE and western blotting are the methods that can be used to determine the electerophoretic pattern of somatic and excretory-secretory proteins of parasites. In this study, after obtaining the plerocercoidal stage of this parasite from the spirlin (Alburnoides bipunctatus), its somatic proteins were prepared using ultrasonicae, and excretory-secretory proteins were prepared using the PBS solution. After protein assay, which included using the Bradford method and then SDS-PAGE on these two antigenic solutions, 5 protein bands of 26, 33, 38, 58, 70kDa in somatic antigens, and 7 bands of 25, 28, 33, 43, 49, 60, 70kDa in excretory-secretory antigens were observed. After western blotting on both antigens and adding the primary antibody (the sera of infected fish) and then the secondary antibody (Rabbit Anti-fish Polyclonal Antibody Conjugated from Abnova Corporation) no band was seen in excretory-secretory antigen. And only in the 55kDa band of somatic antigen, a positive response, in comparison of fish positive serum was observed.

  4. Protection of Human Colon Cells from Shiga Toxin by Plant-based Recombinant Secretory IgA

    PubMed Central

    Nakanishi, Katsuhiro; Morikane, Shota; Ichikawa, Shiori; Kurohane, Kohta; Niwa, Yasuo; Akimoto, Yoshihiro; Matsubara, Sachie; Kawakami, Hayato; Kobayashi, Hirokazu; Imai, Yasuyuki

    2017-01-01

    Shiga toxin is a major virulence factor of food-poisoning caused by Escherichia coli such as O157:H7. Secretory immunoglobulin (Ig) A (SIgA) is supposed to prevent infection of the mucosal surface and is a candidate agent for oral immunotherapy. We previously established a recombinant monoclonal antibody (mAb) consisting of variable regions from a mouse IgG mAb specific for the binding subunit of Shiga toxin 1 (Stx1) and the Fc region of mouse IgA. Here we produced a secretory form of the recombinant IgA (S-hyIgA) with transgenic Arabidopsis thaliana plant. All the S-hyIgA cDNAs (heavy, light, J chain and secretory component) were expressed under the control of a bidirectional promoter of a chlorophyll a/b-binding protein of A. thaliana without using a viral promoter. The plant-based S-hyIgA exhibited antigen binding, and was modified with plant-specific N-linked sugar chains. The Ig heavy chain and secretory components were observed in an intracellular protein body-like structure of the transgenic leaves on immuno-electron microscopy. An extract of the transgenic leaves neutralized the cytotoxicity of Stx1 toward butyrate-treated Caco-2 cells, a human colon carcinoma cell line. These results will contribute to the development of edible therapeutic antibodies such as those for the treatment of mucosal infection. PMID:28368034

  5. Acute insulin responses to glucose and arginine as predictors of beta-cell secretory capacity in human islet transplantation.

    PubMed

    Rickels, Michael R; Naji, Ali; Teff, Karen L

    2007-11-27

    Islet transplantation for type 1 diabetes can enable the achievement of near-normal glycemic control without severe hypoglycemic episodes. How much an islet (beta-cell) graft may be contributing to glycemic control can be quantified by stimulatory tests of insulin (or C-peptide) secretion. Glucose-potentiation of arginine-induced insulin secretion provides a measure of functional beta-cell mass, the beta-cell secretory capacity, as either AIR(pot) or AIR(max), but requires conduct of a hyperglycemic clamp. We sought to determine whether acute insulin responses to intravenous glucose (AIR(glu)) or arginine (AIR(arg)) could predict beta-cell secretory capacity in islet recipients. AIR(arg) was a better predictor of both AIR(pot) and AIR(max) (n=10, r2=0.98, P<0.0001 and n=7, r2=0.97, P<0.0001) than was AIR(glu) (n=9, r2=0.78, P=0.002 and n=6, r2=0.76, P=0.02). Also, the measures of beta-cell secretory capacity were highly correlated (n=7, r2=0.98, P<0.0001). These results support the use of AIR(arg) as a surrogate indicator of beta-cell secretory capacity in islet transplantation.

  6. Protection of Human Colon Cells from Shiga Toxin by Plant-based Recombinant Secretory IgA.

    PubMed

    Nakanishi, Katsuhiro; Morikane, Shota; Ichikawa, Shiori; Kurohane, Kohta; Niwa, Yasuo; Akimoto, Yoshihiro; Matsubara, Sachie; Kawakami, Hayato; Kobayashi, Hirokazu; Imai, Yasuyuki

    2017-04-03

    Shiga toxin is a major virulence factor of food-poisoning caused by Escherichia coli such as O157:H7. Secretory immunoglobulin (Ig) A (SIgA) is supposed to prevent infection of the mucosal surface and is a candidate agent for oral immunotherapy. We previously established a recombinant monoclonal antibody (mAb) consisting of variable regions from a mouse IgG mAb specific for the binding subunit of Shiga toxin 1 (Stx1) and the Fc region of mouse IgA. Here we produced a secretory form of the recombinant IgA (S-hyIgA) with transgenic Arabidopsis thaliana plant. All the S-hyIgA cDNAs (heavy, light, J chain and secretory component) were expressed under the control of a bidirectional promoter of a chlorophyll a/b-binding protein of A. thaliana without using a viral promoter. The plant-based S-hyIgA exhibited antigen binding, and was modified with plant-specific N-linked sugar chains. The Ig heavy chain and secretory components were observed in an intracellular protein body-like structure of the transgenic leaves on immuno-electron microscopy. An extract of the transgenic leaves neutralized the cytotoxicity of Stx1 toward butyrate-treated Caco-2 cells, a human colon carcinoma cell line. These results will contribute to the development of edible therapeutic antibodies such as those for the treatment of mucosal infection.

  7. Calcineurin is universally involved in vesicle endocytosis at neuronal and nonneuronal secretory cells.

    PubMed

    Wu, Xin-Sheng; Zhang, Zhen; Zhao, Wei-Dong; Wang, Dongsheng; Luo, Fujun; Wu, Ling-Gang

    2014-05-22

    Calcium influx triggers and accelerates endocytosis in nerve terminals and nonneuronal secretory cells. Whether calcium/calmodulin-activated calcineurin, which dephosphorylates endocytic proteins, mediates this process is highly controversial for different cell types, developmental stages, and endocytic forms. Using three preparations that previously produced discrepant results (i.e., large calyx-type synapses, conventional cerebellar synapses, and neuroendocrine chromaffin cells containing large dense-core vesicles), we found that calcineurin gene knockout consistently slowed down endocytosis, regardless of cell type, developmental stage, or endocytic form (rapid or slow). In contrast, calcineurin and calmodulin blockers slowed down endocytosis at a relatively small calcium influx, but did not inhibit endocytosis at a large calcium influx, resulting in false-negative results. These results suggest that calcineurin is universally involved in endocytosis. They may also help explain the discrepancies among previous pharmacological studies. We therefore suggest that calcineurin should be included as a key player in mediating calcium-triggered and -accelerated vesicle endocytosis.

  8. Systematic single-cell analysis of Pichia pastoris reveals secretory capacity limits productivity.

    PubMed

    Love, Kerry Routenberg; Politano, Timothy J; Panagiotou, Vasiliki; Jiang, Bo; Stadheim, Terrance A; Love, J Christopher

    2012-01-01

    Biopharmaceuticals represent the fastest growing sector of the global pharmaceutical industry. Cost-efficient production of these biologic drugs requires a robust host organism for generating high titers of protein during fermentation. Understanding key cellular processes that limit protein production and secretion is, therefore, essential for rational strain engineering. Here, with single-cell resolution, we systematically analysed the productivity of a series of Pichia pastoris strains that produce different proteins both constitutively and inducibly. We characterized each strain by qPCR, RT-qPCR, microengraving, and imaging cytometry. We then developed a simple mathematical model describing the flux of folded protein through the ER. This combination of single-cell measurements and computational modelling shows that protein trafficking through the secretory machinery is often the rate-limiting step in single-cell production, and strategies to enhance the overall capacity of protein secretion within hosts for the production of heterologous proteins may improve productivity.

  9. New class of cargo protein in Tetrahymena thermophila dense core secretory granules.

    PubMed

    Haddad, Alex; Bowman, Grant R; Turkewitz, Aaron P

    2002-08-01

    Regulated exocytosis of dense core secretory granules releases biologically active proteins in a stimulus-dependent fashion. The packaging of the cargo within newly forming granules involves a transition: soluble polypeptides condense to form water-insoluble aggregates that constitute the granule cores. Following exocytosis, the cores generally disassemble to diffuse in the cell environment. The ciliates Tetrahymena thermophila and Paramecium tetraurelia have been advanced as genetically manipulatable systems for studying exocytosis via dense core granules. However, all of the known granule proteins in these organisms condense to form the architectural units of lattices that are insoluble both before and after exocytosis. Using an approach designed to detect new granule proteins, we have now identified Igr1p (induced during granule regeneration). By structural criteria, it is unrelated to the previously characterized lattice-forming proteins. It is distinct in that it is capable of dissociating from the insoluble lattice following secretion and therefore represents the first diffusible protein identified in ciliate granules.

  10. Cobra venom contains a pool of cysteine-rich secretory proteins.

    PubMed

    Osipov, Alexey V; Levashov, Mikhail Yu; Tsetlin, Victor I; Utkin, Yuri N

    2005-03-04

    A large family of cysteine-rich secretory proteins (CRISPs) includes proteins of different origin, the function of the majority of CRISPs being unknown. For CRISPs isolated from snake venom, two types of activities were found: two proteins blocked cyclic nucleotide-gated ion channels, several others blocked potassium-stimulated smooth muscle contraction. Thus, snake CRISPs represent potentially valuable tools for studies of ion channels, which makes promising a search for new CRISPs. Here we report on the isolation of several novel CRISPs from the venoms of Asian cobra Naja kaouthia and African cobra Naja haje using a combination of different types of liquid chromatography. Four CRISP variants were identified in N. kaouthia venom and three proteins, one of them acidic, were found in N. haje venom. Acidic CRISP was found in a reptilian venom for the first time. Our data suggest that each cobra venom contains a pool of different CRISPs.

  11. The role of cysteine-rich secretory proteins in male fertility.

    PubMed

    Koppers, Adam J; Reddy, Thulasimala; O'Bryan, Moira K

    2011-01-01

    The cysteine-rich secretory proteins (CRISPs) are a subgroup of the CRISP, antigen 5 and Pr-1 (CAP) protein superfamily, and are found only in vertebrates. They show a strong expression bias to the mammalian male reproductive tract and the venom of poisonous reptiles. Within the male reproductive tract CRISPs have been implicated in many aspects of male germ cell biology spanning haploid germ cell development, epididymal maturation, capacitation, motility and the actual processes of fertilization. At a structural level, CRISPs are composed of two domains, a CAP domain, which has been implicated in cell-cell adhesion, and a CRISP domain, which has been shown to regulate several classes of ion channels across multiple species. Herein, we will review the current literature on the role of CRISPs in male fertility, and by inference to related non-mammalian protein, infer potential biochemical functions.

  12. On psychobiology in psychoanalysis - salivary cortisol and secretory IgA as psychoanalytic process parameters

    PubMed Central

    Euler, Sebastian; Schimpf, Heinrich; Hennig, Jürgen; Brosig, Burkhard

    2005-01-01

    This study investigates the psychobiological impact of psychoanalysis in its four-hour setting. During a period of five weeks, 20 subsequent hours of psychoanalysis were evaluated, involving two patients and their analysts. Before and after each session, saliva samples were taken and analysed for cortisol (sCortisol) and secretory immunoglobuline A (sIgA). Four time-series (n=80 observations) resulted and were evaluated by "Pooled Time Series Analysis" (PTSA) for significant level changes and setting-mediated rhythms. Over all sessions, sCortisol levels were reduced and sIgA secretion augmented parallel to the analytic work. In one analytic dyad a significant rhythm within the four-hour setting was observed with an increase of sCortisol in sessions 2 and 3 of the week. Psychoanalysis may, therefore, have some psychobiological impact on patients and analysts alike and may modulate immunological and endocrinological processes. PMID:19742067

  13. On psychobiology in psychoanalysis - salivary cortisol and secretory IgA as psychoanalytic process parameters.

    PubMed

    Euler, Sebastian; Schimpf, Heinrich; Hennig, Jürgen; Brosig, Burkhard

    2005-03-31

    This study investigates the psychobiological impact of psychoanalysis in its four-hour setting. During a period of five weeks, 20 subsequent hours of psychoanalysis were evaluated, involving two patients and their analysts. Before and after each session, saliva samples were taken and analysed for cortisol (sCortisol) and secretory immunoglobuline A (sIgA). Four time-series (n=80 observations) resulted and were evaluated by "Pooled Time Series Analysis" (PTSA) for significant level changes and setting-mediated rhythms. Over all sessions, sCortisol levels were reduced and sIgA secretion augmented parallel to the analytic work. In one analytic dyad a significant rhythm within the four-hour setting was observed with an increase of sCortisol in sessions 2 and 3 of the week. Psychoanalysis may, therefore, have some psychobiological impact on patients and analysts alike and may modulate immunological and endocrinological processes.

  14. Immunomodulatory effects of Trichinella spiralis-derived excretory-secretory antigens.

    PubMed

    Radovic, Ivana; Gruden-Movsesijan, Alisa; Ilic, Natasa; Cvetkovic, Jelena; Mojsilovic, Slavko; Devic, Marija; Sofronic-Milosavljevic, Ljiljana

    2015-03-01

    Helminth-derived products, either released into the circulation during the course of the infection or isolated after in vitro cultivation of the parasite and applied by the injection, are able to suppress the host immune response to autoantigens and allergens, but mechanisms could differ. Prophylactic application of Trichinella spiralis excretory-secretory muscle larvae (ES L1) products ameliorates experimental autoimmune encephalomyelitis (EAE) with the same success as infection did. However, a shift to the Th2-type response in the periphery and in the central nervous system, accompanied by activation of regulatory mechanisms, had a striking, new feature of increased proportion of unconventional CD4(+)CD25(-)Foxp3(+) regulatory cells both in the periphery and in the central nervous system of animals treated with ES L1 before the induction of EAE.

  15. Excretory-secretory antigens: a suitable candidate for immunization against ocular toxoplasmosis in a murine model.

    PubMed

    Norouzpour Deilami, Kiumars; Daryani, Ahmad; Ahmadpour, Ehsan; Sharif, Mehdi; Dadimoghaddam, Yousef; Sarvi, Shahabeddin; Alizadeh, Ahad

    2014-12-01

    Toxoplasmosis, responsible for ocular impairment, is caused by Toxoplasma gondii. We investigated the effect of Toxoplasma excretory-secretory antigens (ESA) on parasite load and distribution in the eye tissue of a murine model. Case and control groups were immunized with ESA and PBS, respectively. Two weeks after the second immunization, the mice were challenged intraperitoneally with virulent RH strain of Toxoplasma; eye tissue samples of both groups were collected daily (days 1, 2, 3, and the last day before death). Parasite load was determined using real-time quantitative PCR targeted at the B1 gene. Compared to the control group, infected mice that received ESA vaccine presented a considerable decrease in parasite load in the eye tissue, demonstrating the effect of ESA on parasite load and distribution. Diminution of parasite load in mouse eye tissue indicated that ESA might help control disease-related complications and could be a valuable immunization candidate against ocular toxoplasmosis.

  16. Myosin VI small insert isoform maintains exocytosis by tethering secretory granules to the cortical actin.

    PubMed

    Tomatis, Vanesa M; Papadopulos, Andreas; Malintan, Nancy T; Martin, Sally; Wallis, Tristan; Gormal, Rachel S; Kendrick-Jones, John; Buss, Folma; Meunier, Frédéric A

    2013-02-04

    Before undergoing neuroexocytosis, secretory granules (SGs) are mobilized and tethered to the cortical actin network by an unknown mechanism. Using an SG pull-down assay and mass spectrometry, we found that myosin VI was recruited to SGs in a Ca(2+)-dependent manner. Interfering with myosin VI function in PC12 cells reduced the density of SGs near the plasma membrane without affecting their biogenesis. Myosin VI knockdown selectively impaired a late phase of exocytosis, consistent with a replenishment defect. This exocytic defect was selectively rescued by expression of the myosin VI small insert (SI) isoform, which efficiently tethered SGs to the cortical actin network. These myosin VI SI-specific effects were prevented by deletion of a c-Src kinase phosphorylation DYD motif, identified in silico. Myosin VI SI thus recruits SGs to the cortical actin network, potentially via c-Src phosphorylation, thereby maintaining an active pool of SGs near the plasma membrane.

  17. Inhibitory Effect of Orientin on Secretory Group IIA Phospholipase A2.

    PubMed

    Bae, Jong-Sup

    2015-08-01

    It is well known that the expression level of secretory group IIA phospholipase A2 (sPLA2-IIA) is elevated in inflammatory diseases and lipopolysaccharide (LPS) upregulates the expression of sPLA2-IIA in human umbilical vein endothelial cells (HUVECs). Orientin, a C-glycosyl flavonoid, is known to have anxiolytic, anti-oxidative, and anti-inflammatory activity. Here, orientin was examined for its effects on the expression and activity of sPLA2-IIA in HUVECs and mouse. Prior treatment of cells or mouse with orientin inhibited LPS-induced expression and activity of sPLA2-IIA. And orientin suppressed the activation of cytosolic phospholipase A2 (cPLA2) and extracellular signal-regulated kinase (ERK) 1/2 by LPS. Therefore, these results suggest that orientin may inhibit LPS-mediated expression of sPLA2-IIA by suppression of cPLA2 and ERK 1/2.

  18. The Role of Secretory Autophagy in Zika Virus Transfer through the Placental Barrier

    PubMed Central

    Zhang, Zhong-Wei; Li, Zi-Lin; Yuan, Shu

    2017-01-01

    Recent studies indicated that the Zika virus genome could be detected in the amniotic fluid and the fetal brain, which confirms that the virus can cross the placental barrier. Secretory autophagy or exosome pathways may participate in this virus transfer. Autophagy modulators regulate autophagosome formation or membrane fusion with lysosomal vesicles and therefore inhibit viral nucleocapsid releasing or virus transfer to the fetus hypothetically. However, some autophagy modulators may enhance virus replication. Autophagy inhibitors may arrest placental development; while exaggeration of autophagy in human placenta may be associated with the fetal growth restriction. Therefore, autophagy modulators should be used carefully due to their complex clinical effects. Alternatively, exosome-specific inhibitors might be also considered, although their safety of both maternal and fetal conditions must be carefully assessed before any advancement to human clinical trials. PMID:28119857

  19. The role of cysteine-rich secretory proteins in male fertility

    PubMed Central

    Koppers, Adam J; Reddy, Thulasimala; O'Bryan, Moira K

    2011-01-01

    The cysteine-rich secretory proteins (CRISPs) are a subgroup of the CRISP, antigen 5 and Pr-1 (CAP) protein superfamily, and are found only in vertebrates. They show a strong expression bias to the mammalian male reproductive tract and the venom of poisonous reptiles. Within the male reproductive tract CRISPs have been implicated in many aspects of male germ cell biology spanning haploid germ cell development, epididymal maturation, capacitation, motility and the actual processes of fertilization. At a structural level, CRISPs are composed of two domains, a CAP domain, which has been implicated in cell–cell adhesion, and a CRISP domain, which has been shown to regulate several classes of ion channels across multiple species. Herein, we will review the current literature on the role of CRISPs in male fertility, and by inference to related non-mammalian protein, infer potential biochemical functions. PMID:20972450

  20. Secretory breast carcinoma in a 6-year-old girl: mastectomy with sentinel lymph node dissection.

    PubMed

    Soyer, Tutku; Yaman Bajin, İnci; Orhan, Diclehan; Yalçin, Bilgehan; Özgen Kiratli, Pinar; Oğuz, Berna; Karnak, İbrahim

    2015-07-01

    Secretory breast carcinoma (SBC) is a rare type of breast neoplasia that was originally described in children. SBC is an indolent breast tumor with good clinical outcome and rare systemic involvement. Since, majority of studies concerning pediatric SBC have been case reports, it has been difficult to clearly elucidate the characteristics and optimal treatment strategies for SBC in children. Although treatment recommendations vary, surgical excision is the primary mode of treatment. Also, necessity of axillary and/or sentinel lymph node dissection is another matter of discussion in children. We report a 6-year-old girl who was diagnosed as SBC was reported to discuss the use of mastectomy with sentinel lymph node dissection in the treatment of this rare tumor in children.

  1. Syntaxin clusters assemble reversibly at sites of secretory granules in live cells.

    PubMed

    Barg, S; Knowles, M K; Chen, X; Midorikawa, M; Almers, Wolfhard

    2010-11-30

    Syntaxin resides in the plasma membrane, where it helps to catalyze membrane fusion during exocytosis. The protein also forms clusters in cell-free and granule-free plasma-membrane sheets. We imaged the interaction between syntaxin and single secretory granules by two-color total internal reflection microscopy in PC12 cells. Syntaxin-GFP assembled in clusters at sites where single granules had docked at the plasma membrane. Clusters were intermittently present at granule sites, as syntaxin molecules assembled and disassembled in a coordinated fashion. Recruitment to granules required the N-terminal domain of syntaxin, but not the entry of syntaxin into SNARE complexes. Clusters facilitated exocytosis and disassembled once exocytosis was complete. Syntaxin cluster formation defines an intermediate step in exocytosis.

  2. Delayed adrenal insufficiency long after unilateral adrenalectomy: prolonged glucocorticoid therapy reduced reserved secretory capacity of cortisol.

    PubMed

    Kazama, Itsuro; Komatsu, Yasuhiro; Ohiwa, Takafumi; Sanayama, Kyo; Nagata, Mikio

    2005-06-01

    A 51-year-old woman with Cushing's syndrome underwent unilateral adrenalectomy for left adrenal adenoma. After 7 years of prednisolone treatment (with some interruptions), followed by 4 years of total withdrawal from prednisolone treatment, she presented with hypotension, weight loss, general fatigue, nausea, hyponatremia and hypoglycemia. These clinical features together with a low response in the rapid adrenocorticotropic hormone test led to the diagnosis of acute adrenal insufficiency. Relatively low serum adrenocorticotropic hormone levels in the face of increased demand for cortisol during adrenal crisis suggested a disordered hypothalamic-pituitary function, indicating secondary adrenal insufficiency. This patient demonstrated the etiology of acute adrenal insufficiency long after unilateral adrenalectomy in association with subsequent glucocorticoid therapy. A reduction in the reserved secretory capacity of cortisol after prolonged prednisolone treatment was considered to have induced secondary adrenal insufficiency, even after 4 years of total withdrawal from prednisolone.

  3. Mitochondrial calcium in the life and death of exocrine secretory cells.

    PubMed

    Voronina, Svetlana; Tepikin, Alexei

    2012-07-01

    The remarkable recent discoveries of the proteins mediating mitochondrial Ca(2+) transport (reviewed in this issue) provide an exciting opportunity to utilise this new knowledge to improve our fundamental understanding of relationships between Ca(2+) signalling and bioenergetics and, importantly, to improve the understanding of diseases in which Ca(2+) toxicity and mitochondrial malfunction play a crucial role. Ca(2+) is an important activator of exocrine secretion, a regulator of the bioenergetics of exocrine cells and a contributor to exocrine cell damage. Exocrine secretory cells, exocrine tissues and diseases affecting exocrine glands (like Sjögren's syndrome and acute pancreatitis) will, therefore, provide worthy research areas for the application of this new knowledge of the Ca(2+) transport mechanisms in mitochondria.

  4. Sandwich enzyme-linked immunosorbent assay for detection of excretory secretory antigens in humans with fascioliasis.

    PubMed Central

    Espino, A M; Finlay, C M

    1994-01-01

    A sandwich enzyme-linked immunosorbent assay has been developed for the detection of Fasciola hepatica excretory secretory (ES) antigens in stool specimens of infected humans. The assay uses antibodies against F. hepatica ES antigens. A monoclonal antibody (ES78, mouse immunoglobulin G2a) was used to capture ES antigens, and a rabbit polyclonal antibody, peroxidase conjugate, was used to identify ES antigens. Thirteen of 14 patients with parasitological evidence of fascioliasis had a detectable concentration of ES antigens (more than 15 ng/ml). None of the stool specimens from controls and from patients with parasites other than F. hepatica showed a positive reaction, suggesting the absence of cross-reactions in this assay. When the 14 patients were retested 2 months after treatment, all of the specimens from the 11 parasitologically cured patients were negative by the antigen detection assay while the specimens from the 3 patients with persisting F. hepatica eggs in their stools remained positive. PMID:8126178

  5. Expression of Active Fluorophore Proteins in the Milk of Transgenic Pigs Bypassing the Secretory Pathway

    PubMed Central

    Mukherjee, Ayan; Garrels, Wiebke; Talluri, Thirumala R.; Tiedemann, Daniela; Bősze, Zsuzsanna; Ivics, Zoltán; Kues, Wilfried A.

    2016-01-01

    We describe the expression of recombinant fluorescent proteins in the milk of two lines of transgenic pigs generated by Sleeping Beauty transposon-mediated genetic engineering. The Sleeping Beauty transposon consisted of an ubiquitously active CAGGS promoter driving a fluorophore cDNA, encoding either Venus or mCherry. Importantly, the fluorophore cDNAs did not encode for a signal peptide for the secretory pathway, and in previous studies of the transgenic animals a cytoplasmic localization of the fluorophore proteins was found. Unexpectedly, milk samples from lactating sows contained high levels of bioactive Venus or mCherry fluorophores. A detailed analysis suggested that exfoliated cells of the mammary epithelium carried the recombinant proteins passively into the milk. This is the first description of reporter fluorophore expression in the milk of livestock, and the findings may contribute to the development of an alternative concept for the production of bioactive recombinant proteins in the udder. PMID:27086548

  6. Interaction of secretory immunoglobulin A antibodies with Naegleria fowleri trophozoites and collagen type I.

    PubMed

    Shibayama, Mineko; Serrano-Luna, José de Jesús; Rojas-Hernández, Saúl; Campos-Rodríguez, Rafael; Tsutsumi, Victor

    2003-03-01

    In this work, we analyzed the in vitro interaction of human secretory immunoglobulin A (sIgA) antibodies with Naegleria fowleri trophozoites and the capacity of these antibodies to inhibit amoeba adherence to collagen type I. We also studied N. fowleri antigens that are recognized by sIgA, using immunoblot assays. Immunocytochemical analysis of the interaction showed a redistribution of antigens on the surface of trophozoites by sIgA antibodies. Ultrastructural analysis of antibody-amoeba interaction showed that besides the patching and cap formation, parasites were capable of eliminating the antigen-antibody complex produced on the surface. sIgA antibodies were capable of inhibiting the in vitro adhesion of trophozoites to collagen type I. We suggest that nonsymptomatic infections by N. fowleri may stimulate a local specific immunity that prevents trophozoite adhesion and invasion of nasal mucosa.

  7. [Secretory activity of the stomach during modeling of increased filling of abdominal veins].

    PubMed

    Afonin, B V; Goncharova, N P

    2009-01-01

    Determination of the stomach secretory activity and ultrasonic investigation of the gastroduodenal organs and vessels were performed in test-subjects tilted at -12 degrees and -15 degrees. Short HDT(12-24 hrs.) as a hypokinesia factor was minimized which caused enlargement of the parenchymatous and thickening of walls of the hollow organs. Increased filling of the abdominal veins was paralleled by rise in pepsinogen levels in blood and urine, and increase in the intravitelline content in fasting subjects. Elevated tone of the pylorus and retarded evacuation from the stomach indicated active hydrochloric acid secretion. Concurrently, bile and pancreas juices were secreted more profusely and intestinal content in the duodenum increased. It was shown that modeled increase of abdominal vein filling stimulates secretion as by fasting stomach so by the liver and pancreas.

  8. Quantification of pancreatic secretory trypsin inhibitor in colonic carcinoma and normal adjacent colonic mucosa.

    PubMed Central

    Bohe, H; Bohe, M; Jönsson, P; Lindström, C; Ohlsson, K

    1992-01-01

    AIMS: To measure the content of immunoreactive human pancreatic secretory trypsin inhibitor (irPSTI) in colonic carcinoma and adjacent normal colonic mucosa. METHODS: From a stable hybridoma cell line producing monoclonal antibodies specific for human PSTI, a specific enzyme linked immunosorbent assay (ELISA) for human PSTI was developed. In a precipitation assay system these antibodies bound human PSTI in a dose-dependent manner. The specimens were obtained from resectional surgery. RESULTS: The content of irPSTI was 19.9 micrograms/g protein (0.55 micrograms/g tissue wet weight) in colonic carcinoma. In adjacent normal colonic mucosa 43.6 micrograms/g protein (1.12 micrograms/g tissue wet weight) was shown. CONCLUSIONS: The enzymatic degradation of surrounding tissue necessary for tumour cell invasion could be facilitated by this relative deficit of the inhibitor in infiltrative carcinoma. PMID:1479031

  9. Quantification of secretory leukocyte protease inhibitor (SLPI) in oral gargle specimens collected using mouthwash

    PubMed Central

    Pierce Campbell, Christine M.; Guan, Wei; Sprung, Robert; Koomen, John M.; O’Keefe, Michael T.; Ingles, Donna J.; Abrahamsen, Martha; Giuliano, Anna R.

    2014-01-01

    Background Secretory leukocyte protease inhibitor (SLPI) is an innate immunity-associated protein known to inhibit HIV transmission, and is thought to inhibit a variety of infectious agents, including human papillomaviruses (HPVs). We aimed to optimize an established ELISA-based SLPI quantification assay for use with oral gargle specimens collected using mouthwash, and to assess preliminary associations with age, smoking status, and alcohol intake. Methods Oral gargle supernatants from 50 individuals were used to optimize the Human SLPI Quantikine ELISA Kit. Sample suitability was assessed and quality control analyses were conducted. Results Salivary SLPI was successfully recovered from oral gargles with low intra-assay and high inter-individual variability. Initial measurements showed that salivary SLPI varied considerably across individuals, and that SLPI was inversely associated with age. Conclusions This optimized assay can be used to examine the role of SLPI in the acquisition of oral HPV and other infections. PMID:24140751

  10. Dynamic Regulation of Ero1α and Peroxiredoxin 4 Localization in the Secretory Pathway*

    PubMed Central

    Kakihana, Taichi; Araki, Kazutaka; Vavassori, Stefano; Iemura, Shun-ichiro; Cortini, Margherita; Fagioli, Claudio; Natsume, Tohru; Sitia, Roberto; Nagata, Kazuhiro

    2013-01-01

    In the early secretory compartment (ESC), a network of chaperones and enzymes assists oxidative folding of nascent proteins. Ero1 flavoproteins oxidize protein disulfide isomerase (PDI), generating H2O2 as a byproduct. Peroxiredoxin 4 (Prx4) can utilize luminal H2O2 to oxidize PDI, thus favoring oxidative folding while limiting oxidative stress. Interestingly, neither ER oxidase contains known ER retention signal(s), raising the question of how cells prevent their secretion. Here we show that the two proteins share similar intracellular localization mechanisms. Their secretion is prevented by sequential interactions with PDI and ERp44, two resident proteins of the ESC-bearing KDEL-like motifs. PDI binds preferentially Ero1α, whereas ERp44 equally retains Ero1α and Prx4. The different binding properties of Ero1α and Prx4 increase the robustness of ER redox homeostasis. PMID:23979138

  11. Dynamic regulation of Ero1α and peroxiredoxin 4 localization in the secretory pathway.

    PubMed

    Kakihana, Taichi; Araki, Kazutaka; Vavassori, Stefano; Iemura, Shun-ichiro; Cortini, Margherita; Fagioli, Claudio; Natsume, Tohru; Sitia, Roberto; Nagata, Kazuhiro

    2013-10-11

    In the early secretory compartment (ESC), a network of chaperones and enzymes assists oxidative folding of nascent proteins. Ero1 flavoproteins oxidize protein disulfide isomerase (PDI), generating H2O2 as a byproduct. Peroxiredoxin 4 (Prx4) can utilize luminal H2O2 to oxidize PDI, thus favoring oxidative folding while limiting oxidative stress. Interestingly, neither ER oxidase contains known ER retention signal(s), raising the question of how cells prevent their secretion. Here we show that the two proteins share similar intracellular localization mechanisms. Their secretion is prevented by sequential interactions with PDI and ERp44, two resident proteins of the ESC-bearing KDEL-like motifs. PDI binds preferentially Ero1α, whereas ERp44 equally retains Ero1α and Prx4. The different binding properties of Ero1α and Prx4 increase the robustness of ER redox homeostasis.

  12. Glucosidase II and MRH-domain containing proteins in the secretory pathway

    PubMed Central

    D’Alessio, Cecilia; Dahms, Nancy M.

    2015-01-01

    N -glycosylation in the endoplasmic reticulum (ER) consists of the transfer of a pre-assembled glycan conserved among species (Glc3Man9GlcNAc2) from a lipid donor to a consensus sequence within a nascent protein that is entering the ER. The protein-linked glycans are then processed by glycosidases and glycosyltransferases in the ER producing specific structures that serve as signalling molecules for the fate of the folding glycoprotein: to stay in the ER during the folding process, to be retrotranslocated to the cytosol for proteasomal degradation if irreversibly misfolded, or to pursue transit through the secretory pathway as a mature glycoprotein. In the ER, each glycan signalling structure is recognized by a specific lectin. A domain similar to that of the mannose 6-phosphate receptors (MPRs) has been identified in several proteins of the secretory pathway. These include the beta subunit of glucosidase II (GII), a key enzyme in the early processing of the transferred glycan that removes middle and innermost glucoses and is involved in quality control of glycoprotein folding in the ER (QC), the lectins OS-9 and XTP3-B, proteins involved in the delivery of ER misfolded proteins to degradation (ERAD), the gamma subunit of the Golgi GlcNAc-1-phosphotransferase, an enzyme involved in generating the mannose 6-phosphate (M6P) signal for sorting acidic hydrolases to lysosomes, and finally the MPRs that deliver those hydrolytic enzymes to the lysosome. Each of the MRH-containing proteins recognizes a different signalling N-glycan structure. Three-dimensional structures of some of the MRH domains have been solved, providing the basis to understand recognition mechanisms. PMID:25692846

  13. von Willebrand factor multimerization and the polarity of secretory pathways in endothelial cells

    PubMed Central

    Lopes da Silva, Mafalda

    2016-01-01

    The von Willebrand factor (VWF) synthesized and secreted by endothelial cells is central to hemostasis and thrombosis, providing a multifunctional adhesive platform that brings together components needed for these processes. VWF secretion can occur from both apical and basolateral sides of endothelial cells, and from constitutive, basal, and regulated secretory pathways, the latter two via Weibel-Palade bodies (WPB). Although the amount and structure of VWF is crucial to its function, the extent of VWF release, multimerization, and polarity of the 3 secretory pathways have only been addressed separately, and with conflicting results. We set out to clarify these relationships using polarized human umbilical vein endothelial cells (HUVECs) grown on Transwell membranes. We found that regulated secretion of ultra–large (UL)-molecular-weight VWF predominantly occurred apically, consistent with a role in localized platelet capture in the vessel lumen. We found that constitutive secretion of low-molecular-weight (LMW) VWF is targeted basolaterally, toward the subendothelial matrix, using the adaptor protein complex 1 (AP-1), where it may provide the bulk of collagen-bound subendothelial VWF. We also found that basally-secreted VWF is composed of UL-VWF, released continuously from WPBs in the absence of stimuli, and occurs predominantly apically, suggesting this could be the main source of circulating plasma VWF. Together, we provide a unified dataset reporting the amount and multimeric state of VWF secreted from the constitutive, basal, and regulated pathways in polarized HUVECs, and have established a new role for AP-1 in the basolateral constitutive secretion of VWF. PMID:27106123

  14. The Harderian gland, its secretory duct and porphyrin content in the mongolian gerbil (Meriones unguiculatus).

    PubMed Central

    Johnston, H S; McGadey, J; Thompson, G G; Moore, M R; Payne, A P

    1983-01-01

    The Harderian gland, its secretory duct and porphyrin content were examined in the mongolian gerbil (Meriones unguiculatus). The gland consisted of tubules lined by a single layer of epithelial cells and a myoepithelial network. The tubule cells were often binucleate and possessed lipid vacuoles in the apical half of the cell, a corona of granular endoplasmic reticulum surrounding the nucleus, and cytoplasmic 'slashes'. The latter are probably derived from dense membranous couplets and may be precursors of the lipid vacuoles. Holocrine and merocrine secretion was observed. Interstitial cells included plasma cells, mast cells and (predominantly) melanocytes which render the gland black. The gland was surrounded by a collagen capsule and an outer layer of highly attenuated (possibly endothelioid) cells. Within the gland, the secretory duct was lined by a single layer of normal tubule cells. Outside the gland, the duct enlarged to form an ampulla, from which clefts led off to deep crypts. The ampulla and clefts were lined by cells with small dense apical granules and stubby microvilli; some possessed lipid vacuoles. The crypts were lined by serous cells with active Golgi regions. At the duct opening, ampullary cells became squamous and goblet cells occurred. Geometric crystalloid deposits (with a layered structure of 7.6 nm periodicity) occurred at cleft-crypt junctions. Islets of extra-glandular ductal tissue were occasionally found within the gland. Porphyrins were detectable both by chemical assay and fluorescence microscopy. There was a trend for female glands to have a higher content than males. Solid intraluminal accretions of porphyrin and/or lipid were present. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5 Fig. 6 Fig. 7 Fig. 8 Fig. 9 Fig. 10 Fig. 12 Fig. 13 Fig. 14 Fig. 15 Fig. 16 PMID:6654750

  15. Persistent dominant follicle alters pattern of oviductal secretory proteins from cows at estrus.

    PubMed

    Binelli, M; Hampton, J; Buhi, W C; Thatcher, W W

    1999-07-01

    The experimental objective was to compare synthesis of oviductal secretory proteins of dairy cows bearing a persistent dominant follicle (PDF) versus a fresh dominant follicle (FDF) at estrus. On Day 7 after synchronized estrus (Day 0), cows received an intravaginal progesterone device and injection of prostaglandin F2alpha (PGF2alpha). On Day 9, cows received an injection of a GnRH agonist (FDF group; n = 3) or received no injection (PDF group, n = 3). On Day 16, all cows received PGF2alpha, and progesterone devices were removed. At slaughter on Day 18 or Day 19, oviducts ipsilateral and contralateral to the dominant follicle were divided into infundibulum, ampulla, and isthmus regions. Explants from oviductal regions were cultured in minimal essential medium supplemented with [3H]leucine for 24 h. Two-dimensional fluorographs of proteins in conditioned media were analyzed by densitometry. Rate of incorporation of [3H]leucine into macromolecules was greater in the infundibulum, ampulla, and isthmus of FDF cows (p < 0.01). Overall, intensities of radiolabeled secretory protein (P) 2 and P13 were greater for FDF than for PDF. In the ampulla, P14 was more intense for FDF while P7 was more intense for PDF. Abundance of P1 in the isthmus was greater for PDF cows. Across regions, P5, P6, P8, P9, and P11 were more intense for PDF than for FDF in the ipsilateral side. In the contralateral side, P19 was more intense for PDF than for FDF, whereas P6, P8, P9, and P11 were more intense for FDF. Differences in biosynthetic activity and in secreted oviductal proteins from cows bearing a PDF may contribute to the decrease in fertility associated with a PDF.

  16. Mammaglobin and S-100 immunoreactivity in salivary gland carcinomas other than mammary analogue secretory carcinoma.

    PubMed

    Patel, Kalyani R; Solomon, Isaac H; El-Mofty, Samir K; Lewis, James S; Chernock, Rebecca D

    2013-11-01

    Mammary analogue secretory carcinoma (MASC) is a recently described salivary gland tumor that has morphologic features similar to secretory carcinoma of the breast and that also harbors the same ETV6 translocation. Diffuse mammaglobin and S-100 immunoreactivity are used to differentiate MASC from its morphologic mimics, especially acinic cell carcinoma and adenocarcinoma, not otherwise specified. However, the combination of mammaglobin and S-100 immunoreactivity has not been well studied in other types of salivary gland carcinomas that may have focal areas reminiscent of MASC. Here we evaluated mammaglobin and S-100 immunoreactivity in 15 cases each of polymorphous low-grade adenocarcinoma, adenoid cystic carcinoma and mucoepidermoid carcinoma, and also in 2 cases of adenocarcinoma, not otherwise specified, and 1 mucinous adenocarcinoma. Cases with significant co-expression of mammaglobin and S-100 (moderate or strong immunoreactivity in >25% of tumor cells) were further analyzed by fluorescence in situ hybridization using the ETV6 (12p13) break-apart probe. Nine cases (60%) of polymorphous low-grade adenocarcinoma and two (13.3%) of adenoid cystic carcinoma met the criteria for significant co-expression of mammaglobin and S-100. All were negative for the ETV6 translocation by fluorescence in situ hybridization. Although mammaglobin and S-100 positivity was seen in the majority of polymorphous low-grade adenocarcinomas and a minority of adenoid cystic carcinomas, none were positive for the ETV6 translocation characteristic of MASC. This indicates a need for caution in the use of immunohistochemistry for diagnosing MASC, especially in the absence of cytogenetic confirmation.

  17. A novel secretory pathway for interleukin-1 beta, a protein lacking a signal sequence.

    PubMed Central

    Rubartelli, A; Cozzolino, F; Talio, M; Sitia, R

    1990-01-01

    Interleukin 1 (IL-1) is a major soluble mediator of inflammation. Two human IL-1 genes, alpha and beta, have been isolated, which encode polypeptides with only 20-30% amino acid sequence homology. Unlike most secreted proteins, the two cytokines do not have a signal sequence, an unexpected finding in view of their biological role. Here we show that IL-1 beta is actively secreted by activated human monocytes via a pathway of secretion different from the classical endoplasmic reticulum--Golgi route. Drugs which block the intracellular transport of IL-6, of tumour necrosis factor alpha and of other secretory proteins do not inhibit secretion of IL-1 beta. Secretion of IL-1 beta is blocked by methylamine, low temperature or serum free medium, and is increased by raising the culture temperature to 42 degrees C or by the presence of calcium ionophores, brefeldin A, monensin, dinitrophenol or carbonyl cyanide chlorophenylhydrazone. IL-1 beta is contained in part within intracellular vesicles which protect it from protease digestion. In U937 cells large amounts of IL-1 beta are made but none is secreted. In these cells IL-1 beta is not found in the vesicular fraction, and all the protein is accessible to protease digestion. This suggests that intracellular vesicles that contain IL-1 beta are part of the protein secretory pathway. We conclude that IL-1 beta is released by activated monocytes via a novel mechanism of secretion which may involve translocation of intracellular membranes and is increased by stress conditions. Images Fig. 1. Fig. 2. Fig. 3. Fig. 4. Fig. 6. PMID:2328723

  18. Monoclonal Antibodies as Probes for Unique Antigens in Secretory Cells of Mixed Exocrine Organs

    NASA Astrophysics Data System (ADS)

    Basbaum, C. B.; Mann, J. K.; Chow, A. W.; Finkbeiner, W. E.

    1984-07-01

    In the past, it has been difficult to identify the secretory product and control mechanisms associated with individual cell types making up mixed exocrine organs. This report establishes the feasibility of using immunological methods to characterize both the biochemical constituents and regulatory mechanisms associated with secretory cells in the trachea. Monoclonal antibodies directed against components of tracheal mucus were produced by immunizing mice with dialyzed, desiccated secretions harvested from tracheal organ culture. An immunofluorescence assay revealed that of the total 337 hybridomas screened, 100 produced antibodies recognizing goblet cell granules; 64, gland cell granules; and 3, antigen confined to the ciliated apical surface of the epithelium. The tracheal goblet cell antibody described in this report was strongly cross-reactive with intestinal goblet cells, as well as with a subpopulation of submandibular gland cells, but not with cells of Brunner's glands or the ciliated cell apical membrane. The serous cell antibody was not cross-reactive with goblet, Brunner's gland, or submandibular cells, or the ciliated cell apical membrane. The antibody directed against the apical membrane of ciliated cells did not cross-react with gland or goblet cells or the apical membrane of epithelial cells in the duodenum. Monoclonal antibodies, therefore, represent probes by which products unique to specific cells or parts of cells in the trachea can be distinguished. The antibodies, when used in enzyme immunoassays, can be used to quantitatively monitor secretion by individual cell types under a variety of physiological and pathological conditions. They also provide the means for purification and characterization of cell-specific products by immunoaffinity chromatography.

  19. Mammary analogue secretory carcinoma of salivary glands: a clinicopathologic study of 11 cases.

    PubMed

    Din, Nasir Ud; Fatima, Saira; Kayani, Naila

    2016-06-01

    Mammary analogue secretory carcinoma (MASC) is a recently described tumor sharing the histologic, immunohistochemical, and molecular profile of secretory carcinoma of breast. We aimed to evaluate the morphologic and histochemical features needed/required for the diagnosis of MASC without adjunct of molecular analysis. Six retrospective cases suspicious for MASC and 5 prospective cases reported as MASC were included in the study. Molecular analysis of ETV6 by fluorescence in situ hybridization was performed at the University of Pittsburg, USA. The ages of the patients ranged from 9 to 60 years (mean, 27.5 years). Histologically, all tumors showed mixed growth patterns including microcystic, macrocystic, papillary, tubular, and solid, papillary the being most common pattern. The tumor cells showed round to oval vesicular nuclei with small nucleoli, and eosinophilic to vacuolated cytoplasm. All cases demonstrated luminal and cytoplasmic mucin on periodic acid-Schiff with and without diastase digestion and alcian blue stain. ETV6 fusion gene rearrangement by fluorescence in situ hybridization was detected in 10 of 11 tumors. Recurrences occurred in 3 patients, and 1 patient died of disease 5 years after surgery. In conclusion, MASC is a relatively rare salivary gland malignancy exhibiting distinct histologic and histochemical features which can help to differentiate it from other mimics. Histologically, papillary-cystic and microcystic patterns are the main clues to diagnosis. The follicular pattern of acinic cell carcinoma might represent MASC, as 4 cases in our series had this pattern. Two patients in our series were 9 and 9½ years old respectively, which are the youngest ages ever recorded for MASC.

  20. Relationships between levels of lysozyme, lactoferrin, salivary peroxidase, and secretory immunoglobulin A in stimulated parotid saliva.

    PubMed Central

    Rudney, J D; Smith, Q T

    1985-01-01

    Recent studies suggest that salivary lysozyme (Lz), lactoferrin (Lf), peroxidase (Spx), and secretory immunoglobulin A (sIgA) may interact in a common antimicrobial system. A multiple protein approach therefore may be needed to determine the role of this system in oral health and ecology. In the present study we investigate the relationships between levels of Lz, Lf, Spx, and sIgA (adjusted for flow rate and total protein) in stimulated parotid saliva from 44 dental students. Principal components analysis was used to determine major patterns of intercorrelation between variables; cluster analysis was used to identify groups of subjects with similar salivary profiles for Lz, Lf, Spx, and sIgA. Spx tended to vary independently of Lz and Lf, which, in turn, tended to vary together. sIgA showed a weak negative relationship with Spx and a weak positive relationship with Lz and Lf. Six major clusters of subjects with similar antimicrobial protein profiles were found. These were significantly different at P less than 0.0001. Spx was the most important determinant of cluster membership followed (in order of importance) by Lz, Lf, and sIgA. Cluster profiles were Spx-, sIgAmu, Lf-, Lz-; Spx-, sIgA+, Lfmu, Lz+; Spxmu, sIgAmu, Lfmu, Lzmu; Spx+, sIgA-, Lf-, Lz-; Spx+, sIgAmu, Lf+, Lz-; and Spx+, sIgAmu, Lf+, Lz+ (-, mu, and + refer to the position of the cluster mean each variable relative to the overall mean for that variable). Results suggest that clusters may be a product of independent variation in the secretory activity of acinar and intercalated duct cells. PMID:4030086